SSR fingerprint of Cr02 strain of Lentinus edodes, and construction method and application thereof

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

Active Publication Date: 2013-01-02
山东御苑生物科技有限公司
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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 fingerprint of Cr02 strain of Lentinus edodes, and construction method and application thereof
  • SSR fingerprint of Cr02 strain of Lentinus edodes, and construction method and application thereof
  • SSR fingerprint of Cr02 strain of Lentinus edodes, and construction method and application thereof

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Embodiment 1

[0049] (1) Mycelia culture: transfer the mushroom strains to a conical flask containing 100ml of potato dextrose medium PDL, culture in a dark shaking flask at 23°C~25°C, and collect mycelia after 3d~4d;

[0050] (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;

[0051] The CTAB method for extracting the genomic DNA from mycelium includes:

[0052] ① Grind the mycelium of shiitake mushrooms freeze-dried at -20°C into powder, add 2×CTAB extract preheated at 65°C, keep warm at 65°C for 45~60min, and shake gently occasionally;

[0053] ② Centrifuge at 12000rpm, 4°C for 20min, and take the supernatant;

[0054] 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 minute...

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Abstract

The invention relates to an SSR fingerprint of the Cr02 strain of Lentinus edodes, and a construction method and application thereof. The fingerprint is composed of 7 pairs of SSR markers. The construction method comprises the following steps: (1) culture of mycelia; (2) extraction of genome DNA; (3) detection of the SSR molecular markers; and (4) electrophoresis detection. Application of the SSR fingerprint is as follows: SSR marker amplification is carried out on a strain of Lentinus edodes, an obtained banding pattern is contrasted with the fingerprint, and it is determined that the strain of Lentinus edodes is the Cr02 strain of Lentinus edodes if the banding pattern is identical with the fingerprint. Compared with conventional morphological detection, antagonism test and fruiting test, the invention has the advantages of a short detection time, high accuracy and good repeatability, and the Cr02 strain of Lentinus edodes has specificity among collected 25 dominant strains of Lentinus edodes having passed examination in China.

Description

technical field [0001] The invention belongs to the field of detection of shiitake mushroom strains, in particular to a SSR marker fingerprint of shiitake mushroom Cr02 strain and its construction method and application. 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 张丹章炉军巫萍谭琦尚晓冬宋春艳鲍大鹏杨慧
Owner 山东御苑生物科技有限公司
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