• 专利附录
  • 专利说明
  • 权利要求
  • 类似技术
  • 同族专利
  • 引用文献
  • 法律状态
  • 优先权
    • 专利摘要:
    Gegenstand der Anmeldung ist ein Bacillus thuringiensis des Pathotyps C, insbesondere var. tenebrionis. Dieser Stamm zeigt eine insektizide Wirkung, und zwar auf Käferarten (Coleoptera). Ferner wird beansprucht ein aus diesem Stamm erhältliches Toxin gegen Coleoptera. Dieses Toxin erhält man, indem man bei Kulturen eines Bacillus thuringiensis vom Pathotyp C nach der Lyse der Sporangien die Kristalle von den Sporen und Zellresten trennt und die Kristallmasse reinigt. Ein weiterer Gegenstand der Anmeldung ist ein Insektizid, das als aktiven Wirkstoff Bacillus thuringiensis vom Pathotyp C oder ein hieraus erhältliches Toxin enthält.
    公开号:SU1448995A3
    申请号:SU3875505
    申请日:1984-12-19
    公开日:1988-12-30
    发明作者:Криг Алоисиус；Михаэль Хугер Алоис；Шнеттер Вольфганг
    申请人:Берингер Маннхайм Гмбх (Фирма)；
    IPC主号:
    专利说明:

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    The invention of OT IOCHTCH to agricultural and domestic microbiology, in particular to the manufacture of insecticidal preparations, concerns the bacterial strain, with which the insecticidal preparation is obtained.
    The purpose of the invention is to obtain a novel patotype C that allows an insecticidal preparation to be obtained against Coleoptera spp.
    The strain Bacillus thuringiensis varo tenebrionis pathotype C was isolated from the larvae of the mealworm (Tepe-; brio molitor L), deposited in a col-; lectures of Germany under the number DSM 2803,
    The strain has the following cultural and morphological features.
    Vegetative cell has swollen. The sticks are about 1.0x4 in size - 8 mm Grammplographic. The spore cell contains, along with subterminal ellipsoidal spores (0.8x1.4 - 1.6 mm), a pg-spacial crystal of a flat plate-like shape and with outlines of a parallelogram, a rhombus or a square (approx. 0.8-1.5 µm edge length),
    When you strain a strain for 24 hours with a sterile environment (OH01D SM-3) containing, g / l
    M Laboy Lemco 1.0 Extract Yeast Extract 2.0 Peptone5.0
    NaCli.
    Agar15,0
    pH .7,4
    round whitish colonies with a diameter of about 5 mm are formed, rough with uneven edges. Sporulation and formation of parasporic crystals occurs after approximately 48 hours.
    When cultivating the strain on calcium-caseinate agar (MERCK 5409 composition, g / l
    Merka 3.0 peptone extract. 5.00
    Casein by Hainmarsten 2.50 NaCl, 5eoo
    0.15 13.50
    WaterReal
    With the addition of skimmed milk powder of 5.0 g per liter of medium having a pH of 7.0, under these conditions, round whitish bacterial colonies are formed, around which there are transparent halos (proteinase-positive). Abrasion of spores and paraCa (OH); Agar
    O
    five
    0 5
    0
    five
    0
    five
    „
    0
    spore crystals occur after about 48 h.
    On selective agar for Bacillus cereus (OX01D 617), containing, g / l: Peptone1,00
    Mannitol10,00
    NaCl2,00MgS04-0, 10
     40.25
    Na-pyruvic acid 10,00
    Bromthymol
    blue 12
    AgarTs 500
    Water up to 1 l
    at pH 7, .2, with the addition of egg yolk, (sterile) 25.00 ml / l, after 24 hours of incubation, bluish-colored (mannitol-negative) colonies are formed, which is characteristic of the bacteria Bacillus seretus / B , thuringiensis. However, in contrast to V.-ce reus. And also to many other bacteria of Bacillus thurin giensis, around bacterial colonies B, thuringiensis var. tenebrions DSM 2803, there is no precipitated halo (-leitzinase-negative). The sporulation and formation of parasporic crystals take place in approximately 48 hours,
    Physiological and biochemical characteristics, Optional anaerobic, but spore, l chi is carried out strictly under aerobic conditions. The optimum growth temperature of vegetative cells is 25-30 ° C. Autoclaving for 15 minutes at (1 bar overpressure, respectively) completely inactivates the spores and the critical toxin. UV radiation at 254 nm inactivates spores at the appropriate dose, but: non-crystalline toxin,;
    Relation to carbon source: from glucose, mannose and sucrose forms acids and austil methylcarbanal, but not, gas 5 does not metabolize lactose, cellobiose, salicin, arabinose, xylose and manniTo. Hydrolyzes esculin and starch L,
    Relation to the source of nitrogen; as the sole source of nitrogen does not use ammonium sulphate. For growth requires amino acids such as glutamic acid, 0.3%). However, special growth factors are not required.
    Forms catalase, nitrate reductase and proteinase. 1e forms lysine decarbonxnlase, phenylalanine, urease, and lecithinase.
    Serological characteristics of the strain DSM 2803 (respectively
    The toxin is encoded by the gene contained in the plasmid. Toxin is also formed in asporogenic mutants obtained by treatment with methyl methanesulfonate. The ability to form a toxin can be transferred by transferring a plasmid or gene to other Bacillus thuringiensis species or
    serotyping for B, thuringiensip | o other bacterial systems. Insekpo de BARIACSBONNEPOI for H-antigens or SEKIJIMA 4ONO for O-antige. new):
    H-serotype 8a8b
    0-serotype IX.15
    The marker symptom of the strain is DSM 2803 (antibiogram), growth retardation with the following antibiotics: erythromy1din, nalidixic acid nitrofurantoin and tetracycline.
    Strain growth is not inhibited by ampicillin, colistin MS, Co-trimoxale, penicillin G, methicillin
    and sulfadiazine.
    An example. parath receive as follows.
    The strain of bacteria Bacillus thuringiensis var tenebrions DSM 2803 cultivated in Erleinmeyer flasks
    The ticidal action is based on the presence of a plasmid-encoded toxin.
    For use as an insecticide of Bacillus thuringiensis var. tenebrionsis or the insecticidal product derived from them. or the toxin is mixed. in a known manner with conventional additives (fillers, adhesives, wetting agents, etc.) and the translation of TVs is convenient. form for use. An insecticide of this composition can be used in the form of a powder, suspension, granulate, and the like.
    Insecticidal pre-25 It is preferable to apply purified
    sporulated culture in the form of a suspension with the addition of a wetting agent, for example, in the form of a suspension with the addition of a wetting agent, for example, 250 tons containing 50 oil-up to 0.1% Citowet (BASF). Per hectare, including,%; the yeast extract of the treated floor is poured 10, preferably 10 -10 spores and an equivalent amount of toxic crystals.
    The effect of strain on; maggots
    0.5, trypton 0.5, glyco 0.1, 0.08, having a pH of 7.0, at 25-30 ° C before the formation of spores. The biomass is then separated by centrifugation and washed with water or a bery solution. The titer of the spores in the preparation is 3 10 spores / ml. LCj5-5-10 spore / ml for L larvae Leptino tarsa deumeimata.
    The insecticidal active principle of the proposed strain is a crystalline toxin. The crystalline mass can be selected by separating and purifying the crystals from cell residues and spores after lysis of the sporangia, using, for example, the density gradient method.
    The molecular weight was determined by electrophoresis of SOS-polyacrylamide on a gel by the Lemmley method. Found two main bands approx. 6500pOD and 70000D. Smaller bands of lower intensity were in the range of 20000-400000.
    With the help of a toxin, antibodies can be obtained that, in the Ouchterlony immunodiffusion test (Ouchterlony), do not show a cross-reaction with other toxins of pathotype A or B.
    35
    of coleoptera is realized according to the scheme known according to patatnup A: in the confluence of several hours the feeding stops and after a few days
    4Q death occurs depending on the dose. The toxic effect is based on the fact that the midgut acitelium is damaged by crystalline toxin dissolved in the intestinal juice. Each damaged insect intestinal cells are separated from the basement membrane from one another and drift into the intestinal lumen. Due to the breakdown of the intestinal barrier, bacterial cells can penetrate into the hemocoel and complete the disintegration process with septicemia.
    A new insecticide (pathotype C) shows a selective insecticidal action against Coleoptera, for example, against an alder leaf beetle or potato beetle. The effect on the larvae is much stronger than on adult insects. Action not set50
    55
    The toxin is encoded by the gene contained in the plasmid. Toxin is also formed in asporogenic mutants obtained by treatment with methyl methanesulfonate. The ability to form a toxin can be transferred by transferring a plasmid or gene to other Bacillus thuringiensis species or
    other bacterial systems. Insect strain action on; maggots
    35
    of coleoptera is realized according to the scheme known according to patatnup A: in the confluence of several hours the feeding stops and after a few days
    4Q death occurs depending on the dose. The toxic effect is based on the fact that the midgut acitelium is damaged by crystalline toxin dissolved in the intestinal juice. Each damaged insect intestinal cells are separated from the basement membrane from one another and drift into the intestinal lumen. Due to the breakdown of the intestinal barrier, bacterial cells can penetrate into the hemocoel and complete the disintegration process with septicemia.
    A new insecticide (pathotype C) shows a selective insecticidal action against Coleoptera, for example, against an alder leaf beetle or potato beetle. The effect on the larvae is much stronger than on adult insects. Action not set50
    55
    has been seen on Lepidoptera, HangtHMep, mealworm (Ephestia kShniella Zell), cabbage moth (Plutella x Formula of the invention
    Lostella L.) and on Dipteraj for example
    , five
    The bacterial strain Bacillus thuringi-ensis var. tenebrionis DSM (2803
    yellow fly fever (Aedes aegypbi for insecticide production.). The table shows the results.
    biotest.
    Insect parate Coleoptera. spp.
    The toxic effect of the insecticide on various insects
    10 / cm
    10 cm 2.5x10 / cm
    4x10 / cm
    fever
    1/4489956
    权利要求:
    Claims (1)
    [1]
    Invention Formula
    +
    +
    5x107 cm
    类似技术:
    公开号 | 公开日 | 专利标题
    SU1448995A3|1988-12-30|Strain of bacteria bacillus thuringiensis var. tenebriogis for producing insecticide compound against insects coleoptera spp
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    同族专利:
    公开号 | 公开日
    DE3484196D1|1991-04-04|
    JPS60156382A|1985-08-16|
    PL145191B1|1988-08-31|
    HU195724B|1988-07-28|
    ES538984A0|1986-03-16|
    EP0149162A3|1986-12-30|
    PL251010A1|1985-08-27|
    IL73838A|1988-11-30|
    DE3346138A1|1985-07-11|
    HUT36690A|1985-10-28|
    AT60980T|1991-03-15|
    JPH06192027A|1994-07-12|
    CA1268133A|1990-04-24|
    IL73838D0|1985-03-31|
    US4851340A|1989-07-25|
    PL154613B1|1991-08-30|
    JPH0729895B2|1995-04-05|
    US4973676A|1990-11-27|
    EP0149162A2|1985-07-24|
    US4766203A|1988-08-23|
    EP0149162B1|1991-02-27|
    JPH0634708B2|1994-05-11|
    ES8605032A1|1986-03-16|
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    法律状态:
    2005-05-10| REG| Reference to a code of a succession state|Ref country code: RU Ref legal event code: MM4A Effective date: 20031220 |
    优先权:
    申请号 | 申请日 | 专利标题
    DE19833346138|DE3346138A1|1983-12-21|1983-12-21|BACILLUS THURINGIENSIS VAR. TENEBRIONIS AND AN INSECTICIDALLY EFFECTIVE PRESERVATIVE OR AVAILABLE. TOXIN AND THEIR USE FOR COMBATING COLEOPTERA|
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