Specific DNA fragment for identification of thelephora ganbajun, amplification primer, preparation method and application thereof

A specific, dried bacteria technology, applied in the direction of DNA / RNA fragments, microbial-based methods, biochemical equipment and methods, etc., can solve the problems of artificial cultivation of mushrooms, it is difficult to determine the authenticity of isolated strains, and achieve high detection specific effect

Active Publication Date: 2019-04-23
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It is a pity that many precious wild edible fungi cannot be artificially cultivated for the time being. In this case, it is difficult to determine the authenticity of the isolated strains.
The species can be identified by the appearance area of ​​the mature fruiting bodies of D. spp., but not by the hyphae

Method used

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  • Specific DNA fragment for identification of thelephora ganbajun, amplification primer, preparation method and application thereof
  • Specific DNA fragment for identification of thelephora ganbajun, amplification primer, preparation method and application thereof

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preparation example Construction

[0020] The present invention provides a kind of preparation method of described specific DNA fragment, comprises the following steps:

[0021] (1) extracting the genomic DNA of the dried fungus;

[0022] (2) using the genomic DNA extracted in the step (1) as a template, carry out PCR amplification with the primers claimed in claim 2, to obtain the amplified product;

[0023] (3) The amplified product in the step (2) is gel-cut and recovered to obtain a DNA fragment of a specific DNA fragment used for the identification of the dry bacteria.

[0024] The present invention has no special limitation on the extraction method, and any portable or kit known in the art can be used. In the embodiment of the present invention, the extraction method preferably adopts Qiagen's DNA Isolation Kit (product number: 12888-50) was used for DNA extraction according to the instructions.

[0025] The extracted genomic DNA was subjected to quantitative nucleic acid detection. Dilution with buf...

Embodiment 1

[0031] 1. Materials

[0032] The soil samples growing D. spp. were all collected in Wuding County, Yunnan Province (see Table 1). Collection method: In each place where the fungus grows, use the 5-point method to take the soil with a radius of 5 cm and a depth of 14 cm with the fruiting body as the center, put it into a prepared sterile sampling bag, number it, and store the soil sample at -80 ℃ (the soil sample that grows dry bacteria is the mixed fungal sample containing the bacteria pond).

[0033] Table 1 Sample collection information

[0034] serial number

sample name

location description

Types of

g1

Thelephora ganbajμn Zang

Wuding County, Yunnan Province

Soil samples growing D.

g2

Thelephora ganbajμn Zang

Wuding County, Yunnan Province

Soil samples growing D.

g3

Thelephora ganbajμnZang

Wuding County, Yunnan Province

Soil samples growing D.

[0035] 2. Method

[0036] 2.1 DNA extraction

...

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Abstract

The invention provides a specific DNA fragment for identification of thelephora ganbajun, an amplification primer, a preparation method and application thereof, and belongs to the technical field of biotechnology and enzyme engineering. A nucleotide sequence of the specific DNA fragment for the identification of the thelephora ganbajun is shown in an SEQ ID No.1 sequence table. The primer for amplifying the specific DNA fragment includes a 158F primer and a 437R primer. The specific DNA fragment for the identification of the thelephora ganbajun, the amplification primer or the application of the thelephora ganbajun specific DNA fragment obtained by the preparation method in the identification of the thelephora ganbajun can quickly and accurately complete the identification of the thelephora ganbajun.

Description

technical field [0001] The invention belongs to the technical field of strain molecular identification, and in particular relates to a specific DNA fragment, an amplification primer and a preparation method and application thereof for identification of dry bacteria. Background technique [0002] Wild edible fungi are an important forest resource and play an important role in green food development, biopharmaceuticals, environmental protection and other industries, especially some wild edible fungi that grow in forest areas. The genus Basidiomycota, Thelephoraceae, and Thelephora mostly grow in pine forests at an altitude of 600-2300m in central and southern Yunnan, and can co-exist with a variety of Pinaceae plants Ectomycorrhizal fungi. Dried fungus is a characteristic wild edible fungus in Yunnan Province. It has a unique taste, a fragrance similar to beef jerky, rich nutrition, and high nutritional and economic value. It has been reported that new compounds isolated fro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6806C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6806C12Q1/6895C12Q2531/113
Inventor 沙涛杨小雨张亚平王丁卉何志红
Owner YUNNAN UNIV
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