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SSR-labeled fingerprint of shiitake mushroom Minfeng No.1 strain, and application thereof

A technology of fingerprints and shiitake mushrooms, which is applied in the field of detection of shiitake mushroom strains, can solve the problems of affecting cultivation enthusiasm, loss of mushroom farmers, and affecting the rapid development of shiitake mushrooms, and achieves good application prospects, high accuracy, and short detection time.

Active Publication Date: 2013-01-02
SHANGHAI ACAD OF AGRI SCI
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Problems solved by technology

As far as the country is concerned, the phenomenon of "same species with different names and different species with the same name" in the cultivation of shiitake mushrooms has not only brought great losses to mushroom farmers, but also affected their enthusiasm for cultivation, and also greatly affected the rapid development of shiitake mushrooms in China. and, along with the appearance of large-scale industrialized cultivation methods, the requirements for the quality of the cultivated strains of shiitake mushrooms are getting higher and higher, and it is necessary to develop a more simple, fast and accurate strain identification technology to ensure that each batch of species is used. accurate

Method used

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  • SSR-labeled fingerprint of shiitake mushroom Minfeng No.1 strain, and application thereof
  • SSR-labeled fingerprint of shiitake mushroom Minfeng No.1 strain, and application thereof
  • SSR-labeled fingerprint of shiitake mushroom Minfeng No.1 strain, and application thereof

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Experimental program
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Effect test

Embodiment 1

[0025] (1) Mycelia culture: transfer the shiitake mushroom strains to the potato glucose medium, culture in shake flasks in the dark at 23°C~25°C, and collect the mycelia after 3d~4d;

[0026] (2) Genomic DNA extraction: use CTAB (cetyltrimethylammonium bromide) method to extract the genomic DNA of the above hyphae, detect the concentration and purity of the total genomic DNA by ultraviolet spectrophotometry, and adjust the concentration of the sample DNA to be consistent;

[0027] The CTAB method for extracting the genomic DNA from mycelium includes:

[0028] ① Grind the mycelium of shiitake mushrooms freeze-dried in liquid nitrogen into powder, add 2×CTAB extract preheated at 65°C, keep warm at 65°C for 45-60min, and shake gently occasionally to mix well;

[0029] ②Centrifuge at 12000rpm, 4°C for 20min, and take the supernatant;

[0030] ③ Add a mixture of phenol, chloroform and isoamyl alcohol with a volume ratio of 25:24:1 to the above supernatant, mix gently for 10 minut...

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Abstract

The invention relates to an SSR-labeled fingerprint of shiitake mushroom Minfeng No.1 strain, and an application thereof. The fingerprint is composed of 7 pairs of SSR-labeled specific allelic fragments developed according to a shiitake mushroom genomic sequence. The application comprises the steps that: SSR-labeled amplification is carried out upon a shiitake mushroom strain; and an obtained band type is compared with the fingerprint. If the band type is consistent with the fingerprint, the strain is a shiitake mushroom Minfeng No.1 strain. Compared with a conventional morphological detection method, an antagonistic test, and mushroom fruiting trials, the application provided by the invention has the advantages of short detection time, high accuracy, and good repeatability. In 25 collected nationally validated shiitake mushroom main cultivated strains of our nation, the fingerprint has shiitake mushroom Minfeng No.1 strain specificity.

Description

technical field [0001] The invention belongs to the detection field of shiitake mushroom strains, and in particular relates to an SSR marker fingerprint and application of shiitake mushroom Minfeng No. 1 strain. Background technique [0002] The output of shiitake mushrooms in my country has grown rapidly from 19,500 tons in 1983 to 2.42 million tons in 2005, accounting for more than 70% of the total output of shiitake mushrooms in the world. Some experts predict that due to the rapid development of China's shiitake mushroom industry, it will become the edible fungus with the highest output in the world in the past 10 years. Chinese shiitake mushrooms have attracted the attention of people in the world's mushroom industry for their astonishing development speed, high-quality quality and low cost. Chinese shiitake mushrooms have become popular all over the world. [0003] The contribution rate of high-quality strains to the yield and quality of shiitake mushrooms is very impo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
Inventor 宋春艳谭琦尚晓冬鲍大鹏杨慧章炉军张丹巫萍
Owner SHANGHAI ACAD OF AGRI SCI
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