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A kind of biological agent for controlling clubroot of cruciferous crops and its application

A technology of cruciferous crops and biological preparations, applied in the field of applied microorganisms, can solve problems such as inability to increase crop yields, changes in microbial diversity, and poor soil microorganisms, and achieve low bacterial liquid application doses, low application concentrations, and simple operations Effect

Active Publication Date: 2020-08-25
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, using bio-control bacteria alone to control clubroot often has poor control effect and cannot increase the yield of crops, and long-term use of bio-control bacteria will inevitably lead to changes in microbial diversity in the soil and have a negative impact on soil microorganisms

Method used

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  • A kind of biological agent for controlling clubroot of cruciferous crops and its application
  • A kind of biological agent for controlling clubroot of cruciferous crops and its application
  • A kind of biological agent for controlling clubroot of cruciferous crops and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: Bacillus amyloliquefaciens strain Bacillus amyloliquefaciens Screening Isolation of KM 68

[0021] LB medium formula: tryptone 10 g / L, yeast extract 5 g / L, sodium chloride 5 g / L, agarose 15 g / L; pH value is 7.4±0.1; Put it into a Erlenmeyer flask, sterilize it at 121°C under high temperature and high pressure, and pour it into a plate;

[0022] Weigh 1 g of the soil sample collected in Songming County, Yunnan Province, put it into a conical flask with glass beads filled with 50 mL of sterile water, shake it well for 30 min, and then use a pipette to draw 1 g from the conical flask. mL of the supernatant was added to another test tube filled with 9 mL of sterile water and mixed. According to this method, the diluted concentration was 10% of the concentration of the original solution. -3 、10 -4 、10 -5 times the soil solution, and spread 100 uL of the diluted soil solution on the LB medium plate, repeat 3 times for each dilution, culture on a shaker at ...

Embodiment 2

[0023] Embodiment 2: Bacillus amyloliquefaciens strain Bacillus amyloliquefaciens Morphology of KM 68

[0024] Preparation of Bacillus amyloliquefaciens Strains by Gradient Dilution Bacillus amyloliquefaciens For the bacterial suspension of KM 68, draw 10 uL of different dilutions of the bacterial suspension and spread it on the LB medium plate for cultivation. After 8-15 hours, the size, shape, surface, edge, color and properties such as transparency;

[0025] Different dilutions of Bacillus amyloliquefaciens strains were cultured on LB medium Bacillus amyloliquefaciens For KM 68, after culturing for 8-15 hours, draw 10uL of the bacterial suspension to make slices, and observe the strain of Bacillus amyloliquefaciens under an optical microscope Bacillus amyloliquefaciens The growth and morphological characteristics of KM 68;

[0026] Through culture and observation, the bacteria have the following characteristics: the colony is round or oval, wrinkled, slightly ...

Embodiment 3

[0027] Embodiment 3: Bacillus amyloliquefaciens strain Bacillus amyloliquefaciens Identification of 16s rDNA Gene Sequence of KM 68

[0028] After a single colony was inoculated in LB medium and cultured for 24 h, 1 uL was taken for colony PCR verification. The primers used for bacterial liquid PCR are as follows: Forward primer 27F: 5 ， -AGAGTTTGATCMTGGCTCAG-3 ， ; reverse primer 1492R: 5 ， -GGTTACCTTGTTACGACTT-3 ， . PCR reaction system: 10xTaq Buffer 2.5 uL, forward primer 0.5 uL, reverse primer 0.5 uL, bacterial solution 1 uL, dNTP (10 mmol / L) 0.5 uL, Taq DNA polymerase 0.5 U, add water to 25uL, PCR amplification conditions : After pre-denaturation at 94°C for 3 min, perform 35 cycles according to the program of 94°C, 30 s, 57°C, 120 s, 72°C, 90 s, and finally extend at 72°C for 10 min. Agarose gel electrophoresis was used for electrophoresis, the nucleic acid dye was EB, and the DNA Mark length was 2000. Finally, the PCR purified product was sent to Yunnan Shuoqing ...

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Abstract

The invention discloses a biological preparation for preventing and treating clubroot of cruciferous crops and an application thereof, belonging to the field of applied microorganisms. The production strain of biological preparation is Bacillus amyloliquefaciens strain Bacillus amyloliquefaciens KM 68, has been deposited in the China Center for Type Culture Collection, and the deposit number is CCTCC NO:M 2017228. The method of the invention is soil solar heat treatment combined with Bacillus amyloliquefaciens strain Bacillus amyloliquefaciens The KM 68 bacterial solution irrigation root treatment method can effectively prevent and cure clubroot of cruciferous crops, which is beneficial to increase the yield of cruciferous crops, and the control method is simple to operate, low in cost, and easy to produce biological preparations in factories.

Description

technical field [0001] The invention relates to a biological preparation for preventing and treating clubroot of cruciferous crops and an application thereof, belonging to the field of applied microorganisms. Background technique [0002] Vegetable clubroot of cruciferous crops is caused by Plasmodium brassica ( Plasmodiophora brassicae Woron) is a worldwide soil-borne disease. At present, most countries and regions have occurred. Due to the strong infectivity of the pathogen, rapid spread and difficult control, clubroot of cruciferous crops has spread rapidly in most areas of my country in recent years, seriously threatening the production of cruciferous crops. [0003] At present, the methods for controlling clubroot of cruciferous crops mainly include: chemical control, disease-resistant breeding, and biological control. Since Plasmodium brassica is an obligate parasite, disease-resistant breeding is theoretically the most effective way to control clubroot in crucifero...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/22A01P3/00A01G13/00C12R1/07
CPCA01G13/00A01N63/00C12N1/20C12N1/205C12R2001/07
Inventor 伊日布斯李勤超祁腾飞严金平
Owner KUNMING UNIV OF SCI & TECH
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