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Bacillus methylotrophicus, microbial agent containing bacillus methylotrophicus and application

A technology of methylotrophic bacillus, applied in the field of methylotrophic bacillus, including methylotrophic bacillus, can solve the problems of damage, rapid mutation of rice blast bacteria, unstable control effect, etc. , to achieve the effects of increased yield, good colonization ability, and excellent control effect

Inactive Publication Date: 2021-09-21
LIAONING ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The use of biocontrol bacteria to prevent and control plant diseases has attracted more and more attention in production. It has the characteristics of being safe for humans, animals and crops, and does not cause harm to natural enemies and beneficial organisms. However, the control effect of different biocontrol bacteria on target diseases and There are certain differences in the ability to colonize in the host body, especially the rice blast can cause damage to rice at different growth stages, and the mutation speed of the rice blast fungus is relatively fast, so the biological bacteria need to have a good antibacterial effect and be effective in rice. Different parts and growth stages can be colonized to play a good role in prevention and control
[0006] At present, the colonization effect of biological agents for the control of rice blast is poor, and the control effect is unstable, and there are few studies on the effect of biocontrol bacteria on crop yield increase

Method used

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  • Bacillus methylotrophicus, microbial agent containing bacillus methylotrophicus and application
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  • Bacillus methylotrophicus, microbial agent containing bacillus methylotrophicus and application

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preparation example Construction

[0031] In the present invention, the preparation method of the methylotrophic bacillus CB-12 fermentation liquid preferably comprises the following steps:

[0032] 1) Inoculate CB-12 in LBA medium for activation culture to obtain a single colony of CB-12; the method of activation is preferably plate marking method, and in the specific implementation process of the present invention, use a sterile bacterial inoculation loop to dip A small amount of CB-12 strains, evenly streaked in a Z shape on the LBA plate; the temperature of the activation culture is preferably 30-35°C; the time of the activation culture is preferably 20-24h; the activation culture is in a dark condition under;

[0033] 2) pick the single colony of activated CB-12 and transfer it to LB medium for expanded culture to obtain CB-12 seed solution; the temperature of the expanded culture is preferably 30-35°C; the time for the expanded culture is preferably 4-6d; the expanded culture is preferably carried out un...

Embodiment 1

[0043] The separation and screening of embodiment 1 biocontrol bacteria

[0044] The methylotrophic Bacillus CB-12 used in the present invention is sampled from the biogas slurry produced by Dengta Jinkang Planting Co., Ltd. by the Institute of Plant Protection, Liaoning Academy of Agricultural Sciences, and the bacteria suspension is prepared after dilution, and the plate culture method is used to screen the rice. Magnaporthe grisea is an antagonistic biocontrol bacterium. CB-12 colony morphology see figure 1 .

[0045] Separation steps:

[0046] The separation medium used LB medium: 6g of yeast extract, 10g of tryptone, 10g of sodium chloride, dilute to 1L with deionized water, and adjust the pH to 7.

[0047] Separation and screening conditions: mother liquor preparation: take 1ml biogas slurry into 9ml sterile water and mix well. Using the gradient dilution method, the mother liquor was diluted to 10 -4 ~10 -7 Afterwards, spread evenly on the LB medium plate, and set...

Embodiment 2

[0049] The identification of embodiment 2 biocontrol bacteria CB-12

[0050] 1. Physiological and biochemical identification

[0051] Physiological and biochemical identification was searched with reference to the "Common Bacteria System Identification Manual" (Dong Xiuzhu and Cai Miaoying, 2001), and CB-12 was identified as Bacillus. The specific results are shown in Table 1.

[0052] Table 1 Physiological and biochemical identification results of CB-12

[0053]

[0054]

[0055] 2. Molecular identification

[0056] experiment method

[0057] (1) Extraction of CB-12 genomic DNA. Extract genomic DNA using the Bacterial Rapid Extraction Kit.

[0058] (2) PCR amplification and sequence determination of 16SrDNA sequence. 16S rDNA was amplified by PCR using universal primers 27F and 1492R. PCR conditions: 95°C, 5min; 94°C, 1min, 56°C, 2min, 72°C, 2min, 30 cycles; 72°C, 10min. PCR products were sent to Sangon Bioengineering (Shanghai) Co., Ltd. for sequencing.

[0059...

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Abstract

The invention provides bacillus methylotrophicus, a microbial agent containing the bacillus methylotrophicus and application, and belongs to the technical field of microbial agents. The preservation number of the bacillus methylotrophicus is CGMCC (China General Microbiological Culture Collection Center) No. 22734. The bacillus methylotrophicus CB-12 disclosed by the invention has an antagonistic effect on phyricularia grisea, the biological agent containing the bacillus methylotrophicus CB-12 has an excellent prevention and treatment effect on phyricularia grisea, and the bacillus methylotrophicus CB-12 has a good colonization capability in rice. In addition, after the rice is treated by the bacillus methylotrophicus CB-12 disclosed by the invention, the yield of the rice can be obviously improved.

Description

technical field [0001] The invention relates to the technical field of microbial bacterial agents, in particular to a strain of methylotrophic bacillus, bacterial agents including methylotrophic bacillus and applications thereof. Background technique [0002] Rice (Oryza sativa L.) is an important food, feed, industrial and energy crop, and plays an extremely important role in agricultural production. In the world, rice planting area and total output are higher than that of wheat and corn, ranking first. In the past two decades, the rice planting area in my country has fluctuated firstly and then increased, and the total output and per unit yield have shown a growing trend. [0003] Rice blast is a fungal disease caused by Magnaporthe oryzae infection. It is the most serious disease on rice and ranks first among the top ten fungal diseases in the world. Rice blast can lead to a 10% to 30% reduction in rice production, and in severe cases it can reach 40% to 50%, or even no...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01G22/22A01G13/00A01G7/06A01C1/00A01N63/22A01P3/00C12R1/07
CPCC12N1/20A01G22/22A01G13/00A01G7/06A01C1/00A01N63/22
Inventor 褚晋董海玄元虎贾姝闫晗徐晗梅琼缪建锟杨皓李天亚孙倩郑楠杨眉刘兴远安福涛白元俊
Owner LIAONING ACAD OF AGRI SCI
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