Method for efficiently screening verticillium dahliae antagonistic bacteria from soil sample

A technology for Verticillium dahliae and soil samples is applied in the field of efficient screening of Verticillium dahliae antagonistic bacteria, can solve problems such as large workload and low efficiency, and achieve the effects of reducing workload, narrowing scope and saving manpower

Inactive Publication Date: 2017-05-31
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to establish a high-efficiency and rapid antagonistic bacteria screening method from soil samples for the practical problems of traditional antagonistic bacteria screening with large workload and low efficiency.

Method used

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  • Method for efficiently screening verticillium dahliae antagonistic bacteria from soil sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Culture medium preparation

[0019] Recipe 1. Common Bacteria LB Medium

[0020] Peptone 10.0g / L, yeast extract 5.0g / L, NaCl 2.0g / L, agar 15-20g / L, pH 7.0-7.5.

[0021] Formula 2, common fungus PDA medium

[0022] Potato 200g / L, glucose 20g / L, agar 15-20g / L.

Embodiment 2

[0023] Example 2 Screening of antagonistic bacteria in soil samples

[0024] a. Collect field diseased soil in Nanjing City, Jiangsu Province, weigh 10g soil sample and add 100mL sterile water containing glass beads, let it stand for 20min, and shake it in a shaker at 200r / min for 30min, which is the soil suspension. Use a sterile pipette gun to draw 5mL of soil suspension and add it to 45mL of sterile water, and perform serial dilution at 1:10 to obtain 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 The dilution, take the appropriate dilution, use a sterile pipette to draw 10 -4 、10 -5 100 μL of each of the two dilution gradients was evenly spread on LB medium plates with a diameter of 15 cm, and three plates were repeatedly coated for each gradient, and the plates were placed in an incubator at 30°C for 2 days.

[0025] b. Verticillium dahliae V 茄nj Inoculate in PDA medium for activation, take the bacterial block at the edge of the colony and inoculate in PDB liquid medium, cu...

Embodiment 3

[0027] Example 3 Purification of antagonistic bacteria

[0028] After 2-3 days of spray inoculation, the plate is covered with V 茄nj hyphae, obvious inhibition zone appeared in the plate (see figure 1 ), pick out 30 strains of antagonistic bacteria in all inhibition zones, streak on another LB plate, purify, place the plate in a 30°C incubator and culture it for 1 day, so that a single colony grows on the plate, and observe each plate The traits such as the shape, size and color of the single colony above, if there are colonies with inconsistent traits, you need to repeat the streak purification process again until the colonies in each plate have the same traits.

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Abstract

The invention relates to a method for efficiently and rapidly screening verticillium dahliae antagonistic bacteria. The method comprises the following steps: firstly, coating a plate with soil suspensions of different gradients of a sample; directly spraying and inoculating verticillium dahliae spore liquid on the plate after bacteria grow; culturing continually; when verticillium dahliae hyphae grow on the plate fully, observing whether an inhibition zone is formed on the periphery of the bacteria or not, wherein if the inhibition zone is formed, the bacteria in the inhibition zone are antagonistic bacteria; picking out the antagonistic bacteria, and scribing and purifying the antagonistic bacteria; performing a plate antagonistic experiment on the antagonistic bacteria; further verifying the action of the antagonistic bacteria. By adopting the method, all bacteria growing from the coated plate do not need to be picked out one by one, so that the workload is reduced greatly compared with the conventional method, the screening efficiency is increased, the screening of the antagonistic bacteria is facilitated, and the development and utilization of biocontrol bacteria are accelerated; the method has important practical application value.

Description

technical field [0001] The invention relates to a high-efficiency screening method for Verticillium dahliae antagonistic bacteria, belonging to the field of biological control research. Background technique [0002] Verticillium dahliae (Verticillium dahliae) is a kind of widely distributed and highly destructive plant soil-borne pathogenic fungi, which can cause Verticillium wilt in host plants worldwide. The pathogen is not only widely distributed in regions, but also has a wide range of hosts. It can infect more than 400 kinds of plants, including annual and perennial crops, vegetables, flowers, fruits and vegetables, woody ornamental plants and fiber oil crops. continue to expand. Verticillium dahliae can cause economic losses of 50% or more on many crops such as cotton, potatoes, lettuce, and olives; up to 75% in unfumigated strawberry fields; and root-knot nematodes Coexistence can result in greater losses. [0003] Verticillium wilt is a soil-borne vascular disease...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/18C12R1/645
CPCC12Q1/18
Inventor 王卿邓晟林玲
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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