A black fungus strain-specific molecular marker primer, kit and application thereof

A technology of molecular markers and black fungus, applied in the field of microorganisms, can solve the problems of inaccurate identification and achieve specific effects

Active Publication Date: 2021-02-09
GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The invention overcomes the technical problem of inaccurate identification of black fungus strains in the prior art based on morphology, traits and physiological characteristics, and provides a kind of black fungus strain-specific molecular marker primer, kit and application thereof

Method used

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  • A black fungus strain-specific molecular marker primer, kit and application thereof
  • A black fungus strain-specific molecular marker primer, kit and application thereof
  • A black fungus strain-specific molecular marker primer, kit and application thereof

Examples

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Effect test

Embodiment 1

[0020] A molecular marker primer pair, the primer pair is named 3-ty026-3; its upstream primer sequence is: 5'-CGAGAGGCAAGGTGAACTCC-3'; the downstream primer sequence is: 5'-GCTGGCAGTCGATCAAGATC-3'.

Embodiment 2

[0022] Application of primer pair 3-ty026-3 in the identification of black fungus strain Guiyun No.3. Among them, the preservation number of black fungus strain Guiyun No. 3 is CGMCC No.15367.

Embodiment 3

[0024] A method for identifying black fungus strain Guiyun No. 3 from multiple black fungus strains using the primer pair 3-ty026-3, the method is: extracting the total DNA of the black fungus strain to be screened, using the primers 3-ty026-3 was amplified by PCR, and when a 445bp fragment was amplified, it was determined that the strain was the black fungus strain Guiyun No. 3.

[0025] In the method, the reaction procedure of the PCR amplification is: pre-denaturation at 95°C for 5 min, denaturation at 95°C for 30 sec, annealing at 60°C for 30 sec, extension at 72°C for 60 sec, 30 cycles, extension at 72°C for 7 min, and storage at 4°C for 30 min;

[0026] In the method, the PCR reaction system used in the PCR amplification is calculated as 25 μL: 2×EsTaqMasterMix 11.5 μL, ddH2O 10.5 μl, 10 μmol / L upstream primer 1 μl, 10 μmol / L downstream primer 1 μl, black fungus strain to be tested Genomic DNA 1 μL.

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Abstract

The invention discloses a black fungus strain specific molecular marker primer, a test kit and application thereof. The primer pair is named as 3-ty026-3; the sequence of an upstream primer of the primer pair is 5 '-CGAGAGGCAAGGTGAACTCC-3', and the sequence of a downstream primer of the primer pair is 5 '-GCTGGCAGTCGATCAAGATC-3'. The molecular marker primer pair 3-ty026-3 is high in specificity, genome DNA of the black fungus strain Guiyun No.3 can be specifically amplified, the amplification effect on other black fungus strains is avoided, and the black fungus strain Guiyun No.3 can be rapidly and accurately identified through the specific molecular marker primer pair 3-ty026-3.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a black fungus strain-specific molecular marker primer, a kit and an application thereof. Background technique [0002] The black fungus strain Guiyun No. 3 is a new strain isolated, domesticated and cultivated from wild black fungus in Baise. The fruiting body of the strain has a high foaming rate (over 20.0), and is rich in ferrite (above 166mg / Kg) and polysaccharides (4.15g / 100g); the fruiting body of the strain is single-flaky, with no or few veins, The ear pieces are soft and elastic, and the taste is smooth and smooth, which is deeply loved by consumers. The fresh product is yellowish-brown, bright and transparent; the dry product is dark brown on the ventral surface and gray-black on the back, with excellent appearance and broad market prospects. The strain has applied for a patent (patent application number: 20181126406.2). At present, the identification of black...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6895
Inventor 吴圣进陈雪凤韦仕岩王灿琴
Owner GUANGXI ZHUANG AUTONOMOUS REGION ACAD OF AGRI SCI
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