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Molecular mark, detecting method and application of mushroom Xiangjiu strain

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

Inactive Publication Date: 2011-06-01
SHANGHAI ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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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 mark, detecting method and application of mushroom Xiangjiu strain
  • Molecular mark, detecting method and application of mushroom Xiangjiu strain

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Experimental program
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Embodiment 1

[0035] 1. Mycelia culture

[0036] The shiitake mushroom strains preserved at 4°C were transferred to PDA plates and cultured at 25°C in the dark. After 14 days, they were placed in 100 mL PDY medium, 150 rpm, and cultured in shake flasks at 25°C. When the mycelia grow to a certain amount, the mycelia are collected and put into a -20°C refrigerator for freezing.

[0037] 2. Genomic DNA extraction

[0038] Genomic DNA was extracted using a modified CTAB method. The concentration and purity of the total genomic DNA were detected by ultraviolet spectrophotometry, the concentration of the sample DNA was adjusted to be consistent, and the samples were refrigerated for later use.

[0039] 3. Detection of SCAR molecular markers

[0040]Amplification system (total volume 25 μL): 10×PCR buffer 2.5 μL, 25 mmol / LMgCl 2 2μL, 10mmol / LdNTP0.25L, 2.5U / μL Taq DNase 0.5μL, 10μmol / L Xiangjiu F / R each 1μL, template DNA 1μL (concentration 1ng~10ng / μL), ddH 2 O 18.6 μL.

[0041] PCR reactio...

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Abstract

The invention relates to a molecular mark of mushroom Xiangjiu strains, the detecting method and the application thereof. The molecular mark is a molecular specific detecting mark of gene SCAR, and a PCR special primer pair thereof has the sequence that XiangjiuF:5<1>GGTCCATGTATAGTCTG 3<1> and XiangjiuR:5<1>GTCCATTGGTTCAAAC 3<1>. The detecting method comprises the steps that are (1) mycelium culture, (2) the extraction of genome DNA, (3) the detection of the SCAR molecular mark, and (4) electrophoresis detection. The method has the application that the gene SCAR in mushroom strains is expanded, and the mushroom strains with SCAR marks are the mushroom Xiangjiu strains. Compared with the conventional morphological detection, an antagonism test and a fruiting test, the molecular mark of themushroom Xiangjiu strains, the detecting method and the application thereof have the advantages of short detecting time and high accuracy, and have specificity of Xiangjiu strains in the thirty collected major mushroom strains in China.

Description

technical field [0001] The invention belongs to the detection field of shiitake mushroom strains, and in particular relates to a molecular marker of shiitake mushroom strains, a detection method and an 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 i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04C12Q1/68C12N15/11
Inventor 宋春艳谭琦尚晓冬陈明杰
Owner SHANGHAI ACAD OF AGRI SCI