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InDel label fingerprint spectrum of Lentinula edodes (Berk.) Pegler Guangxiang bacterial strain, and construction method of InDel label fingerprint spectrum

A fingerprint and construction method technology, which is applied in the field of detection of mushroom strains, can solve problems such as damage to the intellectual property rights of mushroom breeders, hidden dangers in the development of the mushroom industry, and producer risks, and achieve good application prospects, short detection time, and accuracy high effect

Active Publication Date: 2021-04-09
SHANGHAI ACAD OF AGRI SCI
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Problems solved by technology

In recent years, the shiitake mushroom industry has become an important starting point for targeted poverty alleviation in China. According to statistics, more than 20% of national-level poverty-stricken counties in the country choose shiitake mushrooms as an important industry. The scale of shiitake mushroom cultivation in my country has further expanded. The problem of "same species with different names, different species with the same name" has become increasingly prominent, which not only damages the intellectual property rights of mushroom breeders, but also causes great risks for producers, and creates hidden dangers for the further development of mushroom industry

Method used

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  • InDel label fingerprint spectrum of Lentinula edodes (Berk.) Pegler Guangxiang bacterial strain, and construction method of InDel label fingerprint spectrum
  • InDel label fingerprint spectrum of Lentinula edodes (Berk.) Pegler Guangxiang bacterial strain, and construction method of InDel label fingerprint spectrum
  • InDel label fingerprint spectrum of Lentinula edodes (Berk.) Pegler Guangxiang bacterial strain, and construction method of InDel label fingerprint spectrum

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Experimental program
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Effect test

Embodiment 1

[0063] (1) Mycelia culture: transfer the mushroom mycelium to the potato glucose medium PDA, culture in the dark at 23-22°C, and collect the mycelium after 10 days;

[0064] (2) Extraction of genomic DNA: extract the genomic DNA of the above-mentioned hyphae with the CTAB method, detect the concentration and purity of the total genomic DNA by ultraviolet spectrophotometry, and adjust the concentration of the sample DNA to be 20 to 30 ng / uL;

[0065] The CTAB method for extracting the genomic DNA from mycelium includes:

[0066] ①Take the freeze-dried shiitake mycelia in a 2mL centrifuge tube, and put 2-3 steel balls into it;

[0067] ②Put the centrifuge tube into a multi-sample tissue grinder, set the frequency to 60Hz, and grind for 30s to a uniform powder;

[0068] ③ Add 2×CTAB extract preheated at 62°C for 1 hour, keep warm at 62°C for 60 minutes, and shake gently once every 20 minutes;

[0069] ④ Centrifuge at 12000rpm, 4°C for 20min, dispense the supernatant into two tu...

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Abstract

The invention relates to an InDel label fingerprint spectrum of a Lentinula edodes (Berk.) Pegler Guangxiang culture, and a construction method of the InDel label fingerprint spectrum. The fingerprint spectrum consists of seven pairs of InDel labels developed after Lentinula edodes (Berk.) Pegler is subjected to whole genome sequencing. The InDel label fingerprint spectrum of the Lentinula edodes (Berk.) Pegler Guangxiang culture can be used for specificity identification of the Lentinula edodes (Berk.) Pegler Guangxiang culture, has the advantages of short detection time, high accuracy and good repeatability and has the singleness of the Lentinula edodes (Berk.) Pegler Guangxiang culture in 44 pieces of collected Lentinula edodes (Berk.) Pegler Guangxiang cultures.

Description

technical field [0001] The invention belongs to the detection field of Lentinus edodes strains, and in particular relates to an InDel-marked fingerprint of Lentinus edodes strains and a construction method thereof. Background technique [0002] Mushrooms (Lentinula edodes (Berk.) Pegler) are popular among consumers because of their high nutritional value and medicinal value. China is the first country in the world to carry out the artificial cultivation of shiitake mushrooms, with a cultivation history of more than 800 years. At present, shiitake mushrooms are the most widely cultivated and the highest yield mushroom species in China. According to statistics from the China Edible Fungi Association, the total output of edible fungi in China in 2018 was 38.4204 million tons, of which the total output of shiitake mushrooms was 10.4312 million tons, accounting for 27.12% of the total domestic output of edible fungi and more than 90% of the world's total output of shiitake mushro...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 于海龙章炉军宋春艳尚晓冬谭琦张美彦张丹李巧珍李玉
Owner SHANGHAI ACAD OF AGRI SCI
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