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Method for identifying new variety of Ziyan tea tree by SSR (simple sequences repeat) fingerprint spectrum

A fingerprint and purple tea technology, applied in biochemical equipment and methods, microbe measurement/inspection, etc., can solve problems such as difficult to control transmission, and achieve the effect of wide application range and low cost

Inactive Publication Date: 2018-05-15
SICHUAN AGRI UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

In addition, since tea trees can be rapidly propagated using the "short ear cutting" technology, once excellent varieties enter the market, it will be difficult to control their spread

Method used

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  • Method for identifying new variety of Ziyan tea tree by SSR (simple sequences repeat) fingerprint spectrum
  • Method for identifying new variety of Ziyan tea tree by SSR (simple sequences repeat) fingerprint spectrum
  • Method for identifying new variety of Ziyan tea tree by SSR (simple sequences repeat) fingerprint spectrum

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Embodiment 1

[0059] 1. Reagents and tea varieties used

[0060] For DNA extraction, Beijing Tiangen Plant Genome Rapid Extraction Kit was used; PCR reagents were purchased from Chengdu Qingke Zixi Biotechnology Co., Ltd.; the tested tea varieties are shown in Table 1, and the leaf samples of each variety were collected from Sichuan Famous Camellia Breeding Farm Varieties resource nursery.

[0061] Table 1 30 tea varieties tested

[0062]

[0063]

[0064] 2. Number the SSR marker primers

[0065] The 12 pairs of SSR marker primers were divided into "homozygous" and "heterozygous" groups according to their band pattern in Ziyan, and according to the relative position (band length) of the target band on the electrophoretic gel, from high to Sort by low to get the numbering order of 12 pairs of primers, and the sequence of primers numbered 1-10 is as follows:

[0066] Primer sequence No. 1 TM056:

[0067] Upstream primer: 5'-TTGTTGGGTAGCTTTACTTGC-3' (SEQ ID NO: 1)

[0068] Downstre...

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Abstract

The invention provides a method for identifying a new variety of Ziyan tea tree by SSR (simple sequences repeat) fingerprint spectrum. The method comprises the following steps of extracting total DNA(deoxyribonucleic acid) of a tea sample, using the extracted DNA as a template, using 12 pairs of SSR marker primers to respectively perform PCR (polymerase chain reaction) amplification, performing electrophoresis and dyeing on the PCR amplification product, judging whether a target strip is in a homozygosis state or a heterozygosis state, forming the fingerprint spectrum, and then identifying the Ziyan variety. The method is characterized in that 12 pairs of SSR marker primers are used for performing PCR amplification, electrophoresis and dyeing on the sample, then the strip form is observed, the homozygosis state or the heterozygosis state is judged, and the unrequired and determined Ziyan samples are simultaneously operated as above to compare. The identifying method has the advantagesthat the operation is simple and rapid, and can be completed by conventional laboratory instrument equipment; the identifying result is reliable; the important meaning is realized for the protectionand popularization of Ziyan new variety of special tea tree.

Description

technical field [0001] The invention belongs to the field of tea tree breeding, and in particular relates to a method for identifying a new variety of Ziyan tea tree by using SSR fingerprints. Background technique [0002] Ziyan is a clonal tea tree variety, which belongs to the shrub type, middle leaf class, and late-growing species. It was bred by a single plant from the wild tea tree resources in Lijiashan, Muchuan County, Leshan City by the Tea Science Department of Sichuan Agricultural University and Sichuan Yizhichun Tea Industry Co., Ltd. A variety bred by law. Zi Yan was authorized by the Ministry of Agriculture to protect new varieties of plants in September 2017 (authorization number CNA20120455.2). Reach more than 3.0% of dry weight, significantly higher than other conventional tea varieties (0.01-0.5%). Studies have shown that anthocyanins have strong antioxidant capacity, and have anti-allergic, anti-cancer incidence, prevention of nervous and cardiovascular s...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858
CPCC12Q1/6858C12Q1/6895C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 谭礼强刘琴玲唐茜邹雪余烨颖邹瑶陈盛相李品武王玥胡锦浩周斌
Owner SICHUAN AGRI UNIV
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