A method for constructing Torreya est-ssr primer and variety fingerprint

Abstract

The invention belongs to the technical field of molecular fingerprints, and discloses a Torreya EST-SSR primer and a method for constructing a variety fingerprint. The sequence is: SEQ ID NO: 1-SEQ ID NO: 40. The screened 20 polymorphic EST-SSR markers were used to amplify the DNA of 13 torreya species by PCR; according to the amplification results, the length of the amplified fragment was accurately read, and after statistics, at least three pairs of EST-SSR were used The combination of primers distinguished the 13 Torreya species tested. Based on the transcriptome data of 'Torreya' and 'Torreya' seeds, the present invention analyzes the obtained SSR data, screens out SSR primers with clear amplified bands and high polymorphism, and uses them to construct characteristic varieties of Torreya At the same time, it provides a good basis for the evaluation, preservation and utilization of Torreya germplasm resources.

Description

technical field [0001] The invention belongs to the technical field of molecular fingerprints, in particular to a Torreya EST-SSR primer and a method for constructing a variety fingerprint. Background technique [0002] Torreya (Torreya) was founded by Arnott in 1938, and Florida Torreya (Torreya taxifolia Arn) is the model plant of this genus. According to literature records, there are only 7 species and 2 varieties of Torreya extant, distributed intermittently in East Asia and North America. Among them, Torreya taxifolia (Torreya taxifolia) and Torreya californica Torrey (Torreya californica Torrey) are produced in North America, and Japanese Torreya (Torreya nucifera (L.) Sieb.et Zucc.) is wildly distributed in Japan and the Korean Peninsula. Some cities in my country are introduced as garden trees. cultivation. Torreya fargesii Franch., Torreya jackii Chun, Torreya grandis Fort.ex Lind, Torreya parvifolia Yi, L Yang et Long, Torreya fargesii Franch.Var.yunnanensis) and...

AI Technical Summary

Problems solved by technology

[0003] To sum up, the problems existing in the prior art are: the amount of DNA required for RFLP labeling is large, the steps are many, the cycle is long, radioactive isotopes are used in the preparation of probes and detection, and the cost is high

ISSR markers are mostly dominant markers, and cannot distinguish dominant homozygous genes from heterozygous genotypes. It takes a certain amount of time to explore the optimal conditions for PCR amplification reactions

The cost of AFLP labeling is relatively expensive, and the method requires multi-step operations, statistical analysis is difficult, and there are high requirements for the purity of DNA and the quality of endonucleases

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