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Bacillus thuringiensis YN108 with high toxicity to lepidopteran pests, culture method and application of bacillus thuringiensis YN108

A Bacillus thuringiensis and Lepidoptera technology, applied in the field of microbial pesticides, can solve the problems of limited control and low toxicity, and achieve the effects of improving utilization efficiency and excellent insecticidal activity

Inactive Publication Date: 2018-09-04
YANBIAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it is reported that although the strain is highly toxic to some insects of Lepidoptera, it has very low toxicity to important Noctuidae pests such as beet armyworm and Spodoptera litura
Not only that, this strain is relatively easy to induce the resistance of pests under laboratory and field conditions, thus limiting the wide application of this type of microbial preparations in the control of crop pests

Method used

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  • Bacillus thuringiensis YN108 with high toxicity to lepidopteran pests, culture method and application of bacillus thuringiensis YN108

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Example 1 Isolation of Bacillus thuringiensis subspecies Ayuzawa YN108 bacterial strain (hereinafter referred to as Bacillus thuringiensis YN108)

[0027] The soil for testing comes from the mountainous area of ​​Xicheng Town, Helong City at the foot of Changbai Mountain, and the sample soil is used for the test after collecting 10g at a depth of about 5cm in the soil surface under the tree. Weigh 1 g of the collected soil sample and put it into the test tube, add 9 mL of sterilized water and stir vigorously for 4 to 5 times. In order to screen and remove the miscellaneous bacteria that cannot form spores, the test tube was treated in a constant temperature water tank at 65°C for 30 minutes, and after the heat treatment, it was allowed to stand for 5 minutes. After the soil settled, the supernatant was taken, diluted 1000 times, and spread evenly on the nutrient agar medium (Nutrient agar ), and place the culture medium in a constant temperature incubator at 27°C for 3 ...

Embodiment 2

[0028] Example 2 Serological Identification of Bacillus thuringiensis YN108

[0029] The result of identifying the flagellar antigen of Bacillus thuringiensis YN108 obtained in Example 1 and the method of agglutination reaction with known antiserum, the strain had agglutination reaction with the antiserum of known Bacillus thuringiensis subspecies Ayuzawa, indicating that YN108 The strain belongs to Bacillus thuringiensis subspecies Ayuzawa, see Table 1 for the results.

[0030] Table 1 Serotype classification of Bacillus thuringiensis YN108

[0031] isolate

Embodiment 3

[0032] Example 3 Observation of endotoxin protein of Bacillus thuringiensis YN108

[0033] The results of observing the endotoxin protein of Bacillus thuringiensis YN108 obtained in Example 1 with a phase-contrast microscope are as follows: In order to obtain the endotoxin protein, it was inoculated on NA medium and cultivated at a constant temperature of 27°C for 3 to 5 days until Spores and endotoxin proteins (paraspore crystals) are formed, and the resulting crystal spores are isolated. Smear the isolated strain producing crystal spores on a glass slide, drop a drop of sterilized water, cover the cover glass, and then drop a drop of cedar oil, observe under a 1000-fold phase-contrast microscope. figure 1 It is a phase-contrast micrograph (×1000) of endotoxin protein (C) and spore (S) of Bacillus thuringiensis YN108. From figure 1It can be seen that the paraspore crystals (endotoxin protein) of Bacillus thuringiensis YN108 are rhomboid, which is consistent with the morphol...

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Abstract

The invention belongs to the technical field of microbial insecticides and particularly relates to bacillus thuringiensis YN108 with high toxicity to lepidopteran pests, a culture method and application of the bacillus thuringiensis YN108. The bacillus thuringiensis YN108 is bacillus thuringiensis subsp.aizawai and has a preservation number of CGMCC No. 15140 in the China General Microbiological Culture Collection Center (CGMCC) on January 02, 2018. The bacillus thuringiensis YN108 dislcosed by the invention has high toxicity to the lepidopteran pests, and particularly has a median lethal concentration (LC500) of only 3.25*105 cfu / ml for the 3<rd> instar larva of spodoptera litura, which is smaller than a published YN1-1 bacterial strain and a published CAB109 bacterial strain. Meanwhile,the bacillus thuringiensis YN108 is also better than a commercial bacillus thuringiensis catfish subspecies I product, an SB product and a bacillus thuringiensis catfish subspecies SL product in indoor drug efficacy and field control effect and shows excellent insecticidal activity. In addition, the bacillus thuringiensis YN108 disclosed by the invention has no residue after use, avoids drug resistance and is green and environmentally-friendly.

Description

technical field [0001] The invention belongs to the technical field of microbial insecticides, and in particular relates to a Bacillus thuringiensis YN108 with high toxicity to lepidopteran pests, a cultivation method and application thereof. Background technique [0002] Bacillus thuringensis (Bt), as an entomopathogenic bacterium, is an important factor for biological control in pollution-free agricultural production. Bt is an aerobic Gram-positive bacterium with flagella that forms endospores and parasporal inclusion crystals (PIC) when growth conditions deteriorate. The special endotoxin protein crystals formed in Bt have strong insecticidal activity against larvae of Lepidoptera, Diptera and Coleoptera, and strains that are toxic to nematodes have been found. These endotoxin proteins can be divided into Cry 1-40, Cyt 1-2, and can be divided into different types according to their amino acid homology. Among them, the endotoxin protein is CryI for Lepidoptera, CryII for...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P7/04C12R1/07
CPCA01N63/00C12N1/20C12N1/205C12R2001/075
Inventor 金大勇
Owner YANBIAN UNIV
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