Method for identifying tea tree variety by using DNA bar code

A barcode and variety technology, applied in the field of identification of tea tree varieties, can solve the problems of small differences in identification traits, rich experience requirements of appraisers, large errors, etc., and achieve the effect of simple method

Inactive Publication Date: 2015-09-30
湖南省茶叶研究所
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Problems solved by technology

[0003] Aiming at the current problems such as the long period of identification of tea tree varieties relying on the botanical characteristics of tea trees, large errors, great difficulty in identifying varieti...

Method used

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  • Method for identifying tea tree variety by using DNA bar code
  • Method for identifying tea tree variety by using DNA bar code
  • Method for identifying tea tree variety by using DNA bar code

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Embodiment Construction

[0033] Describe the present invention in detail below in conjunction with accompanying drawing:

[0034] In the present invention, 5 varieties of Quyeqi, Xiangbolv No. 2, Fuding Dabaicha, Yulu and Jianbohuang No. 13 are used as test samples. First, the DNA extraction method was carried out according to the steps provided by the kit, and finally the DNA was dissolved to 100ul. Nanodrop detects the DNA concentration, and the DNA concentration is diluted to a concentration of 10-20ng / ul with redistilled water.

[0035] Next, 4 pairs of SSR primers with clear, stable and reproducible bands were screened. The specific primer information is shown in Table 1 below.

[0036]

[0037] Table 1 Screened primer information

[0038] Then, according to the reagent formula in Table 2, first mix the other 6 kinds of reagents except DNA, add Taq polymerase at the end when mixing, then divide into 5 parts and add them to 20ul PCR test tubes respectively, and finally in each tube test respe...

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Abstract

The present invention relates to a tea tree variety identifying method, specifically to a method for identifying the tea tree variety by using DNA bar code. The method comprises: respectively extracting DNA from a plurality of tea tree varieties; screening SSR primers; respectively preparing PCR reagents from the plurality of the extracted NDA, and carrying out reaction amplification; carrying out gel electrophoresis on each PCR reaction product, staining, and shooting and recording by using a gel imager; and combining the letters corresponding to the variety conversion on the SSR primers to form the DNA bar code. According to the present invention, the molecular marker technology is utilized, the method for identifying the tea tree variety by using the DNA bar code is established through the technical combination of genome DNA extraction, labeling screening, PCR amplification, product analysis, stripe data reading, letter bar code establishing and the like, and compared with the traditional morphology identifying method, the method of the present invention has the following characteristics that: the method is not affected by the environmental conditions, the method is simple, rapid and accurate, and the accurate identifying can be performed on the tea variety authenticity.

Description

technical field [0001] The invention relates to a method for identifying tea tree varieties. Background technique [0002] At present, my country has bred 114 national-level fine varieties and 154 provincial-level fine varieties. How to accurately identify these varieties is of great significance for the protection of breeders' rights and the promotion of new varieties. The DUS (abbreviation for Specificity, Consistency, and Stability) test guideline for new varieties of tea trees is the first DUS test guideline for new plant varieties formulated by China for the International Union for the Protection of New Varieties of Plants (UPOV). And protection provides the technical basis and authorization basis. With the continuous development of molecular marker technology, UPOV has incorporated DNA marker identification into the DUS test content of crop varieties. DNA molecular marker technology not only has the advantages of stability, identifiability of variation among varietie...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6895
Inventor 刘振杨阳赵洋杨培迪成杨宁静
Owner 湖南省茶叶研究所
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