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Bilateral molecular marker identification technology of melon wilt

A technology of melon wilt and molecular markers, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve misjudgment, inaccurate disease resistance of melon wilt, and the influence of temperature and humidity on inoculation and observation time And other issues

Inactive Publication Date: 2013-08-14
邵元健
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AI Technical Summary

Problems solved by technology

[0009]Technical problem to be solved: In order to overcome the inaccurate shortcomings of artificial inoculation to identify the resistance of muskmelon wilt, the identification result is easily affected by the concentration of spores used for inoculation. , inoculation and observation time, temperature and humidity during the identification period, resulting in misjudgment

Method used

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  • Bilateral molecular marker identification technology of melon wilt
  • Bilateral molecular marker identification technology of melon wilt

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Embodiment Construction

[0018] 1. DNA extraction

[0019] DNA was extracted by CTAB method. The DNA extraction effect is better than the SDS extraction method, and the sugar removal effect is obvious. The specific method is as follows:

[0020] (1) Take a small amount of leaves and place them in an Eppendorf tube (about 70-100 mg), and freeze them in liquid nitrogen for 1 min;

[0021] (2) Open the cap of the tube and mash the young leaves with a small wooden stick (note: mash for a while and then put it in liquid nitrogen, then mash it after freezing until it is very broken, generally 3 times); then, put Put it into 3 small steel ball tubes, shake the oscillator 3-4 times;

[0022] (3) Depending on the amount of material, add 600-700 ul of preheated DNA extraction solution (CTAB), mix gently, and place in a 65°C water bath for 25 minutes, shaking 2-3 times during the process;

[0023] (4) Add an equal volume of phenol / chloroform (600-700ul) into the tube, invert it upside down for 5 minutes to m...

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Abstract

The invention relates to a bilateral molecular marker identification technology of a melon wilt resistant variety. By the utilization of polymerase chain reaction and bilateral molecular markers ML430 and MR296 which are closely linked to a melon wilt resistance gene, specific DNA fragments are amplified. The size differences of amplified DNA fragments between a resistant variety and a susceptible variety can be displayed on different locations of a sepharose gel by electrophoretic technique. And the sizes and locations are compared with that of DNA fragments amplified from a standard resistant variety and a standard susceptible variety. Two varieties which have the same size or lie in the same location belong to the variety of the same type. When banding patterns of the two molecular markers both are resistance type, the variety is the resistant variety. The method provided by the invention has advantages of convenient sampling, precise identification, and no influence by plant growth and development and weather conditions. The technology provided by the invention is mainly used for the screening and identification of wilt resistance variety resources, and can also be used for molecular marker assisted breeding of melon.

Description

[0001] Technical field [0002] [0003] The technical patent of this invention belongs to the field of crop biotechnology breeding. This technology uses polymerase chain reaction (PCR technology) to amplify the DNA fragment of a specific segment of the melon genome with the help of double-sided molecular markers ML430 and MR296 closely linked with the Fusarium wilt resistance gene . The size of the DNA fragment amplified by the disease-resistant variety and the susceptible variety is different, which can be shown on the picture by electrophoresis technology, and the disease resistance of the identified variety to Fusarium wilt can be judged according to the size of the fragment. Background technique [0004] Melon wilt (Fusarium oxysperium) is one of the serious diseases of melons, often causing death of plants in large areas of melon fields, or even failure of harvest, and the degree of damage increases year by year. According to investigations, the rate of diseased plan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 邵元健
Owner 邵元健
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