应用AFLP-银染技术,分析黄金茶群体111个株系的遗传多样性与亲缘关系。选用多态性高、分辨力强的引物组合E37M32、E41M33与E41M42分别扩增样品基因组DNA,共得到229条清晰条带,其中多态性条带186条,多态位点百分比为81.22%,反映了样品基因组DNA具有较高的多态性。111个样品得出的平均有效等位基因数(Ne)为1.42±0.35,平均Nei′s基因多样性(H)为0.25±0.18,平均Shannon′s信息指数(I)为0.38±0.25。应用NTSYSpc2.1软件,计算得到111个样品间的相似系数在0.65~0.99之间,平均为0.76。根据UPGMA法,将111个株系分成8大类群,绘制样品AFLP聚类树状图。本研究为黄金茶种质资源的保护及利用,从分子水平提供了一定的依据。
111 tea strains from ‘Huangjincha’ population were analyzed by AFLP-sliver staining protocol for detecting the genetic diversity and relationship. AFLP fingerprinting of 111 samples with primer combinations E37M32, E41M33 and E41M42 revealed a total number of unambiguous 229 bands, of which 186 were polymeric and the polymorphism frequency was 81.22%. The results showed that abundant diversities existed among samples of genomic DNA. The mean of effective number of alleles, Nei′s gene diversity and Shannon′s information index of 111 samples were 1.42±0.35, 0.25±0.18, 0.38±0.25 respectively. As analyzed by NTSYSpc2.1, the similarity coefficient of 111 samples ranged from 0.65 to 0.99, and the mean was 0.76. These tea strains were divided into 8 groups by UPGMA based on the mean of similarity coefficient, and AFLP dendrogram was established. In conclusion, the results provide the basis for the protection and utilization of ‘Huangjincha’ population from molecular level.
[1] 张湘生. 地方珍稀茶树良种——黄金茶的利用现状与前景[J]. 茶叶通讯, 2006, 33(2): 4-6.
[2] Zabeau M, Vos P. Selective restriction fragment amplification: a general method for DNA fingerprinting[P]. European: EP19920402629, 2005-04-27.
[3] 赵超艺, 周李华, 罗军武, 等. 广东茶树种质资源AFLP分析[J]. 茶叶科学, 2006, 26(4): 249-252.
[4] 黄建安, 李家贤, 黄意欢, 等. 茶树品种资源遗传多样性的AFLP研究[J]. 园艺学报, 2006, 33(2): 317-322.
[5] 季鹏章, 汪云刚, 蒋会兵, 等. 云南大理茶资源遗传多样性的AFLP分析[J]. 茶叶科学, 2009, 29(5): 329-335.
[6] 黄建安, 李家贤, 黄意欢, 等. 茶树AFLP分子连锁图谱的构建[J]. 茶叶科学, 2005, 25(1): 7-15.
[7] 黄福平, 梁月荣, 陆建良, 等. 乌龙茶种质资源种群遗传多样性AFLP评价[J]. 茶叶科学, 2004, 24(3): 183-189.
[8] 黄建安, 黄意欢, 罗军武, 等. 茶树基因组DNA的高效提取方法[J]. 湖南农业大学学报: 自然科学版, 2003, 29(5): 402-407.
[9] Instruction manual AFLP®, AFLP® Analysis System II. (Accessed at http://plantbio.berkeley.edu
[10] 黄建安, 黄意欢, 罗军武, 等. 茶树AFLP-银染技术体系与品种指纹图谱的建立[J]. 湖南农业大学学报: 自然科学版, 2005, 31(4): 427-430.
[11] SSR gel and sliver staining protocol. (Accessed at http://dendrome.ucdavis.com.
[12] Yeh F.Population genetic analysis of codominant and dominant markers and quantitative traits[J]. Belgian Journal of Botany, 1997, 129: 157.
[13] Kimura M, Crow JF.The number of alleles that can be maintained in a finite population[J]. Genetics, 1964, 49(4): 725-738.
[14] Nei M.Analysis of gene diversity in subdivided populations[J]. Proceedings of the National Academy of Sciences, 1973, 70(12): 3321-3323.
[15] Lewontin R C.The apportionment of human diversity[J]. Evolutionary biology, 1972(6): 381-398.
[16] Rohlf F.NTSYS-pc:Numerical Taxonomy and Multivariate Analysis System:Version 2.1[M]. New York: Exceter Software, Applied Biostatistics Inc, 2000.
[17] 陈亮, 高其康. 15个茶树品种遗传多样性的RAPD分析[J]. 茶叶科学. 1998, 18(1): 21-27.
[18] 杨阳, 刘振, 赵洋, 等. 利用EST-SSR标记研究黄金茶群体遗传多样性及遗传分化[J]. 茶叶科学, 2009, 29(3): 236-242.
[19] 宁静, 李健权, 董丽娟, 等. “黄金茶”特异种质资源遗传多样性和亲缘关系的ISSR分析[J]. 茶叶科学, 2010, 30(2): 149-156.
[20] 张文彤. SPSS统计分析基础教程[M]. 北京: 高等教育出版社, 2010.
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