DNA barcode, primer, kit, method and application

Abstract

The invention belongs to the field of species and strain identification, and specifically relates to a DNA sequence, a DNA barcode for identifying Blastobotrys adeninivorans, a primer, a kit, a methodand application. The DNA barcode originates from a genome of Blastobotrys adeninivorans TMCC 70007 and comprises the DNA sequence provided by the invention. The length of the DNA barcode is 500bp-2,200bp, preferably 500bp-1,600bp and more preferably 500bp-1,000bp. The DNA barcode can identify the Blastobotrys adeninivorans TMCC 70007 of Pu'er tea fermentation strains precisely, and can distinguish the strain from easily confused strains or other strains of the same species precisely and rapidly.

Description

technical field

[0001] The invention belongs to the field of species and bacterial strain identification, and specifically relates to DNA barcodes, primers, kits, methods and applications. Background technique

[0002] Pu-erh tea is a post-fermented tea with the geographical indication of Yunnan. It uses large-leaf sun-dried green tea as raw material and is made through a series of processes. The traditional production process of Pu'er tea is as follows: the picked fresh tea leaves are rolled, sun-dried, impurity-removed, tide-watered, heaped, dried, sieved, pressed and molded, and packaged for delivery. In the production of Pu-erh tea, the heaping fermentation process is the main factor to form the quality of Pu-erh tea. During this process, the internal components of the tea such as tea polyphenols, caffeine and some polysaccharides have undergone tremendous changes, making Pu-erh tea The special flavor, taste, quality and various health benefits of tea.

[0003] In the ...

Examples

Embodiment 1

[0073] Example 1: Obtaining of BS1R_4124891 gene and DNA barcode

[0074] 1. Using the high-coverage proteome technology, pFind and pAnno software were used to conduct a deep coverage study on the proteome of the Pu-erh tea industrial fermentation fungus Arthrospora adenorivorum TMCC 70007, and to verify the annotated coding genes of its genome, and found that The annotated gene BS1R_4124891 was eliminated. In order to discover new protein coding regions, using the Six Frame Translation strategy in systematic proteomics, a six-frame translation database of the genome data of Arthrospora adenophagous TMCC70007 was obtained, and the six codings of the genome were exhaustively listed. Possibly (+1, +2, +3, -1, -2, -3), and the nucleic acid sequence is called "six-frame translated nucleic acid sequence", and the protein sequence is called "six-frame translated protein sequence". Generally speaking, a six-frame translated nucleic acid sequence is the sequence from one terminator t...

Embodiment 2

[0098] Example 2. Identification of bacterial strains using DNA barcodes

[0099] According to the amplification result of the sample to be tested and the sequence homology with Arthrospora adenophagous TMCC 70007 strain SEQ ID NO.1, it is determined whether the sample to be tested is the industrially applied strain of Arthrospora adenorivorum TMCC 70007 .

[0100] (1) Source of strain

[0101] Table 4 Information about the relevant strains selected

[0102] Sample serial number

scientific name

Strain number

Habitat

1

Arthrospora adenophagous

TMCC 70007

Puer Tea Wodui, China

2

Arthrospora adenophagous

CBS 8244 T

soil, netherlands

3

Arthrospora adenophagous

CBS 7350

Cellar corn feed, Netherlands

4

Arthrospora adenophagous

CBS 7370

soil, south africa

5

Arthrospora adenophagous

CBS 8335

clay, italy

6

Blastobotrys raffinosifermentans

CBS 6800 T

unk...

technical field

[0001] The invention belongs to the field of species and bacterial strain identification, and specifically relates to DNA barcodes, primers, kits, methods and applications. Background technique

[0002] Pu-erh tea is a post-fermented tea with the geographical indication of Yunnan. It uses large-leaf sun-dried green tea as raw material and is made through a series of processes. The traditional production process of Pu'er tea is as follows: the picked fresh tea leaves are rolled, sun-dried, impurity-removed, tide-watered, heaped, dried, sieved, pressed and molded, and packaged for delivery. In the production of Pu-erh tea, the heaping fermentation process is the main factor to form the quality of Pu-erh tea. During this process, the internal components of the tea such as tea polyphenols, caffeine and some polysaccharides have undergone tremendous changes, making Pu-erh tea The special flavor, taste, quality and various health benefits of tea.

[0003] In the ...

Examples

Embodiment 1

[0073] Example 1: Obtaining of BS1R_4124891 gene and DNA barcode

[0074] 1. Using the high-coverage proteome technology, pFind and pAnno software were used to conduct a deep coverage study on the proteome of the Pu-erh tea industrial fermentation fungus Arthrospora adenorivorum TMCC 70007, and to verify the annotated coding genes of its genome, and found that The annotated gene BS1R_4124891 was eliminated. In order to discover new protein coding regions, using the Six Frame Translation strategy in systematic proteomics, a six-frame translation database of the genome data of Arthrospora adenophagous TMCC70007 was obtained, and the six codings of the genome were exhaustively listed. Possibly (+1, +2, +3, -1, -2, -3), and the nucleic acid sequence is called "six-frame translated nucleic acid sequence", and the protein sequence is called "six-frame translated protein sequence". Generally speaking, a six-frame translated nucleic acid sequence is the sequence from one terminator t...

Embodiment 2

[0098] Example 2. Identification of bacterial strains using DNA barcodes

[0099] According to the amplification result of the sample to be tested and the sequence homology with Arthrospora adenophagous TMCC 70007 strain SEQ ID NO.1, it is determined whether the sample to be tested is the industrially applied strain of Arthrospora adenorivorum TMCC 70007 .

[0100] (1) Source of strain

[0101] Table 4 Information about the relevant strains selected

[0102] Sample serial number

scientific name

Strain number

Habitat

1

Arthrospora adenophagous

TMCC 70007

Puer Tea Wodui, China

2

Arthrospora adenophagous

CBS 8244 T

soil, netherlands

3

Arthrospora adenophagous

CBS 7350

Cellar corn feed, Netherlands

4

Arthrospora adenophagous

CBS 7370

soil, south africa

5

Arthrospora adenophagous

CBS 8335

clay, italy

6

Blastobotrys raffinosifermentans

CBS 6800 T

unk...

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