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A kind of method for identification of oyster mushroom strain and special dna barcode fragment

A barcode, DNA molecule technology, applied in DNA/RNA fragments, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of unreliable results and time-consuming, avoid bacterial species errors, shorten identification time, The effect of improving identification accuracy

Active Publication Date: 2018-02-06
LUDONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods are time-consuming, have certain tissue and developmental stage specificity, and the results are not reliable

Method used

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  • A kind of method for identification of oyster mushroom strain and special dna barcode fragment
  • A kind of method for identification of oyster mushroom strain and special dna barcode fragment
  • A kind of method for identification of oyster mushroom strain and special dna barcode fragment

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Embodiment 1

[0039] Embodiment 1, the acquisition of the DNA barcode fragment of Pleurotus ostreatus strain identification and the establishment of the method

[0040] 1. Genomic DNA acquisition of Pleurotus ostreatus strains

[0041] Genomic DNA of Pleurotus ostreatus strains was extracted.

[0042] 2. PCR amplification

[0043] Using the genomic DNA obtained in 1 above as a template, PCR amplification was performed with primer 728F and primer 1567R.

[0044] 728F: 5'-CATCGAGAAGTTCGAGAAGG-3' (SEQ ID NO: 1)

[0045] 1567R: 5'-ACHGTRCCRATACCACCRATCTT-3' (SEQ ID NO: 2)

[0046] The above-mentioned PCR amplification system is as follows in Table 1:

[0047] Table 1 is the composition of the PCR reaction system (25 μL)

[0048]

[0049] The above PCR amplification program is: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, annealing at 63°C for 55 s, 1°C decrease in each cycle, extension at 72°C for 90 s, 10 cycles; denaturation at 94°C for 30 s, annealing at 53°C fo...

Embodiment 2

[0055] Embodiment 2, distinguish flat mushroom bacterial classification

[0056] 1. Specific detection by the method of the present invention

[0057] Genomic DNA of Pleurotus eryngii strains and Pleurotus eryngii strains were extracted respectively.

[0058] Using the genomic DNA of each variety as a template, the above-mentioned primers 728F and 1567R were used to amplify to obtain PCR products of each variety.

[0059] The PCR product of each kind is sent for sequencing, and the results are as follows:

[0060] The nucleotide sequence of the PCR product of the known varieties of Pleurotus ostreatus strains is sequence 3 (DNA barcode fragment for identification of Pleurotus ostreatus strains).

[0061] The nucleotide sequence of the PCR product of the Pleurotus eryngii strain of known species is sequence 4 (the 1st-20th position of sequence 4 is the upstream primer 728F, and the 375th-397th position of sequence 4 is the reverse complementary sequence of the downstream prim...

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Abstract

The invention discloses a method for identifying a pleurotus ostreatus strain, and a special deoxyribonucleic acid (DNA) bar code fragment. The DNA fragment provided by the invention is as shown in (1) or (2): (1) a fragment shown as sequence 3; (2) a fragment having nucleotide sequence homology of more than 99% with the fragment shown in (1). The pleurotus ostreatus DNA bar code fragment provided by the invention can be used for identifying the pleurotus ostreatus strain; after the DNA bar code fragment is used for identifying the pleurotus ostreatus strain, the pleurotus ostreatus strain identification time is facilitated to be shortened, and the identification accuracy is improved; the pleurotus ostreatus strain can be accurately identified at the shortest time of 48 hours by using the method, so that the consequence that the strain is found to be wrong after mushrooms are cultivated is avoided.

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to rapid identification of edible fungus strains by DNA barcodes, in particular to a method for identification of oyster mushroom strains and special DNA barcode fragments. Background technique [0002] Edible fungi are the main economic crops and export agricultural products in my country. After more than 30 years of rapid development, my country's edible fungi industry has produced more than 28 million tons and an output value of more than 200 billion yuan in 2014, and has become the world's largest producer of edible fungi. Among them, the output of oyster mushroom is the largest, reaching 5.6 million tons, ranking first in the world. The quality of strains is very important to the development of edible fungi industry. There is a saying in the edible fungus industry that "one mother species affects one edible fungus industry". Strains play an important and irreplaceable role in the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/04C12N15/11
CPCC12Q1/6895
Inventor 赵鹏纪森鹏周军辉高兴喜
Owner LUDONG UNIVERSITY
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