Bacillus with nematocidal activity, and preparation method and application of bacillus

A technology of the genus Bacillus and Bacillus, applied in nematicides, botany equipment and methods, methods based on microorganisms, etc., can solve problems such as mass production restrictions, and achieve low production costs and simple production processes

Active Publication Date: 2014-05-07
甘肃绿能农业科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, PGPR mainly prevents crop diseases and insect pests by inducing systemic resistance of crops; Pasteurella puncture is a nematode obligate parasite, which has a significant effe

Method used

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  • Bacillus with nematocidal activity, and preparation method and application of bacillus
  • Bacillus with nematocidal activity, and preparation method and application of bacillus
  • Bacillus with nematocidal activity, and preparation method and application of bacillus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1. Utilize the serial gradient dilution method to isolate plant rhizosphere soil bacteria

[0031] 1. Isolation of bacteria from rhizosphere samples of Chaejama chamaejasma in Cuiying Mountain, Lanzhou University, Yuzhong Campus, Lanzhou City, Gansu Province

[0032] Daphne chamaejasma ( Stellera chamaejasmL.) Isolation of strain B-RS-06 from rhizosphere soil. The method is as follows: select the vigorously growing Rhizoma chamaejasme, dig out the whole plant, gently shake off the excess soil on the root system, immerse 3 g of the root system sample in 1 volume of sterile PBS buffer solution, and obtain a 1 / 10 dilution Liquid sample, fully shake the fresh root system for 5 min, wash the soil attached to the surface of the root system into the buffer solution, and then serially dilute it with 10 times the volume until the minimum concentration of 10 -8 Take 0.1 ml of serial dilutions from the dilutions with a concentration gradient of 10-4-10-8, respectively, ...

Embodiment 2

[0039] Embodiment 2. The cultivation of bacteria and the preparation of metabolites

[0040] 1. Primary strain culture: Transfer and activate the strain B-RS-06 stored on the slant medium to the NA plate medium, and culture it in the dark at 28-30°C. After the colonies visible to the naked eye grow, it will be used as a primary strain. grade bacteria.

[0041] 2. Cultivation of secondary strains: inoculate the activated primary strains on NA liquid medium, place them on a shaking table with a temperature of 28-30°C and a rotation speed of 180 rpm, and cultivate them in the dark, observe and record regularly, and wait for cultivation After the liquid became turbid (about 72 h), the shaking culture was stopped, and it was transferred to a 4°C refrigerator for short-term storage.

[0042] 3. Preparation of metabolites: Place the liquid bacterial agent in a low-temperature high-speed centrifuge, set the centrifugation temperature to 4°C, and the rotation speed to 10,000 rpm. Aft...

Embodiment 3

[0044] Example 3. Nematode contact activity of Bacillus B-RS-06 metabolites

[0045] 1. Cultivation of nematodes

[0046] In the present embodiment, D. destructor of potato ( Ditylenchus destructor ) and pine xylophilus ( Bursaphelenchus xylophilus ) was used as the experimental material, and the nematodes were cultured according to conventional methods. After the nematodes have multiplied and covered the entire Petri dish, turn the Petri dish upside down, add 3-5 ml of sterile water to the lid of the Petri dish, and let it stand for more than 6 hours, so that the nematodes enter the sterile water. Check under a microscope, and perform appropriate dilution according to the nematode density, so that when the microscope magnification is 40 times, the number of nematodes per field of view is about 100-200.

[0047] 2. Contact activity of different concentrations of metabolites on nematodes

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Abstract

The invention discloses a bacillus with nematocidal activity, and a preparation method and an application of the bacillus. The name of a strain of the bacillus is B-RS-06, and the preservation number is CGMCCNo.5352 (China General Microbiological Culture Collection Center Number 5352). The strain is separated from rhizosphere soil of a plant, and metabolin of the strain acts on a plant nematode. A raw material of a nematocide of the bacillus is derived from the rhizosphere soil of the plant, and has no adverse effect on an environment and all organisms such as the plant, people and an animal; the production cost is low; a production process is simple; and the environment is not polluted.

Description

technical field [0001] The invention relates to a Bacillus bacterium with nematicide activity and a preparation method and application thereof, belonging to the technical field of microbial pesticides. Background technique [0002] Plant parasitic nematodes are one of the main pathogens of plants and an important class of soil-borne diseases in agricultural production in my country and even in the world. According to the conservative estimate of FAQ, the economic loss caused by nematode damage is about 100 billion US dollars every year in the world. In my country, the annual loss of soybeans caused by nematodes alone reaches 100 million US dollars, and the damage caused by nematodes in various vegetables reaches more than 3 billion US dollars. At present, the area of ​​crops that urgently need nematode control in production is at least 50 million mu. Nematode diseases that seriously threaten my country's agricultural and forestry production include root-knot nematodes ( M...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02A01N63/02A01P5/00C12R1/07
Inventor 秦波金辉刘权张等宏燕志强
Owner 甘肃绿能农业科技股份有限公司
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