Highly active bacillus thuringiensis strain hncs-93 and its application

A technology of Bacillus aureus and spore thuringiensis, which is applied to the highly active Bacillus thuringiensis strain HNCS-93 and its application field, can solve problems such as the loss of control of diamondback moth, achieves good application and promotion prospects, is beneficial for prevention and control, and kills strong poisons effect of action

Inactive Publication Date: 2016-09-07
HUNAN PLANT PROTECTION INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 1992, some experimental studies were carried out, and Bt insecticides were used for field and indoor bioassays on diamondback moths in different areas of Florida. The study found that Bt insecticides lost control of diamondback moths, indicating that diamondback moths have no effect on Bt insecticides. Appear resistance

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Example 1 : strain isolation and culture

[0014] 1. Surface treatment was carried out on abnormally dead diamondback moth larvae collected from the field. The method was as follows: clamp the head of the larvae with tweezers, dipped in 95% ethanol and burned it, then put it in 75% ethanol for 2 s, then put it in 3% ethanol After soaking in sodium hypochlorite solution for 3 min, the larvae were taken out and washed 3 times with sterile water. Put the processed worms into a 1.5 ml centrifuge tube filled with 0.5 ml of sterile water and crush them to make a suspension of worms. Take 0.1ml of the worm suspension and add it into 100ml of sterile water, incubate with shaking at 30°C for 30min, and heat-treat in a water bath at 75-80°C for 14-20min.

[0015] 2. Dilute the culture solution to an appropriate concentration, spread it evenly on the separation medium plate, and place it in a constant temperature incubator at 30°C for cultivation.

[0016] 3. After culturin...

Embodiment 2

[0021] Example 2 : Identification of strains

[0022] According to the implementation method of the above-mentioned embodiment 1, a strain of Bacillus thuringiensis bacterial strain with poisonous effect on Lepidoptera pests, anti-Bt Lepidoptera pests, anti-Cry1Ac toxic protein Lepidoptera pests and Coleoptera pests is obtained, named after Bacillus thuringiensis HNCS-93 (referred to as HNCS-93), the preservation number of the strain: CGMCC No. 8446. The specific identification results are as follows:

[0023] 1. Bacillus thuringiensis Bacillus thuringiensis Insecticidal activity of HNCS-93: the bacterial strain is cultivated in the fermentation medium until the paraspora crystals fall off, and it is effective against the Lepidoptera pest Plutella xylostella ( Plutella xylostella ) relative to the lethal concentration (LC) of sensitive species 50 ) is 5.07mg / L, the lethal medium concentration (LC 50 ) is 6.57mg / L, the lethal medium concentration (LC 50 ) is 4.93mg...

Embodiment 3

[0028] Example 3 : strain HNCS-93 Bioassays on sensitive diamondback moth

[0029] 1. Insects to be tested:

[0030] Plutella xylostella is a relatively sensitive population: it was raised indoors with clean and insect-free radish seedlings and cabbage seedlings, and no pesticides were ever applied during this period. Plutella xylostella populations were reared in isolation indoors, and the pupae were placed in adult cages (R=10cm, L=40cm). The cages were surrounded by 80-mesh gauze. After the adults emerged, the cages were hung and soaked in 10% honey syrup One of the best absorbent cotton balls is used to supplement nutrition for the adults, allowing them to lay eggs on the cabbage seedlings, and then transfer them to fresh cabbage seedlings for rearing after the eggs hatch. The breeding temperature is 25±1°C, the relative humidity is 60%-70%, and the photoperiod is light:dark=16h:8h.

[0031] 2. Determination of strain activity

[0032] A single colony was inocu...

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PUM

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Abstract

The invention discloses a highly active Bacillus thuringiensis HNCS-93 and its application, the preservation number is: CGMCC No. 8446, and the bacterium carries insecticidal protein coding genes cry1Ac and cry2Aa. The strain uses relatively sensitive Plutella xylostella, Btk-resistant Plutella xylostella, and Cry1Ac-resistant Plutella xylostella as test insects, and the virulent strains against Bt and Cry1Ac toxic protein Plutella xylostella are screened through virulence bioassays. The strain is used to produce high-efficiency and broad-spectrum insecticides, which are used to prevent and control the important lepidopteran pest Plutella xylostella. At the same time, it has a good effect on resistant pests, which is beneficial to the control of resistant Plutella xylostella in the field, and improves the economic value. It is harmless to humans, livestock and other animals, does not pollute the environment, has good economic and ecological benefits, and has good application and promotion prospects.

Description

technical field [0001] The invention relates to a highly active Bacillus thuringiensis Bacillus thuringiensis HNCS-93 bacterial strain, and the application of the Bacillus thuringiensis in preventing and controlling Lepidoptera pests or / and resisting Bt Lepidoptera pests. Background technique [0002] Bacillus thuringiensis ( Bacillus thuringiensis , referred to as Bt), is a very widespread Gram-positive bacteria, in the "Bergey's Bacteria Identification Manual" ninth edition, it is classified as a second group of the eighteenth group of Bacillus (Bacillus) species, subspecies are distinguished. Berliner Ernst in 1911 in the Mediterranean mealworm ( Anagastra kuehniella ) is named after its discovery. It was originally isolated from dead silkworms by Japanese scholar Ishiwata, named Bt damping-off subspecies ( Bacillus thuringiensis subsp. scott ) 。 During sporulation, Bacillus thuringiensis can produce various types of parasporal crystal proteins, namely delta-...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/02A01P7/04C12R1/07
Inventor 符伟杨晶张卓方勇刘勇张德咏徐志德彭兆普马明勇成燕清
Owner HUNAN PLANT PROTECTION INST
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