Constructs for Modulating Transpiration in Plants and Uses Therefor

专利摘要:
Disclosed are constructs for controlling stomatal closure, as well as plants, plant parts and plant cells comprising such constructs. The present invention also discloses methods for increasing transpiration in plants and plant parts through use of these contructs. - 126-
公开号:AU2013205472A1
申请号:U2013205472
申请日:2013-04-12
公开日:2014-02-06
发明作者:Mark David KINKEMA；Ian Mark O'hara；Manuel B. Sainz
申请人:University of Queensland UQ；Queensland University of Technology QUT；Syngenta Participations AG；
IPC主号:C12N15-82

专利说明:
Australian Patents Act 1990 - Regulation 3.2A ORIGINAL COMPLETE SPECIFICATION STANDARD PATENT Invention Title "Constructs for modulating transpiration in plants and uses therefor" The following statement is a full description of this invention, including the best method of performing it known to us: H:rrInterwovenNRPortblDCCRR5072887_1 .DOC - 12/4/13 TITLE OF THE INVENTION "CONSTRUCTS FOR MODULATING TRANSPIRATION IN PLANTS AND USES THEREFOR" 100011 This application claims priority to Australian Provisional Application No. 2012903070 entitled "Constructs for Modulating Transpiration in Plants and Uses Therefor" 5 filed 18 July 2012, the contents of which are incorporated herein by reference in their entirety. FIELD OF THE INVENTION 100021 The present invention is directed generally to the field plant engineering. Specifically, this invention relates to constructs for controlling stomatal closure, as well as plants, plant parts and plant cells comprising those constructs. 10 BACKGROUND OF THE INVENTION 100031 Most land plants have the ability to regulate gas exchange and transpiration by opening and closing of the stomatal aperture. The turgor pressure-mediated movement of a pair of special epidermal cells or guard cells, controls the size of the stomatal aperture and so regulates the extent of water loss via transpiration and also regulates CO 2 uptake into the leaf 15 for photosynthetic carbon fixation. 100041 Plants tightly regulate stomatal closure to prevent excess water loss that would be detrimental to growth and development. While uncontrolled water loss would be deleterious, the controlled manipulation of stomatal closure could be used to increase water loss and improve crop plants in at least three distinct ways: 20 10005] First, it could be used to speed up the drying process for crops such as cotton and grain, and could decrease the transport and processing costs associated with crops such as sugar cane. An efficient drying off process prior to harvest is extremely important in crops such as cotton and grain, and controlling stomatal closure may have advantages over the currently used harvest aids for this purpose. Sugar cane is approximately 70% water at 25 harvest, and consequently a substantial amount of energy is required to remove this water during the milling process used to extract sucrose. Further expense is incurred in the transport of this water-laden plant material from the field to the mill. Decreasing the water content of the sugar cane at harvest has the potential to substantially reduce the transport and milling costs. Other crops, such as tobacco and dried fruits such as raisins and prunes, are dried 30 immediately post-harvest. Thus, it would be advantageous to be able to accelerate or control the rate of crop drying. - 1- 100061 A second way of improving plants through increased stomatal opening is to enable plants to take up and store more carbon dioxide in the form of stored carbohydrates such as starch and simple sugars. 100071 Thirdly, controlled increases in transpiration could be used to increase the 5 sugar content of crops such as sugar cane, sweet sorghum, and sugar beet. Plants naturally respond to drought stress by increasing sugar levels, which act to protect cells from desiccation. Therefore, increased sugar production is expected to occur concomitantly with a decrease in plant water content. [00081 Stomatal closure is generally controlled by the hormone abscisic acid 10 (ABA) in response to drought stress. Proteins involved in ABA-mediated stomatal regulation include the Ca 2 -independent ABA-activated protein kinase (AAPK), protein kinase open stomata 1 (OSTl), respiratory burst oxidase homologs (Rboh) RBOHD and RBOHF NADPH oxidases, the vacuolar trafficking pathway v-SNAREs AtVAMP711-14, heterotrimeric GTP binding (G) protein alpha subunit gene (GPA1), ATP-binding cassette (ABC) transporter 15 AtABCG22, ABC transporter AtABCG40, ABC transporter AtMRP4, and phospholipase D alpha I (PLDalphal). Reducing the expression of, or introducing loss of function mutations in, the endogenous genes that code for the above proteins is known to inhibit stomatal closure, resulting in uncontrollable water loss and drought sensitivity. [0009] Several negative regulators of the ABA signaling pathway are also known. 20 For example, reduced activity of the ABA insensitive I (ABII) and ABA insensitive 2 (ABI2) type 2C protein phosphatases leads to enhanced responsiveness to ABA. Also, ectopic expression of the Arabidopsis homeobox-leucine zipper transcription factor ATHB6 and dominant positive mutants of the Arabidopsis SOS2-like protein kinase PKS3 is known to inhibit stomatal closure. In addition, dominant positive mutants of H(+)-ATPase I AHA1 25 (also known as open stomata 2 (OST2)), which have constitutive ATPase activity, are known to inhibit stomatal closure. [00101 Accordingly, it may be possible to increase water loss in plants by modulating the expression of genes that code for the above proteins so as to inhibit stomatal closure. However, unrestrained inhibition of stomatal closure may be detrimental to plant 30 growth and development due to the uncontrolled loss of water and the disruption of abscisic acid signaling pathways that are involved in plant development. By utilizing an inducible gene switch, however, it may be possible to regulate expression of genes encoding these -2proteins for a defined period close to harvesting time. This may enable a reduction in water content and increased sugar content without any negative effects on plant biomass. 100111 Unfortunately, most inducible gene switches currently available are leaky with consequential unwanted gene expression. Additionally, there is a lack of inducible gene 5 switches that are effective in monocotyledonous plants such as sugar cane and sweet sorghum for giving robust, tightly regulated gene expression. SUMMARY OF THE INVENTION [00121 The present invention is predicated in part on the development of an improved inducible gene switch that has enhanced sensitivity and inducibility, as well as 10 being operable in monocotyledonous plants such as sugar cane. The present inventors propose using this gene switch to control the expression of stomatal aperture-modulating genes such as those noted above in order to inhibit or reduce stomatal closure and thereby accelerate or control the rate of crop drying and/or improve the water and sugar content of crops, as described hereafter. 15 100131 Accordingly, in one aspect, the present invention provides constructs for inhibiting stomatal closure. These constructs generally comprise in operable connection: (1) a cis-acting element comprising, consisting or consisting essentially of a nucleotide sequence represented by the sequence GCGGNNCCGC [SEQ ID NO: 1]; (2) a promoter that is operable in a plant cell (e.g., a plant guard cell); and (3) a nucleic acid sequence encoding an 20 expression product that inhibits stomatal closure. [00141 In some embodiments, the cis-acting element comprises, consists or consists essentially of at least one nucleotide sequence (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20) as set forth in SEQ ID NO: 1. In illustrative examples of this type, the at least one nucleotide sequence is represented by the nucleotide sequence 25 n,,GCGGNNCCGCny [SEQ ID NO:2], wherein N or n can be independently any nucleic acid base (A, G, C, or T) and wherein x and y can be independently any number. 100151 In some embodiments, the at least one nucleotide sequence (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20) is selected from the group consisting of the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:2, 30 ATGCATGCGGAACCGCACGAGG [SEQ ID NO:3], GGCCATGCGGAGCCGCACGCGT [SEQ ID NO:4], ACAAGAGCGGCTCCGCTTGACC [SEQ ID NO:5]; TACGTAGCGGAACCGCTGCTCC [SEQ ID NO:6]; TACCATGCGGAACCGCACGTCC -3- [SEQ ID NO:7], ATGCATGCGGTGCCGCACGAGG [SEQ ID NO:8] and TACGTTGCGGAACCGCAGCTCC [SEQ ID NO:9], in any combination, in any orientation, and/or in any order. [00161 In some embodiments, the expression product that inhibits stomatal closure 5 is a stomatal closure-inhibiting polypeptide, illustrative examples of which include negative regulators of stomatal closure (e.g., ATHB6, ABI1, ABI2), mutant forms of ABIl, ABI2, AAPK, PKS3 and AHAI, which have reduced ABA sensitivity and/or which inhibit stomatal closure, and antibodies that are immuno-interactive with a polypeptide that stimulates stomatal closure or that inhibits stomatal opening (e.g., OSTI, AAPK, v-SNAREs 10 AtVAMP711-14, GPAI, AtABCG22, AtABCG40, ABC transporter AtMRP4, RBOHD and RBOHF and PLDalphal). In other embodiments, the expression product that inhibits stomatal closure is a stomatal closure-inhibiting RNA molecule (e.g., siRNA, shRNA, microRNAs, antisense RNA etc.) that inhibits expression of an endogenous nucleotide sequence encoding a polypeptide that stimulates stomatal closure or that inhibits stomatal opening (e.g., OSTI, 15 AAPK, v-SNAREs AtVAMP7I1-14, GPAI, AtABCG22, AtABCG40, ABC transporter AtMRP4, RBOHD and RBOHF and PLDalphal). 100171 In another aspect, the present invention provides a construct system for inhibiting stomatal closure. The construct system generally comprises, consists or consists essentially of a construct as broadly described above ("first construct") and a second construct 20 comprising a nucleotide sequence encoding a transcription factor (e.g., AlcR), which activates in the presence of a compound that induces expression of the alcohol dehydrogenase (ADHI) system of Aspergillus nidulans (e.g., a primary alcohol such as ethanol) and which interacts with the cis-acting element of the first construct to induce expression of the nucleic acid sequence encoding the expression product that inhibits stomatal closure. 25 [00181 Another aspect of the present invention provides transgenic plant cells (e.g., plant guard cells) that comprise a construct or construct system as broadly described above and elsewhere herein. [0019] In yet another aspect, the present invention provides transgenic plants, plant parts or plant organs (e.g., plant leaves) comprising plant cells (e.g., plant guard cells) as 30 broadly described above and elsewhere herein. [00201 Still another aspect of the present invention provides methods for increasing transpiration in a plant, plant part or plant organ (e.g. plant leaf). These methods generally -4comprise expressing in a cell (e.g., a guard cell) of the plant, plant part or plant organ a polynucleotide that comprises a nucleic acid sequence encoding an expression product that inhibits stomatal closure, where the nucleic acid sequence is under the control of a cis-acting element as broadly defined above and elsewhere herein to thereby increase transpiration in the 5 plant, plant part or plant organ. [00211 Suitably, the methods comprise inducing expression of the polynucleotide in the presence of a compound that induces expression of the alcohol dehydrogenase (ADHI) system of Aspergillus nidulans (e.g., a primary alcohol such as ethanol). 100221 The methods suitably comprise co-expressing in the cell a nucleotide 10 sequence encoding a transcription factor (e.g., AlcR), which activates in the presence of a compound that induces expression of the alcohol dehydrogenase (ADH 1) system of Aspergillus nidulans (e.g., a primary alcohol such as ethanol) and which interacts with the cis acting element to induce expression of the nucleic acid sequence encoding the expression product that inhibits stomatal closure. 15 [00231 In a further aspect, the present invention provides methods for increasing transpiration in a plant, plant part or plant organ (e.g., plant leaf) that comprises a construct or construct system as broadly defined above and elsewhere herein. These methods generally comprise exposing the plant, plant part or plant organ to a compound that induces expression of the alcohol dehydrogenase (ADHI) system of Aspergillus nidulans (e.g., a primary alcohol 20 such as ethanol) so as to inhibit stomatal closure and thereby increase transpiration in the plant, plant part or plant organ. 100241 In some embodiments, the methods comprise exposing the plant, plant part, plant organ (e.g., plant leaf) to the compound around the time of harvesting the plant, plant part or plant organ. In illustrative examples of this type, the methods comprise exposing the 25 plant, plant part or plant organ to the compound prior to harvesting the plant, plant part or plant organ. In other illustrative examples, the methods comprise exposing the plant, plant part or plant organ to the compound at the time of harvesting the plant, plant part or plant organ. In still other illustrative examples, the methods comprise exposing the plant, plant part or plant organ to the compound after harvesting the plant, plant part or plant organ. 30 [00251 In some embodiments, the methods further comprise permitting increased transpiration in the plant, plant part or plant organ over a time and under conditions sufficient for the water content of the plant, plant part or plant organ to reduce by at least about 5% -5- (e.g., at least about 6%, 7%, 8%, 9%, 10%, 15% 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%). [00261 In some embodiments of any of the methods described above, the plant is a monocotyledonous plant, illustrative examples of which include sugar cane, corn, barley, rye, 5 oats, wheat, rice, flax, millet, sorghum, grasses (e.g., switch grass, giant reed etc.), banana, onion, asparagus, lily, coconut, and the like. In other embodiments, the plant is a dicotyledonous plant (e.g., tobacco, cotton, dried fruits such as raisins and prunes, nuts, coffee, tea, cocoa, and ornamental goods). [00271 In some embodiments, the plant is selected from energy crops, 10 representative examples of which include: Miscanthus, Erianthus, Pennisetum, Arundo, Sorghum, Poplars, wheat, rice, oats, willows (e.g., Salix species); switch grass (i.e., Panicum virgatum); alfalfa (i.e., Medicago sativa); prairie bluestem (e.g., Andropogon gerardii); maize (i.e., Zea mays); soybean (i.e., Glycine max); barley (i.e., Hordeum vulgare); sugar beet (i.e., Beta vulgaris); hay and fodder crops. 15 BRIEF DESCRIPTION OF THE DRAWINGS [00281 Figure 1 is a graphical representation showing the results for ethanol inducible GUS expression at two days, four days and seven days post treatment of six month old transgenic sugar cane plants containing the different ethanol switch constructs. n=number of independent, single copy transgenic plants analyzed for each construct. Data with different 20 letters are significantly different (**P<0.01; ***P<0.00l). 100291 Figure 2 is a graphical representation showing ethanol inducible expression from promoters containing either 1, 5, or 9 copies of the inverted repeat AlcR binding site. [00301 Figure 3 is a graphical representation showing ethanol inducible expression from promoters containing various modified inverted repeat AlcR binding sites. 25 [00311 Figure 4 is a graphical representation showing ethanol inducible expression from promoters containing five copies of the inverted repeat AlcR binding sites fused to different minimal promoters. 100321 Figure 5 is a graphical representation showing ethanol inducible expression in the TO and TOV I transgenic plants. 30 [0033] Figure 6 is a schematic representation of the 35S-abil binary construct used for constitutive expression of abil in N. benthamiana. -6- [00341 Figure 7 is a schematic representation of the palcA-I-abil binary construct used for ethanol inducible expression of abil in N. benthamiana. 100351 Figure 8 is a schematic representation of the eFMVe35S-ZmUbil-scoabil construct used for constitutive expression of abil in sugar cane. 5 100361 Figure 9 is a schematic representation of the palcA I-scoabil construct used for ethanol inducible expression of abil in sugar cane. [00371 Figure 10 is a photographic representation showing ethanol inducible expression of abil in transgenic N. benthamiana. Expression of abil was assessed using RT PCR for5 independent transgenic plants prior to ethanol treatment and at 12 hours post 10 ethanol treatment. The positive control consists of vector DNA containing the abil gene. For the negative control, water was used to replace the DNA template. The expected PCR product size is 1311 bp. 10038] Figure 1I is a photographic representation showing ethanol inducible expression of abil in N. benthamiana. Representative images are shown for transgenic control 15 (vector backbone only) and ethanol inducible abil plants. Plants were photographed prior to ethanol treatment and at 12, 24, and 36 hours post treatment (h.p.t.). Ethanol treatment consisted of a single 2% ethanol root drench and aerial spray. 100391 Figure 12 is a graphical representation showing relative water content of N. benthamiana leaves following ethanol inducible expression of abil. * indicates a statistically 20 significant difference (P<0.05) within each time point relative to the control at that time point. n = number of independent clones analyzed for each of the abil transgenic events. For the control plants, data from the clones of three independent events containing the vector backbone only were combined. Ethanol treatment consisted of a single 2% ethanol root drench and aerial spray. Relative water content refers to the amount of water present in the 25 leaves compared to the fully hydrated state. [00401 Figure 13 is a photographic representation showing constitutive expression of either scoA B!] or scoabil in transgenic sugar cane. Expression was assessed using RT PCR for 8 independent scoABIl and 7 independent scoabil transgenic plants growing in soil. The expected PCR product size is 460 bp. - 7- [00411 Figure 14 is a graphical representation of stomatal conductance in wild type sugar cane and transgenic sugar cane possessing the constructs eFMVe35S-ZmUbil-scoabil, eFMVe35S-ZmUbil-scoABII, and the pUKN vector alone. [00421 Figure 15 is another graphical representation showing stomatal conductance 5 in transgenic sugar cane possessing constitutive expression of scoabil. Stomatal conductance was assessed in well watered control (vector backbone only) and transgenic scoabil plants. n = number of independenet transgenic plants analyzed. [00431 Figure 16 is a photographic representation showing wilty phenotype of a transgenic sugar cane plant possessing constitutive expression of scoabil (i.e., containing 10 eFMVe35S-ZmUbi]-scoabil) compared to a control plant (vector backbone only). [0044] Some figures and text contain color representations or entities. Color illustrations are available from the Applicant upon request or from an appropriate Patent Office. A fee may be imposed if obtained from a Patent Office. DETAILED DESCRIPTION OF THE INVENTION 15 1. Definitions [00451 Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, preferred 20 methods and materials are described. For the purposes of the present invention, the following terms are defined below. [00461 The articles "a" and "an" are used herein to refer to one or to more than one (i.e. to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element. Thus, for example, the term "cis-acting 25 sequence" also includes a plurality of cis-acting sequences. [0047] As used herein, "and/or" refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative (or). 100481 Further, the term "about," as used herein when referring to a measurable 30 value such as an amount of a compound or agent, dose, time, temperature, activity, level, number, frequency, percentage, dimension, size, amount, weight, position, length and the like, -8is meant to encompass variations of + 20%, ± 10%, ± 5%, ± 1%, ± 0.5%, or even ± 0.1% of the specified amount. [0049] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, 5 between the upper and lower limit of that range and any other stated or intervening value in that stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges is also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either both of those included 10 limits are also included in the invention. 100501 The term "antibody" is used to refer to any antibody-like molecule that has an antigen binding region, and includes antibody fragments such as Fab', Fab, F(ab') 2 , single domain antibodies (DABs), Fv, scFv (single chain Fv), and the like. 100511 The term "antisense" refers to a nucleotide sequence whose sequence of 15 nucleotide residues is in reverse 5' to 3' orientation in relation to the sequence of deoxynucleotide residues in a sense strand of a DNA duplex. A "sense strand" of a DNA duplex refers to a strand in a DNA duplex which is transcribed by a cell in its natural state into a "sense mRNA." Thus an "antisense' sequence is a sequence having the same sequence as the non-coding strand in a DNA duplex. The term "antisense RNA" refers to a RNA 20 transcript that is complementary to all or part of a target primary transcript or mRNA and that blocks the expression of a target gene by interfering with the processing, transport and/or translation of its primary transcript or mRNA. The complementarity of an antisense RNA may be with any part of the specific gene transcript, in other words, at the 5' non-coding sequence, 3' non-coding sequence, introns, or the coding sequence. In addition, as used herein, antisense 25 RNA may contain regions of ribozyme sequences that increase the efficacy of antisense RNA to block gene expression. "Ribozyme" refers to a catalytic RNA and includes sequence specific endoribonucleases. "Antisense inhibition" refers to the production of antisense RNA transcripts capable of preventing the expression of the target protein. 100521 The terms "cis-acting element," "cis-acting sequence" or "cis-regulatory 30 region" are used interchangeably herein to mean any sequence of nucleotides which modulates transcriptional activity of an operably linked promoter and/or expression of an operably linked nucleotide sequence. Those skilled in the art will be aware that a cis-sequence -9may be capable of activating, silencing, enhancing, repressing or otherwise altering the level of expression and/or cell-type-specificity and/or developmental specificity of any nucleotide sequence, including coding and non-coding sequences. [00531 By "coding sequence" is meant any nucleic acid sequence that contributes to 5 the code for the polypeptide product of a gene. By contrast, the term "non-coding sequence" refers to any nucleic acid sequence that does not contribute to the code for the polypeptide product of a gene. 100541 As used herein, "complementary" polynucleotides are those that are capable of hybridizing via base pairing according to the standard Watson-Crick complementarity 10 rules. Specifically, purines will base pair with pyrimidines to form a combination of guanine paired with cytosine (G:C) and adenine paired with either thymine (A:T) in the case of DNA, or adenine paired with uracil (A:U) in the case of RNA. For example, the sequence "A-G-T" binds to the complementary sequence "T-C-A." It is understood that two polynucleotides may hybridize to each other even if they are not completely or fully complementary to each other, 15 provided that each has at least one region that is substantially complementary to the other. The terms "complementary" or "complementarity," as used herein, refer to the natural binding of polynucleotides under permissive salt and temperature conditions by base-pairing. Complementarity between two single-stranded molecules may be "partial," in which only some of the nucleotides bind, or it may be complete when total complementarity exists 20 between the single stranded molecules either along the full length of the molecules or along a portion or region of the single stranded molecules. The degree of complementarity between nucleic acid strands has significant effects on the efficiency and strength of hybridization between nucleic acid strands. As used herein, the terms "substantially complementary" or "partially complementary" mean that two nucleic acid sequences are complementary at least 25 at about 50%, 60%, 70%, 80% or 90% of their nucleotides. In some embodiments, the two nucleic acid sequences can be complementary at least at about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more of their nucleotides. The terms "substantially complementary" and ''partially complementary" can also mean that two nucleic acid sequences can hybridize under high stringency conditions and such conditions are well known in the art. 30 [00551 Throughout this specification, unless the context requires otherwise, the words "comprise," "comprises" and "comprising" will be understood to imply the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step - 10or element or group of steps or elements. Thus, use of the term "comprising" and the like indicates that the listed elements are required or mandatory, but that other elements are optional and may or may not be present. By "consisting of' is meant including, and limited to, whatever follows the phrase "consisting of'. Thus, the phrase "consisting of' indicates that 5 the listed elements are required or mandatory, and that no other elements may be present. By "consisting essentially of' is meant including any elements listed after the phrase, and limited to other elements that do not interfere with or contribute to the activity or action specified in the disclosure for the listed elements. Thus, the phrase "consisting essentially of' indicates that the listed elements are required or mandatory, but that other elements are optional and 10 may or may not be present depending upon whether or not they affect the activity or action of the listed elements. Thus, the term "consisting essentially of' when used in a claim of this invention is not intended to be interpreted to be equivalent to "comprising." Thus, the term "consisting essentially of' (and grammatical variants), as applied to a nucleic acid sequence of this invention, means a polynucleotide that consists of both the recited sequence (e.g., SEQ ID 15 NO) and a total of twenty or less (e.g. , 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20) additional nucleotides on the 5' and/or 3' ends of the recited sequence such that the function of the polynucleotide is not materially altered. The total of twenty or less additional nucleotides includes the total number of additional nucleotides on both ends added together. 20 100561 As used herein the term "constitutively active" refers to a protein that is always active, i.e., the physiological effect of the protein is always obtained even in the absence of an activator of that protein. Thus, for example, when applied to an ATPase, the term "constitutively active" refers to an ATPase that has the ability to catalyze the hydrolysis of ATP to ADP in the absence of an activator of the ATPase. 25 100571 The term "construct" refers to a recombinant genetic molecule including one or more isolated nucleic acid sequences from different sources. As used herein, the term "expression construct," "recombinant construct" or "recombinant DNA construct" refers to any recombinant nucleic acid molecule such as a plasmid, cosmid, virus, autonomously replicating polynucleotide molecule, phage, or linear or circular single-stranded or double 30 stranded DNA or RNA nucleic acid molecule, derived from any source, capable of genomic integration or autonomous replication, comprising a nucleic acid molecule where one or more nucleic acid molecules have been operably linked. An "expression construct" generally includes at least a control sequence operably linked to a nucleotide sequence of interest. In - 1 this manner, for example, plant promoters in operable connection with the nucleotide sequences to be expressed are provided in expression constructs for expression in a plant, plant part, plant organ and/or plant cell. Methods are known for introducing constructs into a cell in such a manner that a transcribable polynucleotide molecule is transcribed into a 5 functional mRNA molecule that is translated and therefore expressed as a protein product. Constructs may also be made to be capable of expressing inhibitory RNA molecules in order, for example, to inhibit translation of a specific RNA molecule of interest. For the practice of the present invention, conventional compositions and methods for preparing and using constructs and host cells are well known to one skilled in the art, see for example, Molecular 10 Cloning: A Laboratory Manual, 3.sup.rd edition Volumes 1, 2, and 3. J. F. Sambrook, D. W. Russell, and N. Irwin, Cold Spring Harbor Laboratory Press, 2000. [00581 By "corresponds to" or "corresponding to" is meant a nucleic acid sequence that displays substantial sequence identity to a reference nucleic acid sequence (e.g., at least about 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 15 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 97, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or even up to 100% sequence identity to all or a portion of the reference nucleic acid sequence) or an amino acid sequence that displays substantial sequence similarity or identity to a reference amino acid sequence (e.g., at least 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 20 97, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or even up to 100% sequence similarity or identity to all or a portion of the reference amino acid sequence). 100591 "Dominant negative" refers to a gene product that adversely affects, blocks or abrogates the function of a normal, wild-type gene product when co-expressed with the wild type gene product within the same cell even when the cell is heterozygous (wild-type and 25 dominant negative). Expression of the dominant negative mutant generally results in a decrease in normal function of the wild-type gene product. 100601 The term "dominant positive" refers to a gene product that partially or fully mimics the function of a normal, wild-type gene product (and thus possesses the same activity) when co-expressed with the wild type gene product within the same cell even when 30 the cell is heterozygous (wild-type and dominant positive). Expression of the dominant positive mutant generally results in an increase in normal function of the wild-type gene product. - 12 - 100611 As used herein, the terms "encode," "encoding" and the like refer to the capacity of a nucleic acid to provide for another nucleic acid or a polypeptide. For example, a nucleic acid sequence is said to "encode" a polypeptide if it can be transcribed and/or translated to produce the polypeptide or if it can be processed into a form that can be 5 transcribed and/or translated to produce the polypeptide. Such a nucleic acid sequence may include a coding sequence or both a coding sequence and a non-coding sequence. Thus, the terms "encode," "encoding" and the like include an RNA product resulting from transcription of a DNA molecule, a protein resulting from translation of an RNA molecule, a protein resulting from transcription of a DNA molecule to form an RNA product and the subsequent 10 translation of the RNA product, or a protein resulting from transcription of a DNA molecule to provide an RNA product, processing of the RNA product to provide a processed RNA product (e.g., mRNA) and the subsequent translation of the processed RNA product. 100621 The term "endogenous" refers to any polynucleotide or polypeptide which is present and/or naturally expressed within a plant or a cell thereof. For example, an 15 "endogenous" nucleic acid refers to a nucleic acid molecule or nucleotide sequence that is naturally found in the cell into which a construct of the invention is introduced. 100631 The term "expression" with respect to a gene sequence refers to transcription of the gene and, as appropriate, translation of the resulting mRNA transcript to a protein. Thus, as will be clear from the context, expression of a coding sequence results from 20 transcription and translation of the coding sequence. Conversely, expression of a non-coding sequence results from the transcription of the non-coding sequence. [0064] As used herein, the terms "fragment" or "portion" when used in reference to a nucleic acid molecule or nucleotide sequence will be understood to mean a nucleic acid molecule or nucleotide sequence of reduced length relative to a reference nucleic acid 25 molecule or nucleotide sequence and comprising, consisting essentially of and/or consisting of a nucleotide sequence of contiguous nucleotides identical or almost identical (e.g., 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 98%, 99% identical) to the reference nucleic acid or nucleotide sequence. Such a nucleic acid fragment according to the invention may be, where appropriate, included in a larger 30 polynucleotide of which it is a constituent. [00651 As used herein, the term "gene" refers to a nucleic acid molecule capable of being used to produce mRNA, antisense RNA, siRNA, shRNA, miRNA, and the like. Genes - 13may or may not be capable of being used to produce a functional protein. Genes can include both coding and non-coding regions (e.g., introns, regulatory elements, promoters, enhancers, termination sequences and 5' and 3' untranslated regions). A gene may be "isolated" by which is meant a nucleic acid molecule that is substantially or essentially free from components 5 normally found in association with the nucleic acid molecule in its natural state. Such components include other cellular material, culture medium from recombinant production, and/or various chemicals used in chemically synthesizing the nucleic acid molecule. 100661 "Genome" as used herein includes the nuclear and/or plastid genome, and therefore includes integration of the nucleic acid into, for example, the chloroplast genome. 10 [0067] The term "guard cell" refers to specialized epidermal cells that regulate the aperture (i.e., the opening and closing) of stomata and by this control the bulk of gas exchange as well as transpiration. These pairs of bean-like shaped cells are characterized by their highly regulated turgor (i.e., pressure-dependent shape), which causes the stomata to close or to open at states of low or high turgor, respectively. Guard cells derive from epidermal cells and are 15 evenly spaced in the epidermis, i.e., the outermost cell layer of plant organs. Guard cells differ from their surrounding epidermal cells not only by shape but also by their ability to photosynthesize. 10068] "Guard cell-specific promoter" as used herein refers to a promoter that transcribes an operably connected nucleic acid sequence in a way that transcription of the 20 nucleic acid sequence in guard-cells contribute to more than 90%, 95%, 99% of the entire quantity of the RNA transcribed from said nucleic acid sequence in the entire plant during any of its developmental stages. [00691 "Guard cell-preferential promoter" in the context of this invention refers to a promoter that transcribes an operably connected nucleic acid sequence in a way that 25 transcription of the nucleic acid sequence in guard-cells contribute to more than 50%, preferably more than 70%, more preferably more than 80% of the entire quantity of the RNA transcribed from said nucleic acid sequence in the entire plant during any of its developmental stages. [00701 The term "heterologous" as used herein with reference to nucleic acids 30 refers to a nucleic acid molecule or nucleotide sequence that either originates from another species or is from the same species or organism but is modified from either its original form or the form primarily expressed in the cell. Thus, a nucleotide sequence derived from an - 14organism or species different from that of the cell into which the nucleotide sequence is introduced, is heterologous with respect to that cell and the cell's descendants. In addition, a heterologous nucleotide sequence includes a nucleotide sequence derived from and inserted into the same natural, original cell type, but which is present in a non-natural state, e.g. 5 present in a different copy number, and/or under the control of different regulatory sequences than that found in the native state of the nucleic acid molecule. The term "heterologous" when used with reference to portions of a nucleic acid indicates that the nucleic acid comprises two or more subsequences that are not found in the same relationship to each other in nature. For instance, a nucleic acid may be recombinantly produced, having two or more sequences from 10 unrelated genes arranged to make a new functional nucleic acid, e.g., a nucleic acid encoding a protein from one source and a nucleic acid encoding a peptide sequence from another source. Similarly, a "heterologous" protein indicates that the protein comprises two or more subsequences that are not found in the same relationship to each other in nature (e.g., a fusion protein). 15 100711 As used herein the term "homology" refers to the level of similarity between two or more nucleotide sequences and/or amino acid sequences in terms of percent of positional identity (i.e., sequence similarity or identity). Different nucleotide sequences or polypeptide sequences having homology are referred to herein as "homologs." The term homolog includes homologous sequences from the same and other species and orthologous 20 sequences from the same and other species. Homology also refers to the concept of similar functional properties among different nucleic acids, amino acids, and/or proteins. 100721 Reference herein to "immuno-interactive" includes reference to any interaction, reaction, or other form of association between molecules and in particular where one of the molecules is, or mimics, a component of the immune system. 25 [00731 "Introducing" in the context of a plant cell, plant part and/or plant organ means contacting a nucleic acid molecule with the plant, plant part, and/or plant cell in such a manner that the nucleic acid molecule gains access to the interior of the plant cell and/or a cell of the plant and/or plant part. Where more than one nucleic acid molecule is to be introduced these nucleic acid molecules can be assembled as part of a single polynucleotide or nucleic 30 acid construct, or as separate polynucleotide or nucleic acid constructs, and can be located on the same or different nucleic acid constructs. Accordingly, these polynucleotides can be introduced into plant cells in a single transformation event, in separate transformation events, - 15 or, e.g., as part of a breeding protocol. Thus, the term "transformation" as used herein refers to the introduction of a heterologous nucleic acid into a cell. Transformation of a cell may be stable or transient. "Transient transformation" in the context of a polynucleotide means that a polynucleotide is introduced into the cell and does not integrate into the genome of the cell. 5 By "stably introducing" or "stably introduced" in the context of a polynucleotide introduced into a cell, it is intended that the introduced polynucleotide is stably incorporated into the genome of the cell, and thus the cell is stably transformed with the polynucleotide. "Stable transformation" or "stably transformed" as used herein means that a nucleic acid molecule is introduced into a cell and integrates into the genome of the cell. As such, the integrated 10 nucleic acid molecule is capable of being inherited by the progeny thereof, more particularly, by the progeny of multiple successive generations. Stable transformation as used herein can also refer to a nucleic acid molecule that is maintained extrachromosomally, for example, as a minichromosome. 100741 An "isolated" nucleic acid molecule or nucleotide sequence or nucleic acid 15 construct or double stranded RNA molecule of the present invention is generally free of nucleotide sequences that flank the nucleic acid of interest in the genomic DNA of the organism from which the nucleic acid was derived (such as coding sequences present at the 5' or 3' ends). However, the nucleic acid molecule of this invention can include some additional bases or moieties that do not deleteriously or materially affect the basic structural and/or 20 functional characteristics of the nucleic acid molecule. [00751 Thus, an "isolated nucleic acid molecule" or "isolated nucleotide sequence" is a nucleic acid molecule or nucleotide sequence that is not immediately contiguous with nucleotide sequences with which it is immediately contiguous (one on the 5' end and one on the 3' end) in the naturally occurring genome of the organism from which it is derived. 25 Accordingly, in some embodiments, an isolated nucleic acid includes some or all of the 5' noncoding (e.g., promoter) sequences that are immediately contiguous to a coding sequence. The term therefore includes, for example, a recombinant nucleic acid that is incorporated into a vector, into an autonomously replicating plasmid or virus, or into the genomic DNA of a prokaryote or eukaryote, or which exists as a separate molecule (e.g., a cDNA or a genomic 30 DNA fragment produced by PCR or restriction endonuclease treatment), independent of other sequences. It also includes a recombinant nucleic acid that is part of a hybrid nucleic acid molecule encoding an additional polypeptide or peptide sequence. The term "isolated" can further refer to a nucleic acid molecule, nucleotide sequence, polypeptide, peptide or fragment -16that is substantially free of cellular material, viral material, and/or culture medium (e.g., when produced by recombinant DNA techniques), or chemical precursors or other chemicals (e.g., when chemically synthesized). Moreover, an "isolated fragment" is a fragment of a nucleic acid molecule, nucleotide sequence or polypeptide that is not naturally occurring as a 5 fragment and would not be found as such in the natural state. "Isolated" does not mean that the preparation is technically pure (homogeneous), but it is sufficiently pure to provide the polypeptide or nucleic acid in a form in which it can be used for the intended purpose. Accordingly, "isolated" refers to a nucleic acid molecule, nucleotide sequence, polypeptide, peptide or fragment that is altered "by the hand of man" from the natural state; i.e., that, if it 10 occurs in nature, it has been changed or removed from its original environment, or both. For example, a naturally occurring polynucleotide or a polypeptide naturally present in a living organism in its natural state is not "isolated," but the same polynucleotide or polypeptide separated from the coexisting materials of its natural state is "isolated," as the term is employed herein. For example, with respect to polynucleotides, the term isolated means that it 15 is separated from the chromosome and/or cell in which it naturally occurs. A polynucleotide is also isolated if it is separated from the chromosome and/or cell in which it naturally occurs in and is then inserted into a genetic context, a chromosome and/or a cell in which it does not naturally occur. In representative embodiments of the invention, an "isolated" nucleic acid molecule, nucleotide sequence, and/or polypeptide is at least about 5%, 10%, 15%, 20%, 20 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99% pure (w/w) or more. In other embodiments, an "isolated" nucleic acid, nucleotide sequence, and/or polypeptide indicates that at least about a 5-fold, 10-fold, 25-fold, 100-fold, 1000-fold, I 0,000-fold, 100,000-fold or more enrichment of the nucleic acid (w/w) is achieved as compared with the starting material. 25 [00761 The term, "microRNA" or "miRNAs" refer to small, noncoding RNA molecules that have been found in a diverse array of eukaryotes, including plants. miRNA precursors share a characteristic secondary structure, forming short 'hairpin' RNAs. The term "miRNA" includes processed sequences as well as corresponding long primary transcripts (pri-miRNAs) and processed precursors (pre-miRNAs). Genetic and biochemical studies have 30 indicated that miRNAs are processed to their mature forms by Dicer, an RNAse III family nuclease, and function through RNA-mediated interference (RNAi) and related pathways to regulate the expression of target genes (Hannon (2002) Nature 418, 244-251; Pasquinelli, et al. (2002) Annu. Rev. Cell. Dev. Biol. 18, 495-513). miRNAs may be configured to permit -17experimental manipulation of gene expression in cells as synthetic silencing triggers 'short hairpin RNAs' (shRNAs) (Paddison et al. (2002) Cancer Cell 2, 17-23). Silencing by shRNAs involves the RNAi machinery and correlates with the production of small interfering RNAs (siRNAs), which are a signature of RNAi. 5 [00771 The term "non-coding" refers to sequences of nucleic acid molecules that do not encode part or all of an expressed protein. Non-coding sequences include but are not limited to introns, enhancers, promoter regions, 3' untranslated regions, and 5' untranslated regions. Thus, the term "5'-non-coding region" shall be taken in its broadest context to include all nucleotide sequences which are derived from the upstream region of a gene. Such regions 10 may include an intron, e.g., an intron. As used herein, the term "3' non-coding region" refers to nucleic acid sequences located downstream of a coding sequence and include polyadenylation recognition sequences (normally limited to eukaryotes) and other sequences encoding regulatory signals capable of affecting mRNA processing or gene expression. The polyadenylation signal (normally limited to eukaryotes) is usually characterized by affecting 15 the addition of polyadenylic acid tracts to the 3' end of the mRNA precursor. [00781 As used herein, the term "nucleotide sequence" refers to a heteropolymer of nucleotides or the sequence of these nucleotides from the 5' to 3' end of a nucleic acid molecule and includes DNA or RNA molecules, including cDNA, a DNA fragment, genomic DNA, synthetic (e.g., chemically synthesized) DNA, plasmid DNA, mRNA, and anti-sense 20 RNA, any of which can be single stranded or double stranded. The terms "nucleotide sequence" "nucleic acid," "nucleic acid molecule," "oligonucleotide" and "polynucleotide" are also used interchangeably herein to refer to a heteropolymer of nucleotides. Nucleic acid sequences provided herein are presented herein in the 5' to 3' direction, from left to right and are represented using the standard code for representing the nucleotide characters as set forth 25 in the U.S. sequence rules, 37 CFR 1.821 - 1.825 and the World Intellectual Property Organization (WIPO) Standard ST.25. [00791 The term "operably connected" or "operably linked" as used herein refers to a juxtaposition wherein the components so described are in a relationship permitting them to function in their intended manner. For example, a control sequence (e.g., a promoter) 30 "operably linked" to a coding sequence refers to positioning and/or orientation of the control sequence relative to the coding sequence to permit expression of the coding sequence under conditions compatible with the control sequence. The control sequences need not be - 18contiguous with the nucleotide sequence of interest, so long as they function to direct the expression thereof. Thus, for example, intervening untranslated, yet transcribed, sequences can be present between a promoter and a coding sequence, and the promoter sequence can still be considered "operably linked" to the coding sequence. Likewise, "operably connecting" 5 a cis-acting sequence to a promoter encompasses positioning and/or orientation of the cis acting sequence relative to the promoter so that (1) the cis-acting sequence regulates (e.g., inhibits, abrogates, stimulates or enhances) promoter activity. 100801 As used herein, "plant" means any plant and thus includes, for example, angiosperms (monocots and dicots), gymnosperms, bryophytes, ferns and/or fern allies. Non 10 limiting examples of monocot plants of the present invention include sugar cane, corn, barley, rye, oats, wheat, rice, flax, millet, sorghum, grasses, banana, onion, asparagus, lily, coconut, and the like. [00811 As used herein, the term "plant part" includes but is not limited to embryos, pollen, ovules, seeds, leaves, flowers, branches, fruit, kernels, ears, cobs, husks, stalks, roots, 15 root tips, anthers, plant cells including plant cells that are intact in plants and/or parts of plants, plant protoplasts, plant tissues, plant cell tissue cultures, plant calli, plant clumps, and the like. 100821 As used herein, "plant cell" refers to a structural and physiological unit of the plant, which comprises a cell wall and also may refer to a protoplast. A plant cell of the 20 present invention can be in the form of an isolated single cell or can be a cultured cell or can be a part of a higher-organized unit such as, for example, a plant tissue or a plant organ. 100831 The term "plant organ" refers to plant tissue or group of tissues that constitute a morphologically and functionally distinct part of a plant. [00841 As used herein , the terms "polynucleotide," "polynucleotide sequence," 25 "nucleotide sequence," "nucleic acid," "nucleic acid molecule," "nucleic acid sequence"s and the like refer to RNA or DNA that is linear or branched, single or double stranded, or a hybrid thereof. The term also encompasses RNA/DNA hybrids. The term typically refers to polymeric form of nucleotides of at least 10 bases in length, either ribonucleotides or deoxynucleotides or a modified form of either type of nucleotide. The term includes single 30 and double stranded forms of RNA or DNA. When dsRNA is produced synthetically, less common bases, such as inosine, 5-methylcytosine, 6- methyladenine, hypoxanthine and others can also be used for antisense, dsRNA, and ribozyme pairing. For example, polynucleotides - 19that contain C-5 propyne analogues of uridine and cytidine have been shown to bind RNA with high affinity and to be potent antisense inhibitors of gene expression. Other modifications, such as modification to the phosphodiester backbone, or the 2'-hydroxy in the ribose sugar group of the RNA can also be made. 5 [00851 "Polypeptide," "peptide," "protein" and "proteinaceous molecule" are used interchangeably herein to refer to molecules comprising or consisting of a polymer of amino acid residues and to variants and synthetic analogues of the same. Thus, these terms apply to amino acid polymers in which one or more amino acid residues are synthetic non-naturally occurring amino acids, such as a chemical analogue of a corresponding naturally occurring 10 amino acid, as well as to naturally-occurring amino acid polymers. This term also includes within its scope two or more complementing or interactive polypeptides comprising different parts or portions (e.g., polypeptide domains, polypeptide chains etc.) of a polypeptide of the present invention, wherein the individual complementing polypeptides together reconstitute the activity of the different parts or portions to form a functional polypeptide. 15 [0086] As used herein, the term "polypeptide that inhibits stomatal closure" refers to polypeptides that interfere, impair, reduce or otherwise inhibit stomatal closure (e.g., ABA induced stomatal closure), or that stimulate or enhance stomatal opening. 100871 As used herein, the term "post-transcriptional gene silencing" (PTGS) refers to a form of gene silencing in which the inhibitory mechanism occurs after transcription. This 20 can result in either decreased steady-state level of a specific RNA target or inhibition of translation (Tuschl el al. (2001) ChemBiochem 2: 239-245). In the literature, the terms RNA interference (RNAi) and posttranscriptional co-suppression are often used to indicate posttranscriptional gene silencing. [00881 As used herein, the term "promoter" refers to a region of a nucleotide 25 sequence that incorporates the necessary signals for the expression of a coding sequence operably associated with the promoter. This may include sequences to which an RNA polymerase binds, but is not limited to such sequences and can include regions to which other regulatory proteins bind, together with regions involved in the control of protein translation and can also include coding sequences. Furthermore, a "promoter" of this invention is a 30 promoter (e.g., a nucleotide sequence) capable of initiating transcription of a nucleic acid molecule in a cell of a plant. -20- 100891 "Promoter activity" refers to the ability of a promoter to drive expression of a nucleic acid sequence operably linked to the promoter. Promoter activity of a sequence can be assessed by operably linking the sequence to a reporter gene, and determining expression of the reporter. 5 100901 The term "recombinant polynucleotide" refers to a polynucleotide that has been altered, rearranged, or modified by genetic engineering. Examples include any cloned polynucleotide, or polynucleotides, that are linked or joined to heterologous sequences. However, it shall be understood that the term "recombinant" does not refer to alterations of polynucleotides that result from naturally occurring events, such as spontaneous mutations, or 10 from non-spontaneous mutagenesis followed by selective breeding. [00911 As used herein, the terms "RNA interference" and "RNAi" refer to a sequence-specific process by which a target molecule (e.g., a target gene, protein or RNA) is downregulated via downregulation of expression. Without being bound to a specific mechanism, as currently understood by those of skill in the art, RNAi involves degradation of 15 RNA molecules, e.g., mRNA molecules within a cell, catalyzed by an enzymatic, RNA induced silencing complex (RISC). RNAi occurs in cells naturally to remove foreign RNAs (e.g., viral RNAs) triggered by dsRNA fragments cleaved from longer dsRNA which direct the degradative mechanism to other RNA sequences having closely homologous sequences. As practiced as a technology, RNAi can be initiated by human intervention to reduce or even 20 silence the expression of target genes using either exogenously synthesized dsRNA or dsRNA transcribed in the cell (e.g., synthesized as a sequence that forms a short hairpin structure). [00921 As used herein, the terms "small interfering RNA" and "short interfering RNA" ("siRNA") refer to a short RNA molecule, generally a double-stranded RNA molecule about 10-50 nucleotides in length (the term "nucleotides" including nucleotide analogs), 25 preferably between about 15-25 nucleotides in length. In most cases, the siRNA is 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleotides in length. Such siRNA can have overhanging ends (e.g., 3-overhangs of 1, 2, or 3 nucleotides (or nucleotide analogs). Such siRNA can mediate RNA interference. [00931 As used in connection with the present invention, the term "shRNA" refers 30 to an RNA molecule having a stem-loop structure. The stem-loop structure includes two mutually complementary sequences, where the respective orientations and the degree of complementarity allow base pairing between the two sequences. The mutually complementary -21 sequences are linked by a loop region, the loop resulting from a lack of base pairing between nucleotides (or nucleotide analogs) within the loop region. 100941 The term "sequence identity" as used herein refers to the extent that sequences are identical on a nucleotide-by-nucleotide basis or an amino acid-by-amino acid 5 basis over a window of comparison. Thus, a "percentage of sequence identity" is calculated by comparing two optimally aligned sequences over the window of comparison, determining the number of positions at which the identical nucleic acid base (e.g., A, T, C, G, I) or the identical amino acid residue (e.g., Ala, Pro, Ser, Thr, Gly, Val, Leu, Ile, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gln, Cys and Met) occurs in both sequences to yield the number of 10 matched positions, dividing the number of matched positions by the total number of positions in the window of comparison (i.e., the window size), and multiplying the result by 100 to yield the percentage of sequence identity. For the purposes of the present invention, "sequence identity" will be understood to mean the "match percentage" calculated by the DNASIS computer program (Version 2.5 for windows; available from Hitachi Software 15 engineering Co., Ltd., South San Francisco, California, USA) using standard defaults as used in the reference manual accompanying the software. Useful methods for determining sequence identity are also disclosed in Guide to Huge Computers (Martin J. Bishop, ed., Academic Press, San Diego (1994)), and Carillo et al. (Applied Math 48:1073(1988)). More particularly, preferred computer programs for determining sequence identity include but are 20 not limited to the Basic Local Alignment Search Tool (BLAST) programs which are publicly available from National Center Biotechnology Information (NCBI) at the National Library of Medicine, National Institute of Health, Bethesda, Md. 20894; see BLAST Manual, Altschul et al., NCBI, NLM, NIH; (Altschul et al., J. Mol. Biol. 215:403-410 (1990)); version 2.0 or higher of BLAST programs allows the introduction of gaps (deletions and insertions) into 25 alignments; for peptide sequence BLASTX can be used to determine sequence identity; and for polynucleotide sequence BLASTN can be used to determine sequence identity. [00951 "Similarity" refers to the percentage number of amino acids that are identical or constitute conservative substitutions as defined in Table A below. Similarity may be determined using sequence comparison programs such as GAP (Deveraux et al. 1984, 30 Nucleic Acids Research 12: 387-395). In this way, sequences of a similar or substantially different length to those cited herein might be compared by insertion of gaps into the alignment, such gaps being determined, for example, by the comparison algorithm used by GAP. - 22 - TABLE A: EXEMPLARY CONSERVATIVE AMINO ACID SUBSTITUTIONS Original Residue Exemplary Substitutions Ala Ser Arg Lys Asn Gin, His Asp Glu Cys Ser Gin Asn Glu Asp Gly Pro His Asn, Gin Ile Leu, Val Leu Ile, Val Lys Arg, Gin, Glu Met Leu, Ile, Phe Met, Leu, Tyr Ser Thr Thr Ser Trp Tyr Tyr Trp, Phe Val Ile, Leu 100961 Terms used to describe sequence relationships between two or more polynucleotides or polypeptides include "reference sequence," "comparison window", 5 "sequence identity," "percentage of sequence identity" and "substantial identity". A "reference sequence" is at least 12 but frequently 15 to 18 and often at least 25 monomer units, inclusive of nucleotides and amino acid residues, in length. Because two polynucleotides may each comprise (1) a sequence (i.e., only a portion of the complete polynucleotide sequence) that is similar between the two polynucleotides, and (2) a sequence 10 that is divergent between the two polynucleotides, sequence comparisons between two (or -23more) polynucleotides are typically performed by comparing sequences of the two polynucleotides over a "comparison window" to identify and compare local regions of sequence similarity. A "comparison window" refers to a conceptual segment of at least 6 contiguous positions, usually about 50 to about 100, more usually about 100 to about 150 in 5 which a sequence is compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. The comparison window may comprise additions or deletions (i.e., gaps) of about 20% or less as compared to the reference sequence (which does not comprise additions or deletions) for optimal alignment of the two sequences. Optimal alignment of sequences for aligning a comparison window may be 10 conducted by computerized implementations of algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package Release 7.0, Genetics Computer Group, 575 Science Drive Madison, WI, USA) or by inspection and the best alignment (i.e., resulting in the highest percentage homology over the comparison window) generated by any of the various methods selected. Reference also may be made to the BLAST family of 15 programs as for example disclosed by Altschul et al., 1997, Nucl. Acids Res.25:3389. A detailed discussion of sequence analysis can be found in Unit 19.3 of Ausubel et al., "Current Protocols in Molecular Biology", John Wiley & Sons Inc, 1994-1998, Chapter 15. [00971 As used herein, the terms "transformed" and "transgenic" refer to any plant, plant cell, callus, plant tissue, or plant part that contains all or part of at least one isolated or 20 recombinant (e.g., heterologous) polynucleotide. In some embodiments, all or part of the isolated or recombinant polynucleotide is stably integrated into a chromosome or stable extra chromosomal element, so that it is passed on to successive generations. [0098] The term "transgene" as used herein, refers to any nucleotide sequence used in the transformation of a plant, animal, or other organism. Thus, a transgene can be a coding 25 sequence, a non-coding sequence, a cDNA, a gene or fragment or portion thereof, a genomic sequence, a regulatory element and the like. A "transgenic" organism, such as a transgenic plant, transgenic microorganism, or transgenic animal, is an organism into which a transgene has been delivered or introduced and the transgene can be expressed in the transgenic organism to produce a product, the presence of which can impart an effect and/or a phenotype 30 in the organism. 100991 As used herein, the term "5' untranslated region" or "5' UTR" refers to a sequence located upstream (i.e., 5') of a coding region. Typically, a 5' UTR is located -24downstream (i.e., 3') to a promoter region and 5' of a coding region downstream of the promoter region. Thus, such a sequence, while transcribed, is upstream of the translation initiation codon and therefore is generally not translated into a portion of the polypeptide product. 5 [01001 The term "3' untranslated region" or "3' UTR" refers to a nucleotide sequence downstream (i.e., 3') of a coding sequence. It generally extends from the first nucleotide after the stop codon of a coding sequence to just before the poly(A) tail of the corresponding transcribed mRNA. The 3' UTR may contain sequences that regulate translation efficiency, mRNA stability, mRNA targeting and/or polyadenylation. 10 [01011 The terms "wild-type," "natural," "native" and the like with respect to an organism, polypeptide, or nucleic acid sequence, that the organism polypeptide, or nucleic acid sequence is naturally occurring or available in at least one naturally occurring organism which is not changed, mutated, or otherwise manipulated by man. [01021 As used herein, underscoring or italicizing the name of a gene shall indicate 15 the gene, in contrast to its protein product, which is indicated in the absence of any underscoring or italicizing. For example, "A B11" shall mean the ABI1 gene, whereas "ABI I" shall indicate the protein product of the "A BI1" gene. 10103] Each embodiment described herein is to be applied mutatis mutandis to each and every embodiment unless specifically stated otherwise. 20 2. Stomata closure-modulating constructs 101041 The present invention is based in part on a novel cis-acting element that confers a pattern of inducibility on a promoter, which is similar to that of the alcohol dehydrogenase (ADH 1) system of A. nidulans. Thus, when operably linked to this cis-acting element, a promoter that is not already inducible by contact with a chemical compound that 25 can induce the expression of the alcohol dehydrogenase system of A. nidulans is made so inducible. In accordance with the present invention, the resulting inducible promoter is useful for expressing polynucleotides that comprise a nucleic acid sequence encoding an expression product that inhibits stomatal closure. 2.1 Cis-acting element 30 [0105] In some embodiments, the cis-acting element comprises one or more nucleotide sequences selected from the group consisting of a nucleotide sequence of SEQ ID - 25 - NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and SEQ ID NO:9 in any combination, in any orientation, and/or in any order, including but not limited to multiples of the same nucleotide sequence. [0106] The cis-acting element, e.g., the nucleotide sequence of SEQ ID NO:1, SEQ 5 ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, or SEQ ID NO:9, comprises, consists or consists essentially of, inverted repeats of the alcR inverted repeat binding sites, or variants thereof, of the A. nidulans alcohol dehydrogenase system (ADH 1). Thus, SEQ ID NO:1 provides the nucleotide sequence GCGGNNCCGC (inverted repeat underlined), which suitably represents the minimal cis 10 acting sequence. [01071 SEQ ID NO:2 provides the nucleotide sequence of nGCGGNNCCGCny. N or n can be independently any nucleic acid base (A, G, C, or T), and x and y can be independently any number, as set forth below. 101081 Additional cis-acting sequences are as follows: 15 ATGCATGCGGAACCGCACGAGG [SEQ ID NO:3], GGCCATGCGGAGCCGCACGCGT [SEQ ID NO:4], ACAAGAGCGGCTCCGCTTGACC [SEQ ID NO:5]; TACGTAGCGGAACCGCTGCTCC [SEQ ID NO:6]; TACCATGCGGAACCGCACGTCC [SEQ ID NO:7], ATGCATGCGGTGCCGCACGAGG [SEQ ID NO:8] and TACGTTGCGGAACCGCAGCTCC [SEQ ID NO:9]. 20 [0109] In some embodiments, the cis-acting elements can comprise one or more nucleotides (i.e., bases) between the inverted repeats (e.g., an intervening sequence). Thus, in certain embodiments, the number of nucleotides comprising the intervening sequence can be two (i.e., N=2) (e.g., SEQ ID NO:1 = nxGCGGNNCCGCny (intervening sequence bolded and underlined)). Non-limiting examples of the intervening sequence include TT, AA, GG, CC, 25 TA, TG, TC, AT, AG, AC, GT,GC, GA, CA, CT, or CG, and the like. Thus, any 2-mer can be used as an intervening sequence in SEQ ID NO: 1 or 2, respectively. 101101 In some embodiments, SEQ ID NO:2 (RGCGGNNCCGCp (flanking sequences bolded and underlined)) of the present invention can comprise zero to 100 or more nucleotides (i.e., bases) that flank the left (i.e., flanking sequence, nx) and/or right side of the 30 inverted repeats (i.e., flanking sequence, ny). The flanking sequences (i.e., nx or ny) can be of any length and/or composition of nucleotides, wherein each nucleotide can be independently adenine, thymine, guanine and/or cytosine (i.e., wherein n=A, T, G, and/or C), in any -26combination and/or in any order. Thus, in some embodiments of the invention, x and y are each independently zero to 100 nucleotides. The number of nucleotides in a flanking sequence can suitably be 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 5 49,50,51,52,53,54,55,56,57,58,59,60,61,62,63,64,65,66,67,68,69,70,71,72,73, 74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97,98, 99, 100, or more, wherein each nucleotide in a flanking sequence can be independently adenine, thymine, guanine and/or cytosine, in any combination and/or in any order. As one of ordinary skill in the art would appreciate, flanking sequences of any length and composition 10 can be produced that are included within SEQ ID NO:2 according to art-known methods. 101111 In some embodiments, the cis-acting element can comprise, consist or consist essentially of one or more of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9. Thus, in some embodiments, the cis-acting element can comprise, consist or consist 15 essentially of, for example, one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty etc., nucleotide sequences of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9, in any combination, in any orientation, and/or in any order. Thus, in some embodiments, the cis 20 acting element can comprise a multimer of any one or more of the nucleotide sequences of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9, wherein the nucleotide sequences of the multimer (e.g., the cis-acting element) can be the same and/or different from one another, in any combination, in any orientation, and/or in any order. 25 101121 In some embodiments, a cis-acting element comprising one nucleotide sequence of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9, does not comprise only one nucleotide sequence of SEQ ID NO:3, SEQ ID NO:4, or SEQ ID NO:5. 10113] In some embodiments, the number of nucleotide sequences in a cis-acting 30 element can be in a range from one nucleotide sequence to about nine nucleotide sequences, from one nucleotide sequence to about ten nucleotide sequences, from one nucleotide sequence to about eleven nucleotide sequences, from one nucleotide sequence to about twelve - 27 nucleotide sequences, from one nucleotide sequence to about thirteen nucleotide sequences, from one nucleotide sequence to about fourteen nucleotide sequences, from one nucleotide sequence to about fifteen nucleotide sequences, from one nucleotide sequence to about sixteen nucleotide sequences, from one nucleotide sequence to about seventeen nucleotide sequences, 5 from one nucleotide sequence to about eighteen nucleotide sequences, from one nucleotide sequence to about nineteen nucleotide sequences, from one nucleotide sequence to about twenty nucleotide sequences, from about two nucleotide sequences to about five nucleotide sequences, from about two nucleotide sequences to about seven nucleotide sequences, from about two nucleotide sequences to about nine nucleotide sequences, from about two 10 nucleotide sequences to about ten nucleotide sequences, from about two nucleotide sequences to about eleven nucleotide sequences, from about two nucleotide sequences to about twelve nucleotide sequences, from about two nucleotide sequences to about thirteen nucleotide sequences, from about two nucleotide sequences to about fourteen nucleotide sequences, from about two nucleotide sequences to about fifteen nucleotide sequences, from about two 15 nucleotide sequence to about sixteen nucleotide sequences, from about two nucleotide sequence to about seventeen nucleotide sequences, from about two nucleotide sequence to about eighteen nucleotide sequences, from about two nucleotide sequence to about nineteen nucleotide sequences, from about two nucleotide sequence to about twenty nucleotide sequences, from about three nucleotide sequences to about five nucleotide sequences, from 20 about three nucleotide sequences to about seven nucleotide sequences, from about three nucleotide sequences to about nine nucleotide sequences, from about three nucleotide sequences to about ten nucleotide sequences, from about three nucleotide sequences to about eleven nucleotide sequences, from about three nucleotide sequences to about twelve nucleotide sequences, from about three nucleotide sequences to about thirteen nucleotide 25 sequences, from about three nucleotide sequences to about fourteen nucleotide sequences, from about three nucleotide sequences to about fifteen nucleotide sequences, from about three nucleotide sequence to about sixteen nucleotide sequences, from about three nucleotide sequence to about seventeen nucleotide sequences, from about three nucleotide sequence to about eighteen nucleotide sequences, from about three nucleotide sequence to about nineteen 30 nucleotide sequences, from about three nucleotide sequence to about twenty nucleotide sequences, from about four nucleotide sequences to about nine nucleotide sequences, from about four nucleotide sequences to about ten nucleotide sequences, from about four nucleotide sequences to about eleven nucleotide sequences, from about four nucleotide sequences to -28about twelve nucleotide sequences, from about four nucleotide sequences to about thirteen nucleotide sequences, from about four nucleotide sequences to about fourteen nucleotide sequences, from about four nucleotide sequences to about fifteen nucleotide sequences, from about four nucleotide sequence to about sixteen nucleotide sequences, from about four 5 nucleotide sequence to about seventeen nucleotide sequences, from about four nucleotide sequence to about eighteen nucleotide sequences, from about four nucleotide sequence to about nineteen nucleotide sequences, from about four nucleotide sequence to about twenty nucleotide sequences, from about five nucleotide sequences to about nine nucleotide sequences, from about five nucleotide sequences to about ten nucleotide sequences, from 10 about five nucleotide sequences to about eleven nucleotide sequences, from about five nucleotide sequences to about twelve nucleotide sequences, from about five nucleotide sequences to about thirteen nucleotide sequences, from about five nucleotide sequences to about fourteen nucleotide sequences, from about five nucleotide sequences to about fifteen nucleotide sequences, from about five nucleotide sequence to about sixteen nucleotide 15 sequences, from about five nucleotide sequence to about seventeen nucleotide sequences, from about five nucleotide sequence to about eighteen nucleotide sequences, from about five nucleotide sequence to about nineteen nucleotide sequences, from about five nucleotide sequence to about twenty nucleotide sequences, from about six nucleotide sequences to about nine nucleotide sequences, from about six nucleotide sequences to about ten nucleotide 20 sequences, from about six nucleotide sequences to about eleven nucleotide sequences, from about six nucleotide sequences to about twelve nucleotide sequences, from about six nucleotide sequences to about thirteen nucleotide sequences, from about six nucleotide sequences to about fourteen nucleotide sequences, from about six nucleotide sequences to about fifteen nucleotide sequences, from about six nucleotide sequence to about sixteen 25 nucleotide sequences, from about six nucleotide sequence to about seventeen nucleotide sequences, from about six nucleotide sequence to about eighteen nucleotide sequences, from about six nucleotide sequence to about nineteen nucleotide sequences, from about six nucleotide sequence to about twenty nucleotide sequences, from about seven nucleotide sequences to about nine nucleotide sequences, from about seven nucleotide sequences to 30 about ten nucleotide sequences, from about seven nucleotide sequences to about eleven nucleotide sequences, from about seven nucleotide sequences to about twelve nucleotide sequences, from about seven nucleotide sequences to about thirteen nucleotide sequences, from about seven nucleotide sequences to about fourteen nucleotide sequences, from about - 29 seven nucleotide sequences to about fifteen nucleotide sequences, from about seven nucleotide sequence to about sixteen nucleotide sequences, from about seven nucleotide sequence to about seventeen nucleotide sequences, from about seven nucleotide sequence to about eighteen nucleotide sequences, from about seven nucleotide sequence to about nineteen 5 nucleotide sequences, from about seven nucleotide sequence to about twenty nucleotide sequences, from about eight nucleotide sequences to about ten nucleotide sequences, from about eight nucleotide sequences to about eleven nucleotide sequences, from about eight nucleotide sequences to about twelve nucleotide sequences, from about eight nucleotide sequences to about thirteen nucleotide sequences, from about eight nucleotide sequences to 10 about fourteen nucleotide sequences, from about eight nucleotide sequences to about fifteen nucleotide sequences, from about eight nucleotide sequence to about sixteen nucleotide sequences, from about eight nucleotide sequence to about seventeen nucleotide sequences, from about eight nucleotide sequence to about eighteen nucleotide sequences, from about eight nucleotide sequence to about nineteen nucleotide sequences, from about eight nucleotide 15 sequence to about twenty nucleotide sequences, from about nine nucleotide sequences to about eleven nucleotide sequences, from about nine nucleotide sequences to about twelve nucleotide sequences, from about nine nucleotide sequences to about thirteen nucleotide sequences, from about nine nucleotide sequences to about fourteen nucleotide sequences, from about nine nucleotide sequences to about fifteen nucleotide sequences, from about nine 20 nucleotide sequence to about sixteen nucleotide sequences, from about nine nucleotide sequence to about seventeen nucleotide sequences, from about nine nucleotide sequence to about eighteen nucleotide sequences, from about nine nucleotide sequence to about nineteen nucleotide sequences, from about nine nucleotide sequence to about twenty nucleotide sequences, and the like. 25 101141 In some embodiments of the present invention, the cis-acting element comprises, consists or consists essentially of at least two nucleotide sequences of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9, in any combination, in any orientation, and/or in any order. In other embodiments of the present invention, the cis-acting element comprises, 30 consists or consists essentially of at least three nucleotide sequences of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9, in any combination, in any orientation, and/or in any order. In still other embodiments, the cis-acting element comprises, consists or consists essentially of at -30least five nucleotide sequences of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9, in any combination, in any orientation, and/or in any order. In additional embodiments, the cis acting element comprises, consists or consists essentially of about two to about nine 5 nucleotide sequences of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9, in any combination, in any orientation, and/or in any order. In further embodiments, the cis-acting element comprises, consists or consists essentially of about three to about nine nucleotide sequences of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, 10 SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9, and/or SEQ ID NO:10, in any combination, in any orientation, and/or in any order. In still further embodiments, the cis acting element comprises, consists or consists essentially of about five to about nine nucleotide sequences of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9, in any 15 combination, in any orientation, and/or in any order. In other embodiments, the cis-acting element comprises, consists or consists essentially of about five to about fifteen nucleotide sequences of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9, in any combination, in any orientation, and/or in any order. 20 10115] As discussed above, the cis-acting element comprising multimers of the nucleotide sequences can comprise, consist or consist essentially of multimers of any of the nucleotide sequences of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9, in any number of copies of a particular nucleotide sequence of the invention and/or in any combination, in any 25 orientation, and/or in any order of the nucleotide sequences of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9. Thus, in some embodiments the isolated nucleic acid molecule can comprise, consist essentially of, or consist of multiple copies of the same nucleotide sequence (e.g., 2 copies, 3 copies, 4 copies, 5 copies, 6 copies, 7 copies, 8 copies, 9 copies, 10 copies, 30 11 copies, 12 copies, 13 copies, 14 copies, fifteen copies, sixteen copies, seventeen copies, eighteen copies, nineteen copies, twenty copies etc.). In other embodiments, the cis-acting element can comprise, consist or consist essentially of multiple nucleotide sequences as defined above each of which are different from one another. In additional embodiments, the -31 cis-acting element can comprise, consist or consist essentially of multimers of the nucleotide sequences of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9, wherein some of the nucleotide sequences are the same (i.e., particular sequences are present in multiple copies) 5 and some of the nucleotide sequences are different from one another, in any combination, in any orientation, and/or in any order. [01161 Thus, non-limiting examples of a multimer of the nucleotide sequences defined above includes (SEQ ID NO: 1)a(SEQ ID NO:2)b(SEQ ID NO:3)c(SEQ ID NO:4)d(SEQ ID NO:5)e(SEQ ID NO:6)(SEQ ID NO:7)g(SEQ ID NO:8)h(SEQ ID NO:9)i, 10 wherein a, b, c, d, e, f, g, h, i are each independently 0 to 9 or more (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, etc.). Further non-limiting examples of multimers of the nucleotide sequences of the present invention include (SEQ ID NO:2) 4 ; (SEQ ID NO:5)(SEQ ID NO:3)(SEQ ID NO:9); (SEQ ID NO:2)(SEQ ID NO:5)(SEQ ID NO:3)(SEQ ID NO:4); (SEQ ID NO:2) 2 (SEQ ID NO:6) 4 (SEQ ID NO:3) 3 ; (SEQ ID NO:2) 2 (SEQ ID NO:6)(SEQ ID NO:3)(SEQ ID NO:4) 2 ; 15 (SEQ ID NO:5) 3 (SEQ ID NO:3)(SEQ ID NO:4); (SEQ ID NO:2)(SEQ ID NO:5) 2 (SEQ ID NO:3) 2 ; (SEQ ID NO:6) 4 (SEQ ID NO:4) 2 ; (SEQ ID NO:2)(SEQ ID NO:6) 2 ; (SEQ ID NO:2)(SEQ ID NO:5)(SEQ ID NO:3)(SEQ ID NO:4) 2 ; (SEQ ID NO:2) 2 (SEQ ID NO:6) 2 (SEQ ID NO:3) 2 ; (SEQ ID NO:2) 2 (SEQ ID NO:3) 3 (SEQ ID NO:9); (SEQ ID NO:3)(SEQ ID NO:4) 3 ; (SEQ ID NO:2)(SEQ ID NO:5) 5 (SEQ ID NO:4); (SEQ ID 20 NO:2)(SEQ ID NO:4)(SEQ ID NO:5)(SEQ ID NO:8)(SEQ ID NO:9); (SEQ ID NO:5) 3 (SEQ ID NO:8); (SEQ ID NO:5) 2 (SEQ ID NO:6)(SEQ ID NO:7)(SEQ ID NO:8) 2 ; (SEQ ID NO:6) 4 ; (SEQ ID NO:5)(SEQ ID NO:4)(SEQ ID NO:6)(SEQ ID NO:9)(SEQ ID NO:8)(SEQ ID:NO:2); (SEQ ID NO:8) 4 (SEQ ID NO:9) 2 ; (SEQ ID NO:3)(SEQ ID NO:5) 2 ; (SEQ ID NO:6)(SEQ ID NO:9); (SEQ ID NO:3)(SEQ ID NO:4) 2 (SEQ ID NO:7) 2 ; (SEQ ID NO:9) 6 ; 25 (SEQ ID NO:3) 3 ; (SEQ ID NO:5) 3 (SEQ ID NO:9) 4 ; (SEQ ID NO:3)(SEQ ID NO:7) 3 ; (SEQ ID NO:1)(SEQ ID NO:4)s(SEQ ID NO:9) 4 ; (SEQ ID NO:3)s, and the like. [0117] In some embodiments, the cis-acting elements comprising multimers of the nucleotide sequences of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9, can further 30 comprise one or more nucleotides (i.e., bases) between each of the inverted repeats (i.e., between the nucleotide sequences of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9) (e.g., a spacer sequence). The spacer sequences can be of any length and composition of - 32 nucleotides, wherein each nucleotide can be independently adenine, thymine, guanine and/or cytosine (i.e., wherein n=A, T, G, or C), in any order and/or in any combination, and x and y are each independently 0-100 nucleotides or more. In some embodiments, the spacer sequences are the same as (i.e., equivalent to) the flanking sequences (i.e., nx, ny) described 5 herein. Thus, in some embodiments, the cis-acting element comprises, consists or consists essentially of spacer sequences (i.e., flanking sequences, n,, ny) that can be the same as one another (i.e., the same as other spacer sequences of the nucleic acid molecule) and/or different than one another, or any combination thereof. A non-limiting example of a multimer of the present invention comprising a spacer sequence is the following: 10 [0118] nnnnnGCGGNNCCGCnnnnnnnnnnnnnnnnGCGGNNCCGCnnnnnnnnn nnnnnnGCGGNNCCGCnnnnn; (SEQ ID NO:2) 3 , wherein the flanking sequences are in order from left to right: x=5; y+6=16; y+x=15; and y=5. In this example, the spacer sequences are bolded and underlined and are shown to be of different lengths. Further non-limiting examples of a multimer comprising a spacer sequence are the following: 15 10119] (1) nnnnnGCGGNNCCGCnnnnnnnnnnGCGGNNCCGCnnnnnnnnnnGC GGNNCCGCnnnnn; (SEQ ID NO:2) 3 , wherein the flanking sequences are in order from left to right: x=5; y+6=10; y+x=10; and y=5; and [01201 (2) nnnnnGCGGNNCCGCnnnnnnnnnnnnnnGCGGNNCCGCnnnnnnnnn nGCGGNNCCGCnnnnn; (SEQ ID NO:2) 3 , wherein the flanking sequences are in order from 20 left to right: x=5; y+6=14; y+x= 10; and y=5. [01211 In the above two examples, the bolded and underlined nucleotides represent both the spacer sequences and the flanking sequences of the nucleotide sequences that comprise the multimer (i.e., in these examples, the spacer sequences are equivalent to the flanking sequences). 25 2.2 Promoters [01221 In accordance with the present invention, the cis-acting elements described above and elsewhere herein are useful for conferring a pattern of inducibility on a promoter, including a promoter that is operable in a plant cell (e.g., a guard cell), which pattern is similar to that of the alcohol dehydrogenase (ADHI) system of A. nidulans. This generally 30 requires operably connecting a cis-acting element with a promoter (e.g., a promoter that is not already inducible by contact with a chemical compound that can induce the expression of the alcohol dehydrogenase system of A. nidulans), to thereby make the promoter inducible by - 33 contact with a chemical compound that can induce the expression of the alcohol dehydrogenase system of A. nidulans. [01231 Any promoter can be made inducible using the cis-acting elements described above and elsewhere herein. In some embodiments, promoters that can be made 5 inducible with the subject cis-acting elements can include chemically inducible promoters that are not naturally or endogenously inducible by the same compounds/chemicals that induce the alcohol dehydrogenase system of A. nidulans. Additionally, promoters useable with the present invention can include those that drive expression of a nucleotide sequence constitutively, those that drive expression when induced, and those that drive expression in a 10 tissue- or developmentally-specific manner, as these various types of promoters are known in the art and can be made inducible by operably linking thereto the cis-acting elements described above and elsewhere herein. [01241 In particular embodiments, a promoter that can be made inducible with the cis-acting elements described above and elsewhere herein includes a minimal promoter. A 15 minimal promoter is a promoter having only the nucleotides/nucleotide sequences from a selected promoter that are required for binding of the transcription factors and transcription of a nucleotide sequence of interest that is operably associated with the minimal promoter including but not limited to TATA box sequences. These portions or sequences from a promoter are generally placed upstream (i.e., 5') of a nucleotide sequence to be expressed. 20 Thus, nucleotides/nucleotide sequences from any promoter useable with the present invention can be selected for use as a minimal promoter. Any promoter may be altered to generate a minimal promoter by progressively removing nucleotides from the promoter until the promoter ceases to function in order to identify the minimal promoter. Thus, the smallest fragment of a promoter which still functions as a promoter is also considered a minimal 25 promoter. 101251 The promoter may be endogenous to the plant. Alternatively, a heterologous promoter may be employed. For example, a promoter can be heterologous when it is operably linked to a polynucleotide from a species different from the species from which the polynucleotide was derived. Alternatively, a promoter can be heterologous to a selected 30 nucleotide sequence if the promoter is from the same/analogous species from which the polynucleotide is derived, but one or both (i.e., promoter and/or polynucleotide) are modified - 34 from their original form and/or genomic locus, or the promoter is not the native promoter for the operably linked polynucleotide. 10126] The choice of promoters useable with the present invention can be made among many different types of promoters. This choice generally depends upon several 5 factors, including, but not limited to, cell- or tissue-specific expression, desired expression level, efficiency, inducibility and/or selectability. For example, where expression in a specific tissue or organ is desired in addition to inducibility, a tissue-specific promoter can be used (e.g., a guard cell specific promoter). In contrast, where expression in response to a stimulus is desired in addition to inducibility via chemical compounds that induce the expression of the 10 alcohol dehydrogenase system of A. nidulans, a promoter inducible by other stimuli or chemicals can be used. Where continuous expression is desired throughout the cells of a plant in addition to inducibility via the chemicals/compounds of the present invention that induce expression of the alcohol dehydrogenase system of A. nidulans, a constitutive promoter can be chosen. 15 101271 Non-limiting examples of constitutive promoters include cestrum virus promoter (cmp) (U.S. Patent No. 7,166,770), the rice actin I promoter (Wang et al. (1992) Mol. Cell. Biol. 12:3399-3406; as well as US Patent No. 5,641,876), CaMV 35S promoter (Odell et al. (1985) Nature 313:810-812), CaMV 19S promoter (Lawton et al. (1987) Plant Mol. Biol. 9:315-324), nos promoter (Ebert et al. (1987) Proc. Nat]. Acad. Sci USA 84:5745 20 5749), Adh promoter (Walker et al. (1987) Proc. Natl. Acad Sci. USA 84:6624-6629), sucrose synthase promoter (Yang & Russell (1990) Proc. Natl. Acad. Sci. USA 87:4144 4148), and the ubiquitin promoter. [0128] Illustrative examples of tissue-specific promoters include those encoding the seed storage proteins (such as p-conglycinin, cruciferin, napin and phaseolin), zein or oil body 25 proteins (such as oleosin), or proteins involved in fatty acid biosynthesis (including acyl carrier protein, stearoyl-ACP desaturase and fatty acid desaturases (fad 2-1)), and other nucleic acids expressed during embryo development (such as Bce4, see, e.g., Kridl et al. (1991) Seed Sci. Res. 1:209-219; as well as EP Patent No. 255378). Thus, the promoters associated with these tissue-specific nucleic acids can be used in the present invention. 30 Additional examples of tissue-specific promoters include, but are not limited to, the root specific promoters RCc3 (Jeong et al. Plant Physiol. 153:185-197 (2010)) and RB7 (U.S. Patent No. 5459252), the lectin promoter (Lindstrom et al. (1990) Der. Genet. 11:160-167; - 35 and Vodkin (1983) Prog. Clin. Biol. Res. 138:87-98), corn alcohol dehydrogenase 1 promoter (Dennis et al. (1984) Nucleic Acids Res. 12:3983-4000), S-adenosyl-L-methionine synthetase (SAMS) (Vander Mijnsbrugge et al. (1996) Plant and Cell Physiology, 37(8):1108-1115), corn light harvesting complex promoter (Bansal et al. (1992) Proc. Natl. A cad. Sci. USA 5 89:3654-3658), corn heat shock protein promoter (O'Dell et al. (1985) EMBO J. 5:451-458; and Rochester et al. (1986) EMBO J. 5:451-458), pea small subunit RuBP carboxylase promoter (Cashmore, "Nuclear genes encoding the small subunit of ribulose-I,5-bisphosphate carboxylase" 29-39 In: Genetic Engineering of Plants (Hollaender ed., Plenum Press 1983; and Poulsen et al. (1986) Mol. Gen. Genet. 205:193-200), Ti plasmid mannopine synthase 10 promoter (Langridge et al. (1989) Proc. Natl. Acad. Sci. USA 86:3219-3223), Ti plasmid nopaline synthase promoter (Langridge et al. (1989), supra), petunia chalcone isomerase promoter (van Tunen et al. (1988) EMBO J. 7:1257-1263), bean glycine rich protein I promoter (Keller et al. (1989) Genes Dev. 3:1639-1646), truncated CaMV 35S promoter (O'Dell et al. (1985) Nature 313:810-812), potato patatin promoter (Wenzler et al. (1989) 15 Plant Mol. Biol. 13:347-354), root cell promoter (Yamamoto et al. (1990) Nucleic Acids Res. 18:7449), maize zein promoter (Kriz et al. (1987) Mol. Gen. Genet. 207:90-98; Langridge et al. (1983) Cell 34:1015-1022; Reina et al. (1990) Nucleic Acids Res. 18:6425; Reina et al. (1990) Nucleic Acids Res. 18:7449; and Wandelt et al. (1989) Nucleic Acids Res. 17:2354), globulin-I promoter (Belanger et al. (1991) Genetics 129:863-872), a:-tubulin cab promoter 20 (Sullivan et al. (1989) Mol. Gen. Genet. 215:431-440), PEPCase promoter (Hudspeth & Grula (1989) Plant Mol. Biol. 12:579-589), R gene complex-associated promoters (Chandler et al. (1989) Plant Cell 1:1175-1183), and chalcone synthase promoters (Franken et al. (1991) EMBO J. 10:2605-2612). Particularly useful for seed-specific expression is the pea vicilin promoter (Czako et al. (1992) Mol. Gen. Genet. 235:33-40; as well as US Patent No. 25 5,625,136). Other useful promoters for expression in mature leaves are those that are switched on at the onset of senescence, such as the SAG promoter from Arabidopsis (Gan et al. (1995) Science 270:1986-1988). 101291 In specific embodiments, the promoter that is operably linked to the cis acting element is one that is specifically or preferentially operable in a plant guard cell. Non 30 limiting examples of guard cell-specific or guard cell-preferential promoters include: Arabidopsis trehalase gene promoter (EP 1111051), potato KSTJ promoter (Plesch et al. (200 1) Plant Journal 28 (4): 455-464), Arabidopsis pGCl (At1g22690) promoter (Yang et al., (2008) Plant Methods 4:6) Arabidopsis KATI (At5g46240) potassium channel promoter -36- (Nakumura et al. (1995) Plant Physiol. 109, 371-374), Arabidopsis AtMYB60 (At l g088 10) promoter (U.S. Pat. Apple. Pub. No. 20080064091), gcPepC promoter (Kopka et al. (1997) Plant J 11, 871-882), Arabidopsis AtCYP86A2 promoter (Francia et al. (2008) Plant Signaling and Behavior 3, 684-686) and Arabidopsis At5g5 8580 promoters designated 5 pSUH305S, pSUH305, pSUH305GB, pSUK132, pSUK134, pSUK136, pSUK342, pSUK344, pSUK132GB, pSUK134GB, pSUK136GB, pSUK342GB, pSUK344GB (U.S. Pat. Apple. Pub. 20060117408). [01301 In addition, promoters functional in plastids can be used. Non-limiting examples of such promoters include the bacteriophage T3 gene 9 5' UTR and other promoters 10 disclosed in U.S. Patent No. 7,579,516. Other promoters useful with the present invention include but are not limited to the S-E9 small subunit RuBP carboxylase promoter and the Kunitz trypsin inhibitor gene promoter (Kti3). [01311 In some instances, inducible promoters that are not inducible by the same compounds that induce expression of the alcohol dehydrogenase system of A. nidulans are 15 useable with cis-acting sequence described above and elsewhere herein. Examples of inducible promoters useable with the present invention include, but are not limited to, tetracycline repressor system promoters, Lac repressor system promoters, copper-inducible system promoters, salicylate-inducible system promoters (e.g., the PRla system), glucocorticoid-inducible promoters (Aoyama et al. (1997) Plant J. 11:605- 612), and 20 ecdysone-inducible system promoters. Other non-limiting examples of inducible promoters include ABA- and turgor-inducible promoters, the auxin-binding protein gene promoter (Schwob et al. (1993) Plant J. 4:423-432), the UDP glucose flavonoid glycosyltransferase promoter (Ralston et al. (1988) Genetics 119:185-197), the MPI proteinase inhibitor promoter (Cordero et al. (1994) Plant J. 6:141-150), the glyceraldehyde-3- phosphate dehydrogenase 25 promoter (Kohler et al. (1995) Plant Mol. Biol. 29:1293-1298; Martinez et al. (1989) J. Mol. Biol. 208:551-565; and Quigley et al. (1989) J. Mol. Evol. 29:412-421) the benzene sulfonamide-inducible promoters (U.S. Patent No. 5,364,780) and the glutathione S transferase promoters. Likewise, one can use any appropriate inducible promoter described in Gatz (1996) Current Opinion Biotechnol. 7:168-172 and Gatz (1997) Annu. Rev. Plant 30 Physiol. Plant Mol. Biol. 48:89-108. 101321 The cis-acting elements described herein are operably linked to a promoter so as to control the activity of the promoter. The activity or strength of a promoter may be - 37 measured in terms of the amount of mRNA or protein accumulation it specifically produces, relative to the total amount of mRNA or protein. Suitably, an operably linked cis-acting element as described herein is placed at a distance from the promoter so that: (1) in the absence of a compound that induces expression of the alcohol dehydrogenase system of A. 5 nidulans, the promoter suitably expresses an operably linked nucleic acid sequence at a level no more than 1%, 0.1%, 0.01%, 0.00 1%, 0.0001% or 0.00001% of the total cellular RNA or protein; and (2) in the presence of a compound that induces expression of the alcohol dehydrogenase system of A. nidulans, the promoter suitably expresses an operably linked nucleic acid sequence at a level greater than 0.0 1%, 0.05%, 0.1%, 0.25%, 0.5%, 0.75%, 1%, 10 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% (w/w) of the total cellular RNA or protein. Positioning of the cis-acting element relative to the promoter can be determined by the skilled person using customary recombination and cloning techniques (e.g., In Maniatis T, Fritsch E F and Sambrook J (1989) Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor Laboratory, Cold 15 Spring Harbor (NY); Silhavy et al. (1984) Experiments with Gene Fusions, Cold Spring Harbor Laboratory, Cold Spring Harbor (NY); Ausubel et al. (1987) Current Protocols in Molecular Biology, Greene Publishing Assoc. and Wiley Interscience; Gelvin et al. (Eds) (1990) Plant Molecular Biology Manual; Kluwer Academic Publisher, Dordrecht, The Netherlands). The cis-acting element may be upstream or downstream of the promoter. In 20 specific embodiments, the cis-acting element is upstream of the promoter. In some embodiments, the distance between the cis-acting element and the promoter is no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, 200, 225, 250, 275, 300, 25 325, 350, 375, 400, 425, 450, 475, 500 nt. Suitably, the distance between the cis-acting element and the promoter is less than 1000, 950, 900, 850, 800, 750, 700, 650, 600, 550 nt. 2.3 Inducer compounds 101331 Non-limiting examples of chemical compounds that can induce the promoters of the present invention (e.g., inducer compound) include a primary alcohol, a 30 primary monoamine, a ketone, a C3 to C9 ketone, a methyl ketone, a hydrolysable ester, an aliphatic aldehyde, ethanol, allyl alcohol acetaldehyde, ethyl methyl ketone, acetone, ethylamine, cyclohexanone, butan-2-ol, 3-oxobutyric acid, propan-2-ol, propan- 1 -ol, butan-2 ol, threonine, and/or any combination thereof. - 38 - 101341 An inducer compound can be provided in any concentration that is not toxic to the plant. Thus, in some embodiments, the concentration of the inducer compound is about 0.01 % to about 10% (v/v) or more. In other embodiments of the present invention, the concentration of the inducer compound is about 0.1 % to about 20% (v/v). In still other 5 embodiments of the invention, the concentration of the inducer compound is about 0.1 % to about 5% (v/v). In further embodiments of the present invention, the concentration of the inducer compound is about 1% to about 2%. Thus, for example, the concentration of the inducer compound can be about 0.01%, about 0.02%, about 0.03%, about 0.04%, about 0.05%, about 0.06%, about 0.07%, about 0.08%. about 0.09%, about 0.1%, about 0.15%, 10 about 0.2%, about 0.25%, about 0.3%, about 0.35%, about 0.4%, about 0.45%, about 0.6%. about 0.65%, about 0.7%, about 0.75%, about 0.8%, about 0.85%, about 0.9%, about 0.95%, about 1%, about 1.5%, about 2%, about 2.5%, about 3%, about 3.5%, about 4%, about 4.5%, about 5%, about 5.5%, about 6%, about 6.5%, about 7%, about 7.5%, about 8%, about 8.5%, about 9%, about 9.5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 15 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 100% (v/v), and the like, and any combination thereof. [0135] The inducer compound can be provided as a root drench, a spray, a mist, a suspension, an emulsion, a powder, a granule, an aerosol, a foam, a paste, a dip, a vapor, a 20 paint, and the like, and combinations thereof. In some embodiments of the invention, when the inducer compound is provided in the form of a vapor, the concentration of the inducer compound can be, for example, in a concentration of about 95% to about 100% (v/v). Thus, for example, the inducer compound, provided in a concentration of about 95% to about 100% (v/v), can be placed in proximity to the plant, plant part, plant organ or plant leaf (e.g., in a 25 container such as a tube, a dish, and the like, or on a cloth, paper, beads, and the like, that is soaked in the inducer compound), thereby exposing the plant, plant part, plant organ or plant leaf to a vapor comprising the inducer compound. In some embodiments of the present invention, the inducer compound is provided in more than one form. Thus, for example, the inducer compound can be provided as a foliar spray and as a root drench. The concentration 30 of the inducer compound when applied in more than one form can be the same or can be different in the different forms provided. Thus, for example, a foliar spray and a root drench can be provided at the same and/or a different concentration than one another. - 39 - 2.4 Expression products for modulating stomatal closure [01361 The constructs of the present invention also comprise an operably connected nucleic acid sequence encoding an expression product that inhibits stomatal closure. In some embodiments, the expression product inhibits or abrogates the activity or function of an 5 endogenous polypeptide of the plant, which stimulates or otherwise facilitates stomatal closure. In other embodiments, the expression product is a negative regulator of stomatal closure. Non-limiting examples of endogenous polypeptides include ABIl, ABI2 (Himmelbach et al. (1998) Philos. Trans. R. Soc. Lond. B Biol. Sci. 353, 1439-1444; Leung et al. (1998) Annu. Rev. Plant Physiol. Plant Mol. Biol. 49, 199-222.), OST I (Mustilli et al. 10 (2002) The Plant Cell 14, 3089-3099; U.S. Patent No. 7,211,436), AAPK (related to OSTI) (Li et al. (2000) Science 287(5451), 300-303), AHAI (also referred to as OST2) (Merlot et al., (2007) EMBO J. 26, 3216-3226), v-SNAREs AtVAMP711-14 (Leshem et al., (2010) J. Exp. Bot. 61, 2615-2622), GPAI (Wang et al. (200 1) Science 292, 2070-2072), AtABCG22 (Kuromori et al. (2011) Plant J. 67, 885-894), AtABCG40 (Kang et al., (2010) Proc. Natl. 15 Acad Sci. USA 107, 2355-2360), AtMRP4 (Klein et al. (2004) Plant J. 219-236), RBOHD and RBOHF (Kwak et al. (2003) EMBO J. 22, 2623-2633), PLDalphal (Zhang et al. (2004) Proc. Natl. Acad. Sci. USA 101, 9508-9513) PKS3 (Guo et al. (2002) Developmental Cell 3, 233-244) and ATHB6 (Himmelbach et al. (2002) EMBO J. 21, 3029-3038; Grill E. (2002) EMBOJ. 21:3029-38). 20 [0137] Amino acid sequences corresponding to the above endogenous polypeptides as well as nucleic acid sequences corresponding to genes that code for these polypeptides are useful for modulating stomatal closure, as described below. [01381 The present invention contemplates the use of any suitable stomatal closure modulating polypeptide and polynucleotide in the practice of the invention. 25 101391 For example, non-limiting ABIl polypeptides comprise the amino acid sequence: [01401 MEEVSPAIAGPFRPFSETQMDFTGIRLGKGYCNNQYSNQDSENGDL MVSLPETSSCSVSGSHGSESRKVLISRINSPNLNMKESAAADIVVVDISAGDEINGSDIT SEKKMISRTESRSLFEFKSVPLYGFTSICGRRPEMEDAVSTIPRFLQSSSGSMLDGRFDP 30 QSAAHFFGVYDGHGGSQVANYCRERMHLALAEEIAKEKPMLCDGDTWLEKWKKA LFNSFLRVDSEIESVAPETVGSTSVVAVVFPSHIFVANCGDSRAVLCRGKTALPLSVD HKPDREDEAARIEAAGGKVIQWNGARVFGVLAMSRSIGDRYLKPSIIPDPEVTAVKR - 40 - VKEDDCLILASDGVWDVMTDEEACEMARKRILLWHKKNAVAGDASLLADERRKEG KDPAAMSAAEYLSKLAIQRGSKDNISVVVVDLKPRRKLKSKPLN [SEQ ID NO:1 1], as set forth for example in GenPept Accession No. NP_194338, or an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 5 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO: 11. [01411 A representative ABIl nucleic acid sequence comprises a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO: 11, or a complement of the nucleotide sequence; or a nucleotide sequence encoding an amino acid sequence having at 10 least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO: 11, a complement of that nucleotide sequence. In illustrative examples, an ABI] nucleic acid sequence comprises the nucleotide sequence: 15 [01421 gaagcaattgttgcattagcctacccatttcctccttctttctctcttctatctgtgaacaaggcacattagaactct tcttttcaacttttttaggtgtatatagatgaatctagaaatagttttatagttggaaattaattgaagagagagagatattactacaccaatttt tcaagaggtcctaacgaattacccacaatccaggaaaccettattgaaattcaattcatttctttctttctgtgtttgtgattttccgggaaata tttttgggtatatgtetetetgttttgettteetttttcataggagtcatgtgtttettettgtettectagettettetaataaagtcettetettgtga aaatetctcgaattttcatttttgttccattggagetatcttatagatcacaaccagagaaaaagatcaaatctttaccgttaatggaggaagt 20 atctccggcgatcgcaggtcctttcaggccattctccgaaacccagatggatttcaccgggatcagattgggtaaaggttactgcaataa ccaatactcaaatcaagattccgagaacggagatctaatggtttcgttaccggagacttcatcatgctctgtttctgggtcacatggttctga atctaggaaagttttgatttctggatcaattctcctaatttaaacatgaaggaatcagcagctgctgatatagtcgtcgttgatatctccgcc ggagatgagatcaacggctcagatattactagcgagaagaagatgatcagcagaacagagagtaggagtttgtttgaattcaagagtgt gcctttgtatggttttacttegatttgtggaagaagacctgagatggaagatgctgtttcgactataccaagattccttcaatettcctctggtt 25 cgatgttagatggtcggtttgatcctcaatccgccgctcatttcttcggtgtttacgacggccatggcggttctcaggtagcgaactattgta gagagaggatgcatttggctttggcggaggagatagetaaggagaaaccgatgctctgcgatggtgatacgtggctggagaagtgga agaaagctettttcaactcgttcctgagagttgactggagattgagtcagttgcgccggagacggttgggtcaacgtcggtggttgcg ttgttttcccgtctcacatcttcgtcgctaactgcggtgactctagagccgttctttgccgcggcaaaactgcacttcattatccgttgacca taaaccggatagagaagatgaagctgcgaggattgaagccgcaggagggaaagtgattcagtggaatggagctcgtgttttcggtgtt 30 ctcgccatgtcgagatccattggcgatagatacttgaaaccatccatcattcctgatccggaagtgacggctgtgaagagagtaaaaga agatgattgtctgattttggcgagtgacggggtttgggatgtaatgacggatgaagaagcgtgtgagatggcaaggaagcggattctctt gtggcacaagaaaaacgcggtggctggggatgcatcgttgctcgcggatgagcggagaaaggaagggaaagatcctgcggcgatg tccgcggctgagtatttgtcaaagctggcgatacagagaggaagcaaagacaacataagtgtggtggtggttgatttgaagcctcgga -41 ggaaactcaagagcaaacccttgaactgaggcagagagggtcctttttcttaatttttaaaatgaatatgggtctctccaagaaaaagtatt tactattattaatttgtgcttatttttttaactaacaagttataaccatatggagataatgaagcttaatgtgttaagctcttttgtcttgactacatt ctaaaaagccccttgtatttttcttcccgggctaattgtaatatggttacaacatacattaagatgtagtattattgtttatgcaattactttcaaa actttacatac [SEQ ID NO:12], as set forth for example in GenBank Accession NM_118741, or 5 a complement thereof; or a nucleotide sequence that has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO:12, or to a complement thereof; or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO:12 or to a complement thereof. 10 101431 Non-limiting ABI2 polypeptides comprise the amino acid sequence: 101441 MDEVSPAVAVPFRPFTDPHAGLRGYCNGESRVTLPESSCSGDGAMK DSSFEINTRQDSLTSSSSAMAGVDISAGDEINGSDEFDPRSMNQSEKKVLSRTESRSLF EFKCVPLYGVTSICGRRPEMEDSVSTIPRFLQVSSSSLLDGRVTNGFNPHLSAHFFGVY DGHGGSQVANYCRERMHLALTEEIVKEKPEFCDGDTWQEKWKKALFNSFMRVDSEI 15 ETVAHAPETVGSTSVVAVVFPTHIFVANCGDSRAVLCRGKTPLALSVDHKPDRDDEA ARIEAAGGKVIRWNGARVFGVLAMSRSIGDRYLKPSVIPDPEVTSVRRVKEDDCLILA SDGLWDVMTNEEVCDLARKRILLWHKKNAMAGEALLPAEKRGEGKDPAAMSAAE YLSKMALQKGSKDNISVVVVDLKGIRKFKSKSLN [SEQ ID NO:13], as set forth for example in GenPept Accession No. CAA70162, or an amino acid sequence having at least 20 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:13. 101451 A representative ABI2 nucleic acid sequence comprises a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO: 13, or a complement of 25 the nucleotide sequence; or a nucleotide sequence encoding an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:13, or a complement of that nucleotide sequence. In illustrative examples, an ABI2 nucleic acid sequence comprises the 30 nucleotide sequence: [0146] tttttgttaaagttcaagaaagttcttttttctttttttttcctcctttaatggacgaagtttctcctgcagtcgctgttcc attcagaccattcactgaccctcacgccggacttagaggctattgcaacggtgaatctagggttactttaccggaaagttettgttctggcg - 42 acggagctatgaaagattcttcetttgagatcaatacaagacaagattcattgacatcatcatcatctgctatggcaggtgtggatatctcc gccggagatgaaatcaacggttcagatgagtttgatccgagatcgatgaatcagagtgagaagaaagtacttagtagaacagagagta gaagtctgtttgagttcaagtgtgttcctttatatggagtgacttcgatttgtggtagacgaccagagatggaagattctgtctcaacgattc ctagattccttcaagtttcttctagttcgttgcttgatggtcgagtcactaatggatttaatcctcacttgagtgctcatttctttggtgtttacgat 5 ggccatggcggttctcaggtagcgaattattgtcgtgagaggatgcatctggctttgacggaggagatagtgaaggagaaaccggagt tttgtgacggtgacacgtggcaagagaagtggaagaaggctttgttcaactcttttatgagagttgactcggagattgaaactgtggctca tgctccggaaactgttgggtctacctcggtggttgcggttgtctttccgactcacatctttgtcgcgaattgcggcgactctagggcggtttt gtgtcgcggcaaaacgccactcgcgttgtcggttgatcacaaaccggatagggatgatgaagcggcgaggatagaagctgccggtg ggaaagtaatccggtggaacggggctcgtgtatttggtgttctcgcaatgtcaagatccattggcgatagataccttaaaccgtcagtaat 10 tccggatccagaagtgacttcagtgcggcgagtaaaagaagatgattgtctcatcttagcaagtgatggtctttgggatgtaatgacaaa cgaagaagtgtgcgatttggctcggaaacggattttactatggcataagaagaacgcgatggccggagaggctttgttccggggag aaaagaggagaaggaaaagatcctgcagcaatgtccgcggcagagtatttgtcgaagatggctttgcaaaaaggaagcaaagacaat ataagtgtggtagtggttgatttgaagggaataaggaaattcaagagcaaatccttgaattgaaaaagaaggtttggaagaaaagtgaaa aaaaaagttttgatggtgggtaaaaattctctttagtgaaaaaagaaagataaaacaacaggtaataattacattgtaatattaatttctgct 15 taaatttgttatttactttctcaaaaa [SEQ ID NO: 14], as set forth for example in GenBank Accession No. Y08965, or a complement thereof; tccaaettcaatttetetcetttetettccaactttgattectgatttgggtttttgttaaagttcaagaaagttcttttttttttttteetcettaatgg acgaagtttctcctgcagtcgctgttccattcagaccattcactgaccctcacgccggacttagaggctattgcaacggtgaattagggt tactttaccggaaagttcttgttctggcgacggagctatgaaagattetttttgagatcaatacaagacaagattcattgacatcatcatc 20 atctgctatggcaggtgtggatatctccgccggagatgaaatcaacggttcagatgagtttgatccgagatcgatgaatcagagtgaga agaaagtacttagtagaacagagagtagaagtctgtttgagttcaagtgtgttcctttatatggagtgacttcgatttgtggtagacgacca gagatggaagattctgtetcaacgattcctagatteettcaagtttettctagttcgttgcttgatggtcgagtcactaatggatttaatcctca cttgagtgctcatttctggtgtttacgatggccatggcggttctcaggtaatgaatcgatttggtttcgatatgatatgatcggaaactgca aaaaettggtttttgacatttgtttttgtgtgtttaggtagcgaattattgtcgtgagaggatgcatctggctttgacggaggagatagtgaag 25 gagaagccggagttttgtgacggtgacacgtggcaagagaagtggaagaaggctttgttcaactcttttatgagagttgactcggagatt gaaactgtggetcatgctccggaaactgttgggtctacctcggtggttgcggttgtctttccgactcacatctttgtcgcgaattgcggcga ctctagggcggtMgtgtcgcggcaaaacgccactcggttgtcggttgatcacaaagtaagtatagatgtttctatgaatgaattagtac aatcatatattcattaggacttgcggtMttgttatggtgttaccaatcatagcatgtttctatagattagttaatgtacttatagcgtttgggtct acatttatgtgtagccggatagggatgatgaagcggcgaggatagaagctgccggtgggaaagtaatccggtggaacggggctcgtg 30 tatttggtgttctcgcaatgtcaagatccattggtaattatttaactttcttgataatgatecatgaatttggttettttagttgtttccttacatta caacccgtcgcgaacaggcgatagataccttaaaccgtcagtaattccggatccagaagtgacttcagtgcggcgagtaaaagaagat gattgtcatttagcaagtgatggttttgggatgtaatgacaaacgaagaagtgtgcgatttggctcggaaacggattttactatggca taagaagaacgcgatggccggagaggctttgcttccggcggagaaaagaggagaaggaaaagatcctgcagcaatgtccgcggca -43gagtatttgtcgaagatggctttgcaaaaaggaagcaaagacaatataagtgtggtagtggttgatttgaagggaataaggaaattcaag agcaaatccttgaattgaaaaagaaggtttggaagaaaagtgaaaaaaaaagttttgatggtggg [SEQ ID NO:15], as set forth for example in GenBank Accession No. Y08966, or a complement thereof; or a nucleotide sequence that has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 5 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO: 14 or 15, or to a complement thereof; or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO:14 or 15, or to a complement thereof. 101471 An illustrative OSTI polypeptide comprises the amino acid sequence: 10 10148] MDRPAVSGPMDLPIMHDSDRYELVKDIGSGNFGVARLMRDKQSNE LVAVKYIERGEKIDENVKREIINHRSLRHPNIVRFKEVILTPTHLAIVMEYASGGELFER ICNAGRFSEDEARFFFQQLISGVSYCHAMQVCHRDLKLENTLLDGSPAPRLKICDFGY SKSSVLHSQPKSTVGTPAYIAPEVLLKKEYDGKVADVWSCGVTLYVMLVGAYPFED PEEPKNFRKTIHRILNVQYAIPDYVHISPECRHLISRIFVADPAKRISIPEIRNHEWFLKN 15 LPADLMNDNTMTTQFDESDQPGQSIEEIMQIIAEATVPPAGTQNLNHYLTGSLDIDDD MEEDLESDLDDLDIDSSGEIVYAM [SEQ ID NO: 16], as set forth for example in GenPept Accession No. CAC87047, or an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence 20 identity to SEQ ID NO:16. [0149] A non-limiting OST1 nucleic acid sequence comprises a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO: 16, or a complement of the nucleotide sequence; or a nucleotide sequence encoding an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 25 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:16, or a complement of that nucleotide sequence. In representative examples, an OST] nucleic acid sequence comprises the nucleotide sequence: 101501 atggatcgaccagcagtgagtggtccaatggatttgccgattatgcacgatagtgataggtatgaactcgtca 30 aggatattggctccggtaattttggagttgcgagattgatgagagacaagcaaagtaatgagcttgttgctgttaaatatatcgagagagg tgagaaggtcagtttattttcttcttgttttttctccttagtccaattttattttatgtttcaattcactaggttcttgaattcaaaagaaatcatttttgt gtgttcatcatccaatgagagatgtgtgtttggttactttatggttatgaatggtttaaccctatagatttacaaaattgattgatttttatggaag - 44 accaaatttgctctgctttgctttatgacttatgttgtatagtgttgttttaaaggctctaggtgtttcttttgttatggaacgtggtattaatggtg ggactttttgtatttgtacagatagatgaaaatgtaaaaagggagataatcaaccacaggtccttaagacatcccaatatcgttagattcaa agaggtttgttttcaactctcttttaagctgttttcttattattaacattttcaggaaaaaaaaggaattctataatttaatcttaggattgttcag gttatattaacaccaacccatttagccattgttatggaatatgcatctggaggagaacttttcgagcgaatctgcaatgcaggccgttcag 5 cgaagacgaggttgttctctcttttttttcttgctttgatgattaatcgaagattcaaaatgctgaggctaatatatcgtttactatcatgtaggc gaggtttttcttccagcaactcatttcaggagttagttactgtcatgctatggtaatgaaaaatgctctctatcaaatcaatgcttcatcctctgt gaaattcagctgacttaagaagttaaattttatgttgtagcaagtatgtcaccgagacttaaagctcgagaatacgttattagatggtagccc ggcccctcgtctaaagatatgtgatttcggatattcaaaggtatctttgaaaacatttcagaaactctgagttagttagtatctctaagtttgt gttttctttttcagtcatcagtgttacattcgcaaccaaaatcaactgttggaactcctgcttacatcgctcctgaggttttactaaagaaagaa 10 tatgatggaaaggtactccattttcatagttcccaaactagtatgataaccatatcttatagaaagaacaatctttgtatttttatcctctgattat agataggggaaacatgctttctcttgatgaaagctcacacaaataaaacaatcttggctcttcaagaattttggtgaagaaagctattaaga gtetgatttgtaaactgataattettgagttttggttgaattagtcaaatgcatectaatgttettettttttttcaggttgeagatgtttggtettgtg gggttactctgtatgtcatgctggttggagcatatcctttcgaagatcccgaggaaccaaagaatttcaggaaaactatacatgtgagcctt tcactttcttcatgcttcaatagttgaaaaatgtaattatggattttattacttgctagctaaactatctgttetcttgtgaaaatatttgtcagag 15 aatcctgaatgttcagtatgctattccggattatgttcacatatctcctgaatgtcgccatttgatctccagaatatttgttgctgaccctgcaa aggtaaaggaaggatcatgaaagctgcatttgttgatttatttgtgaattttctttatagtatgactgaaaagagaactttgcattttggcctc ttggttggtttcttggcagaggatatcaattccagaaataaggaaccatgaatggtttctaaagaatctaccggcagatctaatgaacgata acacgatgaccactcagtttgatgaatcggatcaacegggccaaagcatagaagaaattatgcagatcattgcagaagcaactgttcct cctgcaggcactcagaatctgaaccattacctcacaggtgagacaacacaaaacataaacttttcgatttcttgtettttttaatgcttctcaa 20 tgtttgaaaaacttatcattaatggataaacaggaagcttggacatagatgacgatatggaggaagacttagagagcgaccttgatgatct tgacatcgacagtagcggagagattgtgtacgcaatgtga [SEQ ID NO: 17], as set forth for example in GenBank Accession No. AJ316009, or a complement thereof; or a nucleotide sequence that has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% 25 sequence identity to SEQ ID NO: 17, or to a complement thereof; or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO:17, or to a complement thereof. [01511 Representative AAPK polypeptides comprises the amino acid sequence: 101521 MDMPPPIMHDSDRYDFVRDIGSGNFGVARLMTDKLTKDLVAVKYI 30 ERGDKIDENVKREIINHRSLRHPNIVRFKEVILTPTHLAIVMEYASGGEMSDRISKAGR FTEDEARFFFQQLISGVSYCHSMQVCHRDLKLENTLLDGDPALHLKICDFGYSKSSVL HSQPKSTVGTPAYIAPEVLLKQEYDGKIADVWSCGVTLYVMLVGSYPFEDPDNPKDF RKTIQRVLSVQYSVPDFVQISPECRDIISRIFVFDPAERITIPEIMKNEWFRKNLPADLVN -45- ENIMDNQFEEPDQPMQSMDTIMQIISEATVPAAGSYYFDEFIEVDEDMDEIDSDYELD VDSSGEIVYAI [SEQ ID NO:58], as set forth for example in GenPept Accession No. AAF27340, or an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 5 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:58. [01531 Illustrative AAPK nucleic acid sequence comprises a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO: 58, or a complement of the nucleotide sequence; or a nucleotide sequence encoding an amino acid sequence having at 10 least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:58, or a complement of that nucleotide sequence. In representative examples, an AAPK nucleic acid sequence comprises the nucleotide sequence: 15 [01541 cggcacgagattaaaaaggccacaatgttgcttactctccaacaacaaccgtaatcctctcggaatctccact acgacgccgtttacttccgatctctctccccgccggagcagcagccatggatatgccgccgccgatcatgcacgacagtgaccgttac gacttcgttcgtgatatcggatcgggaaatttcggcgtcgctagactcatgactgataaactcaccaaagaccttgttgtgtcaagtacat cgaacgtggagataagattgatgaaaatgttaagagagaaataatcaatcacaggtctctaagacatcctaatattgttaggtttaaggag gtcattttaacacctactcatctggccattgtaatggaatatgcatctggaggagaaatgtccgatcgaatagcaaaggggggtttta 20 ctgaggatgaggctcgtttcttctttcaacaactcatatccggggtcagctattgtcattcaatgcaagtatgtcatcgagatctgaagttgg aaaacacgttgttggatggagacccagcacttcatctgaagatttgtgattttggatactccaaatcttcggtgttattcacagccaaagt caactgtgggaactcctgcttatattgctccagaagtacttctgaagcaagagtatgatggaaagattgccgatgtctggtcatgtggtgta accttatacgtgatgctagtggggtcatatccttttgaagatcccgataatccgaaggatttccggaagacaattcagagggttetcagtgt ccagtattccgtaccagactttgttcaaatatctcctgaatgtcgcgacattatatcaagaatctttgtttttgaccctgcagagagaatcacc 25 attccagaaataatgaagaacgaatggttccgaaagaatcttcctgctgacttggtgaatgaaaatataatggataaccaatttgaagagc cagatcagcctatgcagagtatggatacgatcatgcagataatttcagaagctaccgtaccagcagctgggagctattattttgacgagtt tatcgaagtggatgaagatatggatgaaatagactctgactatgaacttgatgtagatagcagtggtgagattgtatatgccatataatttaa tcatcatagaggtcacatattgaaaaggaagcaccttatattgagctttatggctttctcagcctcaaagctaaaaaaataaatattctgaga ctattttctgcagactggatgatgcacgaagttcatcatgttgatttatatattgtatgctttcttggaacatgcattgtccacaccatttatagt 30 atcacttttgtgagttgaggcaacatgttttcgaatttgtagggatcttctttattccttaaaaaaagttccacaacttcaatttaggatgtatatt ggcataattttagaacgtggcatggcataattgagattttatatgcatgaaatatggtaacgagctcttgatttcttttcaaaaaaaaaaaaaa aaaa [SEQ ID NO:59], as set forth for example in GenBank Accession No. AF186020, or a complement thereof; or a nucleotide sequence that has at least 70%, 71%, 72%, 73%, 74%, - 46 - 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO:59, or to a complement thereof; or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO:59, or to a complement thereof. 5 [01551 A non-limiting AHAI polypeptide comprises the amino acid sequence: 10156] MSGLEDIKNETVDLEKIPIEEVFQQLKCTREGLTTQEGEDRIVIFGPN KLEEKKESKILKFLGFMWNPLSWVMEAAALMAIALANGDNRPPDWQDFVGIICLLVI NSTISFIEENNAGNAAAALMAGLAPKTKVLRDGKWSEQEAAILVPGDIVSIKLGDIIPA DARLLEGDPLKVDQSALTGESLPVTKHPGQEVFSGSTCKQGEIEAVVIATGVHTFFGK 10 AAHLVDSTNQVGHFQKVLTSIGNFCICSIAIGIAIEIVVMYPIQHRKYRDGIDNLLVLLI GGIPIAMPTVLSVTMAIGSHRLSQQGAITKRMTAIEEMAGMDVLCSDKTGTLTLNKLS VDKNLVEVFCKGVEKDQVLLFAAMASRVENQDAIDAAMVGMLADPKEARAGIREV HFLPFNPVDKRTALTYIDSDGNWHRVSKGAPEQILDLANARPDLRKKVLSCIDKYAE RGLRSLAVARQVVPEKTKESPGGPWEFVGLLPLFDPPRHDSAETIRRALNLGVNVKM 15 ITGDQLAIGKETGRRLGMGTNMYPSAALLGTDKDSNIASIPVEELIEKADGFAGVFPE HKYEIVKKLQERKHIVGMTGDGVNDAPALKKADIGIAVADATDAARGASDIVLTEPG LSVIISAVLTSRAIFQRMKNYTIYAVSITIRIVFGFMLIALIWEFDFSAFMVLIIAILNDGT IMTISKDRVKPSPTPDSWKLKEIFATGIVLGGYQAIMSVIFFWAAHKTDFFSDKFGVRS IRDNNDELMGAVYLQVSIISQALIFVTRSRSWSFVERPGALLMIAFVIAQLVATLIAVY 20 ADWTFAKVKGIGWGWAGVIWIYSIVTYFPQDILKFAIRYILSGKAWASLFDNRTAFTT KKDYGIGEREAQWAQAQRTLHGLQPKEDVNIFPEKGSYRELSEIAEQAKRRAEIARL RELHTLKGHVESVAKLKGLDIDTAGHHYTV [SEQ ID NO: 18], as set forth for example in GenPept Accession No. NP_179486, or an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 25 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:18. [01571 A representative AHA1 nucleic acid sequence comprises a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO: 18, , or a complement of the nucleotide sequence; or a nucleotide sequence encoding an amino acid sequence having at 30 least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO: 18, , or a complement of that - 47 nucleotide sequence. In illustrative examples, an AHA 1 nucleic acid sequence comprises the nucleotide sequence: [0158] caaccattgatgatcaccaaatcataatcaacggtcgaaatgaatgaaaaataaaagtctgaaacggtggag agtttcgtcgttttgagtggttataaaaagagaaggcgtatatctcctctgcaacagaagctcgctttctctctctctctctcgtgtgtgtgtgt 5 gtgatcgagtggtgaaacgacagagaggggcgtcgttttgttgatttcttctgggtgaagatgtcaggtctcgaagatatcaagaacgag accgttgatctggaaaaaattccgattgaggaagttttccagcagctaaaatgtacaagggaaggattgacaacgcaggaaggggaag acaggattgtgatatttggccccaacaagctcgaagagaagaaggaaagcaaaattctgaagtttctggggttcatgtggaatccgcttt catgggttatggaagctgcagctctcatggccattgctttggctaatggtgataatcgacetceggattggcaagattttgtgggtattatct gtctgcttgttatcaactccacaatcagtttcattgaagaaaacaacgccggaaatgctgcagctgctetcatggctggtcttgctcctaaa 10 accaaggttcttagggatggaaaatggagtgaacaagaggctgctatccttgtcccaggtgatattgttagcattaaacttggagacatta tcccagccgatgcccgtcttcttgaaggagatcctttaaaggttgatcagtctgctctaactggagagtcccttcctgtgaccaagcaccc tggtcaagaagttttctctggttcaacttgtaaacaaggagaaatcgaagcggttgttatagccactggagttcacaccttctttggtaaag ctgctcacettgtggacagcactaaccaagttgggcacttccagaaagttcttacatccattggaaacttctgtatctgttctattgtattgg tatagcgattgaaatagtcgtcatgtaccctatccaacaccgaaagtacagagatggaattgacaatctcttggtcctcttgatcggtggta 15 tccccattgctatgcccacggtcttgtctgtgactatggctategggtctcacaggttgtctcagcaaggtgctatcaccaaacgtatgaca gccattgaagaaatggcgggaatggatgtcctttgcagtgacaaaaccgggacactaacccttaataaattgagtgtggataaaaacttg gttgaggttttctgcaagggtgtggagaaagatcaagttctactatttgcagctatggcttctagggtggagaaccaggacgtattgatg cagccatggttggaatgcttgctgatccgaaagaggcccgagctggaatcagagaggttcacttccttccattcaaccctgtggataag agaactgctttgacttacatcgactctgatggtaactggcacagagtcagcaaaggtgctcccgagcagatccttgaccttgccaatgcc 20 aggcctgaccttaggaagaaggtactctcttgtattgacaagtacgctgagcgcggtcttaggtcgttggcagtagctcgtcaggtggta cccgagaaaacaaaagaaagcccaggtggaccatgggaatttgttggcttgttgcctctttttgaccctccaagacacgacagtgccga aaccattcgtagggcgttgaatctaggtgttaatgtgaagatgatcactggtgatcaacttgctattggtaaggaaaccggtcgcaggett ggaatgggaaccaatatgtatccatctgcggetcttctcggtaccgacaaggactcgaacattgcatccatecctgttgaggagttgat gagaaggctgatgggtttgccggcgtctttccagagcacaaatatgaaattgtgaaaaagctgcaggagaggaagcacattgttggtat 25 gaccggtgatggtgttaatgatgcacccgctttgaagaaagcagatatcggtattgctgtggccgatgctacagatgctgctcgtggtgc ttcagatatcgtcctcacggaacctgggctgagtgtcatcatcagtgccgttctcactagcagagtatttccagagaatgaagaactac accatttatgcagtctcaatcacaatccgtattgtgtttggmcatgcttattgctttgatatgggaatttgacttctcagcgttcatggttctgat tattgccatccttaatgatggtactatcatgacaatetcaaaggacagagtcaagccatctcccacacctgatagctggaagctcaaaga aattttcgccactggaattgtgctgggaggctaccaagccattatgagtgttattttcttctgggctgctcacaagaccgactttttctcgga 30 caagttcggtgtgaggtcaatcagggacaataacgatgaactaatgggtgctgtgtatctacaagttagtatcatcagtcaagctctaatct ttgtcaccaggtcaaggagttggtcatttgtcgaacgtcctggggcgcttctgatgattgcattcgtcattgcacaactggttgcaaetttga tcgcagtgtatgccgactggacatttgcaaaggtgaagggtatcggttggggatgggcaggtgtgatttggatttacagtatcgtaacat acttcccacaggacattttgaagtttgccattcggtatatcttgagtggaaaggcttgggccagcttgtttgacaacaggaccgtttcaca -48 accaagaaagattacggtattggagaaagagaagctcaatgggcacaagctcaaaggacattgcacggtctgcagccaaaagaagat gttaatatettcccagagaaaggaagttacagagagctgtctgagatcgcagagcaagccaagagaagggccgagatagctaggctt agggagcttcacacattgaagggacatgtggaatcagtcgcaaagctaaagggattggacattgatacagcaggacatcactacactg tgtagttggagttgcacaacaacacaaacatttaccgaaaaccaaccccatcatgactcatcttttgttttgtctteacaaatctcttttaaaa 5 gtcattacagtagagggaagagaactcttgtgtattgccataactcttccaaaactcttttttctttttgttattgttagtcattgttcttaaaact ctactcgatatgagctttgcaaatttgcctttgaagaaacaaggccctttgcctttgaagagacaaggcactttgctttttaatggcctaatgt gattattctatcactcttttgtctgtttgatcaatttcatcttgaagataatatttaaatttaattctaaaa [SEQ ID NO: 19), as set forth for example in GenBank Accession No. NM_127453, or a complement thereof; or a nucleotide sequence that has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 10 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO: 19, or to a complement thereof; or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO: 19, or to a complement thereof. [01591 Illustrative vSNAREs AtVAMP711-14 polypeptides comprise the amino 15 acid sequence: [01601 MAILYALVARGTVVLSEFTATSTNASTIAKQILEKVPGDNDSNVSYS QDRYVFHVKRTDGLTVLCMAEETAGRRIPFAFLEDIHQRFVRTYGRAVHTALAYAM NEEFSRVLSQQIDYYSNDPNADRINRIKGEMNQVRGVMIENIDKVLDRGERLELLVD KTANMQGNTFRFRKQARRFRSNVWWRNCKLTVLLILLLLVIIYIAVAFLCHGPTLPSC 20 I [SEQ ID NO:20], as set forth for example in GenPept Accession No. NP 194942, or an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:20. 101611 A representative vSNAREs AtVAMP711-14 nucleic acid sequence comprises 25 a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:20, or a complement of the nucleotide sequence; or a nucleotide sequence encoding an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:20, or a complement of 30 that nucleotide sequence. In illustrative examples, a vSNAREs At VAMP711 nucleic acid sequence comprises the nucleotide sequence: - 49 - 101621 ggcgaaaaggtctccagctccagctaaactccaagaggagagagagagaaagagattcaattatatgcag agagaaaaagaagccaattggttctctctctctctctctctctctctgattgattgatccagcgattcgtctggctggctcggtttccgcggg aaaaccttcttcttcgttgaccttacacgagttttgttgattcggcagggggatcgacgacggagatggcgatttgtacgccctcgtgg tcgtggcacggtggttctttctgagttcaccgccacctctacgaatgegagcaccatcgccaaacagatcctcgagaaggtccctggag 5 acaacgacagcaacgtctcctactctcaggatcgttacgtcttccacgttaaacgcaccgatggcctcaccgttctctgtatggcegaag aaaccgccggaaggagaattccttttgcctttttggaggatattcaccagagattcgtacggacttatggcagggctgttcatacagcact agcttatgcaatgaatgaggaattctctagagttctcagtcagcagattgactattactctaatgatcctaatgccgataggattaatcgtatt aagggtgaaatgaatcaggtgcggggtgtcatgatagaaaacattgacaaagtcctagatagaggtgaacgtttggagcttcttgtcgat aaaaccgccaatatgcaggggaatacattccggttcagaaagcaagctcgtcgttttagaagcaacgtctggtggagaaactgcaagc 10 tcacggtcctcttaatactactactactggtgatcatatacattgcagtggcctttctctgccacggacctactetaccatcttgcatttaagtg catctgtctcttaaggaatccatcccgattctgcgcgttgccacgaetttttatctcctcatctgattgtaaaccttgtgttttcctgagcatttt atgtggcttaaaaagcttatgtttccaactcacgtatatgaaaaagtatacttatttatagagcaatgtaaaagctcattgctttgcgaagtgt gtgaatctttgttgacatcatgttctataaattacatacatagtggaaaagtgaacaggcttgctttgtctgatatgttcatacaactctttgaat ctte [SEQ ID NO:2 1], as set forth for example in GenBank Accession No. NM_119367, or a 15 complement thereof; or a nucleotide sequence that has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO:21, or to a complement thereof; or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO:21, or to a complement thereof. 20 101631 A non-limiting GPA1 polypeptide comprises the amino acid sequence: [01641 MGLLCSRSRHHTEDTDENTQAAEIERRIEQEAKAEKHIRKLLLLGAG ESGKSTIFKQIKLLFQTGFDEGELKSYVPVIHANVYQTIKLLHDGTKEFAQNETDSAK YMLSSESIAIGEKLSEIGGRLDYPRLTKDIAEGIETLWKDPAIQETCARGNELQVPDCT KYLMENLKRLSDINYIPTKEDVLYARVRTTGVVEIQFSPVGENKKSGEVYRLFDVGG 25 QRNERRKWIHLFEGVTAVIFCAAISEYDQTLFEDEQKNRMMETKELFDWVLKQPCFE KTSFMLFLNKFDIFEKKVLDVPLNVCEWFRDYQPVSSGKQEIEHAYEFVKKKFEELY YQNTAPDRVDRVFKIYRTTALDQKLVKKTFKLVDETLRRRNLLEAGLL [SEQ ID NO:22], as set forth for example in GenPept Accession No. NP 180198, or an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 30 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:22. - 50 - 101651 A non-limiting GPA1 nucleic acid sequence comprises a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:22, or a complement of the nucleotide sequence; or a nucleotide sequence encoding an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 5 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:22, or a complement of that nucleotide sequence. In illustrative examples, a GPA1 nucleic acid sequence comprises the nucleotide sequence: 101661 gttaacttaatagtatataaaataaaaatgcatataggttccgtaattaatcttctttatcgtcacgagaggcacat 10 cttttttcaacatttgaccactctctctctctctctctcaggacctttcggcgtaatttcgtcttcccctttgcttaacattttctttcttctttttgac caaatattaaaaatatatccatttttattttatttttaattaaattcataatttgcattttatgtatttataataaaaaggagagaataaatccaaaag agtgaagcaaaaacattaaagcggaaagaaagtggtaaaacaataatagaaacaggagaagcagaagtactacttctttttttgct ctcttctcagaccttgttttgtactttcttcttcttcttcttcttettgtttgcgaactccgatatcttcttcactacctttgactccatttctttttttctt caggtgtaggcattgtcttgttatgagaagcaactgtagctggaagctcaagtatttgtttttagctgtggagcttgaatcttgatagttttcg 15 acttctatgttattacctgtggggatatagaaacaatcatgggcttactctgcagtagaagtegacatcatactgaagatactgatgagaat acacaggctgctgaaatcgaaagacggatagagcaagaagcaaaggctgaaaagcatattcggaagcttttgctacttggtgctgggg aatctggaaaatctacaatttttaagcagataaaacttctattccaaacgggatttgatgaaggagaactaaagagctatgttcagtcatt atgccaatgtctatcagactataaaattattgcatgatggaacaaaggagtttgctcaaaatgaaacagattctgtaaatatatgttattt tgaaagtattgcaattggggagaaactatctgagattggtggtaggttagactatccacgtcttaccaaggacatcgctgagggaataga 20 aacactatggaaggatcctgcaattcaggaaacttgtgctcgtggtaatgagcttcaggttcctgattgtacgaaatatctgatggagaac ttgaagagactatcagatataaattatattccaactaaggaggatgtactttatgcaagagttcgcacaactggtgtcgtggaaatacagtt cagccctgtgggagagaataaaaaaagtggtgaagtgtaccgattgtttgacgtgggtggacagagaaatgagaggaggaaatggat tcatctgtttgaaggtgtaacagctgtgatattttgtgctgccatcagcgagtacgaccaaacgctctttgaggacgagcagaaaaacag gatgatggagaccaaggaattattcgactgggtcctgaaacaaccctgttttgagaaaacatccttcatgctgttcttgaacaagttcgac 25 atatttgagaagaaagttcttgacgttccgttgaacgtttgcgagtggttcagagattaccaaccagtttcaagtgggaaacaagagattg agcatgcatacgagtttgtgaagaagaagtttgaggagttatattaccagaacacggcgccggatagagtggacagggtattcaaaatc tacaggacgacggctttggaccagaagcttgtaaagaaaacgttcaagctcgtagatgagacactaagaaggagaaatttactggagg ctggccttttatgaccttattattacatatctctagtaaattacctctccttattattataagaaaaactcgaaaactgaatgaccgtgtaatttat ctttcgggacaaaagacttagcgattcaaaatctaatgtgtctcgatggctacgactagtttctattttatcattgtttttgttaacattcctctgt 30 etttgacttettatttttttttetcatcaaaaacatetcattttgatettgtggggttatattattattaaaatgaggcatecacatcccgaaate [SEQ ID NO:23], as set forth for example in GenBank Accession No. NM 128187, or a complement thereof; or a nucleotide sequence that has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, -51 - 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO:23, or to a complement thereof; or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO:23, or to a complement thereof. [01671 An illustrative AtABCG22 polypeptide comprises the amino acid sequence: 5 101681 MSMEKPPLASGLARTRSEQLYETVAADIRSPHGSMDANGVPATAPA AVGGGGTLSRKSSRRLMGMSPGRSSGAGTHIRKSRSAQLKLELEEVSSGAALSRASS ASLGLSFSFTGFAMPPEEISDSKPFSDDEMIPEDIEAGKKKPKFQAEPTLPIFLKFRDVT YKVVIKKLTSSVEKEILTGISGSVNPGEVLALMGPSGSGKTTLLSLLAGRISQSSTGGS VTYNDKPYSKYLKSKIGFVTQDDVLFPHLTVKETLTYAARLRLPKTLTREQKKQRAL 10 DVIQELGLERCQDTMIGGAFVRGVSGGERKRVSIGNEIIINPSLLLLDEPTSGLDSTTAL RTILMLHDIAEAGKTVITTIHQPSSRLFHRFDKLILLGRGSLLYFGKSSEALDYFSSIGCS PLIAMNPAEFLLDLANGNINDISVPSELDDRVQVGNSGRETQTGKPSPAAVHEYLVEA YETRVAEQEKKKLLDPVPLDEEAKAKSTRLKRQWGTCWWEQYCILFCRGLKERRHE YFSWLRVTQVLSTAVILGLLWWQSDIRTPMGLQDQAGLLFFIAVFWGFFPVFTAIFAF 15 PQERAMLNKERAADMYRLSAYFLARTTSDLPLDFILPSLFLLVVYFMTGLRISPYPFFL SMLTVFLCIIAAQGLGLAIGAILMDLKKATTLASVTVMTFMLAGGFFVKASPLFLDFL CF [SEQ ID NO:24], as set forth for example in GenPept Accession No. NP_001031843, or an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 20 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:24. 101691 A representative AtABCG22 nucleic acid sequence comprises a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO: 24, or a complement of the nucleotide sequence; or a nucleotide sequence encoding an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 25 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:24, a complement of that nucleotide sequence. In illustrative examples, an AtABCG22 nucleic acid sequence comprises the nucleotide sequence: 101701 ttccccaaaggtatcgattctatatcctaagaaaaaatcatacccatcttcttatagtcattcctgtttttttttcttttc 30 tattttatgtetgttcatttgtttatgttecatatatacatacatataagttctatatatgatetegaacgtatatattggttataaattttggetaaag attatgatttttggtcgacagtttttgaaaaggtcaaaatgtcaatggagaagccacctttggcctccggtttagctcgaacacgatggag cagctatatgagacggttgcagcagacataaggtcacctcacggctccatggacgctaatggtgtgcctgcgacggctccagcagcc - 52 gttggaggaggaggaacgttgtcgaggaaatcaagccggaggttgatggggatgtctccggggaggagtagcggcgccggaacac acataaggaagtctaggagcgctcagcttaagctcgagctagaggaagtgagtagcggcgcagctttgagccgtgcgtctagcgcat cgctcggtctttcattttccttcaccgggtttgctatgccgccggaggaaatctccgactctaaaccgttcagcgacgacgagatgatacc cgaagatattgaagcgggaaagaagaagcctaagtttcaagcagaaccaacattgcccatetttctcaagttcagggatgttacatacaa 5 agtggtgatcaagaaattgacttcatctgtggagaaagagatattaactgggataagtgggagtgtgaatcctggtgaagttcttgctctc atgggaccctcagggagtggcaaaacaactcttcttagcttacttgctggtcgaatctetcaatcctctactggaggctctgttacttacaa cgacaagccttactctaaatacttgaaaagcaagattgggtttgtgactcaagatgatgttctgtttcctcatcttaccgtgaaagaaacgct aacctatgctgctcgtctgcgcttacccaagactcttacgagagagcagaagaagcaacgagctttagacgttatccaagagttgggtt agagagatgccaagacactatgattggtggagcattcgtgcgtggtgtatcaggtggagagaggaaaagagtttctattggaaacgag 10 atcatcattaatccttctctattacttcttgatgaaccaacctctggtttagattccaccactgctcttagaaccattctgatgctccatgacatc gccgaggcggggaaaaccgtgatcacaacgatacatcagccctcgagtaggctcttccataggtttgacaagctgattctactaggaa gaggaagtcttctctactttggaaaatcatcagaagctttagattacttctcttccattggatgctctcctcttatgccatgaatcctgcaga gttcttgctcgatcttgccaacggtaacatcaacgatatctctgtaccttctgagttagatgatagagttcaagttggcaattcaggtagaga aactcaaactggcaagccatctcctgctgctgttcatgagtatctagtggaggcctacgagactagggttgcagaacaggagaagaag 15 aaactattggatcctgtgccactcgatgaagaagctaaggccaaaagtacgcgtctaaagcgccaatggggaacgtgctggtgggag caatattgcatactattctgcagaggactcaaagaacggcgacacgaatacttcagttggttgcgtgttacgcaagtttttccacagctgt cattttaggtcttctctggtggcagtcggacattaggactccaatgggactacaagatcaggctggtttgctcttcttcatagcagttttttgg ggattcttccctgttttcacagcgatctttgcgtttccgcaagagcgagcgatgttaaataaggagagagcagcggatatgtacagattaa gcgcatatttcctagctcgaaccacgagtgatctecctctcgacMattctaccttctctcttccttcttgtcgttatttcatgacaggtcttcg 20 gatcagcccatatcccttcttcttgagcatgctcacagttttcctttgcatcatcgcagctcagggactcggacttgcaattggtgccatttta atggatttaaagaaggctacgactttggcttcagtaactgtcatgacattcatgctcgccggaggattcttcgtcaaggcaagtcctctgttt ettgattteetetgtttttaaettcccetgtttettgatttcccetgttttactgattteettttetgtgtetacacagaaagtgecggttttcatategt ggatacgttatctatctttcaattaccacacctacaagccttcttaaagtacaatatcaggacttcgctgtgtccatcaacgggatgagaat agacaacggactaactgaagtagccgcactcgttgtcatgatattcggttatcgcctcctcgegtatctgtctctaaggcaaatgaagat 25 gtaacataacccattttccacacgaagaaatcaaataacatagaagaagcataaaaagagtgcatcagatcttgatgatettgacacgac caatccttgacaatgattaagagattggtcctaagattattcttgcttaattacaaaggtgttgtgagtttgataatgattggatggtgatagat gtcttgattggagataaatatatatgttcaagtttgtaattgtagttegaatctaaaattggttaaagtttattaacaagaagaaccttgtggctg gatctgatcttcttaaattccaatccaattattcttagtat [SEQ ID NO:25], as set forth for example in GenBank Accession No. NM_001036766, or a complement thereof; or a nucleotide sequence that has at 30 least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO:25, or to a complement thereof; or a nucleotide sequence - 53 that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO:25, or to a complement thereof. [0171] Non-limiting AtABCG40 polypeptides comprise the amino acid sequence: 101721 MEGTSFHQASNSMRRNSSVWKKDSGREIFSRSSREEDDEEALRWAA 5 LEKLPTFDRLRKGILTASHAGGPINEIDIQKLGFQDTKKLLERLIKVGDDEHEKLLWKL KKRIDRVGIDLPTIEVRFDHLKVEAEVHVGGRALPTFVNFISNFADKFLNTLH LVPNR KKKFTILNDVSGIVKPGRMALLLGPPSSGKTTLLLALAGKLDQELKQTGRVTYNGHG MNEFVPQRTAAYIGQNDVHIGEMTVRETFAYAARFQGVGSRYDMLTELARREKEAN IKPDPDIDIFMKAMSTAGEKTNVMTDYILKILGLEVCADTMVGDDMLRGISGGQKKR 10 VTTGEMLVGPSRALFMDEISTGLDSSTTYQIVNSLRNYVHIFNGTALISLLQPAPETFN LFDDIILIAEGEIIYEGPRDHVVEFFETMGFKCPPRKGVADFLQEVTSKKDQMQYWAR RDEPYRFIRVREFAEAFQSFHVGRRIGDELALPFDKTKSHPAALTTKKYGVGIKELVK TSFSREYLLMKRNSFVYYFKFGQLLVMAFLTMTLFFRTEMQKKTEVDGSLYTGALFF ILMMLMFNGMSELSMTIAKLPVFYKQRDLLFYPAWVYSLPPWLLKIPISFMEAALTTF 15 ITYYVIGFDPNVGRLFKQYILLVLMNQMASALFKMVAALGRNMIVANTFGAFAMLV FFALGGVV LSRDDIKKWWIWGYWISPIMYGQNAILANEFFGHSWSRAVENSSETLGV TFLKSRGFLPHAYWYWIGTGALLGFVVLFNFGFTLALTFLNSLGKPQAVIAEEPASDE TELQSARSEGVVEAGANKKRGMVLPFEPHSITFDNVVYSVDMPQEMIEQGTQEDRLV LLKGVNGAFRPGVLTALMGVSGAGKTTLMDVLAGRKTGGYIDGNITISGYPKNQQT 20 FARISGYCEQTDIHSPHVTVYESLVYSAWLRLPKEVDKNKRKIFIEEVMELVELTPLR QALVGLPGESGLSTEQRKRLTIAVELVANPSIIFMDEPTSGLDARAAAIVMRTVRNTV DTGRTVVCTIHQPSIDIFEAFDELFLLKRGGEEIYVGPLGHESTHLINYFESIQGINKITE GYNPATWMLEVSTTSQEAALGVDFAQVYKNSELYKRNKELIKELSQPAPGSKDLYFP TQYSQSFLTQCMASLWKQHWSYWRNPPYTAVRFLFTIGIALMFGTMFWDLGGKTKT 25 RQDLSNAMGSMYTAVLFLGLQNAASVQPVVNVERTVFYREQAAGMYSAMPYAFA QVFIEIPYVLVQAIVYGLIVYAMIGFEWTAVKFFWYLFFMYGSFLTFTFYGMMAVAM TPNHHIASVVSSAFYGIWNLFSGFLIPRPSMPVWWEWYYWLCPVA WTLYGLIASQFG DITEPMADSNMSVKQFIREFYGYREGFLGVVAAMNVIFPLLFAVIFAIGIKSFNFQKR [SEQ ID NO:26], as set forth for example in GenPept Accession No. NP_173005, or an 30 amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:26. -54- 101731 A non-limiting AtABCG40 nucleic acid sequence comprises a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:26, or a complement of the nucleotide sequence; or a nucleotide sequence encoding an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 5 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:26, or a complement of that nucleotide sequence. In illustrative examples, an AtABCG40 nucleic acid sequence comprises the nucleotide sequence: [0174] atatcatctttcatctacaatttctctcttgagtttcttttcatgtgttaaagtcttattgtcttgctaaattctttaaatctt 10 tcatctgacacaaacaaaaaaagagagaagaaaaaaaagaagaagactttgatttcttggatacaaaatggagggaactagttttcacc aagcgagtaatagtatgagaagaaactcatcggtgtggaagaaagattcaggaagggagattttctcgaggtcatctagagaagaaga cgatgaagaagctttgagatgggctgctcttgagaagettccacttttgatcgtetcaggaaaggaatcctaatgcctcacatgcgg aggacccatcaacgagatcgatattcagaagcttgggtttcaagatactaagaaactgetagagaggetcatcaaagtcggtgacgatg agcatgagaaactcctctggaaactcaagaaacgtatcgatagagttggaatcgatcttccgacaatagaagttcggtttgatcattaaa 15 agttgaagcagaggttcatgttggaggcagagctttacctacgttcgtcaatttcatctccaattttgctgataagttcctgaatactctgcat cttgttccgaaccgaaagaagaagttcactatactcaacgacgtcagcggaatcgtcaagcctggcaggatggctctgcttttgggtct ccaagttctgggaaaacgaccctcttgcttgccttggcgggaaagcttgatcaagaactaaagcaaactggaagagtgacatacaatg gtcatggaatgaacgagtttgtgccacaaagaacagctgcatatatcggccaaaacgatgttcatatcggtgagatgactgttcgtgaga cttttgcttacgcagctcgcttccaaggtgttggttcgcgttatgacatgttgacagagttggcaagaagagagaaagaagcaaacatca 20 aacctgaccctgatattgatatattcatgaaggcgatgtcaacagcaggtgaaaaaacaaatgtgatgacagattatatcctcaagattt aggacttgaggtctgtgcagacactatggtcggcgatgatatgttgagaggcatctccggaggacaaaagaagcgtgtcactactggt gaaatgctggttggaccgtctagggctctgttcatggatgagatatgactggtttagatagttcaacgacttaccagatagtgaatccct cagaaactatgttcatatcttcaatgggacagctctgatctctctccttcagcctgcgccagagacattcaatctcttcgatgatatcattctc attgcagaaggcgagatcatctacgagggccctcgtgatcacgttgtggagttctttgagaccatgggattcaaatgtctccaagaaaa 25 ggcgttgctgatttecttcaagaagtgacatcaaagaaagaccaaatgcagtactgggcacgacgtgatgagcettacaggttcattaga gtgagagagtttgcagaggcgtttcaatcattccacgttggccggagaatcggagatgagcttgctttgccctttgacaagacaaagag ccatccggctgctctaaccaccaagaaatacggagttgggattaaagaacttgtcaagaccagcttctcaagagaatacttactcatgaa aagaaactcctttgtttactacttcaagtttggacaactgctggtaatggcatttttgacaatgacgttgttctttggacggagatgcaaaag aagactgaggttgatgggagtetctacactggagccttgttcttcatccttatgatgctcatgttcaatggaatgtctgaactttcaatgacca 30 tagcaaaacttcctgtgttttacaaacaaagagatctectcttctaccctgcatgggtgtactctctgcctccttggctcctcaagatacctat aagcttcatggaagccgctctcacaacattcatcacttactatgtcatcggctttgatcccaacgttggaaggctgtttaagcagtatattct cctcgtgctcatgaaccaaatggcttcagcattgtttaagatggtggcagcattgggaagaaacatgatcgttgcaaatacatttggtgca tttgcgatgctcgtcttctttgccttgggtggtgtggtactttcacgagacgacattaagaagtggtggatatggggttactggatctcccca - 55 ataatgtatggacagaacgcgatcctagccaatgagttctttggacacagctggagtcgagctgtcgaaaactcgagegaaacacttgg agttactttccttaagtctcgtgggttcttaccccatgcatactggtactggattggaactggagccttacttgggttcgtcgtgttattcaattt tggtttcacgctggctctgacgtttctgaactccttgggaaagcctcaagctgttattgcagaagagcctgcgagtgatgagacagaactt cagtctgctaggtcagaaggtgtagttgaagctggtgccaataagaaaagagggatggtgcttccatttgagccacattcaattacctt 5 gacaatgttgtatactcagttgacatgccccaggaaatgatagagcaaggcacacaagaagacagacttgtcctgttgaaaggtgtgaa tggtgcattcaggccaggegtgctcacggctctcatgggtgtctctggagetggcaaaaccactctgatggatgttcttgccggaagga aaaccggtggttatattgatggcaacatcaccatttccggttaccctaagaatcaacaaacatttgcccgtatctcaggatactgtgaacaa actgatatccattccccacatgtcactgtttacgagtccttggtttactcagcctggctccgattacctaaagaagttgataaaaacaagag aaagatattcatagaggaagtgatggagctggtggagttaacgccgctgaggcaagcactggttggactacctggtgagagcggtttg 10 tcaacagagcaaagaaagagactgaccattgcggtggagctggttgcaaatccttccatcatattcatggatgaacctacttcaggattg gatgcacgagctgctgccatcgttatgaggactgtaaggaacacagttgacactggtagaacagtcgtctgcaccattcaccagcctag catcgacatctttgaagcctttgatgagttgttcctacttaagcgtggaggtgaggagatatacgttggacctcttggccacgaatcaacc catttgatcaactattttgagagtattcaaggaatcaacaagatcacagaaggatacaacccagcaacctggatgcttgaagtctcaacc acatctcaagaagcggctttaggagtcgatttcgcccaagtctacaaaaattcagaactttacaagagaaacaaggagctaatcaagga 15 gctaagccagccagetccaggatcaaaagatttatatttcccaacacaatactctcaategttcttgacacaatgtatggcttctctatggaa acaacactggtcctactggagaaatcctccttacacagccgtgagattcctcttcacaatcggcattgctcttatgttcggcacaatgtttg ggaccttggaggcaaaacgaaaacgagacaggatttatcgaatgcaatgggttcaatgtacacagctgttctcttcctcggattacaaaa cgcagcttcagtgcaaccagtcgtcaacgtcgaaagaactgtcttttaccgagaacaagccgccggaatgtactccgccatgccttatg ctttcgctcaggttttcatcgagatcccatacgttctcgtgcaagcgatagtgtacggtctcatagtgtacgctatgataggattgagtgg 20 acggcggtgaagttettctggtacctcttctttatgtacggatcattcttaacmcaccttctacggaatgatggctgtagctatgacgccta accaccacatcgcctccgtcgtctcctccgctttctacggcatctggaatctcttctccggcttcctcatccctcgtcccagtatgcctgtgt ggtgggaatggtactactggctttgcccagttgcatggacattgtatggattaatcgcatcacagttcggtgatattacagaacctatggca gatagtaatatgagtgtgaagcaattcattagagaatttatggatatagagaaggtttcttgggtgtggttgcgcatgaacgtcatcttt cctttgctetttgccgttatctttgctatcggaatcaagagtttcaatttccaaaaacgatagacagtttatagttttgcatttatttcatgtaac 25 acaaataaaaagagactttttgtttatatgctattctttctatttttgtaatgccgtattgatattaataaaaggatgatcaacaacactggattag aatg [SEQ ID NO:27], as set forth for example in GenBank Accession No. NM_101421, or a complement thereof; or a nucleotide sequence that has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO:27, or to 30 a complement thereof, or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO:27, or to a complement thereof. 101751 An illustrative AtMRP4 polypeptide comprises the amino acid sequence: - 56 - 101761 MWLLSSSPWLSELSCSYSAVVEHTSSVPVPIQWLRFVLLSPCPQRAL FSAVDFIFLLCFALHKLFSSPSSSSEINGHAEIRKPLIGIRGRTPTRTTAWFKTTVAVTVL LSFCSVVLCVLAFTGKRRTQRPWNLIDPLFWLIHAVTHLVIAVLVLHQKRFAALNHPL SLRIYWISSFVLTSLFAVTGIFHFLSDAATSLRAEDVASFFSFPLTAFLLIASVRGITGLV 5 TAETNSPTKPSDAVSVEKSDNVSLYASASVFSKTFWLWMNPLLSKGYKSPLTLEQVP TLSPEHKAERLALLFESSWPKPSENSSHPIRTTLLRCFWKEILFTAILAIVRLGVMYVGP VLIQSFVDFTSGKRSSPWQGYYLVLILLVAKFVEVLTTHQFNFDSQKLGMLIRSTLITA LYKKGLKLTGSARQNHGVGQIVNYMAVDAQQLSDMMLQLHAIWLMPLQVTVALV LLYGSLGASVITAVIGLTGVFVFILLGTQRNNGYQFSLMGNRDSRMKATNEMLNYM 10 RVIKFQAWENHFNKRILKFRDMEFGWLSKFLYSIAGNIIVLWSTPVLISALTFATALAL GVKLDAGTVFTTTTIFKILQEPIRTFPQSMISLSQAMISLGRLDSYMMSKELSEDAVER ALGCDGNTAVEVRDGSFSWDDEDNEPALSDINFKVKKGELTAIVGTVGSGKSSLLAS VLGEMHRISGQVRVCGSTGYVAQTSWIENGTVQDNILFGLPMVREKYNKVLNVCSL EKDLQMMEFGDKTEIGERGINLSGGQKQRIQLARAVYQECDVYLLDDVFSAVDAHT 15 GSDIFKKCVRGALKGKTVLLVTHQVDFLHNVDCILVMRDGKIVESGKYDELVSSGLD FGELVAAHETSMELVEAGADSAAVATSPRTPTSPHASSPRTSMESPHLSDLNDEHIKS FLGSHIVEDGSKLIKEEERETGQVSLGVYKQYCTEAYGWWGIVLVLFFSLTWQGSLM ASDYWLAYETSAKNAISFDASVFILGYVIIALVSIVLVSIRSYYVTHLGLKTAQIFFRQ LNSILHAPMSFFDTTPSGRILSRASTDQTNVDILIPFMLGLVVSMYTTLLSIFIVTCQYA 20 WPTAFFVIPLGWLNIWYRNYYLASSRELTRMDSITKAPIIHHFSESIAGVMTIRSFRKQ ELFRQENVKRVNDNLRMDFHNNGSNEWLGFRLELVGSWVLCISALFMVLLPSNVIRP ENVGLSLSYGLSLNSVLFFAIYMSCFVENKMVSVERIKQFTDIPSESEWERKETLPPSN WPFHGNVHLEDLKVRYRPNTPLVLKGITLDIKGGEKVGVVGRTGSGKSTLIQVLFRL VEPSGGKIIIDGIDISTLGLHDLRSRFGIIPQEPVLFEGTVRSNIDPTEQYSDEEIWKSLER 25 CQLKDVVATKPEKLDSLVVDNGENWSVGQRQLLCLGRVMLKRSRLLFLDEATASV DSQTDAVIQKIIREDFASCTIISIAHRIPTVMDGDRVLVIDAGKAKEFDSPARLLERPSL FAALVQEYALRSAGI [SEQ ID NO:28], as set forth for example in GenPept Accession No. NP_182301, or an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 30 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:28. 101771 A representative AtMRP4 nucleic acid sequence comprises a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:28, or a complement of -57the nucleotide sequence; or a nucleotide sequence encoding an amino acid; 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:28; or a complement of that nucleotide sequence. In illustrative examples, an AtMRP4 nucleic acid sequence comprises 5 the nucleotide sequence: 101781 ccacttcaaacaaaccgataattcagaggaattctcttcctctctctctctctttatattttttcactgggaaaa atgtggttgctttcgtcttctccatggctctctgagctctcatgttcatattcggctgttgtagaacatacgtcttcagttccagtaccaateca atggctcagatttgttttactctctccttgccctcaacgcgctctcttctccgccgttgattttatattcctcctctgttcgctcttcacaagctct tttcttctccttcttcttcttccgaaatcaacggacacgctgagattaggaaacctcttattggtatccgtggaagaaccccaaccagaaca 10 accgcatggttcaaaacgacggtcgcagtcaccgttctattgtcgttttgctccgtcgtgctctgcgttttggccttcactggcaagcgacg gactcagagaccatggaacctcatagacccgctettttggcttattcacgccgttacacacctagtcatcgccgttctcgtcctccaccag aagagattcgctgctctaaatcatcccttatctctacgaatctactggatttccagtttcgtcctcacgtctctcttcgccgtcactgggatttt ccattttctctccgacgccgccacgagtctgagagcagaagacgtcgcttcattcttctccttccctttaaccgcctttcttetcatcgcctcc gtcagaggaatcaccggcctcgtcacagcggagaccaacagtcctacgaaaccatccgacgccgtttcggtggagaaatccgataac 15 gtctctctctacgcgtctgcttctgttttctcgaaaacgttctggttatggatgaatcctttactcagcaaaggctacaaatctccactgacgc tcgaacaagtccccacgctttctccagagcacaaagcagagaggctcgcgcttctcttcgaatcgagttggcccaaaccgtcggagaa ttctagccaccctatccgtacgactctactccgatgtttctggaaggagatcctcttcaccgcgattctagccatcgtccgtctcggcgtca tgtacgttggtcccgttctcatccagagcttcgtcgatttcacctccggcaagagatcctccccgtggcaaggttattacctcgtcctcatc ctccttgttgccaaattcglcgaggtcttgacgacgcatcagttcaatttcgattcccagaagcttgggatgcttataaggtcaactctaat 20 actgcactctacaagaaaggtttaaagctcacaggctctgcgcgtcagaaccacggcgtaggacaaatcgtgaattacatggccgtag atgcacaacagctctctgacatgatgcttcagctccacgcaatctggctcatgcctttgcaagtcactgttgcactagtgcttctctacggg agcctaggcgcgtctgttataaccgcggttattgggctgactggagtgttcgtcttcatcctcctggggactcagagaaacaacggatac caattcagcttgatgggaaaccgagattctcggatgaaggccaccaacgagatgctcaattacatgcgagtcatcaagtttcaggcttgg gagaatcattttaacaagaggatcctcaaattcagggacatggagtttggttggctatccaagtttctttactccattgctggcaatattattg 25 tcctctggagcacgccagtgcttatctctgctctcaccttcgccaccgcccttgccttgggagtcaagcttgacgctgggactgtgttcac caccacaaccattttcaagatcctgcaagaacccatcaggacgtttcctcagtctatgatttctctctcgcaggcaatgatctctcttggga gactggactcatacatgatgagcaaagagctgtcggaagatgctgtggagagagccctgggttgtgatggtaatactgccgtggaggt cagagatggaagctttagttgggatgatgaggacaacgaacctgctctcagtgatatcaaettcaaggttaagaaaggtgagctcactg cgatagttggaaccgttggttcagggaaatcttctctgttagcttcggttcttggtgaaatgcacagaatctcaggccaggtgagagtttgt 30 gggagcacaggttatgtagctcagacgtcgtggattgaaaacgggacggttcaagacaacatcttgtttggtcttecaatggttagagag aagtacaacaaagttetcaatgtctgttctcttgaaaaagacetacaaatgatggagtttggagataagactgagattggagaacgcgga atcaacctcagcggagggcagaagcaacgtatacagetcgcacgtgctgtctatcaggaatgcgatgtatacttgctcgacgatgttttt agegcagtggatgctcataccggttcagatatattcaagaaatgtgtaagaggagctctgaaaggcaagaccgtattactcgttacccat -58caagtggatttcttgcacaacgtggattgcatcttggtgatgcgggatggaaagattgttgaatcaggaaaatatgacgaattagtcagct ccggattggattttggggaacttgtggctgcacatgagacgtcaatggagctggttgaagccggtgcagactctgcagcagtcgccac atccccaagaacaccaacgtctccccatgcaagctctccgagaacgtcaatggagtctcctcacttaagtgatctaaacgatgagcatat caaatcatttctcggttctcacatcgtagaagatggctcgaagctcatcaaagaagaagaaagggaaaccggacaggttagcttagga 5 gtttacaaacagtactgcactgaggcttatggctggtggggaattgtgcttgttctgttcttetctctgacgtggcagggatctctaatggcc agcgattactggcttgcatacgaaacatcagccaaaaatgcaatatcatttgatgcttccgttttcattetcggatatgtaattattgcacttgt ttccatcgttttggtgagcatccggtcatattacgtcacccacttgggactcaagacggctcagatctttttccgacagattcttaatagtatc ttacacgctcccatgtcattctttgacaccacgccatcgggaagaattctcagtcgggcatcgactgatcagaccaatgtcgatatccttat tccgtttatgctcggacttgtggtctcaatgtacaccactctgctgagcattttcatagttacctgccagtacgcttggccaactgcattctttg 10 tgattccccttggctggcttaacatctggtaccggaactattacctcgcttcttcccgtgaattaacacgcatggactcaatcactaaggctc ccatcatccaccatttctctgaaagtatagctggagtgatgacaatccgatcattcaggaagcaggagttgtttagacaagagaatgtaaa acgtgtaaatgataatctcaggatggacttccacaacaatggctccaacgaatggctcgggtttcggctggagctggttgggagctggg tgctctgcatctcggctttgtttatggtattgttacccagcaacgttatcagaccagagaatgtagggttgtccctgtcgtacggactgtccc tgaactcggttctgttctttgccatatacatgagctgctttgtcgagaacaagatggtttcagttgaaaggatcaaacagttcactgatattcc 15 ctcagaatccgagtgggagagaaaagaaacccttccaccttcgaattggcccttccatggcaatgtacatctcgaagacctcaaggtgc getacagaccgaacactccacttgtgctcaaggggatcactcttgacatcaaaggaggagagaaggttggtgtggttggacggacgg gaagcgggaaatcgacattgatccaagtcttgttcaggcttgtagaaccatcaggagggaagataatcatagacgggattgatataagc actctagggctacatgatctcaggtcaagattcggaatcattccgcaagaacctgtcctctttgaaggaaccgtgagaagcaacatcgac ccgacagagcaatactctgacgaagaaatctggaagagcctggaacggtgtcaactcaaggatgttgtagctaccaagcctgagaag 20 ctcgattctttggtggttgataatggggagaactggagcgtagggcagaggcagcttctatgcttaggcagggttatgttgaaacgcagc agacttctcttcctagacgaagcaactgcatccgttgattcccaaaccgacgccgtgattcagaagatcatcagagaagactttgcgtcg tgcaccatcatcagcatcgcccaccggattcctacagtgatggacggcgatcgagtccttgtcattgatgctgggaaagcgaaagagtt cgatagcccggctcgcttgctggagaggccgtctctgtttgcggcgctggtgcaagagtacgctctccgatctgccggaatatgaatctt ttacgccggcggggactgaaaatattttaaaaccttcttcaaatgagatttgtatcaaagaaaattcctataatctcacatgttttagattcaa 25 aaacgacatcgtaattttagtgcagcaagcaacacagtaaatttttaattcgctcactatacttcaaacaactctctaataacaatccagtaat ctaccaacacgtgaatttttctc [SEQ ID NO:29], as set forth for example in GenBank Accession No. NM_130347, or a complement thereof; or a nucleotide sequence that has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to 30 SEQ ID NO:29, or to a complement thereof; or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO:29, or to a complement thereof. [0179] A non-limiting RBOHD polypeptide comprises the amino acid sequence: - 59 - 101801 MKMRRGNSSNDHELGILRGANSDTNSDTESIASDRGAFSGPLGRPK RASKKNARFADDLPKRSNSVAGGRGDDDEYVEITLDIRDDSVAVHSVQQAAGGGGH LEDPELALLTKKTLESSLNNTTSLSFFRSTSSRIKNASRELRRVFSRRPSPAVRRFDRTS SAAIHALKGLKFIATKTAAWPAVDQRFDKLSADSNGLLLSAKFWECLGMNKESKDF 5 ADQLFRALARRNNVSGDAITKEQLRIFWEQISDESFDAKLQVFFDMVDKDEDGRVTE EEVAEIISLSASANKLSNIQKQAKEYAALIMEELDPDNAGFIMIENLEMLLLQAPNQSV RMGDSRILSQMLSQKLRPAKESNPLVRWSEKIKYFILDNWQRLWIMMLWLGICGGLF TYKFIQYKNKAAYGVMGYCVCVAKGGAETLKFNMALILLPVCRNTITWLRNKTKLG TVVPFDDSLNFHKVIASGIVVGVLLHAGAHLTCDFPRLIAADEDTYEPMEKYFGDQPT 10 SYWWFVKGVEGWTGIVMVVLMAIAFTLATPWFRRNKLNLPNFLKKLTGFNAFWYT HHLFIIVYALLIVHGIKLYLTKIWYQKTTWMYLAVPILLYASERLLRAFRSSIKPVKMI KVAVYPGNVLSLHMTKPQGFKYKSGQFMLVNCRAVSPFEWHPFSITSAPGDDYLSV HIRTLGDWTRKLRTVFSEVCKPPTAGKSGLLRADGGDGNLPFPKVLIDGPYGAPAQD YKKYDVVLLVGLGIGATPMISILKDIINNMKGPDRDSDIENNNSNNNSKGFKTRKAYF 15 YWVTREQGSFEWFKGIMDEISELDEEGIIELHNYCTSVYEEGDARVALIAMLQSLQHA KNGVDVVSGTRVKSHFAKPNWRQVYKKIAVQHPGKRIGVFYCGMPGMIKELKNLA LDFSRKTTTKFDFHKENF [SEQ ID NO:30], as set forth for example in GenPept Accession No. NP_199602, or an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 20 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:30. 101811 A non-limiting RBOHD nucleic acid sequence comprises a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:30, or a complement of the nucleotide sequence; or a nucleotide sequence encoding an amino acid sequence having at 25 least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:30, or a complement of that nucleotide sequence. In illustrative examples, a RBOHD nucleic acid sequence comprises the nucleotide sequence: 30 [01821 atacacaaaaatcaaacaccttttgagagcggttattttttctctatcaactaatacagtaaccttacgggtgtttat ttgtatagatctctgtggttttcttggccaaatctagtgagatctttttggtttctcgaattcgatgaaaatgagacgaggcaattcaagtaacg accatgaacttgggattctacgaggagctaactcggacaccaactcggacacggagagcatcgctagcgaccgtggtgcctttagcg gtccgcttggccggcctaaacgtgcgtccaagaaaaacgcaagattcgccgacgatcttcccaagagaagcaatagtgttgctggcg - 60 gccgtggtgatgacgatgagtacgtggagatcacgctagacatcagggacgactcggtggccgtccatagtgtccaacaagcagctg gaggtggaggccacctggaggacccggagctagcccttcttacgaagaagactctcgagagcagcctcaacaacaccacctccttat ctttcttccgaagcacctcctcacgcatcaagaacgcctcccgcgagctccgccgcgtgttctctagacgtccctccccggccgtggg cggtttgaccgcacgagctccgcggccatccacgcactcaaaggtctcaagttcattgccaccaagacggccgcatggccggccgtc 5 gaccaacgtttcgataaactctccgctgattccaacggcctcttactctctgccaagttttgggaatgcttaggaatgaataaggaattaa agacttcgctgaccagctctttagagcattagctcgccggaataacgtetccggcgatgcaatcacaaaggaacagttaggatatttg ggaacagatctcagacgaaagctttgatgccaaactccaagtcttttttgacatggtggacaaagatgaagatgggcgagtaacagaag aagaggtggctgagattattagtettagtgcttctgcaaacaagctctcaaatattcaaaagcaagccaaagaatatgcggcactgataat ggaagagttggacccagacaatgctgggtttattatgatcgaaaacttggaaatgttgctattacaagcaccaaaccagtcggtgggat 10 gggagacagcaggatacttagtcagatgttaagtcagaagcttagaccggcaaaagagagcaaccctttagtgagatggtcggagaa aatcaaatatttcatacttgacaattggcagagactatggataatgatgttatggcttggcatctgtggtggcctctttacttataaattcattc agtacaagaacaaagctgcctatggtgtcatgggttattgcgtttgtgtcgccaaaggaggcgccgagactctcaaattcaacatggtc tcatattgttgcctgtttgtcgaaacaccatcacttggcttaggaacaagactaagcttggtactgtcgttcettttgatgatagtcttaaettc cacaaggttattgcaagcgggatagtcgtcggtgttttactccatgcgggtgcccatttaacgtgtgattttccacgtttaattgccgcggat 15 gaggacacctatgagccgatggaaaaatactttggggatcaaccgactagctactggtggtttgtgaaaggagtggaaggatggactg gcattgtgatggttgtgctaatggctatagcctttacactcgcgacgccttggttccgacgtaacaagcttaacttacctaacttcctcaaga agcttaccggtttcaacgccttttggtacacccaccatttgttcatcattgtttatgctcttctcattgtccatggtatcaagctctacctcacaa agatttggtatcagaaaacgacatggatgtatcttgctgtacccatccttctatatgcatcggagaggctgctccgtgtttcagatcaagc atcaaaccggttaagatgatcaaggtggctgtttacccogggaacgtgttgtctctacacatgacgaagccacaaggattcaaatacaaa 20 agtggacagttcatgttggtgaactgccgagccgtatctccattcgaatggcatcctttctcaatcacatcagctcccggagacgattacc tgagcgtacatatccgcactctcggtgactggacacgtaagctcaggaccgttttctccgaggtttgcaaacctctaccgccggtaaaa gcggtcttctccgagcagacggaggagatggaaacctcccgtteccgaaggtccttatcgacggtccatacggtgctcccgcacaag actacaagaaatacgacgtggtactcctcgtaggtctcggcattggagccacgcctatgatcagtatcttaaggacatcatcaacaaca tgaaaggtcctgaccgcgacagcgacattgagaacaataacagtaacaacaatagtaaagggtttaagacaaggaaagttatttctac 25 tgggtgactagggaacaaggatcattcgagtggttcaagggaataatggacgagatttcggagttagacgaggaaggaatcatgag cttcacaattattgcacgagtgtgtacgaggaaggtgatgcaagagtggctctcattgccatgcttcagtcgttgaacacgtaagaac ggtgtggatgttgtgtcgggtacacgtgtcaagtcccacttcgctaaacctaactggagacaagtctacaagaagatcgctgttcaacat cccggcaaaagaataggagtcttctactgtggaatgccaggaatgataaaggaattaaaaaatctagctttggatttttctcgaaagacaa ctaccaagtttgacttccacaaagagaacttctagattaattatatacgttgtagaaaaataaaacaagaaacaactatacaaataaatattt 30 attttaaattcttcatttttatgtaaaattatetgagttatetttttttgttettetatatccctaccgttttgttggttactttttcactacttagtttaat gccaaattaagtataagatagtagaagtttatatagttacagtttggtgttgtaaacatgtaatcatggagttatctgtactattattttgtactt atattcgaatattaacaactaaagatagttttg [SEQ ID NO:3 1], as set forth for example in GenBank Accession No. NM_124165, or a complement thereof; or a nucleotide sequence that has at -61 least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO:3 1, or to a complement thereof; or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID 5 NO:31, or to a complement thereof. 101831 An illustrative RBOHF polypeptide comprises the amino acid sequence: 101841 MKPFSKNDRRRWSFDSVSAGKTAVGSASTSPGTEYSINGDQEFVEV TIDLQDDDTIVLRSVEPATAINVIGDISDDNTGIMTPVSISRSPTMKRTSSNRFRQFSQE LKAEAVAKAKQLSQELKRFSWSRSFSGNLTTTSTAANQSGGAGGGLVNSALEARAL 10 RKQRAQLDRTRSSAQRALRGLRFISNKQKNVDGWNDVQSNFEKFEKNGYIYRSDFA QCIGMKDSKEFALELFDALSRRRRLKVEKINHDELYEYWSQINDESFDSRLQIFFDIVD KNEDGRITEEEVKEIIMLSASANKLSRLKEQAEEYAALIMEELDPERLGYIELWQLETL LLQKDTYLNYSQALSYTSQALSQNLQGLRGKSRIHRMSSDFVYIMQENWKRIWVLSL WIMIMIGLFLWKFFQYKQKDAFHVMGYCLLTAKGAAETLKFNMALILFPVCRNTIT 15 WLRSTRLSYFVPFDDNINFHKTIAGAIVVAVILHIGDHLACDFPRIVRATEYDYNRYLF HYFQTKQPTYFDLVKGPEGITGILMVILMIISFTLATRWFRRNLVKLPKPFDRLTGFNA FWYSHHLFVIVYILLILHGIFLYFAKPWYVRTTWMYLAVPVLLYGGERTLRYFRSGS YSVRLLKVAIYPGNVLTLQMSKPTQFRYKSGQYMFVQCPAVSPFEWHPFSITSAPED DYISIHIRQLGDWTQELKRVFSEVCEPPVGGKSGLLRADETTKKSLPKLLIDGPYGAP 20 AQDYRKYDVLLLVGLGIGATPFISILKDLLNNIVKMEEHADSISDFSRSSEYSTGSNGD TPRRKRILKTTNAYFYWVTREQGSFDWFKGVMNEVAELDQRGVIEMFHNYLTSVYEE GDARSALITMVQALNHAKNGVDIVSGTRVRTHFARPNWKKVLTKLSSKHCNARIGV FYCGVPVLGKELSKLCNTFNQKGSTKFEFHKEHF [SEQ ID NO:32], as set forth for example in GenPept Accession No. NP_564821, or an amino acid sequence having at least 25 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:32. 101851 A representative RBOHF nucleic acid sequence comprises a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:32, or a complement of 30 the nucleotide sequence; or a nucleotide sequence encoding an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% - 62 sequence similarity or sequence identity to SEQ ID NO:32, or a complement of that nucleotide sequence. In illustrative examples, a RBOHF nucleic acid sequence comprises the nucleotide sequence: [01861 atgcataaactagaatgaccaacaaagatactctcaaagtctcacttgtctaaaaaacagataaaaatcatttcc 5 aattttgagaaaaggaaaaaaaacaactccaataatactttttgattttcttttcgttgcattaaaataaatgtggcaaaaatccaataatatta ctattataagttaaaaaaactcaaaaaaaaaaaaaagtatattccgtcaactttttctcagtccggttcaatcatctccatcatcgttgatctct ctctctccgtcaacgttctctetataaagcagagagtttcacagcgcgtgaaaaatggcttttcccacttagccacaacgacgcgtttcaat tctccatctctccctctctctctctctttcttcaaagattccaccaacctatacatatacttatatataatctatgctacgtgtatatacttcgatat ccttcaaccaactctttgaattccgactttggatctatgaaaccgttctcaaagaacgatcggcgacggtggtcatttgattcagtttcgcc 10 ggaaaaaccgccgtcggaagtgcatcaacttcaccgggaactgaatactccattaacggtgatcaagagttcgttgaagtcacaatcga tcttcaagacgatgacacaatcgttcttcgtagcgtcgagccagcaaccgccattaatgtcatcggagatatctccgacgacaacaccg gaataatgactccggtttcgatttcgagatctccgacgatgaaacgaacttcatctaatcggttccgacaattctcacaagagcttaaagc cgaagctgtggcgaaagcgaaacagttatctcaggagttgaaacgattctcatggtctcgttctttctcaggtaacttaaccactactagta ccgccgctaatcaaagcggcggtgctggtggtggtttggtgaactcggctttagaagcgcgagcgttgcgaaagcaacgtgctcagtt 15 agatcggactcggtctagtgctcaaagagctcttcgtggtttgagattcattagcaataagcaaaagaacgttgatggttggaacgatgtt caatcaaatttcgaaaaattcgaaaaaaatggttacatctatcgctccgatttcgctcaatgcataggaatgaaagattcgaaagaatttgc attggaactgttcgatgcattgagtagaagaagaagattaaaagtagagaaaatcaatcacgatgagctttatgagtattggtcacaaatc aacgacgagagttttgattctcgtctccagatcttcttcgacatagtggacaagaatgaagatgggagaattacagaagaggaagtaaaa gagataataatgttgagtgcatctgcaaataagctatcaagattaaaggaacaagcagaggaatatgcagctttgattatggaagagtta 20 gatcctgaaagacttggetacatagagctatggcaactagagactttgcttctacaaaaagacacatacctcaattacagtcaagcattga gctatacgagccaagcattgagccaaaaccttcaagggttaaggggaaagagtcgaatacatagaatgagttcggatttcgtctacatta tgcaagagaattggaaaaggatatgggttttatccttatggatcatgatcatgatcggattattcttgtggaaattttccaatacaagcaaa aagatgcatttcatgtgatgggatattgtttactcacagccaaaggagcagctgaaacacttaaattcaacatggctctaatacttttcccag tttgcagaaacaccattacttggcttagatccacaagactctcttacttcgttccttttgatgataatatcaacttccacaagacaattgtgga 25 gccattgtagtagctgtgatccttcatattggagaccatcttgcttgtgatttccctagaattgttagagccaccgaatacgattacaatcggt atctgtttcattactttcaaacaaaacagccaacatacttcgacctcgttaagggacctgaaggaatcactgggattttaatggtcattttgat gattatttcattcacattagcaacaagatggtttaggcgtaacctagtcaagcttcctaagccattgatcgactaaccggtMaacgcctttt ggtattcgcatcatttgttcgtcattgtttatatcttgcttattcttcatggtatcttcctctatttcgcaagccttggtatgttcgtacgacatgg atgtatcttgcagtaccagttttactctatggtggagaaagaacacttaggtacttccgttctggttcttattcggttcgactgcttaaggttgc 30 tatatatcctggtaatgttctaacgctacaaatgtcgaaaccaactcaatttcgttacaaaagcggacaatacatgtttgtccaatgtcctgc ggtttcgccattcgagtggcatccattctcaattacttccgcacctgaagatgattatatcagcattcacattagacaacttggtgattggact caagaactcaaaagagtattctctgaagtttgtgagccaccggttggcggtaaaagcggactttcagagcgacgaaacaacaaaga aaagtttgccaaagctattgatagatggaccgtacggtgcaccagcacaagattataggaaatatgatgttctcttattagttggtttgg - 63 attggtgcaactccatttatcagtatcttgaaagatttgcttaacaacattgttaaaatggaagagcatgcggattcgatctcggatttcagta gatcatcagaatacagcacaggaagcaacggtgacacgccaagacgaaagagaatactaaaaaccacaaatgcttatttctactgggt cacaagagaacaaggctcttttgattggttcaaaggtgtcatgaacgaagttgcagaacttgaccaacggggtgtgatagagatgcata actatttaacaagtgtgtatgaagaaggtgatgctcgttctgctctcattacaatggttcaagctcttaatcatgccaaaaatggtgtcgaca 5 ttgtctctggcactagggtcagaacacactttgcaagacctaattggaagaaggttctcacaaagctaagttccaagcattgcaatgcaa gaataggagtgttttattgcggagtaccggttttagggaaggagcttagcaaactatgcaacacattcaatcaaaaaggttcaaccaagtt tgaatttcacaaggagcatttctaaaagacaagaaggaagaagccaaaagccctetagattctttaatatctcaaatttagccacttatagt ataaaggcaatctcttcactatttaattcaaagtgattaaacgttaacacactgtcaaaagtgagtgtgttaacgtttagtcacacgttcta ggtttatatacaccgaggcatacgtgtaaatatacgagacagaagaaattcaagggggtttgatagaagcatatagtaaaattttaaaattt 10 cttgtatagtaagcaaatgagatggagactctagaagagaggtgaggtttggtgagggatagcgacagtaatacgacgtcgtattgatt gtagaggaagtgcaatacagctgcagggaagataaatgatggaagcaaataatggttgataaggtccatgtgattaggaagatgaca ctctttttgggggatatttttttctattttttttttttagtggagtcggtgaaagagagactagagtttttagaatattggaagaatatttgggaata agggagttataatatttttgtatcattgtattatataactaatgtaaggtgtacaaagcataaaatttgatagttcctttcttct [SEQ ID NO:33], as set forth for example in GenBank Accession No. NM_105079, or a complement 15 thereof; or a nucleotide sequence that has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO:33, or to a complement thereof; or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO:33, or to a complement thereof. 20 101871 Non-limiting PLDalphal polypeptides comprise the amino acid sequence: 101881 MAQHLLHGTLHATIYEVDALHGGGVRQGFLGKILANVEETIGVGKG ETQLYATIDLQKARVGRTRKIKNEPKNPKWYESFHIYCAHLASDIIFTVKDDNPIGATL IGRAYIPVDQVINGEEVDQWVEILDNDRNPIQGGSKIHVKLQYFHVEEDRNWNMGIK SAKFPGVPYTFFSQRQGCKVSLYQDAHIPDNFVPRIPLAGGKNYEPQRCWEDIFDAIS 25 NAKHLIYITGWSVYAEIALVRDSRRPKPGGDVTIGELLKKKASEGVRVLLLVWDDRT SVDVLKKDGLMATHDEETENFFRGSDVHCILCPRNPDDGGSIVQSLQISTMFTHHQKI VVVDSEMPSRGGSEMRRIVSFVGGIDLCDGRYDTPFHSLFRTLDTVHHDDFHQPNFT GAAITKGGPREPWHDIHSRLEGPIAWDVMYNFEQRWSKQGGKDILVKLRDLSDIIITP SPVMFQEDHDVWNVQLFRSIDGGAAAGFPESPEAAAEAGLVSGKDNIIDRSIQDAYIH 30 AIRRAKDFIYVENQYFLGSSFAWAADGITPEDINALHLIPKELSLKIVSKIEKGEKFRVY VVVPMWPEGLPESGSVQAILDWQRRTMEMMYKDVIQALRAQGLEEDPRNYLTFFCL GNREVKKDGEYEPAEKPDPDTDYMRAQEARRFMIYVHTKMMIVDDEYIIIGSANINQ RSMDGARDSEIAMGGYQPHHLSHRQPARGQIHGFRMSLWYEHLGMLDETFLDPSSL - 64- ECIEKVNRISDKYWDFYSSESLEHDLPGHLLRYPIGVASEGDITELPGFEFFPDTKARIL GTKSDYLPPILTT [SEQ ID NO:34], as set forth for example in GenPept Accession No. NP_188194, or an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 5 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:34. [01891 A non-limiting PLDalphal nucleic acid sequence comprises a nucleotide sequence encoding the sequence set forth in SEQ ID NO:34, or a complement of the nucleotide sequence; or a nucleotide sequence encoding an amino acid sequence having at 10 least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:34, or a complement of that nucleotide sequence. In illustrative examples, an PLDalphal nucleic acid sequence comprises the nucleotide sequence: 15 10190] aaagagcttccatcacggaccagatcccgaattcttcttctgaccaccgaacgattgagtttctccgatcagat ctcagtttctgggaataattcgaagtgaaaaaatggcgcagcatctgttgcacgggactttacatgctaccatctatgaagttgatgcctc catggtggtggtgttaggcaaggcttccttggcaagattctggcaaatgtagaagagacgattggtgttggtaaaggagaaacacagtt gtatgcgacgattgatctgcaaaaagctagagttgggagaaccaggaagatcaaaaatgaacctaagaacccaaagtggtatgagtcg tttcatatttactgtgctcacttggcttctgatatcatcttcactgttaaagatgataatcccattggagctacccttatggaagagttacatt 20 cctgttgatcaagtcattaacggcgaggaagtggatcagtgggttgagatcttggataatgacagaaaccctattcagggaggatcaaa gattcatgtcaagettcaataMccatgttgaggaggatcgtaactggaacatgggtatcaaaagtgccaagttccctggagtgccatac acattcttctcgcagagacaaggctgcaaagtttctctgtaccaagatgctcatattccagacaactttgtccctagaattcctctcgtgga gggaagaactatgagcctcaaagatgttgggaggatatttttgatgctattagcaatgcaaaacacttgatctacattactggttggtctgtt tacgctgagattgctttagtgagggactcgaggaggcctaagcctggaggtgatgtgaccattggtgagctactcaagaagaaggta 25 gtgaaggtgtcagggttcttttgettgtttgggatgacagaacttctgttgatgtgctgaagaaagatgggctcatggctactcatgatgaa gagaccgagaatttcttcaggggaagtgatgtccattgtattctgtgccctcgtaaccggatgacggtggtagcatagtcaaagtttg agatctctactatgttcacgcatcatcagaaaatcgttgttgtggacagegagatgccaagcagaggaggatcagaaatgaggagaatt gtgagttttgttggcggtattgatctttgtgatggaagatacgacactccgttccactccttgttcaggacattggaacagtcaccatgat gacttccatcaacetaacttcactggtgctgctatcactaaaggtggtccaagggagccttggcatgacattcactcccgtcttgaaggt 30 caattgcttgggatgtcatgtacaacttcgagcagagatggagcaagcagggtggtaaagacattctggttaagttgagagatcttagtg atattattatcaccccttctcctgttatgttccaagaggaccacgatgtgtggaatgtccaattgtttaggtccattgatggaggagtgtg ctgggtttcccgagtcgcctgaagctgctgcggaagccgggcttgtaagtgggaaagataacatcattgataggagtatccaagatgct tacattcatgcaatcagacgtgctaaggatttcatctacgttgaaaaccagtacttccttgggagttcttttgettgggcagccgatggtatta - 65 ctcctgaggacatcaatgccctgcacttaatcccaaaagagttgtcgctgaagatagttagcaagattgagaaaggagagaagttcagg gtctatgttgtggttccaatgtggccagaaggtctcccagagagtggatcagtgcaagctatattagactggcagaggaggaccatgga gatgatgtacaaggatgtgattcaggctctcagggcccagggtcttgaggaagatccaagaaactatctgacattcttctgtcttggaaac cgtgaggtcaagaaagatggagagtatgagcctgctgagaaaccagaccccgacactgattacatgagggcgcaagaagcacgcc 5 gtttcatgatttacgtccacaccaaaatgatgatcgttgacgatgaatacattatcattgggtctgctaacatcaaccagaggtcaatggac ggtgcaagagactctgagatagcaatgggaggttatcaaccacatcacttgtcccatagacaaccagctcgtggccagatccatgggtt tcgtatgtcactctggtacgaacacctgggaatgctcgatgaaaccttcctcgatccatcaagcttggaatgcattgagaaagttaaccgc atttctgacaagtattgggacttttactcaagtgagtcactcgaacatgaccttcctggtcacttgctccgctacccgatcggtgtagccag cgaaggcgacatcactgagcttccaggatttgaattcttcccggacacaaaggcccgtatcctcggcaccaaatcagactacctgcctc 10 caatccttacaacctaatctcactaagcatgtcaagtaatgatctctctctccetctctgctttgctgctgttgtagctttgaataaaacttgagt gtctacctttagaattaagaagtcaaatggttgttatgatgatgcacttctttacccctttggtttttatattcgtacaatgacgtggtgagagaa tgtagctttgtgatcttgttttgttgttgttatgtacctttggacttatgaatcttatatctgatccttttctttttatttagtgtgtttcacat [SEQ ID NO:35], as set forth for example in GenBank Accession No. NM_112443, or a complement thereof; or a nucleotide sequence that has at least 70%, 71%, 72%, 73%, 74%, 15 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO:35, or to a complement thereof; or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO:35, or to a complement thereof. [01911 Non-limiting examples of PKS3 polypeptides comprise the amino acid 20 sequence: 10192] MEKKGSVLMLRYEVGKFLGQGTFAKVYHARHLKTGDSVAIKVIDK ERILKVGMTEQIKREISAMRLLRHPNIVELHEVMATKSKIYFVMEHVKGGELFNKVST GKLREDVARKYFQQLVRAVDFCHSRGVCHRDLKPENLLLDEHGNLKISDFGLSALSD SRRQDGLLHTTCGTPTYCAPEVISRNGYDGFKADVWSCGVILFVLLAGYLPFRDSNL 25 MELYKKIGKAEVKFPNWLAPGAKRLLKRILDPNPNTRVSTEKIMKSSWFRKGLQEEV KESVEEETEVDAEAEGNASAEKEKKRCINLNAFEIISLSTGFDLSGLFEKGEEKEEMRF TSNREASEITEKLVEIGKDLKMKVRKKEHEWRVKMSAEATVVEAEVFEIAPSYHMV VLKKSGGDTAEYKRVMKESIRPALIDFVLAWH [SEQ ID NO:60], as set forth for example in GenPept Accession No. AAK26842, or an amino acid sequence having at least 30 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:60. - 66 - 101931 A representative PKS3 nucleic acid sequence comprises a nucleotide sequence encoding the amino acid sequence set forth in SEQ ID NO:60, or a complement of the nucleotide sequence; or a nucleotide sequence encoding an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 5 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:60, or a complement of that nucleotide sequence. In illustrative examples, an PKS3 nucleic acid sequence comprises the nucleotide sequence: 101941 atggagaagaaaggatctgtgttgatgetccgttatgaggttgggaagtttctcggtcaaggtacctttgctaa 10 ggtataccatgctaggcatttgaaaactggtgatagtgtagcgattaaggtcatcgacaaagaaagaatcctcaaagttggcatgaccga gcagattaagcgagagatctctgccatgagactcttgaggcatcccaacatcgttgagctccatgaagtcatggccaccaaatctaaaat ctacttcgtcatggaacatgttaagggtggtgagctcttcaacaaagtgtcaactgggaagctgagagaagacgttggagaaagtactt tcagcagcttgtacgcgctgttgacttctgtcacagccgtggagtatgccacagggacctgaagccggagaatctcttgttggatgag atgggaatcttaagatctctgattttggtctcagcgetctttctgactctagaaggcaagacgggttgctgcatactacatggggacccct 15 acatattgtgcaccggaggtgataagcaggaacgggtatgatgggtttaaagcggatgtgtggtcctgtggagtgatattgttcgtcttgc tcgctggatatcttcctttccgtgattccaatctgatggagctgtataagaagataggcaaagctgaagtcaagttccccaactggttgct ccgggggcaaagagattgctcaagaggatcttggatcctaaccccaacacaagggtatcaactgagaaaataatgaagagctcttggt tccgtaaaggcctacaagaggaggtgaaagaatcagttgaggaagagacagaagtggacgcagaggcagagggaaacgcaagtg cagagaaggaaaagaagcggtgtatcaacctgaacgcgtttgagatcatatctctgtccacggggtttgatctctcgggactgttgaga 20 agggagaggagaaggaggagatgaggtttacatcaaacagagaggcatctgagataacagagaagctggtggagattgggaagga ccttaagatgaaagtgaggaagaaggaacacgaatggagggtgaaaatgtcggctgaggctacagtggtggaageggaagtgtttg agattgcgccgagctatcacatggtggtgctaaagaagageggtggagatactgctgagtataagagagtcatgaaggagagtataag accggctttgatcgactttgtattagcttggcactga [SEQ ID NO:6 1], as set forth for example in GenBank Accession No. AF339144, or a complement thereof; or a nucleotide sequence that has at least 25 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO:37, or to a complement thereof; or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO:61, or to a complement thereof. 30 [01951 An illustrative ATHB6 polypeptide comprises the amino acid sequence: 10196] MMKRLSSSDSVGGLISLCPTTSTDEQSPRRYGGREFQSMLEGYEEEE EAIVEERGHVGLSEKKRRLSINQVKALEKNFELENKLEPERKVKLAQELGLQPRQVA - 67 - VWFQNRRARWKTKQLEKDYGVLKTQYDSLRHNFDSLRRDNESLLQEISKLKTKLNG GGGEEEEEENNAAVTTESDISVKEEEVSLPEKITEAPSSPPQFLEHSDGLNYRSFTDLR DLLPLKAAASSFAAAAGSSDSSDSSALLNEESSSNVTVAAPVTVPGGNFFQFVKMEQ TEDHEDFLSGEEACEFFSDEQPPSLHWYSTVDHWN [SEQ ID NO:36], as set forth for 5 example in GenPept Accession No. NP_565536, or an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:36. [01971 A representative A THB6 nucleic acid sequence comprises a nucleotide 10 sequence encoding the amino acid sequence set forth in SEQ ID NO:36, or a complement of the nucleotide sequence; or a nucleotide sequence encoding an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:36, or a complement of that 15 nucleotide sequence. In illustrative examples, an A THB6 nucleic acid sequence comprises the nucleotide sequence: 10198] gttcatatggaaaacgctcatcaacctcaacaatctctctcctctctctctctgtatatagaagaatctccattgtc tttaatttetctccatttctctttctctcttcettcaaagttttctcttttcttgatgggtttaagagagtaaagatcatcaagtactatgcattaaatt gagaagttattaaaaatttcgaaagtaattaaagattgttgatgatgaagagattaagtagttcagattcagtgggtggtctcatcttttatgt 20 cctacaacttccacagatgagcagagtccgaggagatacggtgggagagagtttcagtcgatgcttgaaggatacgaggaagaagaa gaagctatagtagaagaaagaggacacgtgggcttgtcggagaagaagagaaggttaagcattaaccaagttaaagtttggagaag aattttgagttagagaataagettgagcctgagaggaaagttaagttagctcaagaacttggtcttcaacctcgtcaagttgctgtttggttt caaaaccgtcgtgctcggtggaagacaaaacagcttgagaaagattacggtgttcttaaaacccagtacgattctctcegtcataactttg attccctccgccgtgacaatgaatctctccttcaagagattagtaaactgaaaacgaagcttaatggaggaggaggagaagaagaaga 25 agaagagaacaacgcggcggtgacaacggagagtgatatttcggtcaaggaggaagaagtttcgttgccggagaagattacagagg caccgtcgtctcctccacagtttcttgaacattctgatggtcttaattaccggagtttcacagatctacgtgatttettcattaaagggg ggcttcttcattcgccgccgcagctggatcttcagacagtagcgattcaagcgctctgctgaatgaagaaagcagctctaatgtcactgt ggcggctccggtgacggttccaggaggtaatttcttccagtttgtgaaaatggagcagacggaggatcatgaggacttttgagtggag aagaagcttgtgaattcttttccgatgaacaaccgccgtctctacactggtactccaccgttgatcattggaattgaagtgaaaacacca 30 ccggaagagattttggattggattagatgctettttcttctctgtggaaaaaacagaggagggcaaaatgggaataaagttaaaattgaag ggtgaaagggcaattaaggaagggttaagtctgggcgggaataatgatttagggtgaatttgtaattatgctttttctetagtgattttcgtg cacttcttgtaattaaggatcatcagatcaaagagaaattgagaaggggttaacatgtcaaaggcaaaaattcagcatgggcttttaaatt acaattgacccttgaaacctaaattataccacttctgacccttaaatcaatttttgaatcataaaactacagtgtgacttcgtgt [SEQ ID - 68 - NO:37], as set forth for example in GenBank Accession No. NM_127808, or a complement thereof; or a nucleotide sequence that has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO:37, or to a 5 complement thereof; or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO:37, or to a complement thereof. 101991 In some embodiments, the encoded expression product is a dominant negative form of a polypeptide that stimulates or otherwise facilitates stomatal closure, illustrative examples of which include dominant negative forms of AAPK (e.g., 10 AAPKLyS 43 AIa). While not limiting the invention to any one mechanism, these mutant proteins compete with their wild-type counterparts for interacting proteins in the transgenic plant, or poison multimeric complexes that normally recruit the wild-type counterparts. 102001 In other embodiments, the encoded expression product is an antibody that is immuno-interactive with the endogenous polypeptide that stimulates or otherwise facilitates 15 stomatal closure. In non-limiting examples of this type, the endogenous polypeptide is selected from, OST1, AAPK, v-SNAREs AtVAMP711-14, GPA1, AtABCG22, AtABCG40, AtMRP4, RBOHD, RBOHF and PLDalphal. Exemplary antibodies for use in the practice of the present invention include monoclonal antibodies, Fv, Fab, Fab' and F(ab') 2 immunoglobulin fragments, as well as synthetic antibodies such as but not limited to single 20 domain antibodies (DABs), synthetic stabilized Fv fragments, e.g., single chain Fv fragments (scFv), disulfide stabilized Fv fragments (dsFv), single variable region domains (dAbs) minibodies, combibodies and multivalent antibodies such as diabodies and multi-scFv or engineered human equivalents. Techniques for preparing and using various antibody-based constructs and fragments are well known in the art. Means for preparing and characterizing 25 antibodies are also well known in the art. In illustrative examples, antibodies can be made by conventional immunization (e.g., polyclonal sera and hybridomas) with isolated, purified or recombinant peptides or proteins corresponding to at least a portion of an endogenous polypeptide, or as recombinant fragments corresponding to at least a portion of an endogenous polypeptide, usually expressed in Escherichia coli, after selection from phage 30 display or ribosome display libraries (e.g., available from Cambridge Antibody Technology, BioInvent, Affitech and Biosite). Knowledge of the antigen-binding regions (e.g., complementarity-determining regions) of such antibodies can be used to prepare synthetic antibodies as described for example above. - 69 - 102011 In still other embodiments, the expression product inhibits by RNA interference (RNAi) or post-transcriptional gene silencing (PTGS) the expression of a target gene, which encodes a polypeptide that stimulates or otherwise facilitates stomatal closure. In illustrative examples of this type, the expression product is a RNA molecule (e.g., siRNA, 5 shRNA, miRNA, dsRNA etc.) that comprises a targeting region corresponding to a nucleotide sequence of the target gene and that attenuates or otherwise disrupts the expression of the target gene. Non-limiting examples of such target genes include AAPK, OSTI, v-SNAREs AtVA MP711-14, GPA 1, At ABCG22, AtABCG40, AtMRP4, RBOHD, RBOHF, and PLDalphal. 10 [0202] In certain embodiments, the targeting sequence displays at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% identity to a nucleotide sequence of the target gene. In other embodiments, the targeting sequence hybridizes to a nucleotide sequence of the target gene under at least low stringency conditions, more suitably under at least medium 15 stringency conditions and even more suitably under high stringency conditions. Reference herein to low stringency conditions include and encompass from at least about I % v/v to at least about 15% v/v formamide and from at least about 1 M to at least about 2 M salt for hybridization at 42' C, and at least about 1 M to at least about 2 M salt for washing at 420 C. Low stringency conditions also may include 1% Bovine Serum Albumin (BSA), 1 mM 20 EDTA, 0.5 M NaHPO 4 (pH 7.2), 7% SDS for hybridization at 65' C, and (i) 2xSSC, 0.1% SDS; or (ii) 0.5% BSA, 1 mM EDTA, 40 mM NaHPO 4 (pH 7.2), 5% SDS for washing at room temperature. Medium stringency conditions include and encompass from at least about 16% v/v to at least about 30% v/v formamide and from at least about 0.5 M to at least about 0.9 M salt for hybridization at 420 C, and at least about 0.5 M to at least about 0.9 M salt for 25 washing at 420 C. Medium stringency conditions also may include 1% Bovine Serum Albumin (BSA), 1 mM EDTA, 0.5 M NaHPO 4 (pH 7.2), 7% SDS for hybridization at 650 C, and (i) 2 x SSC, 0.1% SDS; or (ii) 0.5% BSA, 1 mM EDTA, 40 mM NaHPO 4 (pH 7.2), 5% SDS for washing at 420 C. High stringency conditions include and encompass from at least about 31% v/v to at least about 50% v/v formamide and from at least about 0.01 M to at least 30 about 0.15 M salt for hybridization at 420 C, and at least about 0.01 M to at least about 0.15 M salt for washing at 420 C. High stringency conditions also may include 1% BSA, 1 mM EDTA, 0.5 M NaHPO 4 (pH 7.2), 7% SDS for hybridization at 650 C, and (i) 0.2 x SSC, 0.1% SDS; or (ii) 0.5% BSA, 1mM EDTA, 40 mM NaHPO 4 (pH 7.2), 1% SDS for washing at a - 70 temperature in excess of 650 C. Desirably, the targeting sequence hybridizes to a nucleotide sequence of the target gene under physiological conditions. 102031 Other stringent conditions are well known in the art. A skilled artisan will recognize that various factors can be manipulated to optimize the specificity of the 5 hybridization. Optimization of the stringency of the final washes can serve to ensure a high degree of hybridization. For detailed examples, see Ausubel et al., supra at pages 2.10.1 to 2.10.16 and Sambrook et al., supra at sections 1.101 to 1.104. [02041 Suitably, the targeting region has sequence identity with the sense strand or antisense strand of the target gene. In certain embodiments, the RNA molecule is 10 unpolyadenylated, which can lead to efficient reduction in expression of the target gene, as described for example by Waterhouse et al in U.S. Patent No. 6,423,885. 102051 Typically, the length of the targeting region may vary from about 10 nucleotides (nt) up to a length equaling the length (in nucleotides) of the target gene. Generally, the length of the targeting region is at least 10, 12, 13, 14, 15, 16, 17, 18, 19, 20, 15 21, 22, 23, 24, 25 nt, usually at least about 50 nt, more usually at least about 100 nt, especially at least about 150 nt, more especially at least about 200 nt, even more especially at least about 500 nt. It is expected that there is no upper limit to the total length of the targeting region, other than the total length of the target gene. However for practical reason (such as e.g., stability of the targeting constructs) it is expected that the length of the targeting region should 20 not exceed 5000 nt, particularly should not exceed 2500 nt and could be limited to about 1000 nt. [02061 The RNA molecule may further comprise one or more other targeting regions (e.g., from about I to about 10, or from about I to about 4, or from about 1 to about 2 other targeting regions) each of which has sequence identity with a nucleotide sequence of the 25 target gene. Generally, the targeting regions are identical or share at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% sequence identity with each other. [02071 The RNA molecule may further comprise a reverse complement of the targeting region. Typically, in these embodiments, the RNA molecule further comprises a 30 spacer sequence that spaces the targeting region from the reverse complement. The spacer sequence may comprise a sequence of nucleotides of at least about 100-500 nucleotides in length, or alternatively at least about 50-100 nucleotides in length and in a further alternative -71 at least about 10-50 nucleotides in length. Typically, the spacer sequence is a non-coding sequence, which in some instances is an intron. In embodiments in which the spacer sequence is a non-intron spacer sequence, transcription of the nucleic acid sequence will produce an RNA molecule that forms a hairpin or stem-loop structure in which the stem is formed by 5 hybridization of the targeting region to the reverse complement and the loop is formed by the non-intron spacer sequence connecting these 'inverted repeats'. Alternatively, in embodiments in which the spacer sequence is an intron spacer sequence, the presence of intron/exon splice junction sequences on either side of the intron sequence facilitates the removal of what would otherwise form a loop structure and the resulting RNA will form a 10 double-stranded RNA (dsRNA) molecule, with optional overhanging 3' sequences at one or both ends. Such a dsRNA transcript is referred to herein as a "perfect hairpin". The RNA molecules may comprise a single hairpin or multiple hairpins including "bulges" of single stranded RNA occurring adjacent to regions of double-stranded RNA sequences. [0208] Alternatively, a dsRNA molecule as described above can be conveniently 15 obtained using an additional polynucleotide from which a further RNA molecule is producible, comprising the reverse complement of the targeting region. In this embodiment, the reverse complement of the targeting region hybridizes to the targeting region of the RNA molecule transcribed from the second polynucleotide. [02091 In another example, a dsRNA molecule as described above is prepared using 20 a second polynucleotide that comprises a duplex, wherein one strand of the duplex shares sequence identity with a nucleotide sequence of the target gene and the other shares sequence identity with the complement of that nucleotide sequence. In this embodiment, the duplex is flanked by two promoters, one controlling the transcription of one of the strands, and the other controlling the transcription of the complementary strand. Transcription of both strands 25 produces a pair of RNA molecules, each comprising a region that is complementary to a region of the other, thereby producing a dsRNA molecule that inhibits the expression of the target gene. [02101 In another example, PTGS of the target gene is achieved using the strategy by Glassman et al described in U.S. Patent Application Publication No 2003/0036197. In this 30 strategy, suitable nucleic acid sequences and their reverse complement can be used to alter the expression of any homologous, endogenous target RNA (i.e., comprising a transcript of the target gene) which is in proximity to the suitable nucleic acid sequence and its reverse -72complement. The suitable nucleic acid sequence and its reverse complement can be either unrelated to any endogenous RNA in the host or can be encoded by any nucleic acid sequence in the genome of the host provided that nucleic acid sequence does not encode any target mRNA or any sequence that is substantially similar to the target RNA. Thus, in some 5 embodiments of the present invention, the RNA molecule further comprises two complementary RNA regions which are unrelated to any endogenous RNA in the host cell and which are in proximity to the targeting region. In other embodiments, the RNA molecule further comprises two complementary RNA regions which are encoded by any nucleic acid sequence in the genome of the host provided that the sequence does not have sequence 10 identity with the nucleotide sequence of the target gene, wherein the regions are in proximity to the targeting region. In the above embodiments, one of the complementary RNA regions can be located upstream of the targeting region and the other downstream of the targeting region. Alternatively, both the complementary regions can be located either upstream or downstream of the targeting region or can be located within the targeting region itself. 15 [02111 In some illustrative examples, the RNA molecule is an antisense molecule that is targeted to a specific region of RNA encoded by the target gene, which is critical for translation. The use of antisense molecules to decrease expression levels of a pre-determined gene is known in the art. Antisense molecules may be designed to correspond to full-length RNA transcribed from the target gene, or to a fragment or portion thereof. This gene silencing 20 effect can be enhanced by transgenically over-producing both sense and antisense RNA of the target gene coding sequence so that a high amount of dsRNA is produced as described for example above (see, for example, Waterhouse et al. (1998) Proc Natl Acad Sci USA 95:13959 13964). [02121 In other embodiments, the expression product that inhibits stomatal closure 25 corresponds to an expression product of the endogenous target gene targeted for repression. In many cases, this "co-suppression" results in the complete repression of the native target gene as well as the transgene. 102131 In still other embodiments, the expression product that inhibits stomatal closure corresponds to an expression product of a negative regulator of the ABA signaling 30 pathway. In illustrative examples of this type, the negative regulator is ATHB6. In some of these examples, the nucleic acid sequence encoding the expression product that inhibits stomatal closure comprises a nucleotide sequence corresponding to the coding sequence of - 73 - A THB6. A non-limiting example of a A THB6 coding sequence is represented by the following sequence: [0214] atgatgaagagattaagtagttcagattcagtgggtggtctcatctetttatgtcctacaacttccacagatgagc agagtccgaggagatacggtgggagagagtttcagtcgatgcttgaaggatacgaggaagaagaagaagctatagtagaagaaaga 5 ggacacgtgggcttgtcggagaagaagagaaggttaagcattaaccaagttaaagctttggagaagaattttgagttagagaataagct tgagcctgagaggaaagttaagttagctcaagaacttggtcttcaacctcgtcaagttgctgtttggtttcaaaaccgtcgtgctcggtgga agacaaaacagcttgagaaagattacggtgttettaaaacccagtacgattctctccgtcataactttgattccctccgccgtgacaatgaa tctctcettcaagagattagtaaactgaaaacgaagcttaatggaggaggaggagaagaagaagaagaagagaacaacgcggcggt gacaacggagagtgatatttcggtcaaggaggaagaagtttcgttgccggagaagattacagaggcaccgtcgtctcctccacagtttc 10 ttgaacattctgatggtcttaattaccggagtttcacagatctacgtgatcttcttccattaaaggcggcggcttcttcattcgccgccgcag ctggatcttcagacagtagcgattcaagcgctctgctgaatgaagaaagcagctctaatgtcactgtggcggctccggtgacggttcca ggaggtaatttcttccagtttgtgaaaatggagcagacggaggatcatgaggactttctgagtggagaagaagcttgtgaattttttcg atgaacaaccgccgtctctacactggtactccaccgttgatcattggaattga [SEQ ID NO:38], as for example set out in GenBank Accession NM_127808, or a nucleotide sequence that has at least 70%, 71%, 15 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to that coding sequence or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to that coding sequence. 102151 Alternatively, any other nucleotide sequence that codes for the amino acid 20 sequence of ATHB6 may be used. A non-limiting example of a ATHB6 amino acid sequence is represented by [SEQ ID NO:36], or an amino acid having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ ID NO:36, as described for example above. 25 102161 In other embodiments, the negative regulator is selected from ABII, ABI2 or mutant forms of ABIl (e.g., ABIIGlyI80Asp) or ABI2 (e.g., ABI 2 Gly1 6 8ASp), which result in reduced ABA sensitivity and/or which inhibit stomatal closure. While not limiting the invention to any one mechanism or mode of operation, these mutant proteins may have reduced susceptibility to ABA inhibition than the wild-type counterparts, or compete with 30 their wild-type counterparts for interacting proteins in the transgenic plant, or poison multimeric complexes that normally recruit the wild-type counterparts. - 74 - [02171 In still other embodiments, the negative regulator is a dominant positive AHAI mutant (e.g., a constitutively active AHA1 polypeptide), illustrative examples of which include AHA1Trp8 75 Leu; AHA1Pro 68 ser; AHAI Leul69Phe; AHAGly8 67 Ser; AHA 1 GiulOAsp AHAITrp8 75 Leu. 5 10218] In still other embodiments, the negative regulator is a dominant positive PKS3 mutant, an non-limiting example of which includes the dominant positive PKS3 deletion mutant disclosed by Guo et al. (2002, supra). 2.5 Other construct elements 102191 In addition to the operably linked cis-acting elements, promoters and nucleic 10 acid sequence encoding an expression product that inhibits stomatal closure described above, the constructs of the present invention, which are suitably expression constructs, can also include other regulatory sequences. As used herein, "regulatory sequences" means nucleotide sequences located upstream (5' non-coding sequences), within or downstream (3' non-coding sequences) of a coding sequence, and which influence the transcription, RNA processing or 15 stability, or translation of the associated coding sequence. Regulatory sequences include, but are not limited to, enhancers, introns, translation leader sequences and polyadenylation signal sequences. [02201 A number of non-translated leader sequences derived from viruses are known to enhance gene expression. Specifically, leader sequences from Tobacco Mosaic 20 Virus (TMV, the "O-sequence"), Maize Chlorotic Mottle Virus (MCMV) and Alfalfa Mosaic Virus (AMV) have been shown to be effective in enhancing expression (Gallie et al. (1987) Nucleic Acids Res. 15:8693-8711; and Skuzeski et al. (1990) Plant Mol. Biol. 15:65-79). Other leader sequences known in the art include, but are not limited to, picornavirus leaders such as an encephalomyocarditis (EMCV) 5' noncoding region leader (Elroy-Stein et al. 25 (1989) Proc. Natl. A cad. Sci. USA 86:6126-6130); potyvirus leaders such as a Tobacco Etch Virus (TEV) leader (Allison et al. (1986) Virology 154:9-20); Maize Dwarf Mosaic Virus (MDMV) leader (Allison et al. (1986), supra); human immunoglobulin heavy-chain binding protein (BiP) leader (Macejak & Samow (1991) Nature 353:90-94); untranslated leader from the coat protein mRNA of AMV (AMV RNA 4; Jobling & Gehrke (1987) Nature 325:622 30 625); tobacco mosaic TMV leader (Gallie et al. (1989) Molecular Biology of RNA 237-256); and MCMV leader (Lommel et al. (1991) Virology 81:382-385). See also, Della-Cioppa et al. (1987) Plant Physiol. 84:965-968. In some embodiments, translational enhancers are - 75 employed such as the overdrive-sequence containing the 5'-untranslated leader sequence from tobacco mosaic virus enhancing the polypeptide per RNA ratio (Gallie et al. (1987) Nucleic Acids Research 15:8693-8711). [02211 An expression construct also can optionally include a transcriptional and/or 5 translational termination region (i.e., termination region) that is functional in plants. A variety of transcriptional terminators are available for use in expression constructs and are responsible for the termination of transcription beyond the heterologous nucleotide sequence of interest and correct mRNA polyadenylation. The termination region may be native to the transcriptional initiation region, may be native to the operably linked nucleotide sequence of 10 interest, may be native to the plant host, or may be derived from another source (i.e., foreign or heterologous to the promoter, the nucleotide sequence of interest, the plant host, or any combination thereof). Appropriate transcriptional terminators include, but are not limited to, the CAMV 35S terminator, the tml terminator, the nopaline synthase terminator and the pea rbcs E9 terminator. These can be used in both monocotyledons and dicotyledons. In addition, 15 a coding sequence's native transcription terminator can be used. A signal sequence can be operably linked to a nucleic acid molecule of the present invention to direct the nucleic acid molecule into a cellular compartment. In this manner, the expression construct will comprise a nucleic acid molecule of the present invention operably linked to a nucleotide sequence for the signal sequence. The signal sequence may be operably linked at the N- or C- terminus of 20 the nucleic acid molecule. Exemplary polyadenylation signals can be those originating from Agrobacterium tumefaciens t-DNA such as the gene known as octopine synthase of the Ti plasmid pTiACH5 (Gielen et al. (1984) EMBO J. 3:835) or functional equivalents thereof, but also all other terminators functionally active in plants are suitable. 102221 The expression construct also can include a nucleotide sequence for a 25 selectable marker, which can be used to select a transformed plant, plant part and/or plant cell. As used herein, "selectable marker" means a nucleotide sequence that when expressed imparts a distinct phenotype to the plant, plant part and/or plant cell expressing the marker and thus allows such transformed plants, plant parts and/or plant cells to be distinguished from those that do not have the marker. Such a nucleotide sequence may encode either a selectable or 30 screenable marker, depending on whether the marker confers a trait that can be selected for by chemical means, such as by using a selective agent (e.g., an antibiotic, herbicide, or the like), or on whether the marker is simply a trait that one can identify through observation or testing, - 76 such as by screening (e.g., the R-locus trait). Of course, many examples of suitable selectable markers are known in the art and can be used in the expression constructs described herein. [02231 Examples of selectable markers include, but are not limited to, a nucleotide sequence encoding neo or nptlI, which confers resistance to kanamycin, G418, and the like 5 (Potrykus et al. (1985) Mol. Gen. Genet. 199:183-188); a nucleotide sequence encoding bar, which confers resistance to phosphinothricin; a nucleotide sequence encoding an altered 5 enolpyruvylshikirmate-3-phosphate (EPSP) synthase, which confers resistance to glyphosate (Hinchee et al. (1988) Biotech. 6:915-922); a nucleotide sequence encoding a nitrilase such as bxn from Klebsiella ozaenae that confers resistance to bromoxynil (Stalker et al. (1988) 10 Science 242:419-423); a nucleotide sequence encoding an altered acetolactate synthase (ALS) that confers resistance to imidazolinone, sulfonylurea or other ALS-inhibiting chemicals (EP Patent Application No. 154204); a nucleotide sequence encoding a methotrexate-resistant dihydrofolate reductase (DHFR) (Thillet et al. (1988) J. Biol. Chem. 263:12500-12508); a nucleotide sequence encoding a dalapon dehalogenase that confers resistance to dalapon; a 15 nucleotide sequence encoding a mannose-6-phosphate isomerase (also referred to as phosphomannose isomerase (PMI)) that confers an ability to metabolize mannose (US Patent Nos. 5,767,378 and 5,994,629); a nucleotide sequence encoding an altered anthranilate synthase that confers resistance to 5-methyl tryptophan; and/or a nucleotide sequence encoding hph that confers resistance to hygromycin. One of skill in the art is capable of 20 choosing a suitable selectable marker for use in an expression construct of this invention. [0224] Additional selectable markers include, but are not limited to, a nucleotide sequence encoding p-glucuronidase or uidA (GUS) that encodes an enzyme for which various chromogenic substrates are known; an R-locus nucleotide sequence that encodes a product that regulates the production of anthocyanin pigments (red color) in plant tissues (Dellaporta 25 et al., "Molecular cloning of the maize R-nj allele by transposon-tagging with Ac" 263-282 In: Chromosome Structure and Function: Impact of New Concepts, 18th Stadler Genetics Symposium (Gustafson & Appels eds., Plenum Press 1988)); a nucleotide sequence encoding p-lactamase, an enzyme for which various chromogenic substrates are known (e.g., PADAC, a chromogenic cephalosporin) (Sutcliffe (1978) Proc. Natl. Acad Sci. USA 75:3737-3741); a 30 nucleotide sequence encoding xylE that encodes a catechol dioxygenase (Zukowsky et al. (1983) Proc. Natl. Acad. Sci. USA 80:1101-1105); a nucleotide sequence encoding tyrosinase, an enzyme capable of oxidizing tyrosine to DOPA and dopaquinone, which in turn condenses to form melanin (Katz et al. (1983) J. Gen. Microbiol. 129:2703-2714); a nucleotide sequence - 77 encoding p-galactosidase, an enzyme for which there are chromogenic substrates; a nucleotide sequence encoding luciferase (lux) that allows for bioluminescence detection (Ow et al. (1986) Science 234:856-859); a nucleotide sequence encoding aequorin which may be employed in calcium-sensitive bioluminescence detection (Prasher et al. (1985) Biochem. 5 Biophys. Res. Comm. 126:1259-1268); or a nucleotide sequence encoding green fluorescent protein (Niedz et al. (1995) Plant Cell Reports 14:403-406). One of skill in the art is capable of choosing a suitable selectable marker for use in an expression construct of this invention. [02251 An expression construct of the present invention also can include nucleotide sequences that encode other desired traits. Such nucleotide sequences can be stacked with any 10 combination of nucleotide sequences to create plants, plant parts or plant cells having the desired phenotype. Stacked combinations can be created by any method including, but not limited to, cross breeding plants by any conventional methodology, or by genetic transformation. If stacked by genetically transforming the plants, the nucleotide sequences of interest can be combined at any time and in any order. For example, a transgenic plant 15 comprising one or more desired traits can be used as the target to introduce further traits by subsequent transformation. The additional nucleotide sequences can be introduced simultaneously in a co-transformation protocol with a nucleotide sequence, nucleic acid molecule, nucleic acid construct, and/or composition of this invention, provided by any combination of expression constructs. For example, if two nucleotide sequences will be 20 introduced, they can be incorporated in separate cassettes (trans) or can be incorporated on the same cassette (cis). Expression of the nucleotide sequences can be driven by the same promoter or by different promoters. It is further recognized that nucleotide sequences can be stacked at a desired genomic location using a site-specific recombination system. See, e.g., Int'l Patent Application Publication Nos. WO 99/25821; WO 99/25854; WO 99/25840; WO 25 99/25855 and WO 99/25853. [02261 In addition to the nucleic acid encoding an expression product that inhibits stomatal closure, the expression construct can include a coding sequence for one or more polypeptides for agronomic traits that primarily are of benefit to a seed company, grower or grain processor. A polypeptide of interest can be any polypeptide encoded by a nucleotide 30 sequence of interest. Non-limiting examples of polypeptides of interest that are suitable for production in plants include those resulting in agronomically important traits such as herbicide resistance (also sometimes referred to as "herbicide tolerance"), virus resistance, bacterial pathogen resistance, insect resistance, nematode resistance, and/or fungal resistance. - 78 - See, e.g., U.S. Patent Nos. 5,569,823; 5,304,730; 5,495,071; 6,329,504; and 6,337,431. The polypeptide also can be one that increases plant vigor or yield (including traits that allow a plant to grow at different temperatures, soil conditions and levels of sunlight and precipitation), or one that allows identification of a plant exhibiting a trait of interest (e.g., a 5 selectable marker, seed coat color, etc.). Various polypeptides of interest, as well as methods for introducing these polypeptides into a plant, are described, for example, in US Patent Nos. 6,084,155; 6,329,504 and 6,337,431; as well as US Patent Publication No. 2001/0016956. See also, on the World Wide Web at lifesci.sussex.ac.uk/home/NeilCrickmore/Bt/. Nucleotide sequences conferring resistance/tolerance to an herbicide that inhibits the growing point or 10 meristem, such as an imidazalinone or a sulfonylurea can also be suitable in some embodiments of the invention. Exemplary nucleotide sequences in this category code for mutant ALS and AHAS enzymes as described, e.g., in U.S. Patent Nos. 5,767,366 and 5,928,937. U.S. Patent Nos. 4,761,373 and 5,013,659 are directed to plants resistant to various imidazalinone or sulfonamide herbicides. U.S. Patent No. 4,975,374 relates to plant cells and 15 plants containing a nucleic acid encoding a mutant glutamine synthetase (GS) resistant to inhibition by herbicides that are known to inhibit GS, e.g., phosphinothricin and methionine sulfoximine. U.S. Patent No. 5,162,602 discloses plants resistant to inhibition by cyclohexanedione and aryloxyphenoxypropanoic acid herbicides. The resistance is conferred by an altered acetyl coenzyme A carboxylase (ACCase). 20 [02271 In specific embodiments, a polynucleotide comprising a nucleotide sequence encoding a transcription factor is expressed in the same cell in which the nucleic acid sequence encoding the expression product that inhibits stomatal closure is expressible. In these embodiments, the transcription factor activates in the presence of a compound that induces expression of the alcohol dehydrogenase (ADH I) system of Aspergillus nidulans 25 (e.g., as broadly described above) and interacts with the cis-acting element to induce expression of the stomatal closure-inhibiting nucleic acid sequence. Illustrative transcription factors comprise an amino acid sequence corresponding to the amino acid sequence of the AlcR transcription factor, as set forth for example in GenPept Accession No. AAQ06627. In non-limiting examples, the ethanol receptor comprises the amino acid sequence: 30 [02281 MADTRRRQNHSCDPCRKGKRRCDAPENRNEANENGWVSCSNCKR WNKDCTFNWLSSQRSKAKGAAPRARTKKARTATTTSEPSTSAATIPTPESDNHDAPP VINSHDALPSWTQGLLSHPGDLFDFSHSAIPANAEDAANVQSDAPFPWDLAIPGDFSM GQQLEKPLSPLSFQAVLLPPHSPNTDDLIRELEEQTTDPDSVTDTNSVQQVAQDGSLW - 79 - SDRQSPLLPENSLCMASDSTARRYARSTMTKNLMRIYHDSMENALSCWLTEHNCPYS DQISYLPPKQRAEWGPNWSNRMCIRVCRLDRVSTSLRGRALSAEEDKAAARALHLAI VAFASQWTQHAQRGAGLNVPADIAADERSIRRNAWNEARHALQHTTGIPSFRVIFAN IIFSLTQSVLDDDEQHGMGARLDKLLENDGAPVFLETANRQLYTFRHKFARMQRRGK 5 AFNRLPGGSVASTFAGIFETPTPSSESPQLDPVVASEEHRSTLSLMFWLGIMFDTLSAA MYQRPLVVSDEDSQISSASPPRRGAETPINLDCWEPPRQVPSNQEKSDVWGDLFLRTS DSLPDHESHTQISQPAARWPCTYEQAAAALSSATPVKVLLYRRVTQLQTLLYRGASP ARLEAAIQRTLYVYNHWTAKYQPFMQDCVANHELLPSRIQSWYVILDGHWHLAAM LLADVLESIDRDSYSDINHIDLVTKLRLDNALAVSALARSSLRGQELDPGKASPMYRH 10 FHDSLTEVAFLVEPWTVVLIHSFAKAAYILLDCLDLDGQGNALAGYLQLRQNCNYCI RALQFLGRKSDMAALVAKDLERGLNGKVDSFL [SEQ ID NO:56]; or 102291 an amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence similarity or sequence identity to SEQ 15 ID NO:56. [02301 Exemplary AlcR-encoding polynucleotides may be selected from: [02311 atggctgacagcatccagaccgcacgtttttggacttctgataccatctatacagatatcactgtcgatattctcc tgcacacagcatggcagatacgcgccgacgccagaatcatagctgcgatccctgtcgcaagggcaagcgacgctgtgatgccccgg aaaatagaaacgaggccaatgaaaacggctgggtttcgtgttcaaattgcaagcgttggaacaaggattgtaccttcaattggctctcat 20 cccaacgctccaaggcaaaaggggctgcacctagagcgagaacaaagaaagccaggaccgcaacaaccaccagtgaaccatcaa cttcagctgcaacaatccctacaccggaaagtgacaatcacgatgcgcctccagtcataaactctcacgacgcgctcccgagctgga tcaggggctactctcccaccccggcgaccttttcgatttcagccactctgctattcccgcaaatgcagaagatgcggcaacgtgcagt agacgcaccttttccgtgggatctagccatccccggtgatttcagcatgggccaacagctcgagaaacctctcagtcgctcagttttca agcagtccttcttccgccccatagcccgaacacggatgacctcattcgcgagctggaagagcagactacggatccggactcggttacc 25 gatactaatagtgtacaacaggtcgctcaagatggatcgctatggtctgatcggcagtcgccgetactgcctgagaacagtctgtgcatg gcctcagacagcacagcacggcgatatgcccgttecacaatgacgaagaatctgatgcgaatctaccacgatagtatggagaatgcac tgtcctgctggctgacagagcacaattgtccatactccgaccagatcagctacctgccgcccaagcagcgggcggaatggggcccga actggtcaaacaggatgtgcatccgggtgtgccggctagatcgcgtatctacctcattacgcgggcgcgccctgagtgcggaagagg acaaagccgcagcccgagccctgcatctggcgatcgtagcttttgcgtcgcaatggacgcagcatgcgcagaggggggctgggta 30 aatgttcctgcagacatagccgccgatgagaggtccatccggaggaacgcctggaatgaagcacgccatgccttgcagcacacgac agggattccatcattccgggttatatttgcgaatatcatcttttctctcacgcagagtgtgctggatgatgatgagcagcacggtatgggtg cacgtctagacaagctactcgaaaatgacggtgcgcccgtgttcctggaaaccgcgaaccgtcagctttatacattcgacataagtttg -80cacgaatgcaacgccgcggtaaggctttcaacaggctcccgggaggatctgtcgcatcgacattcgccggtattttcgagacaccgac gccgtcgtctgaaagcccacagcttgacccggttgtggccagtgaggagcatcgcagtacattaagccttatgttctggctagggatcat gttcgatacactaagcgctgcaatgtaccagcgaccactcgtggtgtcagatgaggatagccagatatcatcggcatctccaccaaggc gcggcgctgaaacgccgatcaacctagactgctgggagcccccgagacaggtcccgagcaatcaagaaaagagcgacgtatggg 5 gcgacctcttcctccgcacctcggactctctcccagatcacgaatcccacacacaaatctctcagccagcggctcgatggccctgcacc tacgaacaggccgccgccgctctctcctctgcaacgcccgtcaaagtcctcctctaccgccgcgtcacgcagetccaaaccctcctcta tcgcggcgccagccctgcccgccttgaagcggccatccagagaacgctctacgtttataatcactggacagcgaagtaccaaccattt atgcaggactgcgttgctaaccacgagctcctcccttcgcgcatccagtcttggtacgtcattctagacggtcactggcatctagccgcg atgttgctagcggacgttttggagagcatcgaccgcgattcgtactctgatatcaaccacatcgaccttgtaacaaagctaaggctegata 10 atgcactagcagttagtgcccttgcgcgctcttcactccgaggccaggagctggacccgggcaaagcatctccgatgtatcgccatttc catgattctctgaccgaggtggcattcctggtagaaccgtggaccgtcgttcttattcactcgtttgccaaagctgcgtatatttgtggac tgtttagatctggacggccaaggaaatgcactagcggggtacctgcagctgcggcaaaattgcaactactgcattcgggcgctgcaatt tctgggcaggaagtcggatatggcggcgctggttgcgaaggatttagagagaggtttgaatgggaaagttgacagctttttgtag [SEQ ID NO: 57] as set forth for example in GenBank Accession No. AF496548; or a 15 nucleotide sequence that has at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to SEQ ID NO:57, or a nucleotide sequence that hybridizes under at least medium stringency or high stringency conditions to SEQ ID NO:57. 10232] Thus, expression of the transcription factor-encoding nucleotide sequence in 20 the host cell permits chemical compounds that induce the expression of the alcohol dehydrogenase system of A. nidulans to activate expression of the stomatal closure-inhibiting nucleic acid sequence. Generally, the transcription factor-encoding nucleotide sequence is operably linked to a promoter that is operable in a plant cell to form a separate construct ("second construct") relative to the construct that comprises the cis-acting element ("first 25 construct"). The promoter of the second construct is suitably selected from constitutive promoters and cell or tissue specific/preferential promoters, as described for example above. Use of a cell or tissue specific/preferential promoter to drive expression of the transcription factor-encoding nucleotide sequence facilitates chemically inducible expression of the stomatal closure-inhibiting nucleic acid sequence in specific cell or tissues or preferentially in 30 specific cells or tissues (e.g., guard cells). Alternatively, use of a constitutive promoter to drive expression of the transcription factor-encoding nucleotide sequence facilitates chemically inducible expression of the stomatal closure-inhibiting nucleic acid sequence - 81 throughout the plant. The first and second constructs may be present on the same vector or on separate vectors. 3. Transgenic plants, plant parts, plant organs and plant cells 102331 The present invention further encompasses plant cells, plant parts, plant 5 organs and plants in accordance with the embodiments of this invention. Thus, in some embodiments, the present invention provides a transformed plant cell, plant part, plant organ and/or plant comprising a nucleic acid molecule, a nucleic acid construct, a nucleotide sequence, a promoter, and/or a composition of this invention. Representative plants include, for example, angiosperms (monocots and dicots), gymnosperms, bryophytes, ferns and/or fern 10 allies. 102341 In some embodiments, the plants are selected from monocotyledonous plants. Non-limiting examples of monocot plants include sugar cane, corn, barley, rye, oats, wheat, rice, flax, millet, sorghum, grasses (e.g., switch grass, giant reed, turf grasses etc.), banana, onion, asparagus, lily, coconut, and the like. In some embodiments, the monocot 15 plants of the invention include plants of the genus Saccharum (i.e., sugar cane, energy cane) and hybrids thereof, including hybrids between plants of the genus Saccharum and those of related genera, such as Miscanthus, Erianthus, Sorghum and others. As used herein, "sugar cane" and "Saccharum spp." mean any of six to thirty-seven species (depending on taxonomic interpretation) of tall perennial grasses of the genus Saccharum. In particular, the plant can be 20 Saccharum aegyptiacum, Saccharum esculentum, Saccharum arenicol, Saccharum arundinaceum, Saccharum barberi, Saccharum bengalense, Saccharum biflorum, Saccharum chinense, Saccharum ciliare, Saccharum cylindricum, Saccharum edule, Saccharum elephantinum, Saccharum exaltatum, Saccharum fallax, Saccharum fallax, Saccharum floridulum, Saccharum giganteum, Saccharum hybridum, Saccharum japonicum, Saccharum 25 koenigii, Saccharum laguroides, Saccharum munja, Saccharum narenga, Saccharum officinale, Saccharum officinarum, Saccharum paniceum, Saccharum pophyrocoma, Saccharum purpuratum, Saccharum ravennae, Saccharum robustum, Saccharum roseum, Saccharum sanguineum, Saccharum sara, Saccharum sinense, Saccharum spontaneum, Saccharum tinctorium, Saccharum versicolor, Saccharum violaceum, Saccharum violaceum, 30 and any of the interspecific hybrids and commercial varieties thereof. 102351 Further non-limiting examples of plants of the present invention include soybean, beans in general, Brassica spp., clover, cocoa, coffee, cotton, peanut, rape/canola, - 82 safflower, sugar beet, sunflower, sweet potato, tea, vegetables including but not limited to broccoli, brussel sprouts, cabbage, carrot, cassava, cauliflower, cucurbits, lentils, lettuce, pea, peppers, potato, radish and tomato, fruits including, but not limited to, apples, pears, peaches, apricots and citrus, avocado, pineapple and walnuts; and flowers including, but not limited to, 5 carnations, orchids, roses, and any combination thereof. [02361 In some embodiments, the plants are selected from energy crops, representative examples of which include: [02371 Miscanthus (e.g., Miscanthus affinis, M boninensis, M brevipilus, M capensis, M changii, M. chejuensis, M chinensis, M chrysander, M condensatus, M 10 coreensis, M cotulhfer, M depauperatus, M ecklonii, M eulalioides, M flavidus, M floribundus, M floridulus, M formosanus, M fuscus, M gossweileri, M hackelii, M hidakanus, M intermedius, M ionandros, M japonicus, M jucundum, M junceus, M kanehirai, M kokusanensis, M littoralis, M longiberbis, M lutarioriparius, M luzonensis, M matsudae, M matsumurae, M miser, M nakaianus, M neo-coreanus, M nepalensis, M 15 nudipes, M ogiformis, M oligostachyus, M oligostachyus, M paniculatus, M polydactylos, M purpurascens, M pycnocephalus, M ridleyi, M rufipilus, M ryukyuensis, M saccariflorus, M sacchaliflorus, M saccharifer, M sacchariflorus, M sieboldi, M sinensis, M sorghum, M szechuanensis, M tanakae, M taylorii, M teretifolius, M tincrorius, M tinctorius, M transmorrisonensis, M violaceus, M wardii, M yunnanensis, M zebrinus 20 Hybrid: M x giganteus); 102381 Erianthus (e.g., Erianthus acutecarinatus, E. acutipennis,E. adpressus, E. alopecuroides, E. angulatus, E. angustifolius, E. armatus, E. articulatus, E. arundinaceus, E. asper, E. aureus, E. bakeri, E. balansae, E. beccarii, E. bengalensis, E. biaristatus, E. bifidus, E. birmanicus, E. bolivari, E. brasilianus, E. brevibarbis, E. capensis, E. chrysothrix, E. 25 ciliaris, E. clandestinus, E. coarctatus, E. compactus, E. contortus, E. cumingii, E. cuspidatus, E. decus-sylvae, E. deflorata, E. divaricatus, E. dohrni, E. ecklonii, E. elegans, E. elephantinus, E. erectus, E. fallax, E. fastigiatus, E. flifolius, E. fischerianus, E. flavescens, E. flavipes, E. flavoinflatus, E. floridulus, E. formosanus, E. formosus, E. fruhstorferi, E. fulvus, E. giganteus, E. glabrinodis, E. glaucus, E. griffithii, E. guttatus, E. hexastachyus, E. 30 hookeri, E. hostii, E. humbertianus, E. inhamatus, E. irritans, E. jacquemontii, E. jamaicensis, E. japonicus, E. junceus, E. kajkaiensis, E. kanashiroi, E. lancangensis, E. laxus, E. longesetosus, E. longifolius, E. longisetosus, E. longisetus, E. lugubris, E. luzonicus, E. -83mackinlayi, E. macratherus, E. malcolmi, E. manueli, E. maximus, E. mishmeensis, E. mollis, E. monstierii, E. munga, E. munja, E. nepalensis, E. nipponensis, E. nudipes, E. obtusus, E. orientalis, E. pallens, E. parviflorus, E. pedicellaris, E. perrieri, E. pictus, E. pollinioides, E. procerus, E. pungens, E. purpurascens, E. purpureus, E. pyramidalis, E. ravennae, E. rehni, 5 E. repens, E. rockii, E. roxburghii, E. rufipilus, E. rufus, E. saccharoides, E. sara, E. scriptorius, E. sesquimetralis, E. sikkimensis, E. smallii, E. sorghum, E. speciosus, E. strictus, E. sukhothaiensis, E. sumatranus, E. teretifolius, E. tinctorius, E. tonkinensis, E. tracyi, E. trichophyllus, E. trinii, E. tristachyus, E. velutinus, E. versicolor, E. viguieri, E. villosus, E. violaceus, E. vitalisi, E. vulpinus, E. wardii, E. williamsii); 10 102391 Pennisetum (e.g., Pennisetum adoense, P. advena, P. alapecuroides, P. albicauda, P. alopecuroides, P. alopecuros, P. americanum, P. amethystinum, P. amoenum, P. ancylochaete, P. angolense, P. angustifolium, P. annuum, P. antillarum, P. araneosum, P. aristidoides, P. arnhemicum, P. articulare, P. arvense, P. asperifolium, P. asperum, P. atrichum, P. aureum, P. bambusiforme, P. baojiense, P. barbatum, P. barteri, P. basedowii, 15 P. beckeroides, P. benthami, P. blepharideum, P. borbonicum, P. brachystachyum, P. breve, P. breviflorum, P. caffrum, P. calyculatum, P. caninum, P. carneum, P. catabasis, P. cauda ratti, P. cenchroides, P. centrasiaticum, P. cereale, P. chevalieri, P. chilense, P. chinense, P. chudeaui, P. ciliare, P. ciliares, P. ciliatum, P. cinereum, P. clandestinum, P. cognatum, P. complanatum, P. compressum, P. cornucopiae, P. corrugatum, P. crinitum, P. crus-galli, P. 20 cupreum, P. cylindricum, P. cynosuroides, P. dalzielii, P. darfuricum, P. dasistachyum, P. dasystachyum, P. davyi, P. densiflorum, P. depauperatum, P. dichotomum, P. dilloni, P. dioicum, P. dispiculatum, P. distachyum, P. distylum, P. divisum, P. domingense, P. dowsonii, P. durum, P. echinurus, P. elatum, P. elegans, P. elymoides, P. erubescens, P. erythraeum, P. exaltalum, P. exiguum, P. exile, P. fallax, P. fasciculatum, P. felicianum, P. flaccidum, P. 25 flavescens, P. flavicomum, P. flavisetum, P. flexile, P. flexispica, P. foermerianum, P. franchetianum, P. frutescens, P. gabonense, P. gambiense, P. geniculatum, P. germanicum, P. gibbosum, P. giganteum, P. glabrum, P. glaucifolium, P. glaucocladum, P. glaucum, P. gossweileri, P. gracile, P. gracilescens, P. grandiflorum, P. griffithii, P. haareri, P. hamiltonii, P. helvolum, P. henryanum, P. hirsutum, P. hohenackeri, P. holcoides, P. 30 hordeiforme, P. hordeoides, P. humboldtianum, P. humile, P. identicum, P. imberbe, P. implicatum, P. inclusum, P. incomptum, P. indicum, P. intectum, P. intertextum, P. italicum, P. jacquesii, P. japonicum, P. javanicum, P. kamerunense, P. karwinskyi, P. kirkii, P. kisantuense, P. lachnorrhachis, P. laevigatum, P. lanatum, P. lanuginosum, P. latifolium, P. - 84 laxior, P. laxum, P. lechleri, P. ledermanni, P. leekei, P. leonis, P. linnaei, P. longifolium, P. longisetum, P. longissimum, P. longistylum, P. macrochaetum, P. macropogon, P. macrostachyon, P. macrostachys, P. macrostachyum, P. macrourum, P. maiwa, P. malacochaete, P. marquisense, P. massaicum, P. megastachyum, P. merkeri, P. mexicanum, 5 P. mezianum, P. mildbraedii, P. molle, P. mollissimum, P. mongolicum, P. monostigma, P. montanum, P. multiflorum, P. mutilatum, P. myosuroides, P. myurus, P. natalense, P. nemorum, P. nepalense, P. nervosum, P. nicaraguense, P. nigricans, P. nigritarum, P. niloticum, P. nitens, P. nodiflorum, P. notarisii, P. nubicum, P. numidicum, P. obovatum, P. occidentale, P. ochrops, P. orientale, P. orthochaete, P. ovale, P. oxyphyllum, P. pallescens, 10 P. pallidum, P. panormitanum, P. pappianum, P. parisii, P. parviflorum, P. paucisetum, P. pauperum, P. pedicellatum, P. pennisetqforme, P. pentastachyum, P. persicum, P. perspeciosum, P. peruvianum, P. petiolare, P. petraeum, P. phalariforme, P. phalaroides, P. pilcomayense, P. pirottae, P. plukenetii, P. plumosum, P. polycladum, P. polygamum, P. polystachion, P. polystachyon, P. preslii, P. prieurii, P. pringlei, P. procerum, P. prolificum, 15 P. proximum, P. pruinosum, P. pseudotriticoides, P. pumilum, P. pungens, P. purpurascens, P. purpureum, P. pycnostachyum, P. qianningensis, P. quartinianum, P. ramosissimum, P. ramosum, P. rangei, P. refractum, P. respiciens, P. reversum, P. richardii, P. rigidum, P. riparioides, P. riparium, P. robustum, P. rogeri, P. rueppelianum, P. rufescens, P. rupestre, P. ruppellii, P. sagittatum, P. sagittifolium, P. saifex, P. sampsonii, P. scaettae, P. scandens, 20 P. schimperi, P. schliebenii, P. schweinfurthii, P. sciureum, P. sclerocladum, P. scoparium, P. secundiflorum, P. sericeum, P. setaceum, P. seligerum, P. setosum, P. shaanxiense, P. sichuanense, P. sieberi, P. sieberianum, P. siguiriense, P. simeonis, P. sinaicum, P. sinense, P. snowdenii, P. somalense, P. sordidum, P. spectabile, P. sphacelatum, P. spicatum, P. squamulatum, P. stapfianum, P. stenorrhachis, P. stenostachyum, P. stolzii, P. stramineum, P. 25 subangustum, P. subeglume, P. swartzii, P. tempisquense, P. teneriffae, P. tenue, P. tenuifolium, P. tenuispiculatum, P. thulinii, P. thunbergii, P. tiberiadis, P. togoense, P. trachyphyllum, P. triflorum, P. trisetum, P. tristachyon, P. tristachyum, P. triticoides, P. typhoides, P. typhoideum, P. uliginosum, P. unflorum, P. unisetum, P. vahlii, P. validum, P. variabile, P. versicolor, P. verticillatum, P. villosum, P. violaceum, P. viride, P. vulcanicum, 30 P. vulpinum, P. weberbaueri, P. yemens); [0240 Saccharum (e.g., as described above including S. ravennae and S. sponteneum); 102411 Arundo (Arundo donax, A. formosana, A. mediterranea, A. pliniana); - 85 - 102421 Sorghum (e.g., Sorghum abyssinicum, S. aethiopicum, S. album, S. andropogon, S. ankolib, S. annuum, S. anomalum, S. arctatum, S. arduini, S. arenarium, S. argenteum, S. arunidinaceum, S. arvense, S. asperum, S. aterrimum, S. australiense, S. avenaceum, S. balansae, S. bantuorum, S. barbatum, S. basiplicatum, S. basutorum, S. 5 bicorne, S. bipennatum, S. bourgaei, S. brachystachyum, S. bracteatum, S. brevicallosum, S. brevicarinatum, S. brevifolium, S. burmahicum, S. cabanisii, S. caffrorum, S. campanum, S. campestre, S. camporum, S. candatum, S. canescens, S. capense, S. capillare, S. carinatum, S. castaneum, S. caucasicum, S. caudatum, S. centroplicatum, S. cernum, S. cernuum, S. chinense, S. chinese, S. cirratum, S. commune, S. compactum, S. condensatum, S. 10 consanguineum, S. conspicuum, S. contortum, S. controversum, S. coriaceum, S. crupina, S. cubanicus, S. cubense, S. deccanense, S. decolor, S. decolorans, S. dimidiatum, S. dochna, S. dora, S. dubium, S. dulcicaule, S. durra, S. elegans, S. elliotii, S. elliottii, S. elongatum, S. eplicatum, S. exaratum, S. exsertum, S. fastigiatum, S. fauriei, S. flavescens, S. flavum, S. friesii, S. fulvum, S. fuscum, S. gambicum, S. giganteum, S. glabrescens, S. glaucescens, S. 15 glaziovii, S. glomeratum, S. glycychylum, S. gracile, S. gracilipes, S. grandes, S. guineence, S. guineense, S. guinense, S. halapense, S. halenpensis, S. halepensis, S. hallii, S. hewisonii, S. hirse, S. hirtiflorum, S. hirtifolium, S. hirtum, S. hybrid, S. incompletum, S. japonicum, S. junghuhnii, S. lanceolatum, S. laterale, S. laxum, S. leicladum, S. leptocladum, S. leptos, S. leucostachyum, S. liebmanni, S. liebmannii, S. lithophilum, S. longiberbe, S. macrochaeta, S. 20 malacostachyum, S. margaritiferum, S. medioplicatum, S. mekongense, S. melaleucum, S. melanocarpum, S. mellitum, S. membranaceum, S. micratherum, S. miliaceum, S. miliiforme, S. minarum, S. mixture, S. mjoebergii, S. muticum, S. myosurus, S. nankinense, S. negrosense, S. nervosum, S. nigericum, S. nigricans, S. nigrum, S. niloticum, S. nitens, S. notabile, S. nubicum, S. nutans, S. orysoidum, S. pallidum, S. panicoides, S. papyrascens, S. parviflorum, 25 S. pauciflorum, S. piptatherum, S. platyphyllum, S. pogonostachyum, S. pohlianum, S. provinciale, S. pugionfolium, S. purpureo-sericeum, S. pyramidale, S. quartinianum, S. repens, S. riedelii, S. rigidifolium, S. rigidum, S. rollii, S. roxburghii, S. rubens, S. rufum, S. ruprechtii, S. saccharatum, S. saccharoides, S. salzmanni, S. sativum, S. scabriflorum, S. schimperi, S. schlumbergeri, S. schottii, S. schreberi, S. scoparium, S. secundum, S. 30 semiberbe, S. serratum, S. setifolium, S. simulans, S. somaliense, S. sorghum, S. spathiflorum, S. splendidum, S. spontaneum, S. stapfli, S. striatum, S. subglabrescens, S. sudanense, S. tataricum, S. technicum, S. technicus, S. tenerum, S. ternatum, S. thonizzi, S. trichocladum, S. trichopus, S. tropicum, S. truchmenorum, S. usambarense, S. usorum, S. verticillatum, S. - 86 verticilh'florum, S. vestitum, S. villosum, S. virgatum, S. virginicum, S. vogelianum, S. vulgare, S. wrightii, S. zeae, S. zollingeri Hybrid S. x almum, S. x almum Parodi, S. bicolor x sudanense, S. x derzhavinii, S. x drummondii, S. x randolphianum); [02431 Poplars (e.g., Populus P. acuminata, P. adenopoda, P. alba, P. afghanica, 5 P. alaschanica, P. amurensis, P. angustifolia, P. baicalensis, P. balsamifera, P. beijingensis, P. candicans, P. cathayana, P. charbinensis, P. ciliata, P. davidiana, P. deltoides, P. dimorpha, P. euphratica, P. flexibilis, P. fremontii, P. grandidentata, P. heterophylla, P. incrassata, P. koreana, P. lasiocarpa, P. laurifolia, P. maximowiczii, P. moskoviensis, P. nigra, P. petrowskiana, P. pruinosa, P. purdomii, P. rasumowskiana, P. sargentii, P. 10 sieboldii, P. simonii, P. suaveolens, P. szechuanica, P. tomentosa, P. tremula, P. tremuloides, P. trichocarpa, P. tristis, P. vernirubens, P. wilsonii, P. woobstii, P. yunnanensisONothospecies: P. x acuminata, P. x berolinensis, P. x brayshawii, P. x canadensis, P. x canescens, P. x generiosa, P. x hinckleyana, P. x jackii); 102441 wheat (e.g., Triticum abyssinicum, T. accessorium, T acutum, T 15 aegilapoides, T. aegilopoides, T. aegilops, T. aesticum, T aestivum, T. aethiopicum, T. affine, T. afghanicum, T agropyrotriticum, T alatum, T. album, T. algeriense, T alpestre, T. alpinum, T. amyleum, T amylosum, T angustifolium, T. angustum, T antiquorum, T. apiculatum, T. aragonense, T. aralense, T. araraticum, T. arenarium, T. arenicolum, T. arias, T. aristatum, T. arktasianum, T. armeniacum, T. arras, T. arundinaceum, T. arvense, T. 20 asiaticum, T asperrimum, T. asperum, T athericum, T. atratum, T attenuatum, T aucheri, T. baeoticum, T barbinode, T. barbulatum, T barrelieri, T batalini, T bauhini, T benghalense, T. bicorne, T. bifaria, T. biflorum, T. biunciale, T. bonaepartis, T. boreale, T. borisovii, T. brachystachyon, T. brachystachyum, T. breviaristatum, T. brevisetum, T. brizoides, T. bromoides, T. brownei, T bucharicum, T. bulbosum, T. bungeanum, T buonapartis, T 25 burnaschewi, T caeruleum, T caesium, T. caespitosum, T. campestre, T candissimum, T caninum, T. capense, T carthlicum, T. caucasicum, T caudatum, T. cereale, T. cerulescens, T. cevallos, T. chinense, T cienfuegos, T. ciliare, T. ciliatum, T. cinereum, T. clavatum, T coarctatum, T cochleare, T. comosum, T. compactum, T. compositum, T. compressum, T. condensatum, T crassum, T. cretaceum, T. creticum, T. crinitum, T. cristatum, T. curvifolium, 30 T. cylindricum, T cynosuroides, T. czernjaevi, T. dasyanthum, T. dasyphyllum, T. dasystachys, T. dasystachyum, T. densiflorum, T densiusculum, T. desertorum, T. dichasians, T. dicoccoides, T dicoccon, T. dicoccum, T. distachyon, T. distans, T distertum, T. distichum, T. divaricatum, T divergens, T diversifolium, T. donianum, T dumetorum, T. duplicatum, T. - 87 duriusculum, T. duromedium, T. durum, T. duvalii, T. elegans, T. elongatum, T. elymogenes, T elymoides, T emarginatum, T. erebuni, T erinaceum, T. farctum, T farrum, T. fastuosum, T. festuca, T. festucoides, T. fibrosum, T. filiforme, T. firmum, T. flabellatum, T. flexum, T. forskalei, T. fragile, T. freycenetii, T fuegianum, T fungicidum, T. gaertnerianum, T. 5 geminatum, T. geniculatum, T. genuense, T. giganteum, T glaucescens, T. glaucum, T. gmelini, T. gracile, T. halleri, T. hamosum, T. hebestachyum, T heldreichii, T. hemipoa, T. hieminfiatum, T hirsutum, T hispanicum, T hordeaceum, T hordeiforme, T hornemanni, T. horstianum, T hosteanum, T. hybernum, T. ichyostachyum, T. imbricatum, T. immaturatum, T. infestum, T infiatum, T. intermedium, T. ispahanicum, T. jakubzineri, T. juncellum, T. 10 junceum, T. juvenale, T. kiharae, T kingianum, T kirgianum, T koeleri, T. kosanini, T. kotschyanum, T kotschyi, T labile, T lachenalii, T laevissimum, T lasianthum, T latiglume, T. latronum, T. laxiusculum, T. laxum, T. leersianum, T. ligusticum, T linnaeanum, T litorale, T litoreum, T. littoreum, T. loliaceum, T. lolioides, T. longearistatum, T longisemineum, T. longissimum, T lorentii, T lutinflatum, T. luzonense, T. macha, T. 15 macrochaetum, T. macrostachyum, T. macrourum, T. magellanicum, T maritimum, T. markgrafli, T martius, T. maturatum, T. maurorum, T. maximum, T. mexicanum, T miguschovae, T missuricum, T. molle, T. monococcum, T. monostachyum, T. multiflorum, T murale, T. muricatum, T nardus, T. neglectum, T. nigricans, T. nodosum, T nubigenum, T obtusatum, T obtusiflorum, T. obtusifolium, T. obtusiusculum, T. olgae, T orientale, T 20 ovatum, T palaeo-colchicum, T palmovae, T. panarmitanum, T paradoxum, T. patens, T. patulum, T. pauciflorum, T pectinatum, T. pectinforme, T. percivalianum, T peregrinum, T. persicum, T. peruvianum, T petraeum, T petropavlovskyi, T phaenicoides, T. phoenicoides, T pilosum, T. pinnatum, T planum, T. platystachyum, T poa, T. poliens, T. polonicum, T. poltawense, :. m polystachyum, i. nticum, T pouzolzii, T. proliferum, T. prostratum, T 25 pruinosum, T. pseudo-agropyrum, T. pseudocaninum, T puberulum, T pubescens, T. pubiflorum, T. pulverulentum, T pumilum, T. pungens, T pycnanthum, T. pyramidale, T. quadratum, T. ramificum, T. ramosum, T. rarum, T recognitum, T. rectum, T. repens, T reptans, T. requlenii, T richardsonii, T rigidum, T rodeti, T roegnerii, T rossicum, T. roitboellia, T rouxii, T. rufescens, T. rufinflatum, T rupestre, T. sabulosum, T. salinum, T 30 salsuginosum, T. sanctum, T sardinicum, T. sartarii, T sativum, T savignionii, T. savignonii, T. scaberrimum, T scabrum, T schimperi, T schrenkianum, T scirpeum, T secale, T secalinum, T secundum, T segetale, T semicostatum, T. sepium, T sibiricum, T. siculum, T siliginum, T. silvestre, T simplex, T. sinaicum, T sinskajae, T. solandri, T. sparsum, T spelta, - 88 - T speltaeforme, T. speltoides, T sphaerococcum, T. spinulosum, T. spontaneum, T squarrosum, T. striatum, T. strictum, T. strigosum, T subaristatum, T. subsecundum, T. subtile, T. subulatum, T. sunpani, T. supinum, T. sylvaticum, T sylvestre, T. syriacum, T. tanaiticum, T tauri, T. tauschii, T. tenax, T. lenellum, T tenue, T tenuiculum, T. teretiflorum, 5 T. thaoudar, T. tiflisiense, T. timococcum, T timonovum, T. timopheevi, . timopheevii, T tomentosum, T. tourneforiii, T. trachycaulon, T. trachycaulum, T. transcaucasicum, T. triaristatum, T. trichophorum, T. tricoccum, T. tripsacoides, T. triunciale, T truncatum, T tumonia, T. turanicum, T turcomanicum, T. turcomanieum, T. turgidum, T tustella, T umbellulatum, T uniaristatum, T. unilaterale, T. unioloides, T. urartu, T. vagans, T. vaginans, 10 T vaillantianum, T variabile, T. variegatum, T varnense, T. vavilovi, T. vavilovii, T. velutinum, T. ventricosum, T. venulosum, T. villosum, T. violaceum, T. virescens, T volgense, T. vulgare, T. youngii, T. zea, T zhukovskyi); 10245] rice (e.g, Oryza abnensis, 0. abromeitiana, 0. alta, 0. angustifolia, 0. aristata, 0. australiensis, 0. barthii, 0. brachyantha, 0. breviligulata, 0. carinata, 0. 15 caudata, 0. ciliata, 0. clandestina, 0. coarctata, 0. collina, 0. communissima, 0. cubensis, 0. denudata, 0. dewildemani, 0. eichingeri, 0. elongata, 0. fatua, 0. filiformis, 0. formosana, 0. glaberrima, 0. glaberi, 0. glaberrima, 0. glaberrina, 0. glauca, 0. glumaepatula, 0. glutinosa, 0. grandiglumis, 0. granulata, 0. guineensis, 0. hexandra, 0. hybrid, 0. indandamanica, 0. jeyporensis, 0. latifolia, O. leersioides, 0. linnaeus, 0. 20 longiglumis, 0. longistaminata, 0. madagascariensis, 0. malampuzhaensis, 0. manilensis, 0. marginata, 0. meijeriana, O. meridionalis, 0. meridonalis, 0. mexicana, 0. meyeriana, 0. mezii, 0. minuta, 0. monandra, 0. montana, 0. mutica, 0. neocaledonica, 0. nepalensis, 0. nigra, 0. nivara, 0. officinalis, 0. oryzoides, 0. palustris, 0. paraguayensis, 0. parviflora, 0. perennis, 0. perrieri, 0. platyphyla, 0. plena, 0. praecox, 0. prehensilis, 0. pubescens, 0. 25 pumila, 0. punctata, 0. repens, 0. rhizomatis, 0. ridleyi, 0. rubra, 0. rubribarbis, 0. rufipogon, 0. sativa, 0. schlechteri, 0. schweinfurthiana, 0. segetalis, 0. sorghoidea, 0. sorghoides, 0. spontanea, 0. stapfii, 0. stenothyrsus, 0. subulata, 0. sylvestris, 0. tisseranti, 0. tisserantii, 0. triandra, 0. triticoides, 0. ubanghensis); 102461 oats (e.g., Avena abietorum, A. abyssinica, A. adsurgens, A. adzharica, A. 30 aemulans, A. aenea, A. affinis, A. agadiriana, A. agraria, A. agraria-mutica, A. agraria sesquialtera, A. agropyroides, A. agrostidea, A. agrostoides, A. airoides, A. alba, A. albicans, A. albinervis, A. algeriensis, A. almeriensis, A. alopecuros, A. alpestris, A. alpina, A. alta, A. altaica, A. altior, A. altissima, A. ambigua, A. americana, A. amethystina, A. anathera, A. -89andropogoides, A. andropogonoides, A. anglica, A. anisopogon, A. antarctica, A. arduensis, A. arenaria, A. argaea, A. argentea, A. argentoideum, A. ariguensis, A. aristeliformis, A. aristidioides, A. aristidoides, A. armeniaca, A. arundinacea, A. arvensis, A. aspera, A. atheranthera, A. atlantica, A. atropurpurea, A. aurata, A. australis, A. azo-carti, A. barbata, 5 A. baregensis, A. baumgartenii, A. beguinotiana, A. bellardi, A. benghalensis, A. besseri, A. bifida, A. bipartita, A. blavii, A. bolivaris, A. borbonia, A. bornmuelleri, A. breviaristata, A. brevifolia, A. brevis, A. bromoides, A. brownei, A. brownii, A. bruhnsiana, A. bulbosa, A. burnoufli, A. byzantina, A. caespitosa, A. caffra, A. calicina, A. callosa, A. calycina, A. canariensis, A. candollei, A. canescens, A. canina, A. cantabrica, A. capensis, A. capillacea, 10 A. capillaris, A. carmeli, A. caroliniana, A. carpatica, A. caryophyllea, A. cavanillesii, A. cernua, A. chinensis, A. chlorantha, A. ciliaris, A. cinerea, A. clarkei, A. clauda, A. coarctata, A. colorata, A. compacta, A. compressa, A. condensata, A. convoluta, A. coquimbensis, A. coronensis, A. corymbosa, A. crassifolia, A. cristata, A. cupaniana, A. cuspidata, A. daenensis, A. dahurica, A. damascena, A. decora, A. delavayi, A. depauperata, A. desertorum, 15 A. deusta, A. deyeuxioides, A. discolor, A. dispermis, A. distans, A. disticha, A. distichophylla, A. dubia, A. dufourei, A. dura, A. editissima, A. elata, A. elatior, A. elegans, A. elephantina, A. elongata, A. eriantha, A. fallax, A. fatua, A. fedtschenkoi, A. fertilis, A. festucaeformis, A. festucoides, A. filifolia, A. filiformis, A. flaccida, A. flava, A. flavescens, A. flexuosa, A. forskolei, A. forsteri, A. fragilis, A. freita, A. fusca, A. fuscoflora, A. gallecica, A. gaudiana, A. 20 gaudiniana, A. geminflora, A. georgiana, A. georgica, A. gigantea, A. glabra, A. glabrata, A. glabrescens, A. glacialis, A. glauca, A. glomerata, A. glumosa, A. gonzaloi, A. gracilis, A. gracillima, A. grandis, A. hackelii, A. haussknechtii, A. heldreichii, A. heteromalla, A. hexantha, A. hideoi, A. hirsuta, A. hirta, A. hirtifolia, A. hirtula, A. hispanica, A. hispida, A. hookeri, A. hoppeana, A. hostii, A. hugeninii, A. hungarica, A. hybrida, A. hydrophila, A. 25 insubrica, A. insularis, A. intermedia, A. involucrata, A. involuta, A. jahandiezii, A. japonica, A. junghuhnii, A. koenigii, A. kotschyi, A. lachnantha, A. laconica, A. laeta, A. laevigata, A. laevis, A. lanata, A. lanuginosa, A. lasiantha, A. latifolia, A. leiantha, A. lejocolea, A. lendigera, A. leonina, A. leptostachys, A. letourneuxii, A. levis, A. lodunensis, A. loefflingiana, A. loeflingiana, A. longa, A. longepedicellata, A. longepilosa, A. longespiculata, A. longifolia, 30 A. longiglumis, A. longipilosa, A. lucida, A. ludoviciana, A. lupulina, A. lusitanica, A. lutea, A. macilenta, A. macra, A. macrantha, A. macrocalycina, A. macrocalyx, A. macrocarpa, A. macrostachya, A. magellanica, A. magna, A. malabarica, A. malzevii, A. mandoniana, A. marginata, A. maroccana, A. matritensis, A. maxima, A. mediolanensis, A. melillensis, A. - 90 meridionalis, A. micans, A. michelii, A. micrantha, A. mirandana, A. mollis, A. mongolica, A. montana, A. montevidensis, A. mortoniana, A. multiculmis, A. muralis, A. muricata, A. muriculata, A. murphyi, A. mutica, A. myriantha, A. mysorensis, A. nana, A. neesii, A. neglecta, A. nemoralis, A. nervosa, A. neumeyeriana, A. newtonii, A. nigra, A. nitens, A. 5 nitida, A. nodipilosa, A. nodosa, A. noeana, A. notarisii, A. nuda, A. nudibrevis, A. nutans, A. nutkaensis, A. occidentalis, A. odorata, A. oligostachya, A. opulenta, A. orientalis, A. ovata, A. ovina, A. palaestina, A. pallens, A. pallida, A. palustris, A. panicea, A. panormitana, A. papillosa, A. paradensis, A. paradoxa, A. parlatorei, A. parlatorii, A. parviflora, A. pauciflora, A. paupercula, A. pendula, A. penicillata, A. pennsylvanica, A. pensylvanica, A. 10 persarum, A. persica, A. peruviana, A. phleoides, A. pilosa, A. planiculmis, A. podolica, A. polonica, A. polyneura, A. ponderosa, A. pourretii, A. praecocioides, A. praecoqua, A. praecox, A. praegravis, A. praeusta, A. pratensis, A. precatoria, A. preslii, A. prostrata, A. provincialis, A. pruinosa, A. pseudolucida, A. pseudosativa, A. pseudoviolacea, A. puberula, A. pubescens, A. pulchella, A. pumila, A. pungens, A. purpurascens, A. purpurea, A. pusilla, 15 A. quadridentula, A. quadriseta, A. quinqueseta, A. racemosa, A. radula, A. redolens, A. riabushinskii, A. rigida, A. rotae, A. rothii, A. roylei, A. rubra, A. rufescens, A. rupestris, A. ruprechtii, A. sarracenorum, A. sativa, A. saxatilis, A. scabriuscula, A. scabrivalvis, A. schelliana, A. scheuchzeri, A. secalina, A. secunda, A. sedenensis, A. segetalis, A. sempervirens, A. sensitiva, A. septentrionalis, A. serrulatiglumis, A. sesquiflora, A. 20 sesquitertia, A. setacea, A. set tfolia, A. sexaflora, A. sexflora, A. shatilowiana, A. sibirica, A. sibthorpii, A. sicula, A. sikkimensis, A. smithii, A. solida, A. spica-venti, A. spicaeformis, A. spicata, A. splendens, A. squarrosa, A. sterilis, A. stipaeformis, A. stipoides, A. striata, A. stricta, A. strigosa, A. subalpestris, A. subcylindrica, A. subdecurrens, A. subspicata, A. subulata, A. subvillosa, A. suffusca, A. sulcata, A. sylvatica, A. symphicarpa, A. syriaca, A. 25 tatarica, A. taygetana, A. tenorii, A. tentoensis, A. tenuiflora, A. tenuis, A. thellungii, A. thorei, A. tibestica, A. tibetica, A. toluccensis, A. tolucensis, A. torreyi, A. trabutiana, A. triaristata, A. trichophylla, A. trichopodia, A. triseta, A. trisperma, A. triticoides, A. truncata, A. tuberosa, A. turgidula, A. turonensis, A. unflora, A. unilateralis, A. valesiaca, A. varia, A. vasconica, A. vaviloviana, A. velutina, A. ventricosa, A. versicolor, A. vilis, A. villosa, A. 30 virescens, A. viridis, A. volgensis, A. wiestii, A. wilhelmsi); [0247] willows (e.g., Salix species); switch grass (i.e., Panicum virgatum); alfalfa (i.e., Medicago sativa); prairie bluestem (e.g., Andropogon gerardii); maize (i.e., Zea mays); -91 - 10248] soybean (i.e., Glycine max); barley (i.e., Hordeum vulgare); sugar beet (i.e., Beta vulgaris); hay and fodder crops. [02491 Procedures for transforming plants are well known and routine in the art and are described throughout the literature. Non-limiting examples of methods for transformation 5 of plants include transformation via bacterial-mediated nucleic acid delivery (e.g., via Agrobacteria), viral-mediated nucleic acid delivery, silicon carbide or nucleic acid whisker mediated nucleic acid delivery, liposome mediated nucleic acid delivery, microinjection, microparticle bombardment, calcium-phosphate-mediated transformation, cyclodextrin-mediated transformation, electroporation, nanoparticle-mediated transformation,, 10 sonication, infiltration, PEG-mediated nucleic acid uptake, as well as any other electrical, chemical, physical (mechanical) and/or biological mechanism that results in the introduction of nucleic acid into the plant cell, including any combination thereof. General guides to various plant transformation methods known in the art include Miki et al. ("Procedures for Introducing Foreign DNA into Plants" in Methods in Plant Molecular Biology and 15 Biotechnology, Glick, B. R. and Thompson, J. E., Eds. (CRC Press, Inc., Boca Raton, 1993), pages 67-88) and Rakowoczy-Trojanowska (Cell. Mol. Biol. Lett. 7:849-858 (2002)). [0250] Thus, in some particular embodiments, the introducing into a plant, plant part, plant organ and/or plant cell is via bacterial-mediated transformation, particle bombardment transformation, calcium-phosphate-mediated transformation, cyclodextrin 20 mediated transformation, electroporation, liposome-mediated transformation, nanoparticle mediated transformation, polymer-mediated transformation, virus-mediated nucleic acid delivery, whisker-mediated nucleic acid delivery, microinjection, sonication, infiltration, polyethylene glycol-mediated transformation, any other electrical, chemical, physical and/or biological mechanism that results in the introduction of nucleic acid into the plant, plant part 25 and/or cell thereof, or a combination thereof. 102511 Agrobacterium-mediated transformation is a commonly used method for transforming plants, in particular, dicot plants, because of its high efficiency of transformation and because of its broad utility with many different species. Agrobacterium-mediated transformation typically involves transfer of the binary vector carrying the foreign DNA of 30 interest to an appropriate Agrobacterium strain that may depend on the complement of vir genes carried by the host Agrobacterium strain either on a co-resident Ti plasmid or chromosomally (Uknes et al. (1993) Plant Cell 5:159-169). The transfer of the recombinant - 92 binary vector to Agrobacterium can be accomplished by a triparental mating procedure using Escherichia coli carrying the recombinant binary vector, a helper E. coli strain that carries a plasmid that is able to mobilize the recombinant binary vector to the target Agrobacterium strain. Alternatively, the recombinant binary vector can be transferred to Agrobacterium by 5 nucleic acid transformation (H6fgen & Willmitzer (1988) Nucleic Acids Res. 16:9877). Transformation of a plant by recombinant Agrobacterium usually involves co-cultivation of the Agrobacterium with explants from the plant and follows methods well known in the art. Transformed tissue is regenerated on selection medium carrying an antibiotic or herbicide resistance marker between the binary plasmid T-DNA borders. 10 [02521 Another method for transforming plants, plant parts and plant cells involves propelling inert or biologically active particles at plant tissues and cells. See, e.g., U.S. Patent Nos. 4,945,050; 5,036,006 and 5,100,792. Generally, this method involves propelling inert or biologically active particles at the plant cells under conditions effective to penetrate the outer surface of the cell and afford incorporation within the interior thereof. When inert particles are 15 utilized, the vector can be introduced into the cell by coating the particles with the vector containing the nucleic acid of interest. Alternatively, a cell or cells can be surrounded by the vector so that the vector is carried into the cell by the wake of the particle. Biologically active particles (e.g., a dried yeast cell, a dried bacterium or a bacteriophage, each containing one or more nucleic acids sought to be introduced) also can be propelled into plant tissue. Thus, in 20 particular embodiments of the present invention, a plant cell can be transformed by any method known in the art and as described herein and intact plants can be regenerated from these transformed cells using any of a variety of known techniques. Plant regeneration from plant cells, plant tissue culture and/or cultured protoplasts is described, for example, in Evans et al. (Handbook of Plant Cell Cultures, Vol. 1, MacMilan Publishing Co.New York (1983)); 25 and Vasil I. R. (ed.) (Cell Culture and Somatic Cell Genetics of Plants, Acad. Press, Orlando, Vol. I (1984), and Vol. 11 (1986)). Methods of selecting for transformed, transgenic plants, plant cells and/or plant tissue culture are routine in the art and can be employed in the methods of the invention provided herein. Likewise, the genetic properties engineered into the transgenic seeds and plants, plant parts, and/or plant cells of the present invention described 30 above can be passed on by sexual reproduction or vegetative growth and therefore can be maintained and propagated in progeny plants. Generally, maintenance and propagation make use of known agricultural methods developed to fit specific purposes such as harvesting, sowing or tilling. A nucleotide sequence therefore can be introduced into the plant, plant part - 93 and/or plant cell in any number of ways that are well known in the art. The methods of the invention do not depend on a particular method for introducing one or more nucleotide sequences into a plant, only that they gain access to the interior of at least one cell of the plant. Where more than one nucleotide sequence is to be introduced, the respective nucleotide 5 sequences can be assembled as part of a single nucleic acid construct/molecule, or as separate nucleic acid constructs/molecules, and can be located on the same or different nucleic acid constructs/molecules. Accordingly, the nucleotide sequences can be introduced into the cell of interest in a single transformation event, in separate transformation events, or, for example, in plants, as part of a breeding protocol. In some embodiments of this invention, the introduced 10 nucleic acid molecule may be maintained in the plant cell stably if it is incorporated into a non-chromosomal autonomous replicon or integrated into the plant chromosome(s). Alternatively, the introduced construct may be present on an extra-chromosomal non replicating vector and be transiently expressed or transiently active. Whether present in an extra-chromosomal non-replicating vector or a vector that is integrated into a chromosome, 15 the nucleic acid molecule can be present in a plant expression construct. 4. Uses of the constructs to control transpiration in plants 102531 The constructs of the present invention are useful for controlling stomatal closure, and therefore can be used to control transpiration in transgenic plants containing the constructs. The constructs taught in this invention are particularly valuable in that expression 20 of the stomatal closure-inhibiting nucleic acid sequence of the construct is regulated effectively. In specific embodiments, the expression of the stomatal closure-inhibiting nucleic acid sequence is found only in guard cells. Such constructs will therefore be particularly useful in crop and horticultural varieties in which reduction of moisture content is important. Examples of such crops include but are not limited to cereal grains such as corn, wheat, rye, 25 oats, barley, and rice, soybeans and other beans, as well as other products such as hay and commercial seed. In most of these cases failure to adequately dry the crop due to weather or other conditions results in substantial losses. In other cases including but not limited to tobacco, dried fruits such as raisins and prunes, nuts, coffee, tea, cocoa, and many ornamental goods, the produce needs to be dried immediately after harvest prior and to use. In these cases 30 again, the mutants of this invention may be of tremendous value to growers who could accelerate or control the rate of crop drying. - 94 - 102541 Expression of the stomatal closure-inhibiting nucleic acid sequence of the construct can be achieved by exposing a plant, plant part, plant organ or plant leaf, which comprises the construct, to a compound that induces the expression of the alcohol dehydrogenase (ADH 1) system of Aspergillus nidulans, as described for example above so as 5 to inhibit stomatal closure and thereby increase transpiration in the plant, plant part, plant organ or plant leaf. The plant, plant part, plant organ or plant leaf may be exposed to the compound prior to harvesting (e.g., no more than one month, three weeks, two weeks, one week, six, days, five days, four days, three days, two days, one day prior to harvesting), at the time of harvesting or after harvesting (e.g., no more than one day, two days, three days, four 10 days, five days, six days, one week, two weeks, three weeks, one month after harvesting), the plant, plant part, plant organ or plant leaf. The compound may be applied to the plant, plant part, plant organ or plant leaf using any suitable technique including vapor, dipping, spraying, spray drenching and the like. [02551 Generally, the time and duration of exposing the plant, plant part, plant 15 organ or plant leaf to the compound are chosen to permit increased transpiration in the plant, plant part, plant organ or plant leaf so that the water content of the plant, plant part, plant organ or plant leaf reduces by at least about 5% (e.g., at least about 6%, 7%, 8%, 9%, 10%, 15% 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%). 102561 In some embodiments, the reduced water content results in other beneficial 20 phenotypes, including for example increased stored carbohydrates such as starch and simple sugars (e.g., sucrose, fructose, glucose etc.). 102571 Alternatively, or in addition, the increased transpiration provided by the present invention can be used to dry out the growth medium (e.g., soil) in which transgenic plants of the invention are grown. Illustrative applications of these embodiments include 25 drying out sporting fields with transgenic turf grass, and drying out fields containing transgenic crops (e.g., to permit more facile harvesting of crops). [02581 In order that the invention may be readily understood and put into practical effect, particular preferred embodiments will now be described by way of the following non limiting examples. - 95 - EXAMPLES EXAMPLE I PRODUCTION OF NON-OPTIMIZED CONSTRUCTS AND TRANSFORMATION INTO SUGAR CANE [02591 A ZmUbi-GS construct was constructed comprising the maize ubiquitin 5 promoter (ZmUbi), the GUS reporter gene containing a synthetic intron (GS), and the nopaline synthase (nos) terminator. The GS sequence consists of a 232 bp I" exon, 84 bp synthetic intron, and 1580 bp 2 nd exon. The GS sequence was amplified from p35S-GS by polymerase chain reaction (PCR) using the forward primer 5'-CCCGGGATCCTAAACCATGGTCCGTCCTGTAGAAACCC-3' [SEQ ID NO: 39] and 10 reverse primer 5'-TCATTGTTTGCCTCCCTGCTG-3' [SEQ ID NO:40] and KAPAHiFi DNA polymerase (Geneworks). The resulting PCR product was cloned into pGEM-T (Promega) and sequence verified. The GS sequence was excised from pGEM-T using SmaI and NotI, treated with T4 DNA polymerase (Promega) to blunt the NotI ends and then ligated into the Smal site of a pBluescript (pBS) vector containing the maize Ubi I promoter and nos 15 terminator (ZmUbi-nos/pBS) to generate ZmUbi-GS. [02601 A ZmUbi-AlcR AlcA-GS construct was constructed comprising the AlcR coding sequence and nos under the control of ZmUbi, as well as GS with nos under the control of the AlcA promoter. The AlcR sequence was amplified from 35S-AlcR-AlcA Rep/pUC by PCR using the forward primer 20 5'-TTACTTCTGCAGCCCTAAACCATGGCAGATACGCGCCGACGCCAG3' [SEQ ID NO:41] and reverse primer 5'-TGTTTGAACGATCCCCTACAAAAAGCTGTCAACTTTCCCA -3' [SEQ ID NO:42] and KAPAHiFi DNA polymerase. The resulting PCR product was cloned into the SmaI site of ZmUbi-nos/pBS using the Clontech In Fusion* PCR Cloning System to generate 25 ZmUbi-AlcR, and the PCR product was sequence verified. To generate ZmUbi-AlcR AlcA GS, the AlcA-GS-nos sequence was subeloned from 35S-AlcR AlcA-GS/pUC into ZmUbi AlcR using HindIII. [02611 The ZmUbi-GS and ZmUbi-AlcR AlcA-GS constructs were transformed into sugar cane using microprojectile bombardment of callus material. To generate the callus, 30 sugar cane (cultivar QI 17) "tops" were obtained from The Bureau of Sugarcane Experimental Stations (BSES) LTD Meringa Queensland, Australia. Calli were initiated as described by Franks and Birch (1991, Australian J. Plant Physiol. 18: 471-480) using MSC 3 media consisting - 96 of: 4.43g/L MS basal salts with vitamins (PhytoTechnology Laboratories@, Shawnee Mission, KS, USA ), 500mg/L casein hydrolysate (Merck), 13.6[LM 2,4 Dichlorophenoxyacetic acid (2, 4-D; Phytotechnology laboratories), 1 00ml/L young coconut juice ("Cock" brand, Thailand), 3% (w/v) sucrose and 8 g/L agar (Research organics 10020). 5 Petri dishes used for tissue culture media were 90mm x 25mm high vented type and sealed with micropore surgical tape. Calli were maintained for nine weeks in the dark at 26*C and subcultured every 14 days. [02621 Microprojectile bombardment of callus was performed according to the method of Bower et al (Molecular Breeding 1996 2: 239-249). For callus transformation, 10 sugar cane calli were transferred from MSC 3 media to MSO media (MSC 3 media with the addition of 190mM sorbitol (Sigma) and 190 mM mannitol (Sigma) and arranged in a 3cm diameter circle four hours prior to microprojectile bombardment. The plasmid pUKN (ZmUbi driving expression of the neomycin phosphotransferase II gene) was co-bombarded with each of the GS expression constructs to allow for selection of transformed cells using geneticin. 15 For microprojectile bombardment, a 2 pL aliquot of a 1:1 mixture of pUKN (1 pg/pL) and the GS expression construct DNA (1 pg/pL) was added to approximately 3mg of 1 pm gold particles (Bio-Rad). The solution was vortexed briefly and 25 PL of I M CaCl 2 and 5 pL of 0.1 M spermidine were added simultaneously. The mixture was iced and vortexed for 15 seconds every minute for a total of 5 minutes. The mixture was then allowed to settle on ice 20 for 10 minutes, after which 22 pL of supernatant was removed. The remaining DNA coated gold solution was mixed and 5 pL was used per bombardment. A particle inflow gun (PIG) was used to deliver the DNA to the target tissue. A baffle utilizing stainless steel mesh screen with an aperture of 500 im was positioned approximately 1.5 cm above the target tissue within the PIG chamber. The microflight distance of the DNA from the tip of the swinny to 25 the leaf explant was 10.5 cm. The PIG chamber was vacuum evacuated to -90 kPa and a 10 ms pulse of helium at 1500 psi was used to accelerate the microprojectiles. The vacuum was released immediately following bombardment and each sample plate was rotated 180 degrees and subjected to a second bombardment. [02631 Following bombardment, the callus remained on MSO media for four hours. 30 The callus was subsequently transferred to MSC 3 medium for 4-6 days before being transferred to selection media consisting of MSC 3 and 50 mg/L G418 (Geneticin; Roche). Following microprojectile bombardment, the callus remained on selection media for four weeks in the dark with fortnightly subculturing after which it was transferred to regeneration - 97 medium with selection, consisting of MSC3 with the 2,4-D replaced by 4.4 LM 6 Benzylaminopurine (BAP; Sigma). The callus was maintained at 270 C, under a 16 hour light, and 8 hour dark cycle with fortnightly subculturing. Individual plants were separated and one plant from each clump of callus was retained. After ten weeks of regeneration with BAP, the 5 plants were transferred to rooting medium with selection (the same as regeneration medium, however BAP is replaced with 10.7 pM 1-Naphthalene Acetic Acid (NAA; Sigma). The plants were grown until roots of approximately 1 cm in length had developed after which the plants were transferred to soil for acclimatization in a growth cabinet under the above mentioned lighting and temperature conditions. After approximately six weeks the plants 10 were transferred to soil in 20cm pots, and grown in the greenhouse. EXAMPLE 2 CHARACTERIZATION OF TRANSGENIC PLANTS CONTAINING NON-OPTIMIZED CONSTRUCTS. [02641 Plants were verified to contain the ZmUbi-GS and ZmUbi-AlcR AlcA-GS 15 constructs by TaqMan@ analysis for the GUS transgene. Approximately 15 week-old transgenic plants were analyzed for ethanol inducible GUS reporter gene expression. Leaf samples consisting of five standard hole punches were taken from the first fully unfurled leaf of transgenic plants just prior to ethanol induction. Ethanol treatment was carried out by using a 10% ethanol root drench and aerial spray until runoff. After treatment, the plants were 20 enclosed using plastic sheeting to maximize their exposure to the ethanol vapor. After ethanol treatment, first unfurled leaf samples were taken at three and six days post treatment. All leaf samples were collected on ice and used for GUS histochemical analysis (Jefferson et al. (1987) EMBO J 6: 3901-3907) within four hours of collection. Histochemical analysis was carried out by the addition of GUS staining buffer (50 mM NaPO4 pH 7, 0.1 % Triton, 25 0.5 mM potassium ferrocyanide, 0.5 mM potassium ferricyanide, 10 mM EDTA, and 1 mM X-Gluc), vacuum infiltration of the tissue for 45 minutes, and incubation at 370 C for 48 hours. After 48 hours, the GUS staining buffer was removed and replaced with 100% ethanol to clear the tissue of chlorophyll. Visual inspection of the cleared leaf discs was used to assess GUS expression. 30 102651 No ethanol inducible gene expression was detected in any of the leaf samples taken from 21 independent transgenic sugar cane plants containing the ethanol switch construct ZmUbi-AlcR AlcA-GS. In the transgenic sugar cane plants containing the ZmUbi - 98 - GS positive control construct, leaf samples from six of 13 independent events showed visible GUS staining in both the uninduced and induced leaf samples. 102661 Ten days after the plants had been treated with 10% ethanol, the transgenic I plants were retreated with 2% ethanol using both a root drench and aerial spray until runoff. 5 Leaf samples consisting of five standard hole punches were taken at three and five days post treatment for GUS histochemical staining. Again, none of the leaf samples taken from the 21 transgenic sugar cane plants containing the ethanol switch construct ZmUbi-AlcR AlcA-GS showed any detectable GUS staining. EXAMPLE 3 10 PRODUCTION OF OPTIMIZED ETHANOL SWITCH CONSTRUCTS 102671 To create ethanol switch constructs that may be capable of giving reliable ethanol inducible gene expression in sugar cane, the following modifications were made to the original constructs: [02681 The AlcR and GUS coding sequences were optimized for expression in 15 sugar cane (Geneart optimization). A different Kozak sequence, 5'-gcggccgcc-3' [SEQ ID NO:42] was placed immediately upstream of the scoAlcR and scoGUS coding sequences. The TMV Q translational enhancer sequence 5'-gtatttttacaacaattaccaacaacaacaaacaacaaacaacattacaattactatttacaattaca-3' [SEQ ID NO:43] was placed immediately upstream of the Kozak sequence. 12 different AlcA promoter 20 variants were created as follows: 102691 (1) The SC 12 construct has an unmodified AlcA promoter sequence identical to the sequence present in ZmUbi-AlcR AlcA-GS. This AlcA promoter sequence is identical to the original AlcA promoter sequence used in plants as reported in Caddick et al. (1998, Nature Biotechnology 6: 177-180), and consists of the Aspergillus AlcA promoter 25 (from -349 to -112 relative to the translational start site and lacking the upstream direct repeat AlcR binding site) fused to the CaMV 35S minimal promoter (-32 to +3 relative to the CaMV 35S transcriptional start site) at the TATA box (the TATA box is identical in the AlcA and CaMV 35S promoters). [02701 (2) The SC 13 construct consists of the Aspergillus AlcA promoter 30 sequence (-349 to - 25 112) fused to a longer CaMV 35S minimal promoter element (-73 to +7 relative to the CaMV 35S transcriptional start site). - 99 - 102711 (3) The SC 14 construct consists of an Aspergillus AlcA promoter including the upstream direct repeat AlcR binding site (-400 to -112 relative to the translational start site) fused to the original CaMV 35S minimal sequence (-32 to +3). [02721 (4) The SC 15 construct consists of an Aspergillus AlcA promoter 5 including the upstream direct repeat AlcR binding site (-400 to -112) fused to the longer CaMV 35S minimal sequence (-73 to +7). 102731 (5) The SC16 construct consists of an Aspergillus AlcA promoter (-400 to -1 12) in which the upstream direct repeat AlcR binding site region has been changed from "5'-cgtccgcatcggcatccgcagc-3"' [SEQ ID NO:44] to "5'-tatccgcatgggtatccgcatg-3"' [SEQ ID 10 NO:45] and fused to the original CaMV 35S minimal sequence. [02741 (6) The SC 17 construct consists of an Aspergillus AlcA promoter (-400 to -112) in which the upstream direct repeat AlcR binding site region has been changed from "5'-cgtccgcatcggcatccgcagc-3"' [SEQ ID NO:46] to "5'-tatccgcatgggtatccgcatg-3"' [SEQ ID NO: 47] and fused to the longer CaMV 35S minimal sequence. 15 102751 (7) The SC 18 construct consists of five tandem repeats of the inverted repeat AlcR binding site region sequence "5'-atgcatgcggaaccgcacgagg-3"' [SEQ ID NO:3] at the 5' end of the Aspergillus AlcA promoter including the upstream direct repeat AlcR binding site fused to the original CaMV 35S minimal sequence. 102761 (8) The SC 19 construct consists of five tandem repeats of the inverted 20 repeat AlcR binding site region sequence "5'-atgcatgcggaaccgcacgagg-3"' [SEQ ID NO:3] at the 5' end of the Aspergillus AlcA promoter including the upstream direct repeat AlcR binding site fused to the longer CaMV 35S minimal sequence. [02771 (9) The SC20 construct consists of five tandem repeats of the inverted repeat AlcR binding site region sequence "5'-atgcatgcggaaccgcacgagg-3"' [SEQ ID NO:3] 25 (without any additional AlcA promoter sequence) fused to the longer CaMV 35S minimal sequence. [02781 (10) The SC21 construct consists of the Aspergillus AlcA promoter including the upstream direct repeat AlcR binding site fused to the original CaMV 35S minimal sequence with the addition of the maize Adhl intron (Callis et al. (1987) Genes & 30 Dev. 1:1183-1200) at the 3' end. - 100- 102791 (11) The SC22 construct consists of the Aspergillus AlcA promoter including the upstream direct repeat AlcR binding site fused to the long CaMV 35S minimal sequence with the addition of the maize Adh1 intron at the 3' end. 102801 (12) The SC23 construct consists of the native Aspergillus AlcA promoter 5 (-400 to -64 relative to the translational start site). 102811 The sequences for the AlcA promoter variants, sugar cane optimized GUS (scoGUS; with TMV Q and Kozak) with nos terminator, sugar cane optimized AlcR (scoAlcR; with TMV 0 and Kozak), and the Adhl intron (with 15 bp of 5' exon and 6 bp of 3' exon) were made synthetically and included restriction enzyme sites at each end for cloning. 10 The scoAlcR was excised using the flanking, engineered Hpal sites and cloned into the Smal site of ZmUbi-nos/pBS to generate ZmUbi-scoAlcR. ZmUbi-scoAlcR was digested with EcoRV and Spel, and the ZmUbi-AlcR-nos region was subcloned into the AvrII (blunted using Klenow) and Spel sites of the binary construct UbinptI[Nos(S) to generate scoAlcRnptIl. The scoGUS gene with nos was subcloned into pBluescript using PstI and 15 SaclI to generate scoGUS/pBS. All of the AlcA promoter variants were subsequently cloned into scoGUS/pBS using HindIl and Pstl. For constructs SC21 and SC22, the Adhl intron was cloned into the PstI site located between the AlcA promoter and TMV Q . To generate the final ethanol switch constructs (SC12, SCl3, and SC16-20), the AlcA promoter variant, scoGUS, and nos sequences were subcloned into scoAlcRnptll using HindlIl and Ascl. The 20 final ethanol switch constructs (SC 14, SC15, and SC21-23) were made by subcloning the AlcA promoter variant, scoGUS, and nos sequences into scoAlcRnptlI using AscI alone. 102821 An optimized, constitutive ZmUbi-scoGUS expression construct was generated as follows: ZmUbi was PCR amplified (adding a HindIII site at the 5' end and a Pstl site at the 3' end), cloned into pGEM-T, and sequence verified. ZmUbi was subsequently 25 subcloned into scoGUS/pBS using HindIll and PstI to generate ZmUbi-scoGUS. The ZmUbi scoGUS and nos sequence was subcloned into the binary construct UbinptlINos(S) using HindIII and AscI. 102831 The binary constructs were transferred into Agrobacterium strain AGLI using a standard heat shock transformation method. Agrobacterium containing each of the 30 binary constructs were used to transform sugar cane using following methods (see, Example 4). - 101 - EXAMPLE 4 AGROBACTERIUM-MEDIATED TRANSFORMATION OF SUGAR CANE Plant source and material: 102841 Leaf whorl material from field grown sugar cane plants was collected and 5 initiated on EM3 medium (see below). Transverse sections (approximately 20) of immature leaf whorl between 1-3 mm in thickness were taken from just above the meristem and placed in the top-up orientation. Cultures were maintained in the dark at 250 C for 28 to 42 days. Callus utilized for transformation was within 4-10 days of the last subculture. Callus was selected on morphological characteristics such as compact structure and yellow color. Yellow 10 embryogenic calli were selected wherever possible, as they provided good regeneration, consistent transformation, and fragmented in small clusters (2-4 mm) Infection and co-cultivation: [02851 Callus tissue was heat shocked at 45*C for 5 minutes by adding 50 mL of pre-warmed 1/2 strength MS medium (without sucrose) and then maintaining the callus in a 15 water bath at 450 C. MS medium was then drained from the callus tissue, and 25 mL of the Agrobacterium inoculation suspension was added to each vessel and mixed gently. The callus/Agrobacterium mixture was vacuum-infiltrated by placing it into a vacuum chamber for 10 min at -27.5 mmHg of vacuum. The callus/Agrobacterium mixture was then rested for 5 10 min the dark. The Agrobacterium inoculation suspension was then drained from the callus, 20 and the remaining callus culture was blotted dry to remove excess Agrobacterium inoculation suspension. Plant tissues were blotted on filter paper such as Whatman Grade I paper, until the Agrobacterium inoculation suspension was substantially removed. The callus was then transferred for co-cultivation to 90 x 25 mm petri dishes containing no co-culture medium or containing dry filter papers or filter papers wet with sterile water, and sealed with 25 NESCOFILM@, MICROPORE TM tape (3M; Minneapolis, MN) or similar material. The dishes were incubated in the dark at 22' C for 2-3 days. Post-transformation: 102861 After co-cultivation, the callus tissue was transferred to MS I medium (see below) containing with 200 mg/L of timentin ("resting" medium) and kept in the dark at 25*C 30 for 4 days. The first selection step was made in MS 2 medium (see below) containing 50 mg/L of geneticin and 200 mg/L of timentin for 14-15 days in the dark at 25* C. - 102- Regeneration and rooting: 102871 Regeneration was conducted on MS 3 medium (see below) supplemented with 50 mg/L of geneticin and 200 mg/L of timentin at 25'C in 16 hr. light. Gradual increases in light intensity were required. For the first week, the culture was left on a laboratory bench 5 under normal room lighting, and for the next 3 weeks, the culture was grown at moderate light intensity. 102881 Shoot formation was seen between 2-4 weeks. When the first leaves appeared, the shoots were transferred to MS 4 medium (see below) until the plants grew to 4 5 cm in height. Transformed plants were initially moved from tissue culture and placed in 10 seedling trays containing soil and incubated in a growth chamber. Plants were initially characterized for ethanol inducible GUS expression at 4-7 weeks of age. At approximately eight weeks of age, the plants were moved to 20 cm diameter pots and maintained in a greenhouse. At approximately seven months of age, plants were transferred into 30cm pots until maturity. 15 Media: 102891 The components within the media referred to above are as follows: 102901 EM3: MS salts and vitamins; 0.5 g/L casein hydrolysate; 100 mI/L coconut water; 20 g/L sucrose and 3 mg/l 2,4-D. [02911 LB basic: 10 g/L NaCl; 5 g/L yeast extract; and 10 g/L tryptone. 20 [02921 LB solid: LB basic with 15 g/L of agar. [0293] AB: The following salts were autoclaved and added: 2g/L (NH 4 )2SO 4 ; 6 g/L Na2HPO4; 3 g/L KH2PO4; and 3 g/L NaCl. The following compounds were filter sterilized: 0.1 mM CaCl2; 1.0 mM MgCl 2 0.003 mM FeCI 3 ; and 5 g/L glucose. [02941 MS basic: MS medium salts and vitamins, with 25 g/L sucrose. 25 [02951 MS 1: MS basic supplemented with 3.0 mg/L 2,4- D and 200 mg/L Timentin. [02961 MS2: MS basic supplemented with 3.0 mg/L 2,4- D and 50 mg/L Geneticin and 200mg/L Timentin. - 103 - 102971 MS3: MS basic supplemented with 40 ml of coconut water filter sterilized and 1.0- 2.0 mg/L BAP (cultivar dependent, thus not required for all cultivars) and 50mg/L Geneticin and 200mg/L Timentin. 102981 MS4: MS basic supplemented with 1.0 g/L charcoal and 1.0 mg IBA 5 (indole-3- butyric acid, not required for all cultivars and 50 mg/L Geneticin. 102991 CoCult: Media co-cultivation media as described for banana in Khanna et al. (2004) Molecular Breeding 14(3): 239-252. EXAMPLE 5 CHARACTERIZATION OF TRANSGENIC PLANTS CONTAINING OPTIMIZED CONSTRUCTS 10 103001 Plants were screened for the presence of the nptlI, scoAlcR, and scoGUS genes using TaqMan@ analysis. Plants that contained at least one copy of each gene of interest were subsequently characterized for ethanol inducible expression using GUS histochemical staining. [0301] After the transgenic sugar cane plants had been in soil for 4-7 weeks, a leaf 15 sample of approximately 3cm in length was taken from each plant just prior to ethanol treatment and analyzed for GUS expression by histochemical staining as described above. After sampling, the plants were treated with 2% ethanol using a daily root drench and aerial spray for four days. At five days post treatment, a leaf sample of approximately 3cm in length was taken from each plant and analyzed for GUS expression by histochemical staining. 20 [03021 Prior to ethanol treatment, only a small number of the ethanol switch plants showed detectable GUS expression while three out of seven ZmUbi-scoGUS plants were found to be GUS positive (Table 1). [03031 Following the 2% ethanol treatment, GUS expression was detected for 11 of the 12 optimized ethanol switch constructs with between 6% and 83% of the plants containing 25 these constructs showing detectable GUS expression (Table 1). Visual inspection of the intensity of staining suggested that constructs SC 15, SC 18, SC 19, SC20, and SC22 gave the highest ethanol inducible expression. -104- Table 1: Results from the histochemical GUS staining of transgenic sugar cane plants containing the different ethanol switch constructs in response to the continuous 2% ethanol treatment. GUS Expression Total Plants Construct (TaqMan Positive) Uninduced Induced % Induced SC12 12 0 2 17% SC13 30 0 6 20% SC14 17 0 1 6% SC 15 16 0 8 50% SC16 12 1 10 83% SC17 20 0 4 20% SC 18 10 0 8 80% SC 19 27 0 12 44% SC20 32 0 12 38% SC21 6 1 0 0% SC22 30 2 3 10% SC23 18 0 10 56% pUbi-scoGUS 7 3 3 NA 5 [03041 An independent group of transgenic sugar cane plants were subsequently analyzed for ethanol inducible expression using a less robust ethanol treatment. After these transgenic plants had been in soil for six weeks, a leaf sample of approximately 3cm in length was taken from each plant just prior to ethanol treatment and analyzed for GUS expression by 10 histochemical staining. After sampling, the plants were treated with 1% ethanol using a single root drench (800ml/seedling tray) and aerial spray until runoff. At five days post treatment, a leaf sample of approximately 3cm in length was taken from each plant and analyzed for GUS expression by histochemical staining. - 105 - 103051 Prior to ethanol treatment, leaky GUS expression was visible in a subset of the plants for some of the various constructs (Table 2). Following the 1% ethanol treatment, GUS expression was detected for six of the 12 optimized ethanol switch constructs with between 4% and 65% of the plants containing these constructs showing detectable GUS 5 expression (Table 2). Plants containing constructs SC 18, SC 19 and SC20 had the highest proportion of inducible plants, and visual inspection of the intensity of staining suggested that these plants also had the highest ethanol inducible expression. Table 2: Resultsfor the histochemical GUS staining of transgenic sugar cane plants 10 containing the different ethanol switch constructs in response to the 1% ethanol treatment. GUS Expression Total Plants Construct (TaqMan Positive) Uninduced Induced % Induced SC 12 13 4 0 0% SC 13 14 0 0 0% SC 14 14 2 0 0% SC 15 51 12 2 4% SC 16 7 1 2 29% SC17 20 0 0 0% SC 18 34 16 22 65% SC19 30 2 19 63% SC20 32 2 13 41% SC21 38 13 0 0% SC22 16 0 0 0% SC23 15 2 1 7% [03061 For the quantitative analysis of ethanol inducible expression in more mature plants, only the single copy plants for each construct were selected and transferred to the greenhouse. When the transgenic sugar cane plants had been in soil for approximately six 15 months they were assessed to verify that there was no residual GUS expression from the -106previous ethanol treatment that was carried out at 4-7 weeks of age. To do this, a tissue sample was taken from the first fully unfurled leaf of each plant and analyzed for GUS expression by histochemical staining. No residual GUS expression from the original ethanol treatment was detected in these transgenic plants. 5 103071 Subsequently, these plants were reanalyzed for ethanol inducible expression. A tissue sample roughly equivalent to one standard hole punch was taken from the top, middle, and bottom of the first fully unfurled leaf of each plant. These three leaf samples were combined and placed into one well of a 96 well sample block on ice. Each plant was sampled in duplicate from the same first unfurled leaf. Samples were then frozen at -80* C and freeze 10 dried. After sampling, the plants were treated with 5% ethanol using a single root drench (700 mL / pot) and aerial spray until runoff. At two, four, and seven days post treatment, a tissue sample roughly equivalent to one standard hole punch was taken from the top, middle, and bottom of the same first fully unfurled leaf of each plant. These three leaf samples were combined and placed into one well of a 96 well sample block on ice. Each plant was sampled 15 in duplicate from the same first unfurled leaf at each timepoint. Samples were frozen at 80*C, freeze-dried, and GUS expression was subsequently quantitated by ELISA. For the GUS ELISA, high-binding 96-well plates (Nunc Maxisorp@) were coated at 4' C overnight with 2 pg/mL rabbit anti-GUS IgG (Sigma G5545) in 25 mM borate, 75 mM NaCl, pH 8.5 (100 pL/well). Plates were washed three times with 10 mM Tris, pH 8.0 containing 0.05% 20 Tween-20 and 0.2% NaN3. Samples or standards (GUS Type VII-A, Sigma G7646) were added to the plate (100 FL/well), incubated for I hr at room temperature with shaking, and washed five times. 100 ptL/well of 2 pg/mL HRP-labeled rabbit anti-GUS IgG (Invitrogen A5790 conjugated to HRP) was then added to the plate, incubated for lhr at room temperature with shaking, and washed as before. The HRP-conjugated antibody was detected by adding 25 100 pL/well tetramethylbenzidine (TMB, Sigma T0440) and developing for 30min at room temperature. The reaction was stopped by the addition of 100 pL/well of 0.IN HCL. The absorbance was measured at 450 nm with 620 as a reference using a microplate reader (Tecan Sunrise TM, Research Triangle Park, NC). The GUS standard curve uses a 4-parameter curve fit. The curve is plotted linear vs. log with a range from 0 to 320 ng/mL. Results from the 30 GUS ELISA indicated that there was no detectable GUS expression in any of the transgenic sugar cane plants prior to ethanol treatment. Following ethanol treatment, the highest, most consistent ethanol inducible expression was detected from those plants containing the constructs with multiple copies of the inverted repeat AlcR binding site (SC18, SC19, and - 107- SC20; Fig. 1). Little or no ethanol inducible expression was detected in plants containing the other nine constructs. These results indicate that multiple copies of the inverted repeat AlcR binding site can substantially improve ethanol inducible gene expression. EXAMPLE 6 5 PRODUCTION AND TESTING OF ADDITIONAL INDUCIBLE PROMOTER CONSTRUCTS WITH VARYING NUMBERS OF THE INVERTED REPEAT ALcR BINDING SITE. [03081 Nucleic acid constructs having one copy or nine copies of the inverted repeat AlcR binding site were generated as follows: 10309] (1) The SC35 construct consists of one copy of the inverted repeat AlcR 10 binding site region sequence "5'-atgcatgcggaaccgcacgagg-3'" [SEQ ID NO:3] (without any additional AlcA promoter sequence) fused to the longer CaMV 35S minimal sequence (-73 to +7 relative to the CaMV 35S transcriptional start site). 103101 (2) The SC36 construct consists of nine tandem repeats of the inverted repeat AlcR binding site region sequence "5'-atgcatgcggaaccgcacgagg-3"' [SEQ ID NO:3] 15 (without any additional AlcA promoter sequence) fused to the longer CaMV 35S minimal sequence. [03111 The SC35 and SC36 promoter sequences were made synthetically and included restriction enzyme sites at each end for cloning. These sequences were cloned into scoGUS/pBS using HindlIl and Pstl. To generate the final ethanol switch constructs the 20 promoter variant, scoGUS, and nos sequences were subcloned into scoAlcRnptII using HindIl and AscI. The binary SC35 and SC36 constructs were transferred into Agrobacterium strain AGLI using a standard heat shock transformation method. Agrobacterium containing each of the binary constructs were used to transform sugar cane as described above. 103121 Plants were screened for the presence of the npt/I, scoAlcR, and scoGUS 25 genes using TaqMan@ analysis. Plants that contained at least one copy of each gene of interest were subsequently characterized for ethanol inducible expression using GUS ELISA as described above. After the transgenic sugar cane plants were in soil for approximately six weeks, a leaf sample of approximately 3cm in length was taken from the first fully unfurled leaf of each plant just prior to ethanol treatment and placed into one well of a 96 well sample 30 block on ice. Each plant was sampled in duplicate from the same first unfurled leaf. Samples were frozen at -80' C and freeze dried. After sampling, the plants were treated with 2% - 108ethanol using a single root drench (800 mL/seedling tray) and aerial spray until runoff. At four days post treatment, a leaf sample of approximately 3cm in length was taken from the same leaf previously sampled, and placed into one well of a 96 well sample block on ice. Each plant was sampled in duplicate from the same first unfurled leaf. Samples were frozen at 5 -80* C, freeze-dried, and GUS expression was subsequently quantitated by ELISA as described above. Prior to ethanol treatment, little or no GUS expression was detected in the leaves of the transgenic sugar cane plants (Table 3, Figure 2). Following ethanol treatment, robust ethanol inducible expression was detected from those plants containing the constructs with either five or nine copies of the inverted repeat AlcR binding site (SC20 and SC36; 10 Table 3 and Fig. 2). Only low levels of ethanol inducible expression were detected from plants containing the construct with one copy of the inverted repeat AlcR binding site (SC35; Table 3 and Fig. 2). Furthermore, a substantially higher percentage of the transgenic plants containing the constructs with multiple inverted repeat AlcR binding sites (SC20 and SC36) showed ethanol inducible expression compared to those plants containing a single copy of the 15 inverted repeat AlcR binding site (SC35; Table 3). Table 3: Ethanol inducible expressionfrom promoters containing either 1, 5, or 9 copies of the inverted repeat AIcR binding site. Construct Uninduced Induced Plants showing detectable ID Expression (ng Expression (ng expression (% of TaqMan@ GUS/mg protein) GUS/mg protein) positive plants showing expression) SC35 3.3±1.7 6.6±2.3 3 (23%) SC20 0 36.1±11.8 5(100%) SC36 0 73.2±23.0 17 (71%) 20 EXAMPLE 7 PRODUCTION AND TESTING OF ADDITIONAL INDUCIBLE PROMOTER CONSTRUCTS IN SUGAR CANE [0313] To identify the minimal sequence within the inverted repeat AlcR binding site that is necessary for ethanol inducible expression, the following modified inverted repeat 25 AlcR binding sites were generated: - 109- [03141 (1) The SC38 construct modifies the inverted repeat AlcR binding site region sequence "5'-atgcatgcggaaccgcacgagg-3"' [SEQ ID NO:3] to "5'-tacgtagcggaaccgctgctcc-3"' [SEQ ID NO:6]. 103151 (2) The SC39 construct modifies the inverted repeat AlcR binding site 5 region sequence "5'-atgcatgcggaaccgcacgagg-3"' [SEQ ID NO:3] to "5'-tacgttgcggaaccgcagctcc-3"' [SEQ ID NO:9]. 103161 (3) The SC41 construct modifies the inverted repeat AlcR binding site region sequence "5'-atgcatgcggaaccgcacgagg-3"' [SEQ ID NO:3] to "5'-atgcatgeggtgccgcacgagg-3"' [SEQ ID NO:8]. 10 103171 (4) The SC42 construct modifies the inverted repeat AlcR binding site region sequence "5'-atgcatgcggaaccgcacgagg-3"' [SEQ ID NO:3] to "5'-atgcatgcggaatgcaaccgcacgagg-3'" [SEQ ID NO:10]. [03181 The above sequences were synthesized as pentamers fused with the long 35S minimal sequence. These modified promoter variants were cloned as described above 15 (Example 5). The binary SC38, SC39, SC41 and SC42 constructs were transferred into Agrobacterium strain AGLI using a standard heat shock transformation method. Agrobacterium containing each of the binary constructs were used to transform sugar cane as described above. Ethanol inducibility from these promoters was compared to that of construct SC20 using GUS ELISA as described above. 20 103191 Prior to ethanol treatment, little or no GUS expression was detected in the leaves of the transgenic sugar cane plants (Table 4, Fig. 3). Following ethanol treatment, robust ethanol inducible expression was detected from those plants containing the constructs with the various point mutations to the inverted repeat AlcR binding site (SC38, SC39, and SC41; Table 4 and Fig. 3). In addition, 100% of the transgenic plants containing these 25 constructs were found to exhibit ethanol inducible expression (Table 4). No ethanol inducible expression was detected from transgenic plants containing the construct that alters the length of the 2 bp spacer between the inverted repeat AlcR binding sites (SC42; Table 4 and Fig. 3). From this data, the sequence of "GCGGnnCCGC" [SEQ ID NO:I] (with n representing any nucleotide) can be defined as a minimal sequence necessary for ethanol inducible expression. -110- Table 4: Ethanol inducible expressionfrom promoters containing various modified inverted repeat AlcR binding sites. Construct Uninduced Induced Plants showing detectable ID Expression (ng/mg Expression expression (% of TaqMan@ protein) (ng/mg protein) positive plants showing expression) SC20 0 36.1±11.8 5/5(100%) SC38 4.4±3.5 125.8+64.3 7/7 (100%) SC39 0 105.1±126.8 2/2(100%) SC41 2.2±2.2 137.3±105.4 3/3 (100%) SC42 0 0 0/3 (0%) 5 EXAMPLE 8 PREPARATION AND TESTING OF A MAIZE PROMOTER IN COMBINATION WITH THE ALcR "B" INVERTED REPEAT BINDING SITES. [03201 The minimal maize AdhI promoter, as described by Walker et al. (1987, Proc. Natl. Acad. Sci. USA 84: 6624-6628), can be tested for its ability to be made inducible 10 using the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, and/or SEQ ID NO:9. The AdhI nucleotide sequence (SEQ ID NO:48), below, was identified by successive 5' deletions of the Adhl promoter, and shown to give only background levels of expression: [03211 5'-ccacaggcggccaaaccgcaccctccttcccgtcgtttcccatctcttcctcctttagagctaccactatat 15 aaatcagggctcattttctcgctcctcacaggctcatctcgctttggatcgattggtttcgtaactggtgaaggactgagggtctcggagtg gatgatttgggattctgttcgaagatttgcggaggggggca-3' +106 [SEQ ID NO:48]. [03221 The maize AdhI minimal promoter sequence was fused with five tandem repeats of the inverted repeat AlcR binding site region sequence "5'-atgcatgcggaaccgcacgagg-3'" [SEQ ID NO:3] (replacing the long 35S minimal promoter 20 sequence as described in Example 3, construct SC20). Ethanol inducibility of this promoter (SC37) was compared to that of construct SC20 using GUS ELISA as described above. -111- [03231 Prior to ethanol treatment, little or no GUS expression was detected in the leaves of the transgenic sugar cane plants (Table 5 and Fig. 4). Following ethanol treatment, robust ethanol inducible expression was detected from those plants containing the construct with the five copies of the inverted repeat AlcR binding sites fused to the maize AdhI 5 minimal promoter (Table 5 and Fig. 4). Table 5: Ethanol inducible expression from promoters containing five copies of the inverted repeat AlcR binding sites fused to different minimal promoters. Construct Uninduced Induced Plants showing detectable ID Expression (ng/mg Expression expression (% of TaqMan@ protein) (ng/mg protein) positive plants showing expression) SC20 0 36.1±11.8 5/5(100%) SC37 2.5±4.3 107.9±19.4 11 (69%) 10 EXAMPLE 9 CHARACTERIZATION OF ETHANOL INDUCIBLE EXPRESSION IN THE VEGETATIVE PLANTINGS OF THE PRIMARY TRANSGENIC SUGAR CANE PLANTS 103241 Single-bud setts from stems of selected mature, TO transgenic plants were planted in soil and maintained in a glasshouse. After the TOV I transgenic sugar cane plants 15 were in soil for approximately four months (at which time they were comparable in size to the six month old TO plants describe above), a leaf sample of approximately 3 cm in length was taken from the first fully unfurled leaf of each plant just prior to ethanol treatment and placed into one well of a 96 well sample block on ice. Each plant was sampled in duplicate from the same first unfurled leaf. Samples were frozen at -80' C and freeze dried. After sampling, the 20 TOV 1 plants were treated with 5% ethanol using a single root drench (700 mL/pot) and aerial spray until runoff. At four days post treatment, a leaf sample of approximately 3cm in length was taken from the same leaf previously sampled, and placed into one well of a 96 well sample block on ice. Each plant was sampled in duplicate from the same first unfurled leaf. Samples were frozen at -80' C, freeze-dried, and GUS expression was subsequently 25 quantitated by ELISA as described above. - 112 - [0325] Prior to ethanol treatment, little or no GUS expression was detected in the leaves of the TOV 1 transgenic sugar cane plants (data not shown). Following ethanol treatment, robust ethanol inducible expression was detected for all of the TOV I transgenic plants (Fig. 5), indicating that ethanol inducibility from these constructs is maintained in the 5 vegetatively-propagated material. EXAMPLE 10 PRODUCTION OF TRANSGENE CONSTRUCTS FOR NICOTIANA BENTHAMIANA [0326] To clone the ABI1 and abil genes, PCR was carried out on cDNA made from RNA that was isolated from leaves of wild-type and abil mutant Arabidopsis plants, 10 respectively. The PCR primers were designed to the ABIl GenBank sequence with the accession identifier, AY142623.1. The forward primer, 5' CCCCGGATCCCAACAATGGAGGAAGTATCTCCGGC-3' [SEQ ID NO:49] incorporates a BamHl restriction enzyme site and the Kozak sequence CAACA. The reverse primer, 5'-CCCCGTCGACTCAGTTCAAGGGTTTGCTCT-3' [SEQ ID NO:50] incorporates a Sall 15 restriction enzyme site. The engineered BamHI and SalI restriction enzyme sites were used for sub-cloning the ABI] and abil genes into the plant expression constructs. Constitutive promoter constructs [03271 The 35S-abil plant expression construct consists of the constitutive CaMV 35S promoter driving expression of the Arabidopsis abil gene. The abil gene was sub-cloned 20 into the 35S-GS construct using the restriction enzyme sites BamHI and Sall, replacing the GUS syntron (GS) coding sequence with abil, and generating the plasmid 35S-abil. The 35S abil-35S cassette was excised from the 35S-abil plasmid using EcoRI, and cloned into the EcoRI site of the binary vector pBINPLUS to generate the 35S-abil binary (Fig. 6) construct used for transformation of N. benthamiana. 25 [03281 The 35S-ABIJ plant expression construct consists of the constitutive CaMV 35S promoter driving expression of the Arabidopsis wild type ABIl gene. The ABI1 gene was sub-cloned into the 35S-GS construct using the restriction enzyme sites BamHI and Sall, replacing the GUS syntron (GS) coding sequence with A BI1, and generating the plasmid 35S A BI. The 35S-ABII-35S cassette was excised from the 35S-ABIJ plasmid using AseI and 30 HindIl, and cloned into the AseI and HindIll sites of the binary vector pBINPLUS to generate the 35S-ABI1 binary construct used for transformation of N. benthamiana. -113alc gene switch promoter constructs [03291 To improve plant expression of the AlcR transcription factor that is required for functionality of the alc gene switch, an optimized alcR gene (scoalcR) was synthesised by Geneart (Regensburg, Germany). To generate the alc gene switch constructs for 5 transformation into N. benthamiana, the scoalcR gene was excised from construct 0919814 scoALCR-pMK-RQ using the restriction enzymes NotI and AscI, and the DNA ends were blunted using Klenow DNA polymerase. The scoalcR was subsequently sub-cloned into the blunted BamHI site of the 35S-GS plasmid to generate the 35S-scoalcR plasmid. The 35S scoalcR-35S cassette was excised from the 35S-scoalcR plasmid using EcoRI, and cloned into 10 the EcoRI site of the binary vector pBINPLUS to generate the 35S-scoalcR binary construct. The 35S-scoalcR binary construct was used to create the alc gene switch constructs described below. [03301 The palcA 0-abil plant expression construct consists of the original alc gene switch promoter (palcA 0) driving expression of the Arabidopsis abil gene. This palcA 15 0 promoter sequence is identical to the original alcA promoter sequence used in plants by Caddick et al. (Nature Biotechnology 1998 16:177-180), and consists of the Aspergillus alcA promoter (from -349 to -112 relative to the translational start site and lacking the upstream direct repeat AlcR binding site) fused to the CaMV 35S minimal promoter (-32 to +3 relative to the CaMV 35S transcriptional start site) at the TATA box (the TATA box is identical in the 20 alcA and CaMV 35S promoters). [03311 The palcA 0 sequence was cloned from the construct A IB] -scoG US scoALCR-nptl using PCR with the forward primer 5' CCCCCCATGGCTGCAGGCATGCAAGCTTAG-3' [SEQ ID NO: 51] and the reverse primer 5'-CCCCGGATCCAATACCTGCAGGTCCTCTC-3' [SEQ ID NO: 52] that 25 incorporate an Ncol and BamHI restriction enzyme site, respectively. The palcA 0 PCR product was amplified using KAPAHiFi DNA polymerase (Geneworks) and A-tailed using Taq polymerase. The resulting PCR product was cloned into pGEM-T (Promega) to generate palcA O-pGEM-T, and the palcA 0 promoter was sequence verified. The palcA 0 promoter was excised from palcA 0-pGEM-T using NcoI and BamHl, and cloned into the NcoI and 30 BamHI sites of the 35S-abil plasmid to generate the plasmid palcA 0-abil. The palcA 0 abil-35S cassette was excised from the palcA 0-abil plasmid using HindIII, and cloned into -114the HindIII site of the 35S-scoalcR binary construct to generate the palcA 0-abil binary construct used for transformation of N. benthamiana. [03321 The palcA I-abil plant expression construct consists of an improved alc gene switch promoter (palcA I) driving expression of the Arabidopsis abil gene. This palcA I 5 promoter sequence consists of five tandem repeats of the inverted repeat AlcR binding site region sequence "5'-atgcatgcggaaccgcacgagg-3"' [SEQ ID NO:53] at the 5' end of the Aspergillus alcA promoter (which includes the upstream direct repeat AlcR binding site) fused to the CaMV 35S minimal promoter (-32 to +3 relative to the CaMV 35S transcriptional start site) at the TATA box (the TATA box is identical in the a/cA and CaMV 35S 10 promoters). 103331 The palcA I sequence was cloned from the construct pAlcA A1B4-scoGUS using PCR with the forward primer 5'-CCCCCCATGGGTATCGATAAGCTTAGCTAGC-3' [SEQ ID NO:54] and the reverse primer 5'-CCCCGGATCCTGCAGGTCCTCTCCAAATG-3' [SEQ ID NO: 55] that incorporate an 15 Ncol and BamHI restriction enzyme site, respectively. The pacA I PCR product was amplified using KAPAHiFi DNA polymerase (Geneworks) and A-tailed using Taq polymerase. The resulting PCR product was cloned into pGEM-T (Promega) to generate palcA I-pGEM-T, and the palcA I promoter was sequence verified. The pacA I promoter was excised from palcA I-pGEM-T using NcoI and BamHI, and cloned into the Ncol and BamHI 20 sites of the 35S-abil plasmid to generate the plasmid palcA I-abil. The palcA I-abil-35S cassette was excised from the palcA 1-abil plasmid using HindIll, and cloned into the HindIll site of the 35S-scoalcR binary construct to generate the palcA I-abil binary construct (Fig. 7) used for transformation of N. benthamiana. 103341 The palcA I-A BIJ plant expression construct consists of the palcA I 25 promoter driving expression of the Arabidopsis wild type ABI1 gene. The ABI] gene was excised from the 35S-ABIJ plasmid using BamHI and Sall, and sub-cloned into the BamHI and SalI restriction enzyme sites of the palcA I-abil plasmid (thereby replacing abil with ABI]) to generate the palcA I-A BI] plasmid. The palcA I-ABI1-35S cassette was excised from the palcA I-A BII plasmid using AseI and Hindill, and cloned into the Ase and HindIII sites of 30 the 35S-scoalcR binary construct to generate the palcA 1-ABII binary construct used for transformation of N. benthamiana. - 115- EXAMPLE 11 PRODUCTION OF TRANSGENE CONSTRUCTS FOR SUGAR CANE 103351 A sugar cane-optimized Arabidopsis abil (scoabil) sequence (including the nopaline synthase terminator, tNos) was synthesised by Geneart (Regensburg, Germany), 5 introducing a NotI restriction enzyme site at the 5' end and an AscI restriction enzyme site at the 3' end. To generate a sugar cane-optimized sequence of the ABIJ wild-type gene (scoABI), scoabil was used in a PCR site-directed mutagenesis to change nucleotide 554 from an "A" to "G" using the forward primer 5'-ATGGCCACGGTGGTTCC-3' [SEQ ID NO:56) and the reverse primer 5'-GGAACCACCGTGGCCAT-3' [SEQ ID NO:57]. 10 Constitutive promoter constructs 103361 The eFMVe35S-ZmUbil-scoabil plant expression construct consists of the figwort mosaic virus/CaMV 35S dual enhancer (eFMVe35S), and the maize polyubiquitin-1 promoter (ZmUbil) with TMV omega translational enhancer sequence (gtatttttacaacaattaccaacaacaacaaacaacaaacaacattacaattactatttacaattaca) driving expression of 15 scoabil. The scoabil gene with tNos was sub-cloned into the plasmid eFMVe35S-ZmUbil scoGUS using the restriction enzyme sites NotI and AscI, replacing the scoGUS gene and tNos with scoabil and tNos, and generating the plasmid eFMVe35S-ZmUbil-scoabil (Fig. 8) used for transformation of sugar cane. [03371 The eFMVe35S-ZmUbil-scoABIJ plant expression construct consists of 20 the eFMVe35S dual enhancer and ZmUbil promoter with TMV omega translational enhancer sequence driving expression of scoABi. The scoABIl gene and tNos was sub-cloned into the plasmid eFMVe35S-ZmUbiI-scoGUS using the restriction enzyme sites NotI and Ascl, replacing the scoGUS gene and tNos with scoABIl and tNos, and generating the plasmid eFMVe35S-ZmUbil-scoAB11 used for transformation of sugar cane. 25 alc gene switch promoter constructs [03381 The palcA I-scoabil plant expression construct consists of the palcA I promoter with TMV omega translational enhancer driving expression of scoabil, and the ZmUbil promoter with TMV omega translational enhancer driving expression of scoalcR. The scoabil gene and tNos were excised from the 1107897_abil-tNos_pMK-RQ plasmid 30 (Geneart) using NotI and Ascl, and sub-cloned into the NotI and AscI restriction enzyme sites of the pAlcA A 2B4-scoGUS plasmid (thereby replacing scoGUS and tNos with scoabil and tNos) to generate the palcA I-scoabil intermediate plasmid. The ZmUbil-scoAlcR-tNos -116cassette in the A 1B1-scoGUS-scoAlcR-nptI binary vector was excised using the restriction enzyme KpnI, and cloned into the KpnI restriction enzyme site in the intermediate plasmid palcA I-scoabil to generate the palcA I-scoabil construct (Fig. 9) used for transformation of sugar cane. 5 [0339] The palcA I-scoA BI) plant expression construct consists of the palcA 1 promoter with TMV omega translational enhancer driving expression of scoABI], and the ZmUbil promoter with TMV omega translational enhancer driving expression of scoalcR. The scoA BI] gene and tNos were excised from the scoABIl-pGEM-T plasmid using NotI and Ascl, and sub-cloned into the NotI and Ascl restriction enzyme sites of the pAlcA A2B4 10 scoGUS plasmid (thereby replacing scoGUS and tNos with scoA BI] and tNos) to generate the palcA 1-scoABIl intermediate plasmid. The ZmUbil-scoAlcR-tNos cassette in the AIB] scoGUS-scoAlcR-npt/I binary vector was excised using the restriction enzyme KpnI, and cloned into the Kpnl restriction enzyme site in the intermediate plasmid palcA I-scoA BI] to generate the palcA I-scoA BI] construct used for transformation of sugar cane. 15 10340] The palcA I-scoabil-tNos cassette will be subcloned into the scoalcRnptII binary vector using HindIII and AscI to generate the palcA I-scoabil binary construct used for Agrobacterium-mediated transformation of sugar cane. EXAMPLE 12 PRODUCTION OF TRANSGENIC N. BENTHAMIANA 20 [0341] The binary constructs were transferred into Agrobacterium strain LBA4404 using electroporation. Agrobacterium containing each of the binary constructs were used to transform N. benthamiana using Agrobacterium-mediated transformation as described by Horsch et al. (1985, Science 227:1229-1231). Leaf explants were harvested from N. benthamiana and infected via immersion and incubation with transformed Agrobacterium. 25 After infection of the leaf explants with Agrobacterium, the explants were blotted dry and maintained on selection-free, shoot-induction media consisting of MS salts plus vitamins (Phytotechnology Laboratories), sucrose (30 g/L), 6-benzylaminopurine (BAP) (1 mg/mL), naphthalene acetic acid (NAA) (0.1 mg/mL) and 0.8 % agar for two to three days. The explants were then transferred to media containing Kanamycin (200 pg/mL) and Timentin 30 (200 pg/mL) and subcultured twice weekly. After four to six weeks, the concentration of BAP and NAA in the media was reduced to 0.25 mg/mL and 0.025 pg/mL, respectively. When well defined stems were visible, the emerging shoots were excised and transferred to MS - 117media consisting of MS salts plus vitamins, sucrose (30 g/ L) and 0.8 % agar. Soon after plants had visible roots, they were transferred to soil and acclimatized at 250 C with a 16 hr. photoperiod. EXAMPLE 13 5 PRODUCTION OF TRANSGENIC SUGAR CANE 103421 Transgenic sugar cane plants were regenerated from sugar cane callus that was transformed by microprojectile bombardment (MPB) as described by Finer et al. (1992, Plant Cell Reports 11:323-328) and Bower et al. (1996, Molecular Breeding 2:239-249). To generate the callus, sugar cane (cultivar KQ228) "tops" were obtained from The Bureau of 10 Sugar cane experimental stations (BSES) LTD Meringa Queensland. Calli were initiated as described by Franks and Birch (1991, Australian Journal of Plant Physiology 18:471-480) using MSC 3 media consisting of 4.43 g/L MS basal salts with vitamins (Phytotechnology laboratories Shawnee Mission, KS, USA), 500 mg/L Casein Hydrolysate (Merck), 13.6 pM 2, 4-Dichlorophenoxyacetic acid (2, 4-D; Phytotechnology laboratories), 100 ml/L young 15 coconut juice ("Cock" brand, Thailand), 3 % (w/v) sucrose and 8 g/L agar (Research organics). Calli were maintained for seven weeks in the dark at 26 *C and subcultured every 14 days. [0343] The plasmid pUKN (possessing a selectable marker for geneticin resistance) was co-bombarded with each of the constructs when transforming callus to allow for selection 20 of transformed cells. For MPB, a 2 pL aliquot of a 1:1 mixture of pUKN (1 pg/pL) and the experimental construct DNA (1 pg/pL) was added to approximately 3 mg of 1 pm gold particles (Bio-Rad). The solution was mixed briefly and 25 pL of 2.5 M CaCl 2 and 5 pL of 0.1 M spermidine were added simultaneously. The mixture was iced and mixed for 15 seconds every minute for a total of five minutes. The mixture was then allowed to settle on ice 25 for 10 minutes, after which 22 pL of supernatant was removed. The remaining DNA-coated gold solution was mixed and 5 pL was used per bombardment. [03441 A particle inflow gun (PIG) was used to deliver the DNA to the target tissue. A screen utilizing stainless steel mesh with an aperture of 500 pm was positioned approximately 1.5 cm above the target tissue within the PIG chamber. The distance of the 30 DNA-coated particles to the leaf explant was 10.5 cm. The PIG chamber was vacuum evacuated to -90 kPa and a 10 ms pulse of helium at 1500 psi was used to accelerate the DNA-coated particles. The vacuum was released immediately following the MPB and each - 118sample plate was rotated 180 degrees and subjected to a second MPB. The callus remained on MSO media for four hours post MPB. After four hours the callus was transferred to MSC 3 medium for 4-6 days before being transferred to selection media consisting of MSC 3 and 50 mg/L G418 (Geneticin) (Roche). Non-transformed callus used for the regeneration of wild 5 type plants was transferred to MSC 3 without selection. [0345] Following MPB, the callus remained on selection media for four weeks in the dark with fortnightly subculturing after which it was transferred to regeneration medium with selection, consisting of MSC 3 with the 2,4-D replaced by 4.4 PM 6-Benzylaminopurine (BAP; Sigma). The callus was maintained at 270 C, under a 16 hour light, and 8 hour dark 10 cycle with fortnightly subculturing. Individual plants were separated and one plant from each clump of callus was retained. After 10 weeks of regeneration with BAP, the plants were transferred to rooting medium with selection (the same as regeneration medium, however BAP is replaced with 10.7 ptM a-Naphthalene Acetic Acid (NAA; Sigma)). The plants were grown until roots of approximately 1 cm in length had developed, after which the plants were 15 transferred to soil for acclimatization in a growth cabinet under the above mentioned lighting and temperature conditions. EXAMPLE 14 CHARACTERIZATION OF TRANSGENIC N. BENTHAMIANA 10346] Plants are verified to contain the transgene constructs using PCR to screen 20 for the presence of either the ABI] or abil transgene. Confirmed transgenic plants are subsequently screened using reverse transcriptase PCR (RT-PCR) to identify plants with detectable levels of ABI1/abil transgene expression. RT-PCR is carried out using cDNA generated from leaf RNA. For transgenic plants possessing the alc gene switch constructs, leaf samples are taken just prior to ethanol induction and at 4-48 hours post ethanol treatment. 25 Ethanol treatment is carried out using a single 2% ethanol root drench and aerial spray until runoff. RNA is extracted from N. benthamiana leaf samples using Tri Reagent (Sigma) according to the manufacturer's instructions. Transgenic plants showing either constitutive or ethanol inducible expression of abil (along with the relevant control plants) are characterized for any phenotypic effects that are known to be associated with reduced stomatal closure, in 30 particular a wilty phenotype. 103471 To further assess whether expression of abil is reducing stomatal closure in N. benthamiana, stomatal conductance is measured using the LI-6400XT portable -119photosynthesis system (LI-COR biosciences). Stomatal conductance is measured in plants grown under well-watered conditions, in drought-stressed plants, and in plants treated with the hormone abscisic acid. Transgenic plants are also characterized for relative water content and for the levels of sugar accumulation using High Performance Liquid Chromatography 5 (HPLC). Transgenic plants with constitutive expression of abil (and the relevant control plants) are assessed at various times over the course of development. Analysis of transgenic plants showing ethanol inducible abil expression are also carried out at various times over the course of development with data collected just prior to ethanol treatment and approximately 12-24 hours or 1-4 weeks following ethanol treatment. Ethanol treatment is carried out using 10 either a single treatment or multiple treatments of a 2% root drench and aerial spray. Characterization of the transgenic plants is carried out under well watered conditions and under varying levels of drought stress. [03481 Transgenic N. benthamiana plants displaying ethanol inducible expression of abil were identified (Fig. 10). The expression of abilin these plants was readily detected 15 following ethanol treatment, while little or no expression was detected prior to ethanol treatment (Fig. 10). Constitutive expression of abil is known to be detrimental to N. benihamiana growth and development, with plants displaying stunted growth and smaller leaves, as well as a strong propensity to wilt when removed from tissue culture and placed in soil (Armstrong et al., Proc. Natl. Acad Sci. USA. 92:9520-9524 (1995)). Plants containing 20 the ethanol inducible abil construct did not display these aberrant phenotypes and looked similar to the wild type (Fig. 11, 0 timepoint). 103491 Ethanol induced expression of abil resulted in visible wilting (Fig. 11) and significant water loss (Fig. 12) in the leaves following ethanol treatment. The ethanol treatment used had no detectable effect on control plants (Figs. 11 and 12). These data 25 demonstrate that the regulation of abil expression with the alc gene switch can be used to control water loss in plants. The data also show that the alc gene switch can be used to generate healthy transgenic plants possessing genes that are detrimental to plant growth and development when under the control of a constitutive expression system. EXAMPLE 15 30 CHARACTERIZATION OF TRANSGENIC SUGAR CANE [03501 Plants are verified to contain the transgene constructs using PCR to screen for the presence of either the scoABIl or scoabil transgene. Confirmed transgenic plants are - 120subsequently screened using reverse transcriptase PCR (RT-PCR) to identify plants with detectable levels of scoABIl/scoabil transgene expression. RT-PCR is carried out using cDNA generated from leaf RNA. Leaf samples are taken from the first fully unfurled leaf of each transgenic plant and frozen until use. For transgenic plants possessing the alc gene 5 switch constructs, leaf samples are taken just prior to ethanol induction and at 4-48 hours post ethanol treatment. Ethanol treatment is carried out using a single 4-5% ethanol root drench and aerial spray. RNA is extracted from sugar cane leaf samples using Tri Reagent (Sigma) according to the manufacturer's instructions. [03511 Transgenic plants showing constitutive and ethanol inducible expression of 10 scoabil are characterized for any phenotypic effects that are known to be associated with reduced stomatal closure, in particular a wilty phenotype. To further assess whether expression of scoabil is reducing stomatal closure in sugar cane, stomatal conductance is measured using the LI-6400XT portable photosynthesis system (LI-COR biosciences). Stomatal conductance is measured in plants grown under well-watered conditions, in drought 15 stressed plants, and in plants treated with the hormone abscisic acid. Transgenic plants (scoabil and the relevant control plants) are also characterized for the levels of sugar accumulation in their stem using HPLC, as well as leaf and stem water content. Transgenic sugar cane plants with constitutive expression of scoabil are assessed at various times over the course of development (along with the relevant control plants), except for sugar 20 accumulation which is characterized in approximately 6-9 month old plants. Analysis of stomatal closure in transgenic plants showing ethanol inducible scoabil expression is also carried out at various times over the course of development, with data collected just prior to ethanol treatment and for up to approximately four weeks following ethanol treatment. Sugar accumulation and water content in the stem of the ethanol inducible scoabil transgenic sugar 25 cane (along with the relevant control plants) is assessed in approximately 6-9 month old plants at approximately 12-24 hours or 1-5 weeks after the start of the ethanol treatment. Ethanol treatment is carried out using either a single treatment or multiple treatments of a 4 5% root drench and aerial spray. Characterization of the transgenic plants is carried out under well watered conditions and under varying levels of drought stress. 30 103521 Transgenic sugar cane plants displaying constitutive expression of either scoABIl or scoabil were identified (Fig. 13). Characterization of transgenic sugar cane having constitutive expression of scoabil revealed that abil is able to reduce stomatal closure in sugar cane (Figs. 14 and 15). Constitutive scoabil expression significantly (P<0.05) - 121 increased the stomatal conductance (Fig. 15) and could generate a wilty phenotype (Fig. 16) in transgenic sugar cane grown under well-watered conditions. These data demonstrate that expression of abil is able to control stomatal function in monocotyledonous plants like sugar cane in addition to dicotyledonous plants. 5 [0353] The disclosure of every patent, patent application, and publication cited herein is hereby incorporated herein by reference in its entirety. 103541 The citation of any reference herein should not be construed as an admission that such reference is available as "Prior Art" to the instant application. 10 [03551 Throughout the specification the aim has been to describe the preferred embodiments of the invention without limiting the invention to any one embodiment or specific collection of features. Those of skill in the art will therefore appreciate that, in light of the instant disclosure, various modifications and changes can be made in the particular embodiments exemplified without departing from the scope of the present invention. All such 15 modifications and changes are intended to be included within the scope of the appended claims. - 122-

权利要求:
Claims (26)
[1] 1. A construct for inhibiting stomatal closure, comprising in operable connection: (1) a cis-acting element comprising, consisting or consisting essentially of a nucleotide sequence represented by the sequence GCGGNNCCGC [SEQ ID NO:1]; (2) a promoter that is operable 5 in a plant cell (e.g., a plant guard cell); and (3) a nucleic acid sequence encoding an expression product that inhibits stomatal closure.
[2] 2. A construct according to claim 1, wherein the a cis-acting element comprises, consists or consists essentially of at least one nucleotide sequences (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20) as set forth in SEQ ID NO: 1. 10
[3] 3. A construct according to claim 2, wherein the at least one nucleotide sequence is represented by the nucleotide sequence nxGCGGNNCCGCny [SEQ ID NO:2], wherein N or n can be independently any nucleic acid base (A, G, C, or T) and wherein x and y can be independently any number.
[4] 4. A construct according to claim 2, wherein the at least one nucleotide sequence 15 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20) is selected from the group consisting of the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:2, ATGCATGCGGAACCGCACGAGG [SEQ ID NO:3], GGCCATGCGGAGCCGCACGCGT [SEQ ID NO:4], ACAAGAGCGGCTCCGCTTGACC [SEQ ID NO:5]; TACGTAGCGGAACCGCTGCTCC [SEQ ID NO:6]; TACCATGCGGAACCGCACGTCC 20 [SEQ ID NO:7], ATGCATGCGGTGCCGCACGAGG [SEQ ID NO:8] and TACGTTGCGGAACCGCAGCTCC [SEQ ID NO:9], in any combination, in any orientation, and/or in any order.
[5] 5. A construct according to any one of claims I to 4, wherein the expression product that inhibits stomatal closure is a stomatal closure-inhibiting polypeptide. 25
[6] 6. A construct according to claim 5, wherein the stomatal closure-inhibiting polypeptide is selected from: ATHB6; mutant forms of ABII or ABI2, which result in reduced ABA sensitivity and/or which inhibit stomatal closure; dominant negative forms of AAPK; dominant positive forms of PKS3 and AHA 1; or antibodies that are immuno interactive with a polypeptide that stimulates stomatal closure or that inhibits stomatal 30 opening.
[7] 7. A construct according to any one of claims I to 4, wherein the expression product that inhibits stomatal closure is a stomatal closure-inhibiting RNA molecule that inhibits - 123 - expression of an endogenous nucleotide sequence encoding a polypeptide that stimulates stomatal closure or that inhibits stomatal opening.
[8] 8. A construct according to claim 7, wherein the endogenous nucleotide sequence encodes a polypeptide selected from OSTI, AAPK, v-SNAREs AtVAMP711-14, GPAl, 5 AtABCG22, AtABCG40, AtMRP4, RBOHD and RBOHF, and PLDalphal.
[9] 9. A construct system for inhibiting stomatal closure, the construct system comprising, consisting or consisting essentially of a first construct as defined in any one of claims I to 8 and a second construct comprising a nucleotide sequence encoding a transcription factor, which activates in the presence of a compound that induces expression of 10 the alcohol dehydrogenase (ADH 1) system of Aspergillus nidulans and which interacts with the cis-acting element of the first construct to induce expression of the nucleic acid sequence encoding the expression product that inhibits stomatal closure.
[10] 10. A transgenic plant cell that comprise a construct as defined in any one of claims I to 8 or a construct system as defined in claim 9. 15
[11] 11. A transgenic plant cell according to claim 10, which is a plant guard cell.
[12] 12. A transgenic plant, plant part or plant organ comprising a plant cell according to claim 10 or claim 11.
[13] 13. A method for increasing transpiration in a plant, plant part, plant organ or plant leaf, the method comprising expressing in a cell of the plant, plant part, plant organ a 20 polynucleotide that comprises a nucleic acid sequence encoding an expression product that inhibits stomatal closure, wherein the nucleic acid sequence is under the control of a cis acting element as defined in any one of claims I to 8, to thereby increase transpiration in the plant, plant part or plant organ .
[14] 14. A method according to claim 13, comprising inducing expression of the 25 polynucleotide in the presence of a compound that induces expression of the alcohol dehydrogenase (ADHI) system of Aspergillus nidulans.
[15] 15. A method according to claim 13 or claim 14, comprising co-expressing in the cell a nucleotide sequence encoding a transcription factor, which activates in the presence of a compound that induces expression of the alcohol dehydrogenase (ADH1) system of 30 Aspergillus nidulans and which interacts with the cis-acting element to induce expression of the nucleic acid sequence encoding the expression product that inhibits stomatal closure.
[16] 16. A method for increasing transpiration in a plant, plant part, plant organ or plant leaf comprising a construct according to any one of claims I to 8, or a construct system - 124- according to claim 9, the method comprising exposing the plant, plant part or plant organ to a compound that induces the expression of the alcohol dehydrogenase (ADH I) system of Aspergillus nidulans so as to inhibit stomatal closure and thereby increase transpiration in the plant, plant part or plant organ. 5
[17] 17. A method according to claim 16, comprising exposing the plant, plant part or plant organ to the compound around the time of harvesting the plant, plant part or plant organ.
[18] 18. A method according to claim 16, comprising exposing the plant, plant part or plant organ to the compound prior to harvesting the plant, plant part or plant organ .
[19] 19. A method according to claim 16, comprising exposing the plant, plant part or 10 plant organ to the compound at the time of harvesting the plant, plant part or plant organ.
[20] 20. A method according to claim 16, comprising exposing the plant, plant part or plant organ to the compound after harvesting the plant, plant part or plant organ .
[21] 21. A method according to any one of claims 16 to 20, further comprising permitting increased transpiration in the plant, plant part or plant organ over a time and under conditions 15 sufficient for the water content of the plant, plant part or plant organ to reduce by at least about 5%.
[22] 22. A method according to any one of claims 13 to 21, wherein the plant is a monocotyledonous plant.
[23] 23. A method according to claim 22, wherein the monocotyledonous plant is selected 20 from sugar cane, corn, barley, rye, oats, wheat, rice, flax, millet, sorghum, grasses, banana, onion, asparagus, lily, coconut, and the like.
[24] 24. A method according to any one of claims 13 to 21, wherein the plant is a dicotyledonous plant.
[25] 25. A method according to claim 24, wherein the dicotyledonous plant is selected 25 from tobacco, cotton, plants or plant parts that are dried for consumption such as dried fruits (e.g., raisins and prunes), nuts, coffee, tea, cocoa, and ornamental goods.
[26] 26. A method according to any one of claims 13 to 21, wherein the plant an energy crop selected from the group consisting of: Miscanthus, Erianthus, Pennisetum, Arundo, Sorghum, Poplars, wheat, rice, oats, willows, switch grass, alfalfa, prairie bluestem, maize, 30 soybean, barley, sugar beet, hay and fodder crops. - 125-

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同族专利:

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公开号 | 公开日

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WO2014012145A1|2014-01-23|

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AU2013205472B2|2015-01-29|

引用文献:

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公开号 | 申请日 | 公开日 | 申请人 | 专利标题

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JPH08504327A|1992-12-10|1996-05-14|ギストブロカデスナムローゼフェンノートシャップ|Heterologous protein production in filamentous fungi|

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WO2011003800A1|2009-07-07|2011-01-13|Basf Plant Science Company Gmbh|Plants having modulated carbon partitioning and a method for making the same|US20210285007A1|2016-08-05|2021-09-16|Biogemma|Constructs and methods for controlling stomatal closure in plants|

法律状态:
2013-08-08| DA3| Amendments made section 104|Free format text: THE NATURE OF THE AMENDMENT IS: AMEND THE NAME OF THE INVENTOR TO READ: KINKEMA, MARK; O HARA, IAN MARK AND SAINZ, MANUEL BENITO. |

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2015-05-28| MK25| Application lapsed reg. 22.2i(2) - failure to pay acceptance fee|

优先权:

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申请号 | 申请日 | 专利标题

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AU2012903070A|AU2012903070A0||2012-07-18|Constructs for modulating transpiration in plants and uses therefor|

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AU2012903070||2012-07-18||

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AU2013205472A|AU2013205472B2|2012-07-18|2013-04-12|Constructs for Modulating Transpiration in Plants and Uses Therefor|AU2013205472A| AU2013205472B2|2012-07-18|2013-04-12|Constructs for Modulating Transpiration in Plants and Uses Therefor|

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PCT/AU2013/000799| WO2014012145A1|2012-07-18|2013-07-18|Constructs for modulating transpiration in plants and uses therefor|