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SSR labeled fingerprint of flammulina velutipes F3 strain and applications thereof

A technology of fingerprints and Flammulina velutipes, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems affecting the development of Flammulina velutipes industry, affecting the enthusiasm of cultivation, and the loss of mushroom farmers, etc., achieving good application prospects and short detection time , high accuracy effect

Active Publication Date: 2014-11-05
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 labeled fingerprint of flammulina velutipes F3 strain and applications thereof
  • SSR labeled fingerprint of flammulina velutipes F3 strain and applications thereof
  • SSR labeled fingerprint of flammulina velutipes F3 strain and applications thereof

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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] ②Centrifuge 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, ce...

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Abstract

The invention relates to a SSR label fingerprint of a flammulina velutipes F3 strain and applications thereof. The fingerprint is composed of six pairs of SSR labeled specific allelic sections which are developed on the basis of the genomic sequence of flammulina velutipes. The fingerprint can be used to identify the flammulina velutipes F3 strain, and the identification comprises the following steps: carrying out SSR labeling amplification on a flammulina velutipes strain, and then comparing the obtained fingerprint with the fingerprint of flammulina velutipes F3 strain, if the two fingerprints are identical, the flammulina velutipes strain is the F3 strain. Compared to the conventional morphology detection, antagonism tests, and fruiting experiments, the fingerprint identification method has the advantages of short detection time, high accuracy, and good repeatability, and has the specificity on F3 strain in collected 24 flammulina velutipes strains which are commonly cultured in China.

Description

technical field [0001] The invention belongs to the detection field of Flammulina velutipes strains, and in particular relates to an SSR marker fingerprint and application of Flammulina velutipes F3 strains. 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 importa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/6895C12Q2600/156
Inventor 尚晓冬陆欢王瑞娟章炉军徐珍张丹谭琦
Owner SHANGHAI ACAD OF AGRI SCI
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