不同品种(系)凤凰单丛茶DNA指纹图谱的构建

Construction of DNA Fingerprints in Fenghuang Dancong Tea of Different Cultivars(or Lines)

为能够快速、准确地对不同品种（系）凤凰单丛茶进行鉴定,建立不同品种（系）凤凰单丛茶的DNA指纹图谱,本研究利用ISSR分子标记技术,以4份凤凰单丛茶品种为材料,对UBC802~UBC899等58条ISSR引物进行筛选,得到了12条多态性好的ISSR引物。在确定引物最佳退火温度后,利用这12条ISSR引物对39份凤凰单丛茶进行了扩增,结果显示,从其中筛选出的6条核心引物共扩增出84条带,平均每个引物扩增出14条带,其中多态性条带78条,多态性条带比例为92.86%,多态信息量平均为0.9456,其中引物UBC843多态性最为丰富,品种相似性系数在0.4063-0.9836之间;为将39份凤凰单丛茶品种全部区分开,本研究对6条核心引物进行了组合,发现其中UBC827/UBC846的组合鉴别效率最高;通过综合品种名称、国家地区编号、采样地点、高效核心引物组合名称以及ISSR数据代码,本研究建立了39种凤凰单丛茶的DNA指纹代码。之后,通过整合软件录入如品种指纹图谱代码、香型、海拔、ISSR-PCR扩增数据图片等更多相关信息,形成指纹图谱二维编码,构成凤凰单丛茶的DNA指纹图谱,为凤凰单丛茶品种权益保护及信息平台的建立提供数据支持。

The aim of this study is to quickly and accurately identify different cultivars（or lines） of Fenghuang Dancong tea in which would construct a DNA fingerprint in different cultivars（or lines） of Fenghuang Dancong tea based on ISSR molecular marker. Firstly, 58 ISSR primers were screened with 4 cultivars, indicating 12 of which are polymorphic. After screening the optimal annealing temperature, 12 ISSR primers were used to amplify 39 Fenghuang Dancong tea resources. The results showed that by using 6 reliable ISSR primers, 84 DNA fragments, among which 78 DNA bands were polymorphic, of all 39 tea accessions were amplified. And the polymo rphic proportion of DNA bands（PPB） was 92.86%, including the primer UBC843 with the richest polymorphic, and the average number of DNA bands amplified by each primer was 14. The average polymorphism information content（PIC） was 0.9456, and the genetic similarity coefficient of the 39 accessions ranged from 0.4063 to 0.9836. In order to identify all 39 accessions, it indicated that the combination efficiency of UBC827/UBC846 was the highest by making combination of the 6 core ISSR primers. By connecting the name of cultivars, the national area code, the sampling sites, the name of the highest efficient primer combination and the ISSR data code, the DNA fingerprint codes of 39 Fenghuang Dancong tea were built. And then, the relevant information of the sample such as its flavor type, altitude, ISSR-PCR amplified picture and so on, were all typed in the software to generate the two-dimensional code fingerprint. The construction of the DNA fingerprint in different cultivars（or lines） of Fenghuang Dancong tea could offer data support for the protection of the cultivars rights and interests.