中文摘要：

在转录组基础上批量开发地黄特异EST-SSR标记序列,同时通过优化DNA模板浓度、引物浓度、dNTP浓度、Taq酶浓度,结合正交优化试验建立地黄EST-SSR的PCR最佳扩增体系,为进一步鉴定和开发地黄多态性SSR提供重要的基础资料。结果表明：在地黄87665条转录本序列中,共筛选了1812条不小于18bp的SSR序列,其中二核苷酸数量最多,共有721个（70.8%）,其次为三核苷酸和六核苷酸,分别有486、578个。而在所有SSR类型中AG/CT（457个）为最多的重复类型,其次为AC/GT（256个）。EST-SSR体系优化结果表明：最佳反应体系为15μL体系中含80ng模板DNA、0.1μmol/L引物、150μmol/Ld NTPs和1.5U TaqDNA聚合酶,在退火温度55℃进行30个循环。

英文摘要：

In this study,specific EST-SSR markers sequences of Rehmannia glutinosa were exploited on the base of transcriptome,and at the same time,optimal amplification system of R.glutinosa EST-SSR though optimizing concentration ofDNA template,primers,Taq polymerase and dNTP was established by orthogonal experiment.The results indicated that 1 812 EST-SSR（≥18 bp）were screened from the 87 665 transcription sequences of R.glutinosa.Of which,dinucleotide（70.8%）content was the most（70.8%）,followed by three nucleotides（486）and six nucleotides（578）.In all types of SSR,AG/GT（457） was the maximum repeat type,then the AC/GT（256）.The results of optimization of EST-SSR system suggested that,the best amplification system of 15 μL contained 80 ngDNA template,0.1 μmol/L primers,150 μmol/L d NTPs and 1.5 U TaqDNA polymerase,at 55℃ for 30 cycles.