Simple sequence repeat (SSR) marker fingerprint for mushroom strain 7402, establishment method and application of SSR marker fingerprint
A technology of fingerprint spectrum and construction method, applied in the field of detection of Lentinus edodes strains, can solve the problems of affecting cultivation enthusiasm, loss of mushroom farmers, affecting the rapid development of Lentinus edodes, etc., and achieves the effects of good application prospects, high accuracy and short detection time.
Active Publication Date: 2017-06-06
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
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Embodiment 1
[0043] (1) Mycelia culture: transfer the mushroom strains to the Erlenmeyer flask equipped with 100ml potato dextrose medium PDB, culture in a shaker flask in the dark at 23-25°C, and collect the mycelia after 3-5 days;
[0044] (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;
[0045] The CTAB method for extracting the genomic DNA from mycelium includes:
[0046] a. 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;
[0047] b Centrifuge at 12000rpm, 4°C for 20min, and take the supernatant;
[0048] c. Add a mixture of phenol, chloroform and isoamyl alcohol with a volume ratio of 25:24:1 to the above supernatant, ...
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The invention relates to a simple sequence repeat (SSR) marker fingerprint for a mushroom strain 7402, an establishment method and application of the SSR marker fingerprint. The SSR marker fingerprint is formed by eight pairs of SSR markers. The establishment method comprises the steps of (1) carrying out mycelium culture; (2) extracting genomic deoxyribonucleic acid (DNA); (3) detecting the SSR markers; (4) carrying out electrophoresis detection. The application comprises the steps of carrying out SSR marker amplification on the mushroom strain, comparing the obtained banding pattern with the fingerprint, and determining that the banding pattern is the mushroom strain 7402 if the banding pattern is consistent with the fingerprint. Compared with the conventional morphological detection, antagonism tests and fruiting experiments, the SSR marker fingerprint has the advantage of being short in detection time, high in accuracy and good in repeatability; the SSR marker fingerprint, established by the method, for the strain 7402 has specificity in the main forty cultivation species which are collected in China through the years.
Description
technical field [0001] The invention belongs to the field of detection of mushroom strains, and in particular relates to an SSR-marked fingerprint of mushroom strain 7402 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 7.66 million tons in 2015, 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 ...
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IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 徐珍王瑞娟尚晓冬宋春艳张美彦于海龙章炉军谭琦姜宁董慧李玉周峰
Owner SHANGHAI ACAD OF AGRI SCI