中国中药杂志

2011, v.36(03) 289-293

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丹参新的EST-SSR分布规律及分子标记的建立
New analysis of EST-SSR distribution and development of EST-SSR markers in Salvia miltiorrhiza

王学勇;周晓丽;高伟;崔光红;黄璐琦;刘春生;
WANG Xueyong1,ZHOU Xiaoli1,GAO Wei3,CUI Guanghong2,HUANG Luqi2,LIU Chunsheng1(1.Beijing University of Chinese Medicine,Beijing 100102,China;2.Institute of Chinese materia medica,China Academy of Chinese Medical Scineces,Beijing 100700,China;3.School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China)

摘要(Abstract):

目的:建立新近较高覆盖度丹参EST-SSR分子标记,为不同产地来源丹参居群的遗传变异分析奠定基础。方法:对截止2010年11月Genbank下载的丹参EST序列结合HMPL实验室获取的共计1 408条EST序列,进行处理,获取高质量EST序列;经SSRIT软件搜索、分析EST序列中SSR位点的分布规律及类型;在此基础上,运用Websat软件设计EST-SSR引物,经对不同产地丹参样本DNA模板的PCR筛选和条件优化,建立最新EST-SSR分子标记。结果:丹参的EST-SSR种类丰富、覆盖度较大,平均每5.8 kb就检出1条SSR。各种类型出现的频率相差很大,主要重复类型为二核苷酸、三核苷酸重复,分别占SSR总数的63.0%,35.5%,其次为四核苷酸和五核苷酸重复。在重复基元类型中,二核苷酸重复以CT/AG为主,三核苷酸重复以GAA/TCC为主。所优选的36对引物中有29对引物有扩增产物,有效率达80.5%,所选引物对不同产地丹参居群的分子标记出现了多态性。结论:建立的丹参最新EST-SSR分子标记具有较高的覆盖度和多态性,可用于不同产地丹参居群的遗传变异分析。
Objective: To establish the new EST-SSR markers for analyzing the genetic variation of different population of Salvia miltiorrhiza.Method: It was dealt with ESTs newly downloaded from Genbank and that of acquired from HMPL lab EGassembler software,and then carried out SSR loci search and SSR type analysis by SSRIT software.After that,it was designed the EST-SSR primer pairs for PCR amplification condition optimization.Result: Abundant and high coverage of SSR loci distribution were found in S.miltiorrhiza with having one SSR per 5.8 kb ESTs.Among them,the occurrences of different repeat units were mainly the di-(63.0%) and tri-(35.5%).The CT/AG was the most frequent motif in dinucleotide motif type and the GAA/TCC was the most frequent motif in trinucleotide repeats.Out off 36 primer pairs,29 primer pairs(80.5%) were successfully amplified in all samples of S.miltiorrhiza while the rest failed to give PCR products at various annealing temperature and Mg2+ concentrations.The selected primer pairs also showed the polymorphism in samples from different S.miltiorrhiza populations.Conclusion: The newly establishment of EST-SSR markers showed high SSR loci coverage and genetic polymorphisms in S.miltiorrhiza population.It could be used for genetic variation analysis.

关键词(KeyWords): 丹参;EST-SSR;分子标记
Salvia miltiorrhiza;EST-SSR;molecular markers

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基金项目(Foundation): 国家自然科学基金项目(30801517);; 北京市自然科学基金项目(7092056)

作者(Author): 王学勇;周晓丽;高伟;崔光红;黄璐琦;刘春生;
WANG Xueyong1,ZHOU Xiaoli1,GAO Wei3,CUI Guanghong2,HUANG Luqi2,LIU Chunsheng1(1.Beijing University of Chinese Medicine,Beijing 100102,China;2.Institute of Chinese materia medica,China Academy of Chinese Medical Scineces,Beijing 100700,China;3.School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China)

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