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SSR marker fingerprint spectrum of needle mushroom Qinghe strain and application thereof

A technology of fingerprinting and Flammulina velutipes, which is applied in the direction of microorganism-based methods, measurement/inspection of microorganisms, microorganisms, etc., can solve problems that affect the enthusiasm of cultivation, affect the development of Flammulina velutipes industry, and the loss of mushroom farmers, and achieve good application prospects and detection time Short, accurate effects

Active Publication Date: 2014-11-19
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, different species with the same name" in the cultivation of Flammulina velutipes not only brought great losses to mushroom farmers, but also affected their enthusiasm for cultivation, and also greatly affected the development of China's Flammulina velutipes industry ; And, along with the appearance of large-scale industrialized cultivation mode, the requirement to the quality of Flammulina velutipes cultivation strain is more and more higher, need to develop more simple, fast, accurate bacterial strain identification technology, to guarantee that each batch uses kind accurate

Method used

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  • SSR marker fingerprint spectrum of needle mushroom Qinghe strain and application thereof
  • SSR marker fingerprint spectrum of needle mushroom Qinghe strain and application thereof
  • SSR marker fingerprint spectrum of needle mushroom Qinghe strain and application thereof

Examples

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Embodiment 1

[0024] (1) Mycelia culture: Transfer Flammulina velutipes strains to potato glucose medium, culture in shake flasks in the dark at 19°C to 21°C, and collect mycelia after 3d to 4d;

[0025] (2) Extraction of genomic DNA: extract the genomic DNA of the above-mentioned hyphae with the CTAB (cetyltrimethylammonium bromide) method, detect the total genomic DNA concentration and purity by ultraviolet spectrophotometry, and adjust the concentration of the sample DNA to be consistent;

[0026] The CTAB method for extracting the genomic DNA from mycelium includes:

[0027] ①Grind the mycelium of Flammulina velutipes 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;

[0028] ② Centrifuge at 12000rpm, 4°C for 20min, and take the supernatant;

[0029] 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 minutes, c...

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Abstract

The invention relates to an SSR marker fingerprint spectrum of a flammulina velutipes Qinghe strain and application thereof. The SSR marker fingerprint spectrum of the flammulina velutipes Qinghe strain is combined by six pairs of peculiar allelic fragments, developed on the basis of a flammulina velutipes genomic sequence and marked with SSR. Application of the SSR marker fingerprint spectrum of the flammulina velutipes Qinghe strain comprises that SSR marker amplification is carried out on a flammulina velutipes strain, the obtained banding pattern is compared with the fingerprint spectrum, and if the banding pattern is consistent with the fingerprint spectrum, the flammulina velutipes strain is the flammulina velutipes Qinghe strain. Compared with the conventional morphological detection, the conventional antagonistic test and the conventional fruiting test, the SSR marker fingerprint spectrum of the flammulina velutipes Qinghe strain has the advantages of short detection time, high accuracy and good repeatability and has Qinghe strain specificity in 24 collected major flammulina velutipes strains in China.

Description

technical field [0001] The invention belongs to the detection field of Flammulina velutipes strains, in particular to an SSR marker fingerprint and application of Flammulina velutipes sp. Background technique [0002] Flammulina velutipes (Fr.) Sing.], also known as Flammulina velutipes (Fr.) Sing.], has another name called Flammulina velutipes, Pleurotus velutipes, Structural bacteria, Blue barella, and Trichomonas velutipes. Flammulina velutipes is widely distributed and rich in nutrition. It is an edible and medicinal fungus with high economic value, and it is also one of the natural health foods that are very popular in the market. Flammulina velutipes is one of the main cultivated edible fungi in China and one of the main edible fungi in modern factory production. In 2013, the domestic factory production reached 2,700 tons per day. [0003] The contribution rate of high-quality strains in the yield and quality of Flammulina velutipes is very important, which determines...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12R1/645
CPCC12Q1/6895C12Q2600/156
Inventor 张丹章炉军王瑞娟陆欢徐珍尚晓冬谭琦
Owner SHANGHAI ACAD OF AGRI SCI
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