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A strain of Bacillus thuringiensis with high poisonous activity against Noctuidae pests and its application

A Bacillus and pest technology, applied in Bacillus thuringiensis and application fields

Active Publication Date: 2021-10-08
HUBEI BIOPESTICIDE ENG RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no officially registered Bt insecticides against such pests in China, but there are patents showing that Bt engineering strains expressing Vip protein (patent application number 202011206877) can be used, or Vip pure protein expressed by Escherichia coli can be used for grassland insecticide. Prevention and control of noctuid (patent authorization number CN110622998B)

Method used

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  • A strain of Bacillus thuringiensis with high poisonous activity against Noctuidae pests and its application
  • A strain of Bacillus thuringiensis with high poisonous activity against Noctuidae pests and its application
  • A strain of Bacillus thuringiensis with high poisonous activity against Noctuidae pests and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Obtaining Bacillus thuringiensis NBI-616 Strain

[0025] The applicant took soil samples from Yingcheng, Hubei Province, and used the sodium acetate-antibiotic separation method to isolate Bacillus thuringiensis. The specific process is as follows: 1 g of soil sample is weighed, put into a shaking flask equipped with 10 ml of sodium acetate medium, and penicillin sodium salt and gentamicin sulfate are added respectively (final concentration is 400 ug / mL). The shaker flask was placed on a shaker, and cultured with shaking at 30° C. and 200 rpm / min for 4 h. After the shaking culture, take the soil suspension, put it into a sterile centrifuge tube, centrifuge at 3000r / min for 15min, then take 1mL of the upper cloudy solution in the centrifuge tube and place it in a water bath at 65°C for 15min, take 0.1mL of the heat-treated cloudy solution and apply Luria- Bertani agar plate (tryptone 10g, yeast powder 5g, sodium chloride 10g, agar powder 20g, dissolved in 1L of distille...

Embodiment 2

[0055] Determination of Insecticidal Activity of Bacillus thuringiensis NBI-616 Strain

[0056] Use an inoculation loop to pick the single colony isolated and purified on the Luria-Bertani solid medium plate into the liquid medium, the medium formula: 2.0% starch, 3.0% peanut meal, 0.5% industrial peptone, 2.0% corn steep liquor, sterilized The former pH is 7.5, and the volume of liquid in a 500mL Erlenmeyer bottle is 50mL.

[0057]The temperature is 30-32°C, the speed is 220rpm, and the shaker is shaken for 34 hours until 40% of the spores and crystals are separated. Spodoptera larvae, beet armyworm larvae, cotton bollworm larvae, diamondback moth larvae, housefly larvae, housefly adults and German cockroach adults were used as test insects, and the toxicity of Bacillus thuringiensis NBI-616 fermentation liquid was measured, and the results As shown in Table 6-Table 13, the LC50 value of Bacillus thuringiensis NBI-616 fermentation broth to Spodoptera frugiperda was 0.28μL / mL...

Embodiment 3

[0076] Fermentation of Bacillus thuringiensis NBI-616 strain and preparation of high content technical medicine

[0077] Preparation of seed culture medium: the volume of the shake flask of the seed medium is 50mL / 500mL, the rotation speed is 220rpm, the culture temperature is 30-32°C, the fermentation period is 31-35h, and the unit cells are obtained when the spores and crystals are 40% separated by microscope inspection suspension.

[0078] The formulations of the seed medium and the fermentation medium in this embodiment are: 3.0% corn flour, 4.0% soybean meal, 1.0% peptone, 2.5% corn steep liquor, pH 8.0.

[0079] Adopt primary fermentation: the filling volume of the fermentation medium in the 5000L fermenter is 3500L, and the pH before elimination is 8.0. Sterilization condition 121℃, 1kg / cm 2 , 30min. When the tank temperature drops to 35°C, inoculate with a suspension of Bacillus thuringiensis NBI-616 spores. The inoculum volume is 200mL unit cell suspension, the ve...

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Abstract

The invention belongs to the field of biopesticides, and relates to a strain of Bacillus thuringiensis with high toxicity to Noctuidae pests and its application. The present invention relates to Bacillus thuringiensis NBI-616 with high toxicity to various Noctuidae pests The strain has been preserved in China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: M 2020771. Bioassays show that Bacillus thuringiensis NBI‑616 has high poisonous activity against the larvae of Spodoptera frugiperda, Spodoptera litura, Spodoptera litura, Cotton bollworm and Diamondback moth. The high-content raw powder of Bacillus thuringiensis NBI‑616 was prepared by conventional fermentation-centrifugal concentration-spray drying process. The bacterial strain involved in the invention can be used to develop microbial pesticides for preventing and controlling Noctuidae pests.

Description

technical field [0001] The invention belongs to the field of biological pesticides, and relates to a bacillus thuringiensis strain with high poisonous activity against Noctuidae pests and its application. Background technique [0002] Noctuidae is the largest family of Lepidoptera. There are about 35,000 species in the world, of which about 3,751 species are known in China (Feng Botao et al., 2019). Spodoptera frugiperda, Spodoptera exigua, Spodoptera litura Fabricius, and Heli coverpa armigera belong to the Lepidoptera Noctuidae and are important agricultural and forestry pests in my country (Chen Xiuhong et al. , 2020). [0003] In recent years, fall armyworm has caused the most serious damage. Fall armyworm is one of the important migratory pests on maize and other major commercial crops (Cruz et al., 2012, Nagoshi et al., 2015, Young and McMillian, 1979). It has the characteristics of a wide range of hosts, a wide range of suitable habitats, fast migration speed, stron...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/23A01N63/50A01P7/04C12R1/07
CPCA01N63/23A01N63/50
Inventor 王开梅廖先清周荣华王月莹张志刚刘芳
Owner HUBEI BIOPESTICIDE ENG RES CENT
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