Bacillus thuringiensis for controlling heterodera glycines and meloidogyne incognita as well as preparation and applications

A technology for southern root-knot nematode and soybean cyst nematode, which can be applied in the directions of nematicides, applications, botanical equipment and methods, etc., and can solve problems such as pesticide residues, threats to human health and safety in the ecological environment, and poor control effects of conventional chemical pesticides. , to achieve a significant effect

Active Publication Date: 2018-05-25
HUAZHONG AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the extensive use of highly toxic chemical pesticides, the resistance of nematodes to pesticides continues to increase, and the actual control effect of conventional chemical pesticides is getting worse and worse.

Method used

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  • Bacillus thuringiensis for controlling heterodera glycines and meloidogyne incognita as well as preparation and applications
  • Bacillus thuringiensis for controlling heterodera glycines and meloidogyne incognita as well as preparation and applications
  • Bacillus thuringiensis for controlling heterodera glycines and meloidogyne incognita as well as preparation and applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Isolation and identification of Bacillus thuringiensis and detection of its insecticidal crystal protein

[0026] 1. Isolation of Bacillus thuringiensis

[0027] Soil sample collection: In August 2010, the soil at a distance of 0-15 cm from the ground surface was collected in the suburbs of Lanzhou City, Gansu Province. After removing dead branches, stones and other sundries, it was dried at room temperature, ground and crushed, and passed through a 40-mesh sieve. Store in soil sample bottles at room temperature for later use.

[0028] Separation of Bt: Weigh 1 gram of soil sample into a liquid medium containing 20ml LB+sodium acetate (according to 1L medium: peptone 10g; yeast powder 5g; sodium chloride 10g; sodium acetate 34.02g, add distilled water to 1L; adjust pH to 7.0) in a 250ml Erlenmeyer flask, at 30°C, 220rpm, and cultivate for 4 hours; heat-treat the culture in water at 80°C for 3 minutes; take 100ul of the treated culture solution and use LA soli...

Embodiment 2

[0050] Example 2 Preparation of Bacillus thuringiensis preparation WP-HAN055

[0051] 1. Strain preparation: First, inoculate the preserved Bacillus thuringiensis strain HAN055 onto the slope (BP medium 1L: peptone 5g; beef extract 3g; sodium chloride 5g; add distilled water to 1L; adjust pH to 7.0), 30°C Cultivate for 3-5 days (form powerful spores); then wash the bacteria with 5 mL of sterile water, and incubate on a water bath at 75-80°C for 15-20 minutes; take the spore suspension and inoculate it into 100 mL of TP liquid medium ( Calculated on the basis of 1L culture medium: tryptone 20g; dextrose 2g; disodium hydrogen phosphate 2.5g; sodium chloride 5g; add distilled water to 1L; adjust pH to 7.0) in a 500mL Erlenmeyer flask, culture on a shaker (30°C, 220rpm ) for 6-8 hours, which is the first-class seed; then transfer 1000 mL of new TP medium with 2% volume inoculum to obtain the second-class seed; then use the second-class seed to inoculate the seed tank with 0.5% ino...

Embodiment 3

[0057] Example 3 Bioassay experiment of Bacillus thuringiensis preparation WP-HAN055 on root-knot nematode incognita and soybean cyst nematode

[0058]1. Bioassay test of Bacillus thuringiensis preparation WP-HAN055 on root-knot nematode incognita

[0059] (1) Preparation of 2nd instar larvae of M. incognita: M. incognita egg masses were picked from tomato roots infected with M. 2 o 2 After being sterilized with 0.25% KI disinfectant, it was placed in an incubator at 25° C. for 3-5 days, and the egg masses hatched out of the root-knot nematode incognita, namely the 2nd instar larvae, were preserved for future use.

[0060] (2) Determination of biological activity on a 96-well plate: the Bacillus thuringiensis preparation WP-HAN055 was diluted to 5 concentrations, a blank control was set with sterile deionized water, and each treatment was repeated 4 times. Inhale about 40 2nd instar larvae per well, add samples of different concentrations to the corresponding treatment, make...

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Abstract

The invention belongs to the technical field of microbial pesticides and particularly relates to bacillus thuringiensis for controlling heterodera glycines and meloidogyne incognita as well as a preparation and applications. The bacillus thuringiensis HAN055 having high toxicity for the heterodera glycines and meloidogyne incognita is obtained through separation. The preparation WP-HAN055 preparedfrom the strain has significant control effects on heterodera glycines and meloidogyne incognita damage. The bacillus thuringiensis HAN055 is collected in CCTCC (China Center for Type Culture Collection) with the collection number of CCTCC NO: M2015608.

Description

technical field [0001] The invention belongs to the technical field of microbial pesticide preparation. Specifically, it relates to bacillus thuringiensis for preventing and controlling soybean cyst nematode and root-knot nematode incognita, as well as its preparation and application. Background technique [0002] Plant pathogenic nematodes belong to the phylum Nematoda, class Secernentea, and most of them belong to the order Thyrocystis. The main path of pathogenicity is that the larvae invade the plant from the root tip of the plant, the pathogen is in the plant, plunders the plant nutrition, and causes damage to the plant such as nutrient deficiency, physical damage, chemical pathogenicity and compound infection (Sharon et al., 2001). The general symptoms are short stature, wilting, deformity, abnormal color, early wilting, etc. In severe cases, the whole plant will wither and die. [0003] The serious harm brought by plant pathogenic nematodes to the development of ag...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P5/00C12R1/07
CPCA01N63/00C12N1/205C12R2001/075
Inventor 孙明聂相涛张争名彭东海阮丽芳
Owner HUAZHONG AGRI UNIV
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