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Characteristic nucleotide sequence, primer, kit and method for authenticating Flammulina fennae

A technology of nucleotide sequences and kits, applied in biochemical equipment and methods, measurement/inspection of microorganisms, DNA/RNA fragments, etc., can solve the problem of no Fenna velutipes, difficult specificity, and rapid detection of Fenna Flammulina velutipes etc.

Inactive Publication Date: 2016-08-24
GUANGDONG INST OF MICROORGANISM +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There are no identification methods, characteristic nucleotide sequences, primers and kits for Fenna Flammulina velutipes in the prior art, therefore, there is a problem that it is difficult to detect Fenna Flammulina velutipes specifically and quickly

Method used

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  • Characteristic nucleotide sequence, primer, kit and method for authenticating Flammulina fennae
  • Characteristic nucleotide sequence, primer, kit and method for authenticating Flammulina fennae
  • Characteristic nucleotide sequence, primer, kit and method for authenticating Flammulina fennae

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Embodiment 1

[0085] Embodiment 1: the cultivation method of Fenna Flammulina velutipes new strain CCTCC M 2016179 and the fruiting body and mycelia obtained

[0086] 1. The cultivation method is as follows:

[0087] 1. Culture medium production:

[0088] (1) The separation medium is: 20% of potato, 2% of glucose, 2% of agar, 0.5% of peptone, 0.3% of potassium dihydrogen phosphate, 0.15% of magnesium sulfate, trace amount of vitamin B1 according to the mass percentage, and the rest is water;

[0089] (2) The mother seed medium is: according to the mass percentage, 20% of potatoes, 2% of glucose, 2% of agar, 0.3% of potassium dihydrogen phosphate, 0.15% of magnesium sulfate, trace amounts of vitamin B1, and the rest is water;

[0090] (3) The original seed material is: 40% cottonseed hulls, 38% sawdust, 20% bran, 2% calcium carbonate according to mass percentage;

[0091] The preparation process of the original seed bag is as follows: Weigh the cottonseed hull, soak it in water overnight, ...

Embodiment 2

[0101] Embodiment 2: The characteristic nucleotide sequence of Fenna Flammulina velutipes and the acquisition of characteristic nucleotide primers

[0102] 1. Extraction of Fenna Flammulina velutipes genome DNA template

[0103] The new strain CCTCC M 2016179 of Fenna Flammulina velutipes was inoculated on ordinary PDA medium (200g potato, 20g glucose, 20g agar, 1000ml water), cultured in dark at 25°C for 7-10 days, collected mycelia, ground into powder and used Ezup The column type fungal genomic DNA extraction kit (Shanghai Sangong) was used for total DNA extraction to obtain the total genomic DNA of Fenna Flammulina velutipes.

[0104] 2. The rDNA ITS sequence amplification and sequencing of Flammulina velutipes

[0105] The total genomic DNA of Flammulina velutipes was used as a template, and the universal primer pair ITS1: 5'-TCCGTAGGTGAACCTGCGG-3' and ITS4: 5'-TCCTCCGCTTATTGATATGC-3' were used as primers, and the reaction system was 50 μl. Total volume: 2 × TaqPCR Maste...

Embodiment 3

[0112] Embodiment 3: the identification method of Fenna Flammulina velutipes

[0113] (1) Experimental samples:

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Abstract

The invention relates to the technical field of needle mushroom authentication, in particular to a characteristic nucleotide sequence, primer, kit and method for authenticating Flammulina fennae. The characteristic nucleotide sequence basically comprises SEQ ID NO.1 or part of SEQ ID NO.1 or basically comprises SEQ ID NO.1 with mutation rate not larger than 5% or part of SEQ ID NO.1. Specificity is high, the method is simple, and the characteristic nucleotide sequence, primer, kit and method have great significance in commercialization and utilization of Flammulina fennae.

Description

technical field [0001] The invention relates to the technical field of identifying Flammulina velutipes, in particular to a characteristic nucleotide sequence, primers, a kit and a method for identifying Flammulina velutipes. Background technique [0002] Flammulina velutipes is one of the widely cultivated edible fungi at present, and it is also a kind of edible and medicinal fungus with high economic value. Flammulina velutipes contains a variety of nutrients, among which the 8 kinds of amino acids needed for human growth are relatively high, especially lysine and arginine are particularly rich in content, which can promote the healthy growth and intellectual development of children, so it is often called "increase". Zhi Mushroom". Flammulina velutipes contains pectin, which has significant anti-cancer functions. Regular consumption of enoki mushrooms can also prevent high blood pressure, treat hepatitis and gastrointestinal ulcers and other diseases. [0003] DNA is the...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895
Inventor 胡惠萍谭武平黄龙花刘远超黄志勇吴丽霞
Owner GUANGDONG INST OF MICROORGANISM
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