SRAP 分子标记技术分析鱼腥草居群的遗传多样性

钟军1,3, 王坤2, 仇萍3, 曾维军3, 熊兴耀4,*
1 湖南农业大学农学院, 长沙 410128; 2 湖南农业大学生命科学与技术学院, 长沙 410128; 3 湖南正清制药集团股份有限公司,湖南怀化 418000; 4 湖南农业大学园林园艺学院, 长沙 410128

通信作者:熊兴耀;E-mail: xiongxingyao@yahoo.cn;Tel: 0731-84635 295

摘 要:

在正交设计对鱼腥草SRAP-PCR反应体系进行优化的基础上, 分析鱼腥草居群的遗传多样性的结果表明: 最佳的SRAP- PCR 反应体系为每 10 μL 溶液中含有 0.2 mmol•L-1 dNTPs、20 ng 模板 DNA、30 ng•μL-1 引物、0.5 U Taq 聚合酶和 2 mmol•L-1 MgCl2; 最佳的复性温度和循环次数为 53 ℃和 35 次; 从 340 个引物组合中筛选出条带清晰、多态性好的 118 个引物组合,并扩增出7 582 个谱带, 多态性谱带有6 590 个, 多态率为86.92%; 在这些谱带中发现19 条特异性的谱带, 其中居群ZY42占31.58%; 聚类分析结果显示鱼腥草居群存在非常丰富的遗传变异。

关键词:鱼腥草; 居群; SRAP; 遗传多样性

收稿:2009-12-07   修定:2010-01-08

资助:科技部国家基础条件平台建设基金(2004DKA30430)和湖南农业大学人才引进基金(2003YJ007)

An Analysis of Genetic Diversity of Houttuynia cordata Thunb. Population by SRAP Molecular Markers

ZHONG Jun1,3, WANG Kun2, QIU Ping3, ZENG Wei-Jun3, XIONG Xing-Yao4,*
1College of Agriculture, Hunan Agricultural University, Changsha 410128, China; 2College of Bioscience and Technology, Hunan Agricultural University, Changsha 410128, China; 3Hunan Zhengqing Pharmaceutical Co., Ltd., Huaihua, Hunan 418000, China; 4College of Horticulture and Gardening, Hunan Agricultural University, Changsha 410128, China

Corresponding author: XIONG Xing-Yao; E-mail: xiongxingyao@yahoo.cn; Tel: 0731-84635 295

Abstract:

On the basis of optimized orthogonally SRAP-PCR amplification system, genetic diversity of Houttuynia cordata population was studied. The results showed that the optimum SRAP-PCR reaction system contained 0.2 mmol·L-1 dNTPs, 20 ng template DNA, 30 ng·μL-1 primers, 0.5 U Taq DNA polymerase and 2 mmol·L-1MgCl2 in a total volume of 10 μL and the optimal annealing temperature and cycling times was 53 ℃ and 35 times. 118 effective primer-combinations were screened from 340 primer-combinations. A total of 7 582 bands were detected with those primers and 6 590 of them were polymorphic. The percentage of polymorphy was 86.92%. Among those bands, 19 bands had specificity and ZY42 population was 31.58%. The cluster results showed that the genetic differentiation was very abundant, and the SRAP molecular marker was used to identify the genetic differences of H. cordata population.

Key words: Houttuynia cordata; population; SRAP; genetic diversity

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