Method for screening bio-control bacteria of cucumber fusarium wilt

A technology for cucumber fusarium wilt and biocontrol, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, horticultural methods, etc. disease capacity, the effect of reducing the incidence of

Inactive Publication Date: 2013-10-30
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, not all plants can obtain tissue culture seedlings, and the cultivation of tissue culture seedlings is time-consuming and laborious, and the nutrient substrate involved is different according to different plants, which is not suitable for wide application

Method used

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  • Method for screening bio-control bacteria of cucumber fusarium wilt
  • Method for screening bio-control bacteria of cucumber fusarium wilt
  • Method for screening bio-control bacteria of cucumber fusarium wilt

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Preparation of culture medium

[0024] 1. MS solid medium

[0025] MS solid medium (pH 6.2) is obtained by adding 0.45 g agar per 100 mL of MS liquid medium.

[0026] MS liquid medium is obtained by dissolving the solute and diluting to 1L with water; see Table 1 for the solute and its concentration.

[0027] Table 1 The concentration of each solute in MS liquid medium

[0028]

[0029]

[0030] 2. Preparation of PDA liquid medium

[0031] Cut 200g potato into small pieces and add water to boil it to soften without rotten. Filter the filtrate with four layers of gauze, then add 20g glucose, and finally distilled water to make the volume to 1000ml to obtain PDA liquid medium (natural pH).

[0032] 3. PDA solid medium

[0033] Add 15-20 grams of agar per liter of PDA liquid medium to form PDA solid medium.

Embodiment 2

[0034] Example 2. Obtaining the bacteria to be tested

[0035] In June 2011, soil was taken from the cucumber test field of China Agricultural University, and 400 purely cultured strains were obtained by using the gradient plate dilution method. Fifty-one strains were randomly selected as the bacteria to be tested, and screened in Example 3 respectively.

Embodiment 3

[0036] Example 3 Screening of biocontrol bacteria for cucumber fusarium wilt

[0037] The name of the cucumber seed variety is Xintaimi thorn, which was purchased from Xintaimi thorn cucumber plantation in Shandong Province.

[0038] 1. Preparation of pathogenic spore suspension

[0039] 1. Use a hole punch with a diameter of 0.5 cm to beat the Fusarium oxysporum cucumber specialization type cultured on the PDA solid medium into a bacterial cake, inoculate 5-8 slices of the bacterial cake into 250mL PDA liquid medium, 26℃ Cultivate with shaking for 4 days, filter with three layers of gauze under sterile conditions, and collect the filtrate (i.e. spore liquid, relatively uniform sickle-shaped megaspores can be observed under microscope).

[0040] 2. Centrifuge the filtrate of step 1 (5000rpm for 5 minutes), collect the precipitate (ie spores), wash the spores with sterile water, and then count the spores with sterile water to a concentration of 2×10 8 CFU ml -1 The mother liquid of pat...

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Abstract

The invention discloses a method for screening bio-control bacteria of cucumber fusarium wilt, and provides a method for identifying the bio-control nature of to-be-identified bacteria to pathogenic bacteria of the cucumber fusarium wilt including experimental treatment. The experiment treatment sequentially includes following steps of 1), accelerating sprouting of sterilized cucumber seeds to obtain the sprouting seeds; 2), subjecting the sprouting seeds to soaking in suspension liquid of the to-be-identified bacteria prior to cultivating the sprouting seeds till radicle length reaches by 1.5-2.0cm; 3), inoculating cucumber seedlings to suspension liquid of the pathogenic bateria of the cucumber fusarium wilt; 4), transferring the cucumber seedlings to MS solid media to cultivate, and identifying the bio-control nature of the to-be-identified bacteria to the patheogenic bacteria of the cucumber fusarium wilt by surveying disease indexes and/or observing disease phenotypes. The method is applicable to screening the bio-control bacteria of the cucumber fusarium wilt. In the method, the cucumber seedlings used as samples can truly reflect disease prevention effect of the bio-control bacteria, and can screen bacterial strains of various bio-control mechanisms.

Description

Technical field [0001] The invention relates to a method for screening cucumber wilt biocontrol bacteria. Background technique [0002] Choosing an appropriate screening method for bacteria is the key to successfully obtaining biocontrol strains that can effectively control plant diseases. Ideally, the screening of biocontrol bacteria on plants is more effective than the biocontrol strains obtained by cell culture or other screening methods in vitro. However, large-scale screening on a large number of plants is a waste of time and labor. Therefore, the traditional dual culture method has always been one of the very popular screening methods. In this method, strains with antagonistic activity can form an obvious inhibition zone with pathogens on the PDA plate, but those strains that have no obvious antagonistic activity but may have better control effects in the actual control process pass this screening The method is not available. [0003] Sterile seedlings have been used in t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01G1/00A01G7/06A01G31/00C12N1/20C12R1/125C12R1/77
Inventor 王琦李丽华李燕付学池
Owner CHINA AGRI UNIV
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