Mushroom 45 bacteria molecular specific mark and its obtaining method and uses

A technology of specific markers and molecular markers, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve the problems of affecting cultivation enthusiasm, loss of mushroom farmers, and affecting the rapid development of shiitake mushrooms, etc.

Inactive Publication Date: 2007-12-12
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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  • Mushroom 45 bacteria molecular specific mark and its obtaining method and uses

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Embodiment 1

[0024] Mycelia culture and extraction of genomic DNA

[0025] The 45 strains of Lentinus edodes preserved in low temperature were transferred to the 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 20 ng / μL, and stored in a -20°C refrigerator for later use.

[0026] SCAR-PCR amplification

[0027] The total volume of the amplification system is: 25μL, 10×PCR buffer 2.5μL, 25mmol / L MgCL 2 2μL, 10mmol / LdNTP 0.25L, 2.5U / μL Taq DNase 0.5μL, 10μmol / L 45 strain detection primer pair (5'GCCCTTCAGCTAACCCAAA 3' and 5'CTTCCCGTCGTACACTCG 3') each 1μL, template DNA 1μL (concentration 1ng~10ng / μL), ddH 2 O 18.6 μL.

[0028] PCR reaction conditions: 94°C 1min; 94°C 15second, 61°C 15second, 72°C 1min, 30 cycles; 72°C 5min.

[0029] In...

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Abstract

The invention relates to a molecular special mark for mushroom 45 bacterial and the preparation method and application. The section of molecular marked DNA of mushroom 45 bacterial is 604 bp, the specific PCR augmentation primer squence is 5'GCCCTTCAGCTAACCCAAA 3'and 5'CTTCCCGTCGTACACTCG 3'. The method comprises following steps: culturing bacterial filament and extracting genetic DNA; getting specific DNA section of mushroom 45 bacterial; getting specific PCR augmentation primer; SCAR- PCR augmentation; and ionophresis detection. The molecular mark can be used for fast determination and detection for mushroom 45 bacterial.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a molecular specific marker of mushroom 45 strains, an obtaining method and application thereof. 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 very important, whi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12N15/10
Inventor 宋春艳谭琦陈明杰尚晓冬潘迎捷
Owner 上海市农业科学院食用菌研究所
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