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Bacillus cereus, microbial inoculum, preparation method and application thereof

A technology of Bacillus cereus and bacterial agents, applied in the field of microorganisms, can solve problems such as damage to the soil ecological environment, high toxicity of chemical pesticides, and difficulty in implementation and application, and achieve high-efficiency control effects, significant control effects, and rapid reproduction

Active Publication Date: 2019-05-14
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of the above technical problems, the present invention provides a Bacillus cereus capable of effectively preventing and treating various plant nematode diseases, a culture method, a bacterial agent and its preparation method and application, so as to solve the problem of high toxicity of chemical pesticides in the prevention and treatment of existing plant nematode diseases. , high cost, difficult implementation and application, damage to the soil ecological environment and other deficiencies

Method used

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  • Bacillus cereus, microbial inoculum, preparation method and application thereof
  • Bacillus cereus, microbial inoculum, preparation method and application thereof
  • Bacillus cereus, microbial inoculum, preparation method and application thereof

Examples

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Embodiment 1

[0030] Example 1: Isolation and screening of Bacillus cereus

[0031] 1. Preparation of Medium

[0032] The formula of ordinary beef extract peptone agar medium is: beef extract 5g; peptone 10g; sodium chloride 5g; agar powder 20g; distilled water, 1L. Adjust the pH to 7.2, and sterilize at 121°C for 20 minutes.

[0033] The formula of enriched beef extract peptone medium is: beef extract 10g; peptone 20g; sodium chloride 5g; distilled water, 1L; high temperature sterilization at 121°C for 20min.

[0034] 2. Isolation and screening of Bacillus cereus

[0035]Take by weighing 500g wheat cyst nematode disease field soil soil sample and put in the basin, rinse with tap water, stir, upper layer water is poured into cover sieve (40 mesh sieves of upper floor, 60 mesh sieves of lower floor), operation repeats five times. Wash the soil sample retained by the 60-mesh sieve into a beaker filled with a small amount of water. After mixing, pour it into a porous plastic dish with filt...

Embodiment 3

[0079] The cultivation of embodiment 3 Bacillus cereus BCCY22

[0080] Two experimental groups were set up with optimized culture conditions and conventional culture conditions. The optimal culture conditions were adjusted to 6.5 for the initial pH value of enriched beef extract peptone liquid medium, the inoculum size was 3%, and the temperature was 32.5°C; the conventional culture conditions were beef extract The initial pH value of the peptone liquid medium was adjusted to 7.2, the inoculum size was 1%, and the temperature was 28°C; the test was carried out in a 250mL triangular flask with a filling volume of 20%, and cultured at 140rpm / min for 3 days. Three bottles were taken out at 6h, 12h, 24h, 36h, 48h, 60h, 66h, and 72h for different treatments as samples to be tested, and stored at 4°C. After the sampling is completed, the sample to be tested is serially diluted to 10 with sterilized water according to a 10-fold gradient. -7 、10 -8 、10 -9 , take 0.1mL of the diluti...

Embodiment 4

[0082] The preparation of embodiment 4 Bacillus cereus BCCY22 inoculum

[0083] Add enriched beef extract peptone liquid medium into a fully automatic mechanically stirred and ventilated fermenter with a filling factor of 65%, sterilize at 121°C for 30 minutes, insert 3% of the inoculum into the seed solution that has been cultivated for 16-18 hours, 250r / min , fermented at 32.5°C, using soybean oil as the defoamer to automatically control defoaming; during the fermentation process, the ventilation rate (V / V·min) was 1:1.2, the pH was controlled at 6.5, and the fermentation was 36 hours. At the end of fermentation, the number of viable bacteria was counted by dilution coating method, and the number of BCCY22 in the fermentation broth was 5.3~7.5×10 10 CFU / mL.

[0084] Adjust the BCCY22 fermentation broth to 1 × 10 with sterile water 9 CFU / mL to get BCCY22 liquid inoculum, add BCCY22 liquid inoculum at 1:1 (V:W) to sterilized bran for adsorption, and then get BCCY22 solid ino...

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Abstract

The invention discloses a bacillus cereus BCCY22, the preservation number of which is CGMCC No. 16672, and further discloses a culture method of the strain, a formed microbial inoculum, a preparationmethod of the microbial inoculum and an application of the microbial inoculum. The bacillus cereus colony is milky white, approximately circular and slightly long, has uneven edges, uniform and opaquetexture, and the bacillus cereus BCCY22 is rod-shaped, has the length of 2.86-4.44um and the width of 0.80um-1.21um, and is in chain arrangement or exists independently; the bacillus cereus BCCY22 produce spore, wherein the spore is generated in the middle and is ellipse; cysts do not expand, and Gram staining reaction is positive; the bacillus cereus can utilize glucose and sucrose, and cannot utilize lactose. The bacillus cereus is a broad-spectrum nematode antagonistic strain, and the prepared microbial inoculum has high-efficiency prevention and treatment effects on plant nematode diseases, has certain effects of promoting crop growth and increasing yield, and is favorable for protecting environment.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a bacillus cereus, a bacterial agent and a preparation method and application thereof. Background technique [0002] Plant parasitic nematodes are one of the important pathogens causing plant infectious diseases, and have become the second largest type of diseases in agricultural production, causing serious economic losses. For example, peanut root-knot nematode is the most common and most harmful type of nematode. The damaged peanuts generally reduce production by 20% to 30%, and serious cases can reach more than 70%; the grain loss caused by wheat cyst nematode is 15% to 96%. %; root-knot nematodes lead to a reduction in the yield of vegetable crops, with a general loss of 10%-20%, a loss of 30%-50% of susceptible vegetables, and a loss of more than 75% in severely diseased plots. The annual loss caused by vegetable root-knot nematode in my country is about 5 billion yuan...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/00A01P5/00C12R1/085
Inventor 咸洪泉赵洪海李雅华杨亚楠张启航
Owner QINGDAO AGRI UNIV
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