Method for separating wheat stem base rot bacteria

A separation method, a technology of stem rot, which is applied in the field of separation of wheat stem rot pathogens, can solve the problems of tissue separation time-consuming, unsuitable for the isolation of pathogenic bacteria of wheat stem rot diseased plants, and high operating requirements, so as to inhibit the growth of bacteria Effect

Inactive Publication Date: 2014-10-08
HENAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the characteristics of time-consuming, laborious, high cost and high operation requirements, the tissue separation method is not suitable for the isolation of pathogenic bacteria from a large number of wheat stem rot diseased plants.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] A kind of separation method of wheat stalk rot pathogen, its operation steps are:

[0027] Step 1. Collect wheat plants with white ear symptoms and brown leaf sheaths or stems at the first and second stem nodes at the base of the stem, and set aside;

[0028] Step 2. Spread the collected wheat plants for 1 day under ventilated and cool conditions indoors; then cut the 1.8cm stem section from the browned part of the first and second stem nodes at the base of the placed wheat plant; Rinse the stem section with clean water, then disinfect it with 75% alcohol for 15 seconds, rinse it twice with sterile water, and set aside;

[0029] Step 3. Put the clipped stems into a Petri dish covered with sterile filter paper at the bottom of the dish, add 1-2mL sterile water to the Petri dish, then place it in a biochemical incubator, and cultivate it in the dark at 25°C 1 day, so that the moisture on the surface of the plant evaporates; until the mycelium grows on the stem, set aside...

Embodiment 2

[0035] A kind of separation method of wheat stalk rot pathogen, its operation steps are:

[0036] Step 1. Collect wheat plants with white ear symptoms and brown leaf sheaths or stems at the first and second stem nodes at the base of the stem, and set aside;

[0037] Step 2, spread out the collected wheat plants in a ventilated and cool room for 2 days; then cut the 2.2 cm stems from the browned parts of the first and second stem nodes at the base of the placed wheat plants; Rinse the stem section with clean water, then disinfect it with 75% alcohol for 15 seconds, rinse it with sterile water for 3 times, and set aside;

[0038] Step 3. Put the clipped stems into a Petri dish covered with sterile filter paper at the bottom of the dish, add 1-2mL sterile water to the Petri dish, then place it in a biochemical incubator, and cultivate it in the dark at 25°C 2 days, so that the water on the surface of the plant evaporates; until the mycelia grow on the stem, set aside;

[0039]S...

Embodiment 3

[0044] A kind of separation method of wheat stalk rot pathogen, its operation steps are:

[0045] Step 1. Collect wheat plants with white ear symptoms and brown leaf sheaths or stems at the first and second stem nodes at the base of the stem, and set aside;

[0046] Step 2, spread out the collected wheat plants for 2 days under ventilated and cool conditions indoors; then cut 2.0 cm stem segments from the browned parts of the first and second stem nodes at the base of the placed wheat plants; Rinse the stem section with clean water, then disinfect it with 75% alcohol for 15 seconds, rinse it twice with sterile water, and set aside;

[0047] Step 3. Put the clipped stems into a Petri dish covered with sterile filter paper at the bottom of the dish, add 1-2mL sterile water to the Petri dish, then place it in a biochemical incubator, and cultivate it in the dark at 25°C 1.5 days, in order to volatilize the water on the surface of the plant; until the mycelium grows on the stem, ...

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PUM

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Abstract

The invention relates to a method for separating wheat stem base rot bacteria. The method comprises the following steps of firstly collecting wheat plants, spreading out and standing for 1-2 days under the ventilated and shade conditions, and cutting a stem of which the length is 1.8-2.2cm from the base where the first and second stem nodes become brown; rinsing with clean water, then sterilizing with 75% alcohol for 15 seconds, and then rinsing with sterile water; then placing in a culture dish of which the bottom is covered with sterile filter paper and adding 1-2mL of sterile water, then placing in a biochemical incubator, and culturing at 25 DEG C under a dark condition until mycelia grow on stems; picking mycelia, inoculating the mycelia on a PSA medium added with lactic acid, and carrying out dark culture in the biochemical incubator of which the temperature is 25 DEG C; picking a small bacterial colony from the edge of a bacterial colony with an inoculation needle, transferring the small bacterial colony onto the PSA slant culture medium, placing in the biochemical incubator, culturing at 25 DEG C under a dark condition for 3 days and carrying out strain preservation, namely, separating the wheat stem base rot bacteria out. The method disclosed by the invention has the advantage of high success rate of separation, the mycelia can quickly grow from the treated stem, typical colonies of the wheat stem base rot bacteria can quickly grow after the mycelia are transferred to the PSA and the success rate reaches up to above 95%.

Description

technical field [0001] The invention relates to a method for isolating a pathogen, in particular to a method for isolating a pathogen of wheat stem rot. Background technique [0002] In theoretical research work such as the morphology, physiology, ecology and pathogenicity of pathogenic fungi, as well as in applied research work such as identification of variety disease resistance and screening of agents that have inhibitory effects on pathogenic bacteria, it is often necessary to Pure culture of pathogenic fungi. The fungal mycelium in the diseased plant tissue can generally resume growth and reproduction if given suitable environmental conditions. However, under natural conditions, pathogenic bacteria are usually mixed with other miscellaneous bacteria, and pathogenic bacteria are isolated from diseased tissues or other substrates, which is called the isolation of plant pathogenic fungi. The isolation of plant pathogenic fungi also plays a very important role in the res...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12R1/77C12R1/645
Inventor 侯颖徐建强夏彦飞范倩倩范利峰
Owner HENAN UNIV OF SCI & TECH
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