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Lysinibacillus macrolides, microbial agent, biocontrol agent and preparation method and application thereof

A technology of Bacillus lysinus and microbial agent, which is applied in the field of microorganisms, can solve the problems of high cost and difficulty, and achieve the effects of simple and convenient application, reduced pollution, and good control of plant root-knot nematode activity

Active Publication Date: 2020-10-13
HUNAN PLANT PROTECTION INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention provides a long-form lysine bacillus, microbial agent, biocontrol agent and its preparation method and application, in order to solve the existing technical problems of difficulty and high cost in preventing and treating plant nematode diseases

Method used

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  • Lysinibacillus macrolides, microbial agent, biocontrol agent and preparation method and application thereof
  • Lysinibacillus macrolides, microbial agent, biocontrol agent and preparation method and application thereof
  • Lysinibacillus macrolides, microbial agent, biocontrol agent and preparation method and application thereof

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preparation example Construction

[0049] S4. Preparation of fermented liquid: collecting the fermented liquid in the fermentation culture in step S3 to obtain the microbial bacterial agent of Bacillus longum lysinus;

[0050] In this embodiment, the temperature of the slant seed cultivation in step S1 is 25°C-30°C, and the cultivation time is 30h-40h; the temperature of the liquid seed cultivation in step S2 is 25°C-30°C, and the oscillation speed is 180r / min ~250r / min, the cultivation time is 20h~30h; the temperature of the fermentation culture in step S3 is 25℃~30℃, the oscillation speed is 180r / min~250r / min, the cultivation time is 65h~75h, the inoculation amount of seed culture solution 2% to 5%.

[0051] According to another aspect of the present invention, there is also provided a biocontrol agent for Bacillus longum. The above-mentioned microbial agent of Bacillus longum is centrifuged, and the supernatant is collected and filtered to obtain a long-form Biocontrol agent for Bacillus lysinus. Centrifug...

Embodiment 1

[0081] Cultivate overnight with Escherichia coli (JM109) preserved in the laboratory, collect the bacteria, spread on the NGM medium, use slender and soft feathers to spread Caenorhabditis elegans on the NGM medium with Escherichia coli, place Cultivate in an incubator at 20°C to 25°C for 3d to 5d. Before treatment, sterilize Caenorhabditis elegans with 1mg / mL streptomycin sulfate for 15min-20min, and wash with sterile deionized water for 3 times.

[0082] NGM medium: A. Configure the following reagents: 1M KPO 4 Buffer: 108.3g KH 2 PO 4 , 35.6g K 2 HPO 4 Add water to 1 L (autoclave), 1M MgSO 4 (autoclaved), 1M CaCl 2 (autoclaved), 5mg / ml cholesterol (prepared with absolute ethanol, filtered after preparation, and stored at 4°C); B. Weigh 3g NaCl, 2.5g peptone and 20g Agar into a conical flask, add 975mlH 2 O, autoclave, 121°C for 20min; C. When the medium is cooled to 55°C, add 25ml 1M KPO 4 Buffer, 1ml 1MCaCl 2 , 1ml 1M MgSO4, 1ml 5mg / ml cholesterol, mix well.

[0...

Embodiment 2

[0091] Add the biocontrol agent of Bacillus longum C1 into the 24-well plate, add 900 μl to each well, then take 100 eggs of Meloidogyne incognita and mix it evenly with the biocontrol agent of Bacillus longum C1, repeat three times , set beef extract peptone liquid medium as a control; place the 24-well plate in a 25°C incubator, observe and count under a stereomicroscope after 5 days, and calculate the hatching rate and relative inhibition rate of Meloidogyne incognita eggs. In the following formula, root-knot nematode incognita is referred to as nematode for short.

[0092] Egg hatching rate (%) = average number of hatched nematodes ÷ average number of eggs × 100

[0093] Relative inhibition rate (%) = (average number of hatched nematodes in the control group - average number of hatched nematodes in the treatment group) ÷ average number of hatched nematodes in the control group × 100

[0094] The hatching of M. incognita eggs by Lysinacillus longum C1 biocontrol agents is ...

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Abstract

The invention discloses Lysinibacillus macrolides, a microbial agent, a biocontrol agent and a preparation method and application thereof, and the strain is identified as Lysinibacillus macrolides with the strain name of C1. The strain is preserved in the China General Microbiological Culture Collection Center (CGMCC), the preservation number is CGMCC No.18349, and the preservation date is July 29, 2019. The Lysinibacillus macrolides provided by the invention is a novel Lysinibacillus macrolides C1, and the Lysinibacillus macrolides C1 has good activity of preventing and treating plant root-knot nematodes and lays a foundation for research and development of biological prevention and treatment of the plant root-knot nematodes. Moreover, the strain has a good characteristic of promoting crop growth, and provides a basis for the application of microbial fertilizers.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a lysine bacillus longum. In addition, the present invention also relates to a microbial agent comprising the above-mentioned Bacillus longum lysinus, a biocontrol agent, and a preparation method and application thereof. Background technique [0002] Plant-parasitic nematodes (Plant-parasitic nematodes) are a class of obligate plant-parasitic pathogenic nematodes, which commonly occur in the world and cause huge economic losses to the production of agricultural economic crops. The annual economic losses caused by plant-parasitic nematodes worldwide About $150 billion. Root-knot nematode is considered to be the most harmful plant pathogenic nematode. It can parasitize more than 3000 kinds of plants including vegetables, food crops, cash crops, fruit trees, ornamental crops and weeds. After the disease occurs, the yield is generally reduced by 10 % or so, seriously as high as 75%. ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/20A01P5/00A01P21/00C05F11/08C05G3/60C12R1/01
CPCC12N1/20A01N63/20C05F11/08C05G3/60C12R2001/01C12N1/205
Inventor 程菊娥
Owner HUNAN PLANT PROTECTION INST
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