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Method for extracting wheat powdery mildew DNA from catching band of spore catcher

A technology of wheat powdery mildew and trap, which is applied in the field of plant diseases, can solve the problems of not being able to meet the requirements of accurate and rapid disease monitoring, and can not use powdery mildew resistance monitoring, etc., to achieve strong practicability, wide application and accurate results Effect

Inactive Publication Date: 2013-08-21
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the conventional method of identification and counting of spore species is based on the morphological characteristics of the spores under a microscope. Judging only based on morphological characteristics is prone to misjudgment and cannot meet the accurate and rapid requirements of disease monitoring.
In addition, wheat powdery mildew is an obligate parasitic fungus, which can only be preserved and cultivated on live wheat, so the conidia of wheat powdery mildew collected by spore traps cannot be used for monitoring the resistance of powdery mildew
At present, there is no research report on the extraction of conidia DNA of powdery mildew wheat from the capture zone of the spore trap

Method used

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  • Method for extracting wheat powdery mildew DNA from catching band of spore catcher
  • Method for extracting wheat powdery mildew DNA from catching band of spore catcher

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: The traditional CTAB method extracts the DNA of wheat powdery mildew on the spore trap

[0026] 1) Sample preparation

[0027] The strips retrieved from field spore traps were cut into 48 mm segments.

[0028] 2) Capture the elution and wall breaking of powdery mildew spores on the belt

[0029] Cut the capture band into 6 sections, put it into a 2ml centrifuge tube, add 400μl Nonidet P-40 and 0.2g glass beads with a diameter of 400-600μm, and incubate at 65°C for 30min. exist Vibrate twice for 40s at 6m / s on the instrument, place it on ice for 2min, then vibrate twice for 40s at 6m / s, the conidia of powdery mildew can be eluted from the capture zone, and the spores of the broken bacteria can be obtained suspension.

[0030] 3) DNA extraction and purification

[0031] The eluted and broken spore suspension was transferred to another centrifuge tube, and the DNA of powdery mildew conidia was extracted and purified according to the traditional CTAB method...

Embodiment 2

[0036] Embodiment 2: DNA extraction kit method extracts spore catcher to capture the DNA of wheat powdery mildew on the belt

[0037] 1) Sample preparation

[0038] The strips retrieved from field spore traps were cut into 48 mm segments.

[0039] 2) Capture the elution and wall breaking of the powdery mildew conidia on the belt

[0040] Cut the capture band into 6 sections, put it into a 2ml centrifuge tube, add 400μl Nonidet P-40 and 0.2g glass beads with a diameter of 400-600μm, and incubate at 65°C for 30min. exist Vibrate twice for 40s at 6m / s on the instrument, place it on ice for 2min, then vibrate twice for 40s at 6m / s, the conidia of powdery mildew can be eluted from the capture zone, and the spores of the broken bacteria can be obtained suspension.

[0041] 3) DNA extraction and purification

[0042] The eluted and broken spore suspension was transferred to another centrifuge tube, and the DNA extraction and purification of powdery mildew conidia were carried o...

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Abstract

The invention relates to a method for extracting wheat powdery mildew DNA from the catching band of a spore catcher, and belongs to the plant disease prevalence monitoring field, wherein the method is used for extracting the DNA of powdery mildew conidiospore collected by the spore catcher. The method comprises the following steps: eluting and wall-breaking wheat powdery mildew conidiospores collected on the catching band of the spore catcher, and extracting and purifying DNA of the wall-broken conidiospores to develop the disease outbreak and prevalence monitoring service through utilizing a molecular biological quantitative research technology. The DNA extracted in the invention can be used for accurately and quantitatively detecting the wheat powdery mildew conidiospores collected with the spore catcher, and can also be directly used for the drug resistance monitoring. The method can be popularized in various regions as a disease monitoring technology.

Description

(1) Technical field [0001] The "method for extracting wheat powdery mildew DNA from the capture zone of a spore trap" of the present invention can be used to extract the DNA of powdery mildew conidia collected by the spore trap, and belongs to the field of plant diseases. (2) Background technology [0002] Wheat powdery mildew caused by the obligate parasitic fungus Blumeria graminis f.sp.tritici is one of the important diseases in wheat production in my country. Improving the prediction level of this disease can effectively control the prevalence of the disease and reduce the use of pesticides. Wheat powdery mildew is a typical airborne fungal disease. The number or density of powdery mildew conidia in the air is an important driving variable for the occurrence and prevalence of the disease. There is a very important relationship between the popular speed, popular length and distribution. Wheat powdery mildew can generally re-infect for 5 to 6 generations in spring. The sp...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12R1/645
Inventor 曹学仁周益林骆勇段霞瑜陈万权
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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