Molecular specific detection marker and detection method of 0229 straw mushroom strain

A technology for detection of markers and detection methods, applied in the direction of DNA / RNA fragments, recombinant DNA technology, etc., can solve the problems of narrow genetic background, differences in agronomic traits, single varieties, etc., achieve high accuracy, short detection time, and speed up research and development Effect

Active Publication Date: 2017-10-31
SHANGHAI ACAD OF AGRI SCI
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Problems solved by technology

[0003] Our country has a wide area of ​​straw mushroom cultivation, and its output once ranked fifth among edible fungi, but the variety is single. After testing the mating gene types of the main cultivated straw mushroom strains, it was found that most of the existing strains originated from wild grasses in the 1970s. Mushroom isolate strain V23, obtained through systematic breeding, has a relatively narrow genetic background, but has great differences in agronomic traits

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  • Molecular specific detection marker and detection method of 0229 straw mushroom strain
  • Molecular specific detection marker and detection method of 0229 straw mushroom strain

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Embodiment 1

[0023] DNA extraction:

[0024] Inoculate the PDA plate medium with the strain of straw mushroom, culture it at 32°C for 5 days, scrape the surface mycelia and put it into a sterile centrifuge tube, store it in a -70°C refrigerator, and then use the improved CTAB method to extract genomic DNA (the method is briefly described as follows: Grind the freeze-dried mycelia into powder, add 2×CTAB extract preheated at 65°C, dispense into 2mL centrifuge tubes, keep warm at 65°C for 45min, and shake gently occasionally; centrifuge at 4°C, 12000rpm for 20min, and take the supernatant Add an equal volume of chloroform: isoamyl alcohol (24:1) mixture, mix gently, and centrifuge at 12000rpm for 20min at 4°C after 15min. Take the supernatant and transfer it to a new centrifuge tube, add 2 / 3 volume -20°C Pre-cooled isopropanol, shake gently, centrifuge at 4°C, 8000rpm for 10min after 5min. Discard the supernatant, wash the pellet with 75% ethanol (containing 10mmol / L NaAc) three times, centr...

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Abstract

The present invention relates to a molecular-specific detection marker and a detection method thereof for the straw mushroom strain 0229, which is characterized in that: the specific detection marker is a specific primer, wherein the primer sequence is: F: CTTTCAGGAAGAGCCCAGC; R: TGAAGGGCTTGAAGGAGAAA. The detection method includes: inoculate the PDA plate medium with the straw mushroom strain, culture at 32°C for 5 days, scrape the surface mycelium and put it into a sterile centrifuge tube, store it in a refrigerator at -70°C, and then use the improved CTAB method to extract genomic DNA to obtain DNA: the above DNA is used as a template, and the specific detection marker primer is used as an amplification primer, and PCR amplification is performed to obtain a PCR amplification product, which is then detected by electrophoresis. The invention has the advantages of short detection time and high accuracy, uses the obtained molecular markers to identify the straw mushroom strain 0229, is beneficial to the development of the straw mushroom breeding work, and accelerates the research and development of new varieties of the straw mushroom.

Description

technical field [0001] The invention belongs to the field of detection marks and detection methods of straw mushroom strains, and in particular relates to a molecular-specific detection mark of straw mushroom strain 0229 and a detection method thereof. Background technique [0002] Straw mushroom (Volvariella volvacea) cultivation originated in China, has a cultivation history of more than 300 years so far, and is widely cultivated in southern my country. Straw mushrooms are delicious, not only rich in nutritional value, but also have an important role in health care, and are deeply loved by consumers. Straw mushroom is a high-temperature cultivated edible fungus. Compared with other edible fungi, its remarkable feature is that its growth rate is extremely fast. It usually only takes 2 weeks from sowing to harvesting. Its annual output has always ranked first in my country's edible fungi industry. Top ten. [0003] Our country has a wide area of ​​straw mushroom cultivation...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 汪虹王莹陈明杰鲍大鹏冯爱萍
Owner SHANGHAI ACAD OF AGRI SCI
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