SSR (simple sequence repeat) marker fingerprint chromatogram of shiitake Shenxiang-12 strain, and construction method and application thereof
A fingerprint and construction method technology, which is applied in the field of detection of mushroom strains, can solve the problems of affecting the enthusiasm of cultivation, affecting the rapid development of mushrooms, and the loss of mushroom farmers, and achieves good application prospects, short detection time and high accuracy.
Inactive Publication Date: 2017-05-31
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] 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, m...
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The invention relates to an SSR (simple sequence repeat) marker fingerprint chromatogram of a shiitake Shenxiang-12 strain, and a construction method and application thereof. The fingerprint chromatogram is composed of 8 pairs of SSR markers. The construction method comprises the following steps: (1) mycelium culture; (2) genome DNA (deoxyribonucleic acid) extraction; (3) SSR molecular marker detection; and (4) electrophoresis detection. The application comprises the following steps: performing SSR marker amplification on a shiitake strain, and comparing the obtained banding pattern with the fingerprint chromatogram, wherein the shiitake strain is a shiitake Shenxiang-12 strain in case of conforming to the fingerprint chromatogram. Compared with the conventional morphological detection, antibiosis tests and fruiting tests, the method provided by the invention has the advantages of short detection time, high accuracy and good repeatability; and the Shenxiang-12 strain SSR fingerprint chromatogram constructed by the invention is specific for the collected 40 shiitake varieties mainly cultivated in China for years.
Description
technical field [0001] The invention belongs to the detection field of mushroom strains, and in particular relates to an SSR-marked fingerprint of mushroom strain No. 12 Shenxiang 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...
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IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/156
Inventor 谭琦尚晓冬宋春艳章炉军董慧张美彦于海龙姜宁李玉周峰
Owner SHANGHAI ACAD OF AGRI SCI