Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

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

A technology for protein content and hairy mushrooms, applied in recombinant DNA technology, DNA/RNA fragments, determination/inspection of microorganisms, etc. Stable reliability, overcoming the effects of time-consuming and labor-intensive, short detection cycle

Pending Publication Date: 2022-03-15
杨满军
View PDF0 Cites 1 Cited by
  • 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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • DNA bar code for screening total soluble protein content index of champignon luteo-virens
  • DNA bar code for screening total soluble protein content index of champignon luteo-virens
  • DNA bar code for screening total soluble protein content index of champignon luteo-virens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] The construction of the DNA barcode of embodiment 1 yellow-green mushroom

[0053] Genome sequencing was carried out on the samples of Dangxiong County in Tibet Autonomous Region, Qilian County in Qinghai Province, and Shiqu County in Sichuan Province. The SSR loci in the genome sequences were analyzed using the MISA program.

[0054] Design primers for PCR amplification of these SSR loci, retain primers that can amplify the corresponding fragments, and discard invalid primers.

[0055] The content of total soluble protein was determined by selecting samples from Damxung County of Tibet Autonomous Region, Qilian County of Qinghai Province and Shiqu County of Sichuan Province.

[0056] The samples from the above three origins were respectively amplified using effective primers and detected by capillary electrophoresis. The simple sequence repeat (SSR) site corresponding to the total soluble protein content was established through analysis. Finally, 3 pairs of primers (...

Embodiment 2

[0059] Example 2 Amplification of the total soluble protein content of Pleurotus volvulus strain SSR-specific primers

[0060] (1) Extraction of total soluble protein

[0061] Collect the fruiting bodies of Dangxiong County, Tibet Autonomous Region, Qilian County, Qinghai Province, and Shiqu County, Sichuan Province, and dehydrate them by vacuum freeze-drying, crush them and pass through a 50-mesh sieve, add 20 mL of double-distilled water to 1 gram of dry powder, Use 300W ultrasonic wave to assist extraction for 30min, then centrifuge at 5000r / min for 30min and take the supernatant to prepare soluble total protein extract. The total soluble protein content was measured with the BCA protein concentration assay kit (enhanced) (product number P0010S) of Biyuntian Biotechnology Co., Ltd., and converted into milligrams per gram. It is 175.44 (± 2.69) milligrams per gram, is determined as a test example, and the total soluble protein content is 147.47 (± 2.91) milligrams per gram ...

Embodiment 3

[0090] Example 3 Screening and verification of the total soluble protein content of yellow-green mushrooms

[0091] Validation of DNA barcodes for the total soluble protein content of Pleurotus chinensis through blind experiments.

[0092] In the first blind test, samples from Damxung County, Tibet Autonomous Region whose total soluble protein content was higher than or equal to 175.44 mg / g were used as the test group, and samples from Shiqu County and The samples from Qilian County, Qinghai Province were the comparison group 1 and the comparison group 2, each with 16 copies and a total of 48 samples for blind test;

[0093] In the second test step, primers (SEQ ID NO: 1 and SEQ ID NO: 2, SEQ ID NO: 5 and SEQ ID NO: 6, SEQ ID NO: 9 and SEQ ID NO: 10) were used to amplify and perform capillary electrophoresis . Primer sets can be used in combination of one or more pairs to amplify and distinguish blinded samples with the total soluble protein content DNA barcode signature;

...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a DNA bar code and a primer group for screening total soluble protein content indexes 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 invention relates to the technical field of edible fungus germplasm resource screening, and more specifically relates to a DNA barcode, a primer set and an application for screening indicators of the total soluble protein content of Pleurotus chinensis. Background technique [0002] Yellow-green mushroom, golden in color, also known as chanterelle and golden mushroom, is a high-quality edible fungus with unique flavor, which cannot be cultivated artificially at present. Wild yellow-green mushrooms are mainly distributed in the Qinghai-Tibet Plateau. The main production areas are Damxung County in the Tibet Autonomous Region, Qilian County in Qinghai Province, and Shiqu County in Sichuan Province. The quality of these three main production areas is also the best. The main indicators for evaluating the nutritional value, flavor and biological activity of the yellow-green mushroom include: high content of total soluble protein, total soluble amino aci...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2531/113C12Q2563/107C12Q2563/185C12Q2565/125C12N15/1065C40B40/10C12Q1/68
Inventor 杨满军
Owner 杨满军
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com