Method for rapidly screening hybrid strains of straw mushroom by using molecular marker

A hybrid strain and molecular marker technology, applied in the directions of biochemical equipment and methods, microbial determination/inspection, etc., can solve the restriction on the development of straw mushroom hybrid breeding work, the lack of methods for screening hybrid strains, and the lack of locks on the straw mushroom mycelium. In order to achieve good application prospects, speed up research and development work, and shorten the detection time

Inactive Publication Date: 2013-04-24
SHANGHAI ACAD OF AGRI SCI
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Problems solved by technology

[0003] In classical genetic studies, straw mushroom is the only basidiomycete that is identified as belonging to the primary homophyletic reproduction mode among edible fungi, but there are still many controversies about the genetic research of straw mushroom, especially the research on the reproduction mode. The lack of basic theoretical research has restricted the development of straw mushroom hybrid breeding, and brought many difficult...

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  • Method for rapidly screening hybrid strains of straw mushroom by using molecular marker
  • Method for rapidly screening hybrid strains of straw mushroom by using molecular marker
  • Method for rapidly screening hybrid strains of straw mushroom by using molecular marker

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Experimental program
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Effect test

Embodiment 1

[0028] (1) Primer synthesis

[0029] The specific primers designed for the gene sequence of the mating type A site of the monokaryotic strain A1 of volvulus volvulus are A1F:

[0030] AGGGCATTCCAACCTATTCGCTTTC;

[0031] A1R: AATGTGAACAGTTTGAGCGGAGT.

[0032] (2) Straw mushroom mycelium culture

[0033] The monosporus strain of volvoxin was inoculated on PDA plate medium and cultured at 32°C; the composition of PDA plate medium was: 200g of sliced ​​potato juice filtrate was added, 20g of glucose and 20g of agar were added, and the volume was made up to 1000mL with water.

[0034] (3) Determine the mating genotype of monokaryotic strains

[0035] Pick the hyphae of V. volvulus, and use the Phire Plant Direct PCR Kit (FINNZYMES) kit to directly detect the genotype of the mating type of the strain.

[0036] (4) Straw mushroom hybridization test

[0037] Inoculate the monospore strains of the mating type genotype on the PDA plate medium respectively, culture at 32°C for 5 day...

Embodiment 2

[0043] (1) Primer synthesis

[0044] The specific primer designed for the gene sequence of the mating type A locus of the monokaryotic strain A2 of volvulus is A2F: CACTATCTCCTTGCGTTTGTTATG;

[0045] A2R: ATAACCAAACGCCAGAAACACTA.

[0046] (2) Straw mushroom mycelium culture

[0047] The monosporus strain of volvoxin was inoculated on PDA plate medium and cultured at 32°C; the composition of PDA plate medium was: 200g of sliced ​​potato juice filtrate was added, 20g of glucose and 20g of agar were added, and the volume was made up to 1000mL with water.

[0048](3) Determine the mating genotype of monokaryotic strains

[0049] Pick the hyphae of V. volvulus, and use the Phire Plant Direct PCR Kit (FINNZYMES) kit to directly detect the genotype of the mating type of the strain.

[0050] (4) Straw mushroom hybridization test

[0051] Inoculate the monospore strains of the mating type genotype on the PDA plate medium respectively, culture at 32°C for 5 days, punch holes in the ...

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Abstract

The invention relates to a method for rapidly screening hybrid strains of a straw mushroom by using a molecular marker. The method comprises the following steps: (1) synthesis of a primer; (2) mycelium culture of the straw mushroom; (3) determination of the type of a mating type gene of the mononuclear strain; (4) hybridization test of the straw mushroom; and (5) PCR amplification and determination. Compared with methods like conventional morphological detection, antagonism test and fruiting test, the method provided by the invention has the advantages of simple and convenient operation, a short period of detection time and high accuracy; the method can use the molecular marker of the mating type gene to screen the hybrid strains, which is beneficial for crossbreeding of the straw mushroom, so research and development on a novel variety of the straw mushroom are accelerated, and the method has a good application prospect.

Description

technical field [0001] The invention belongs to the field of screening straw mushroom hybrid strains, in particular to a method for quickly screening straw mushroom hybrid strains with molecular markers. 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] In classical genetic studies, straw mushroom is the only basidiomycete that is identified as belonging to th...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 汪虹陈明杰鲍大鹏冯爱萍薛承琴
Owner SHANGHAI ACAD OF AGRI SCI
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