DNA (deoxyribonucleic acid) barcode reference gene of termitomyces clypeatus and application of DNA barcode reference gene of termitomyces clypeatus

A standard gene, the technology of Gallus firsum, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of less habitat environment, human factors, over-harvesting, etc., and achieve low integrity requirements and improve identification efficiency , the effect of easy comparison

Inactive Publication Date: 2016-01-13
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in recent years, due to the increase in market demand, the natural population of Gallus firji is facing the threat of being over-harvested; in addition, with the destruction of the ecological environment, the habitat of Gallus firji is becoming less and less. Germplasm resources have been seriously threatened, and the protection of wild Gallium firsum is imminent
On the other hand, due to the lack of systematic cognition of the germplasm resources of chicken fir, there are mixed species of fish in the market of chicken fir, with different advantages and disadvantages.
According to the color of chicken fir, the folks are divided into many types such as black skin, white skin, yellow skin, flower skin, etc. The taste is best with black skin (green skin). The best one is garlic fir, followed by Kaiji, and then torched fir, but these morphological characteristics are mostly affected by the development degree and growth environment of chicken fir, with great human factors
In addition, because the public lacks the ability to accurately identify wild Gallus pilosula, deaths from eating poisonous mushrooms have occurred from time to time

Method used

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  • DNA (deoxyribonucleic acid) barcode reference gene of termitomyces clypeatus and application of DNA barcode reference gene of termitomyces clypeatus
  • DNA (deoxyribonucleic acid) barcode reference gene of termitomyces clypeatus and application of DNA barcode reference gene of termitomyces clypeatus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: sample to be tested and pointed shield termite umbrella ( Termitomycesclypeatus) Homology identification of DNA barcode identification standard gene fragments

[0031] 1. Collection of Gallinarum albumen specimens: 8 samples were collected from Kunming, Pu'er and other places in Yunnan Province. Fresh fruiting bodies were directly cut out of tissue or dried with silica gel for later use. See Table 1 for sample and source details.

[0032] Table 1 Sample and source information

[0033]

[0034] 2. Extraction of Genomic DNA from Gallus pilosulae: using the improved CTAB method or using BiomigaEZgene TM Genomic DNA was extracted with the FungalgDNA MiniprepKit kit, and the DNA concentration of the sample was diluted to 0.5 μg / μl with sterilized deionized water.

[0035] 3. Amplify the DNA fragment and carry out polymerase chain reaction (PCR), that is, amplify with specific primers. The primer pair sequence is forward primer RPB2B-f: 5'-CCGCAAAGGCTGGTGT...

Embodiment 2

[0038] Embodiment 2: Using the phylogenetic tree method to set up identification rules to identify species

[0039] Using the steps described in Example 1 to obtain the sequence of the sample to be tested, the standard gene sequence and the sample sequence to be identified were combined and MEGA was used to repeat 1000 times to construct a phylogenetic tree based on the Kimura-2-parameter model. The sample to be tested (BS3-7 and BS3-8) DNA barcode sequences clustered with standard gene sequences, with T. bulborhizus, T. eurrhizus and T. heimii The sequences showed obvious divergence, indicating that the sample to be identified could be identified as the termite ( Termitomycesclypeatus ).

[0040] Compared with the traditional morphological identification method, the gene sequence obtained and the method used in the present invention are beneficial to realize the rapid and accurate identification of the Gallus albuminosa.

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Abstract

The invention belongs to the field of fungus species identification and relates to a reference gene for molecular identification of Termitomyces clypeatus and a molecular identification method. Sequences of adopted primers are RPB2B-f: 5'-CCGCAAAGGCTGGTGTATC-3' and RPB2B-r: 5'-TTCGAATGAGTTCAAGTGT-3', and the primers can be used for amplification of large-subunit gene (RPB2) of RNA (ribonucleic acid) polymerase II of the termitomyces clypeatus to obtain a reference detection gene. The termitomyces clypeatus can be quickly and accurately identified according to RPB2 gene DNA (deoxyribonucleic acid) sequence differences. The RPB2 gene overcomes the defect of difficulty in identification of traditional termitomyces clypeatus forms and has the advantages of universality and easiness in amplification and comparison, reliability and accuracy in identification are greatly improved, and a powerful research implement is provided for excavation, protection and utilization of genetic resources of the termitomyces clypeatus.

Description

technical field [0001] The invention relates to the field of species identification, in particular to a standard gene of a DNA barcode of Gallus arborvitae and an application thereof. Background technique [0002] Chicken fir belongs to the fungus Basidiomycetes Agaricaceae Agaricaceae. We often say that the chicken fir fungus is a kind of fungus that is specifically symbiotic with certain species of the subfamily Macrotermitinae (Macrotermitinae), so it is also called ant nest umbrella or termite umbrella. Called the king of wild edible fungus, it has thick meat, white filaments, crisp and refreshing taste, delicious fragrance, and rich nutrition. It contains calcium, magnesium, iron, protein and other nutrients. It is rich in phosphorus and contains More than 20 kinds of amino acids. In addition, the Gallus pilosula also contains effective ingredients for treating diabetes, which has a significant effect on lowering blood sugar. Chicken fir fungus is the first choice fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12Q1/68C12Q1/04
Inventor 张颖汤晓召徐建平余泽芬张克勤
Owner YUNNAN UNIV
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