中文摘要：

对重要大宗药用植物丹参尚没有高效特异分子标记开发应用研究,为有效利用丹参现有EST资源开发功能性分子标记,本文以生物信息学方法对GenBank的dbEST数据库中公开的10288条丹参（Salvia miltiorrhiza）EST进行整理,共获得非冗余性EST4192条。从中搜索到159个SSR位点,出现频率为3.79%,平均相隔12.74kb出现1个SSR位点。在1bp～6bp核苷酸重复基序中,二核苷酸重复基序SSR出现频率最高（77个、48.43%）,其次是三核苷酸（41个、25.79%）,四核苷酸出现频率最低（6个,3.77%）。在检出的47类重复基序中,出现频率超过5%的6种重复基序依次为：GA/CT（16.35%）、AG/TC（15.09%）、TCA/AGT（10.69%）、AT/TA（6.29%）、GAAAAG/CAAAAC（6.29%）、TA/AT（5.03%）。针对这些序列,设计开发了83对EST-SSR引物,对13个不同居群丹参样品及10种鼠尾草属物种进行了扩增效率、多态性及通用性检测,发现其中72对引物在供试材料中可进行特异性扩增,共产生279个位点,平均每对引物产生3.88个位点,且所有引物均显示多态性扩增。可有效扩增引物在鼠尾10个异源物种中的可转移率为60%～100%,平均85%。依据扩增结果进行遗传相似性分析,结果表明EST-SSR分析可揭示供试样品不同水平的遗传多样性,并可明确区分丹参及其他同属植物。上述工作为进行丹参及其同属物种的基因组差异性分析、遗传图谱构建及比较基因组等研究奠定了基础。

英文摘要：

Despite Salvia miltiorrhiza being one of the most important medicine plants in China,there is a limited availability of genomic resources,especially of the expressed sequence tag-based markers.In this study,we selected and characterized functional markers in S.miltiorrhiza,which consisted of 4 192 non-redundant expressed sequence tags（ESTs） from 10 288 identified S.miltiorrhiza ESTs in dbEST data bank.Among them,159 simple sequence repeats（SSR） were detected,which amounted to 3.79% of the non-redundant starting sequence population.This incidence was equivalent to one EST-SSR in every 12.74 kb of S.miltiorrhiza ESTs.Among the different motifs ranging from 1 bp to 6 bp,di-nucleotide repeat motif was the most abundant（77,48.43%）,followed by tri-nucleotide（41,25.79%）,hexa-nucleotide（23,14.47%）,penta-nucleotide（12,7.55%） and tetra-nucleotide（6,3.77%）.In 47 identified motif types,the detected frequency above 5% were GA/CT（16.35%）,AG/TC（15.09%）,TCA/AGT（10.69%）,AT/TA（6.29%）,GAAAAG/CAAAAC（6.29%） and TA/AT（5.03%）.Based on flank sequence of detected SSR,a total of 83 EST-SSR primer pairs were designed and tested for the amplification efficiency,polymorphism and transferability in thirteen S.mihiorrhiza samples and other ten species from the genus Salvia.The results showed that 72 primer pairs were successfully amplified in S.miltiorrhiza samples to yield and 279 loci with an average of 3.88 loci per primer pair.The cross-transferability of S.miltiorrhiza EST-SSR markers to other ten Salvia plants was very high,ranging from 60% to 100% with an average of 85%.Further analysis of the genetic similarity based on the polymorphic bands showed the EST-SSR could detect the genetic diversity on different levels among the whole test samples and distinguish the S.miltiorrhiza from other Salvia plants effectively.It is expected that the potential markers described here would add to the repertoire of DNA markers needed for genetic analysis,linkage mapping and comparative genomics studi