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A strain of Bacillus cereus for controlling plant nematodes and its application

A technology of Bacillus cereus and plants, applied in the fields of application, nematicides, pest control, etc., can solve the problems of few types of chemical nematicides, serious pollution, high toxicity, etc.

Active Publication Date: 2021-09-21
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the control of plant parasitic nematodes is still dominated by chemical control, but there are few varieties of chemical nematicides, and most of them are highly toxic and high residue, which seriously pollute and damage the environment

Method used

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  • A strain of Bacillus cereus for controlling plant nematodes and its application
  • A strain of Bacillus cereus for controlling plant nematodes and its application
  • A strain of Bacillus cereus for controlling plant nematodes and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1. Isolation, purification and identification of bacterial strain MH61-63-03

[0047] 1. Isolation and purification of strain MH61-63-03

[0048] Strain MH 61-63-03 was isolated from soil samples collected in Mohe, Heilongjiang. The specific steps are as follows: Weigh 5 g of soil sample, add it into a conical flask containing 45 mL of sterile water, shake the conical flask for 30 min in a shaker with a rotating speed of 200 rpm / min, and prepare 10 -1soil dilution. Step-by-step dilutions were carried out in an ultra-clean workbench to prepare a series of soil dilutions with corresponding gradients. Use a pipette gun to draw an appropriate amount of soil dilution on the LB plate, and then spread it evenly with a spreader, and repeat each concentration 3 times. Place it in a constant temperature incubator at 28°C for 24 hours, observe the growth of the colony on the plate, pick a single colony on the plate, name it MH 61-63-03 strain, and purify it on a solid L...

Embodiment 2

[0057] The preparation of embodiment 2, MH61-63-03 bacterial agent

[0058] 1. Culture of plate strains

[0059] The MH61-63-03 strain isolated in Example 1 was inoculated on an LB solid medium plate, cultured at 28° C. for 24 hours, and a single colony was picked to obtain a plate strain.

[0060] 2. Liquid fermentation culture

[0061] Inoculate the plate strains obtained in step 1 into a 250mL Erlenmeyer flask containing 100mL of fermentation medium, and ferment at 28°C for 72h at a shaking speed of 160r / min to obtain MH61-63-03 and its metabolites For the fermentation broth, centrifuge the fermentation broth at 10000rpm / min for 5min to obtain the supernatant, and filter it with an organic filter membrane with a pore size of 0.22μm to obtain the fermentation filtrate, which is named MH61-63-03 bacterial agent.

[0062] The prepared MH61-63-03 bacterial agent was diluted 5 times with sterile water to obtain a 5-fold dilution of the MH61-63-03 bacterial agent.

Embodiment 3

[0063] Embodiment 3, the application of MH61-63-03 bacterial agent in the prevention and treatment of plant nematodes

[0064] 1. Nematicidal activity test of MH61-63-03 bacterial agent on second instar larvae of plant parasitic nematodes

[0065] 1. Obtaining the second instar larvae of the root-knot nematode Meloidogyne incognita

[0066] Take water spinach roots inoculated with second instar larvae of Meloidogyne incognita for 60-90 days, clean the root soil with tap water, pick mature and plump egg masses with tweezers under a magnifying glass and place them in the hatching sieve, and place the hatching sieve containing a large number of egg masses Place in a petri dish, add distilled water to the petri dish until the eggs are slightly submerged, and hatch in a constant temperature incubator at 25°C. In order to ensure the complete rate of hatched nematodes, the second instar larvae hatched on the first day are discarded; The activity of the second-instar larvae after hat...

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Abstract

The invention discloses a bacillus cereus for preventing and controlling plant nematodes and its application. The name of Bacillus cereus provided by the present invention is MH61‑63‑03, and the preservation number is CGMCC No.15616. The experimental results show that: the fermentation filtrate of Bacillus cereus MH61‑63‑03 has a high poisonous activity against the second instar larvae of plant parasitic nematodes; The number of wheat root nematodes treated with the filtrate decreased, and the population reduction rate was 26.82%. After 65 days of inoculation with the second instar larvae of cereal cyst nematodes, the average number of cysts per plant of wheat treated with MH61‑63‑03 fermentation filtrate also decreased. significantly reduced. The biological agent of the invention has good application and development prospects in the control of plant parasitic nematodes.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a bacillus cereus for preventing and controlling plant nematodes and its application. Background technique [0002] Among the more than 25,000 known species of nematodes, there are more than 4,100 species of plant parasitic nematodes, accounting for about 15% of the total number of known nematodes, and the annual losses to the world's agricultural production are estimated to reach 157 billion US dollars. There are many types of plant-parasitic nematodes, mainly including Meloidogyne, Anguina, Ditylenchus, Heterodera, Aphelenchoides and root nematodes. Ectoparasitic nematodes, etc. Among them, root-knot nematodes and cyst nematodes are two types of plant parasitic nematodes that cause serious damage in agricultural production, and their disease on vegetables and food crops will cause relatively significant economic losses. [0003] At present, the control of plant parasitic nematode...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P5/00A01P21/00C12R1/085
CPCA01N63/00A01N63/10C12N1/205C12R2001/085
Inventor 刘倩简恒文洋卢修亮王新新
Owner CHINA AGRI UNIV
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