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DNA bar code for screening total polyphenol content index of champignon luteo-virens

A technology for total polyphenols and C. pilofus Overcome the effects of time-consuming and labor-intensive, stable and reliable results, and short detection cycle

Pending Publication Date: 2022-03-04
杨满军
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the existing DNA barcoding technology, ITS (ribosomal RNA internal transcriptional spacer) and the non-coding region or conserved gene sequence in mitochondria are mainly used for species identification; restriction fragment length polymorphism (restriction fragment length polymorphism, RFLP) operations are very complicated, the reliability and repeatability of the results are poor, random amplified polymorphic DNA (RAPD) is susceptible to interference, and requires a high level of operator skills. In assisted breeding work Difficult to promote; single nucleotide polymorphism (singlenucleotide polymorphism, SNP) requires high equipment and high cost
[0004] Therefore, in view of the shortcomings of traditional breeding methods that are not accurate enough to be time-consuming and labor-intensive, how to provide a DNA barcode that can accurately and quickly identify the strains that belong to the yellow-green mushroom, and at the same time realize high-quality breeding, is cost-effective. Low cost, high efficiency, easy operation, stable results, high reliability and good repeatability are problems that need to be solved urgently by those skilled in the art

Method used

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  • DNA bar code for screening total polyphenol content index of champignon luteo-virens
  • DNA bar code for screening total polyphenol content index of champignon luteo-virens
  • DNA bar code for screening total polyphenol content index of champignon luteo-virens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1 yellow green embodiment Curly barcode DNA construct mushroom

[0063] Damxung collection TAR, Qinghai Qilian County, yellow mushroom samples curl Shiqu County, Sichuan Province genome sequencing program using the MISA genomic sequence of SSR loci analyzed.

[0064] These primers were designed for PCR amplification SSR locus, can retain the amplified fragments corresponding to the primers, primer discard invalid.

[0065] Select Damxung Tibet, Qinghai Qilian County, Sichuan Province Serxu yellow-green mushroom curl total polyphenols content of the sample was measured.

[0066] Effective use of the above primers were amplified and origin of the three samples by capillary electrophoresis. The analysis of simple sequence repeats establishment (simple sequence repeat, SSR) site corresponding to the total polyphenol content. Ultimately, four pairs of primers (see Table 1), using the four primer pairs amplified sample genomic fragment polymorphism resulting assisted s...

Embodiment 2

[0069] EXAMPLE 2 Curly mushroom total polyphenol content of yellow-strain specific primers Amplification SSR

[0070] (1) Extraction of total polyphenol

[0071] Damxung collection TAR, Qinghai Qilian County, Sichuan Province Serxu yellow-green curly mushroom fruiting bodies pulverized and passed through a 50 mesh sieve after vacuum freeze-drying dehydration, 1 g of dry powder was added 20mL double distilled water, 30min with 300W ultrasound-assisted extraction, and then after 5000r / min centrifugal 30min the supernatant was prepared as a total polyphenol extract. Curly mushroom fruiting bodies of yellow-green extract total polyphenol content was determined by Lowry method, with particular reference to Wang Jiadan like (Wang Jiadan, total polyphenol content determination method of Xu Ting, Han Wei. Mushroom preferred [J]. Nanjing University (Natural Science), 2017,39 (02): 113-120), and in terms of milligrams per gram. Qilianensis County Qinghai yellow-curl mushroom total polyphe...

Embodiment 3

[0109] Example 3 Total Polyphenol Content Index Screening Verification of Muscular Mushroom

[0110] The DNA barcode of the total polyphenol content of the yellow-green curling mushroom was verified by a blind test.

[0111] The first step is a blind test, with a total polyphenol content higher than or equal to 7.67 mg of Qilian County sample as a test group, and Xixixi and Sichuan, a Tibet Autonomous Region, a Tibet Autonomous Region below 91.1% (significant P <0.05). The provincial stone counties sample is compared 1 group and comparison of 2 groups, and 16 copies of 48 samples were taken;

[0112] The second step is tested, using primers (SEQ ID NO: 1 and SEQ ID NO: 6, SEQ ID NO: 9 and SEQ ID NO: 10, SEQ ID NO: 15 and SEQ ID NO: 16) Amplifies and performs capillary electrophoresis. The primer group can use a pair of or multi-pair combination amplification to distinguish between the experimental DNA barcode characteristics;

[0113] The third step is revealed, and the results ar...

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Abstract

The invention discloses a DNA bar code and a primer group for screening a total polyphenol content index of champignon luteo-virens and application, and belongs to the technical field of screening of germplasm resources of edible mushrooms. Compared with a traditional breeding method and other existing DNA bar code technologies, the method has the advantages of being time-saving, labor-saving, money-saving, accurate and efficient, plays a positive role in identification of the high-quality virens luteo-virens origin and genetic breeding, and also provides an effective method for identification and protection of germplasm resources.

Description

Technical field [0001] The present invention relates to a technical field of edible fungus germplasm resources screening, more particularly, to a green and yellow indicator total polyphenol content screening curl mushroom DNA barcodes, and primer set application. Background technique [0002] Curly mushroom yellow and green, the color of golden brown, also known as yellow mushroom, golden mushroom, is a unique flavor of high quality edible fungus, is currently not cultivated. Green and yellow curly wild mushrooms are mainly distributed in the Qinghai-Tibet Plateau, the main producing areas of Damxung County of Tibet Autonomous Region, Qinghai and Qilian County Shiqu County, Sichuan Province, but also the best quality of the three main producing areas. Evaluation of yellow-green Curly mushroom flavor and nutritional value of the main indicators of biological activity include: total soluble protein, total soluble amino acids, total polyphenols, high total polysaccharides and total ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2531/113C12Q2565/125C12Q2563/185C12Q2525/151C12Q2563/107C12Q1/686C12Q1/6888
Inventor 杨满军
Owner 杨满军
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