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Method for extracting DNA of cyst parasites of heterodera glycines

A soybean cyst nematode and extraction method technology, which is applied in the field of DNA extraction of soybean cyst nematode cyst parasites, can solve the problems of loss and difficulty in providing accurate information, and achieve the effect of simple steps, easy operation, and less cyst volume

Inactive Publication Date: 2015-12-16
NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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  • Application Information

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Problems solved by technology

[0005] The present invention aims to solve the problems that there is a loss in the DNA extraction process of soybean cyst nematode cyst parasites, it is difficult to accurately provide information on the community structure and diversity of parasites, and the cysts are used in large amounts and the extraction time is long. A kind of DNA extraction method of soybean cyst nematode cyst parasite

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  • Method for extracting DNA of cyst parasites of heterodera glycines
  • Method for extracting DNA of cyst parasites of heterodera glycines

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specific Embodiment approach 1

[0016] Specific embodiment 1: The method for extracting DNA from soybean cyst nematode cyst parasites of this embodiment is carried out according to the following steps:

[0017] 1. Take 100-150 fresh soybean cyst nematode cysts, disinfect the surface with NaClO, and put them into a sterilized 2mL centrifuge tube, soak them in sterilized deionized water at 4℃ for 18-24 hours. Obtain cyst samples;

[0018] 2. Use the soil DNA extraction kit to extract DNA from the cyst samples. Refer to the instruction manual of the soil DNA extraction kit for specific operations to complete the DNA extraction of soybean cyst nematode cyst parasites;

[0019] The parasitic rate of soybean cyst nematode cysts in step one is more than 50%.

[0020] The soil DNA extraction kit used in step 2 of this embodiment is E.Z.N.A. Soil DNA Kit, Omega Biotek, Inc., USA.

[0021] The specific operation of DNA extraction in this embodiment:

[0022] 1. Take 0.5 g of glass beads and place them in a centrifuge tube conta...

specific Embodiment approach 2

[0032] Specific embodiment two: this embodiment is different from specific embodiment one in that in step one, 105 fresh soybean cyst nematode cysts are taken. Other steps and parameters are the same as in the first embodiment.

specific Embodiment approach 3

[0033] Specific embodiment three: this embodiment is different from specific embodiment one in that 110 fresh soybean cyst nematode cysts are taken in step one. Other steps and parameters are the same as in the first embodiment.

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Abstract

The invention relates to a method for extracting DNA of cyst parasites of heterodera glycines and belongs to the nucleic acid extraction technique. According to the method, the problems that the community structure and the diversity information related to the parasites cannot be accurately provided because of loss in the current extraction process of DNA of the cyst parasites of heterodera glycines, the consumption of cysts is high, and the extraction time is long are mainly solved. The method comprises the following steps: (1) preparing fresh cysts of heterodera glycines, carrying out surface sterilization, and soaking by virtue of deionized water, so as to obtain a cyst sample; and (2) extracting DNA of the cyst sample by virtue of a soil DNA extraction kit. The extraction method has the beneficial effects that the consumption of the cysts is low, the steps are simple, the operation is easy, the use is convenient, the DNA is rapidly, accurately and efficiently extracted, and the extraction method is convenient to popularize and apply; the extracted DNA can be applied to microbial diversity analysis such as denaturing gel gradient electrophoresis and can be further applied to the extraction of DNA of similar parasitic microbes, so that the diversity and the types of the cyst parasites can be comprehensively understood, and the problem that microorganisms which are difficult to culture cannot be authenticated is solved.

Description

Technical field [0001] The invention belongs to nucleic acid extraction technology, and specifically relates to a method for extracting DNA from soybean cyst nematode cyst parasites. Background technique [0002] Soybean cyst nematode (Heteroderaglycines) is a soil-borne obligate parasitic nematode, belonging to the genus Heterodera. Due to the formation of cysts, it is sometimes called cyst nematode and is the main pathogen of soybean cyst nematode. It is a common disease in soybean production and one of the main limiting factors in soybean production. Soybean cyst nematode control methods mainly include planting resistant varieties, reasonable crop rotation, chemical control and biological control measures. At present, due to the improvement of people's awareness of environmental protection, biological control has gradually become a research hotspot. The use of natural enemies of soybean cyst nematode microorganisms to control the disease has become a new trend and new trend i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 宋洁许艳丽姚钦
Owner NORTHEAST INST OF GEOGRAPHY & AGRIECOLOGY C A S
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