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利用NCBI公共数据库下载获得的3 306条茶树EST,去除其中低质量和冗余的序列,得到全长为635.46 kb的1 335条无冗余EST,在这些序列中共发现216个SSR,分布于185个EST中,出现的频率为16.18%,平均每2.94 kb的表达序列中有1个SSR。茶树的EST-SSR类型丰富,其中二核苷酸重复出现的频率最高,占主导地位。利用Primer 5.0软件从185条含有SSR的EST中设计引物100对,从中随机选取60对引物检测EST-SSR在云南茶树资源中的多态性。结果表明,有30对引物在供试的63份茶树资源中得到有效扩增,并具有多态性;共检测到等位基因数71个,每对引物检测出的等位基因数2~8个,平均4.9个;多态信息量(PIC)变幅为0~0.731,平均为0.499;观察杂合度变幅为0~0.970,平均0.392。EST-SSR标记分析表明云南茶树资源具有较高的遗传多样性。
A total of 3,306 non-redundant ESTs with 635.46 kb in length were obtained by downloading 3 306 ESTs from the NCBI public database. A total of 216 SSRs were found in these sequences, ranging from 185 Among ESTs, the frequency of occurrence was 16.18%, with an average of 1 SSR in every 2.94 kb expression sequence. Tea tree EST-SSR type rich, in which dinucleotide recurrence frequency of the highest, dominant. Primer 5.0 software was used to design 100 pairs of primers from 185 SSR-containing ESTs. 60 pairs of primers were randomly selected to detect the polymorphism of EST-SSR in Yunnan tea plant resources. The results showed that 30 pairs of primers were effectively amplified in 63 accessions of tea trees and had polymorphism. A total of 71 alleles were detected, and the number of alleles detected per pair was 2-8 With an average of 4.9. The polymorphism information (PIC) amplitude ranged from 0 to 0.731 with an average of 0.499. The observed heterozygosity ranged from 0 to 0.970 with an average of 0.392. EST-SSR marker analysis showed that Yunnan tea tree resources have high genetic diversity.