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Bacillus velezensis microbial agent and application thereof

A technology of Bacillus velesi and microbial inoculants, applied in the directions of microorganisms, biocides, microorganism-based methods, etc., can solve the problems of poor effect of cotton verticillium wilt, insufficient microbial strains, etc., and achieve difficult drug resistance, Good drug effect persistence, no effect of environmental pollution

Active Publication Date: 2021-03-09
THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims at technical problems such as the different types and continuous evolution of pathogenic microorganisms, the poor effect of existing microorganisms on cotton Verticillium wilt, and the insufficient abundance of applicable microbial strains. Bacteria and its application

Method used

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  • Bacillus velezensis microbial agent and application thereof
  • Bacillus velezensis microbial agent and application thereof
  • Bacillus velezensis microbial agent and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Isolation, screening and identification of Bacillus velezensis TK2019

[0031] (1) Separation and purification

[0032] From the soil sample, weigh 1.0g of soil sample and add 100mL of sterile water into a 250mL sterilized Erlenmeyer flask, put it on a shaker, vibrate at 150rpm for 30min, let it stand for 2h, and take 10mL of the supernatant into a sterilized centrifuge tube , 80°C constant temperature water bath for 30min, then take 1mL and add 9mL sterile water to make 10mL 10 -3 times the soil microbial suspension, continue to use sterile water for gradient dilution, respectively from 10 -4 、10 -5 、10 -6 200 μL of each of the three dilutions was spread on the NA medium plate, and placed in a constant temperature incubator at 30°C for 1-3 days. Pick a single colony from the plate, inoculate it on a new separation plate for purification, and repeat 3 times for each concentration. The purified Bacillus velezensis TK2019 was inoculated on the medium, cult...

Embodiment 2

[0047] Example 2: Determination of Physiological and Biochemical Properties of Bacillus velezensis TK2019 CGMCC No: 20810

[0048] For the colony morphology and physiological and biochemical characteristics of the strain TK2019 on the NA medium, please refer to the attached figure 2 And shown in Table 1.

[0049] The bacterial strain TK2019 was cultured on NA medium and was rod-shaped. After 12 hours of culture, it produced spores. The spores were middle-grown, oval, and the cysts were not enlarged. The acid-fast staining was negative. Gram-positive bacteria were identified by Ram's staining, and the strain TK2019 under the microscope was a Gram-positive bacillus [(1-1.5) μm×(3-5) μm], arranged in single or double or short chains. On the nutrient agar plate, the colony is milky white at the initial stage of culture, the surface is rough, the edge is irregular, opaque, and wrinkled and raised; when cultured on the slope of the nutrient agar, it is linear; when it is cultured ...

Embodiment 3

[0054] Embodiment 3: the preparation of Veles bacillus (Bacillus velezensis) TK2019 microbial bacterial agent

[0055] The present embodiment is on the basis of embodiment 1-2, and the preparation method of Bacillus velezensis TK2019 microbial bacterial agent specifically comprises the steps:

[0056] (1) Activate Bacillus velezensis TK2019 stored at low temperature on the NA plate medium, pick a single colony on the NA slant medium, and culture it at 20-37°C for 6-12 hours to obtain the activated shellfish Bacillus velezensis TK2019 strain.

[0057] (2) Scrape a ring with a sterile inoculation loop Step (1) Inoculate the activated Bacillus velezensis TK2019 strain into 100mL NA liquid medium, and inoculate at 20~37°C with a shaker speed of 120~ Cultivate for 6-12 hours under the condition of 180 rpm to obtain Bacillus velezensis TK2019 seed solution.

[0058] (3) The Bacillus velezensis TK2019 seed solution obtained in step (2) is inserted into the corn flour soybean flour ...

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Abstract

The invention discloses a bacillus velezensis TK2019 microbial agent and application thereof. The bacillus velezensis TK2019 is separated from rhizosphere soil of the cotton field of the 44 regiment in the Tumxuk city of Xinjiang, and the bacillus velezensis TK2019 microbial agent is used for preparing the bacillus velezensis TK2019. When the microbial agent is applied to prevention and treatmentof cotton verticillium wilt, cotton fusarium wilt, botrytis cinerea and red date black spots, the inhibition rate of the strain TK2019 on verticillium dahliae of cotton reaches 94.37 percent, the inhibition rate of the strain TK2019 on four tested pathogenic fungi is 84.83-93.73 percent, and the stock solution and 50-time diluted solution of the bacillus velezensis TK2019 microbial agent have thehighest prevention effects which are 92.86 percent and 91.97 percent respectively. Field tests prove that the prevention effect the TK2019 microbial agent on cotton verticillium wilt is 92.69 percent,which indicates that the Bacillus velezensis TK2019 and the microbial agent thereof provided by the invention have the advantages of high efficiency, wide bactericidal spectrum and the like and havewide values for expanding the application field of microbial strains.

Description

technical field [0001] The invention relates to the technical field of microorganisms and their applications, in particular to a Bacillus velezensis TK2019 microbial bacterial agent and the technical field of their applications. Background technique [0002] Cotton Verticillium wilt is a cotton soil-borne disease caused by Verticillium dahliae Kleb. Symptoms generally begin at the stage of 3-5 true leaves, and a large number of diseases occur in the field after budding and flowering of cotton in the middle and late stages of growth. Cotton verticillium wilt was first reported in Virginia, USA in 1914, and then spread to dozens of countries and regions including China, causing huge losses to cotton production and becoming a major obstacle to high and stable cotton production. Cotton verticillium wilt seriously affects the growth and development of cotton. After the onset, cotton buds and bolls are often scarce, and cotton bolls crack in advance, resulting in reduced producti...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/22A01P3/00C12R1/07
CPCC12N1/20A01N63/22
Inventor 史应武杨红梅楚敏牛新湘林青娄恺王宁包慧芳詹发强龙宣杞杨蓉
Owner THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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