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Molecular specific mark of centinus lepideus 0820 bacterial and its obtaining method and application

A technology of leopard skin shiitake mushrooms and specific markers, which is applied in biochemical equipment and methods, microbiological measurement/inspection, DNA/RNA fragments, etc., can solve the problems of mushroom farmers’ losses, influence on cultivation enthusiasm, and affect the rapid development of shiitake mushrooms, etc., and achieve The effect of short detection time and high accuracy

Inactive Publication Date: 2007-12-19
上海市农业科学院食用菌研究所
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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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  • Molecular specific mark of centinus lepideus 0820 bacterial and its obtaining method and application

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Embodiment 1

[0023] (1) Mycelia culture and genomic DNA extraction: the cryopreserved Leopard skin mushroom 0820 strain was transferred to a PDA slant, and cultured and activated at 25°C. Two weeks later, they were placed in 100mL PD liquid medium, cultured with shaking at 100r / min at 25°C for two weeks, mycelia were collected, and stored in a -20°C refrigerator. Genomic DNA was extracted by the improved CTAB method, the DNA was diluted to 20ng / μL, and stored in a -20°C refrigerator for later use.

[0024] (2) Obtaining the specific DNA fragment of Leopard-skin Lentinus 0820 strain: Using PCR technology, after a large number of screening tests, the specific DNA fragment of Leopard-skin Lentinus 0820 was obtained using simple repeat sequences as primers, and the fragment was cloned and sequenced.

[0025] (3) Specific PCR amplification primers: Based on the obtained DNA sequence, design specific PCR amplification primers. The sequences of the primer pair are as follows: 5'AGGTCTATGTTCAGGAAG...

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Abstract

The size of molecule label DNA fragment of the pantherine shii-take 0820 spawn is 400bp, and its specific PCR magnification primer couple sequence is 5, TGGCATGAGCATCTGGTCTG3 and 5, CGTATCGTTTAGGAAGGGTG3, The procedure for obtaining the same are: hyphae culture and genomic DNA extraction; obtaining specific DNA fragment of spawn of the pantherine shii-take 0820; obtaining magnification primer of specific PCR; magnification of SCAR-PCR; and electrophoretic testing. This inventive molecular labeling method can be used for short-term identification and test of pantherine shii-take 0820.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a molecular specific marker of leopard skin shiitake mushroom 0820 strain and its obtaining 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 ...

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11C12N15/10
Inventor 谭琦陈明杰尚晓冬宋春艳秦莲花潘迎捷
Owner 上海市农业科学院食用菌研究所
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