广西莪术EST-SSR标记开发

Development of EST-SSR Markers in Curcama kwangsinesis

目的利用NCBI公布的姜黄EST序列,挖掘SSR位点,开发近缘种广西莪术EST-SSR标记。方法下载NCBI公布的姜黄EST序列,利用SSR-FINDER搜索SSR位点,采用Primer5.0设计SSR引物,挑选30个表现型差异较大的广西莪术进行有效性及多态性检测。结果 12 678条EST序列含有SSR位点1 243个,其中二核苷酸、三核苷酸重复序列最多,分别占50.36%,31.54%,以AT/TA和CT/GA出现频率最高。利用Primer5.0设计引物共325对,PCR检测表明,104对引物可以扩增出稳定清晰的带型;在至少30份不同种质广西莪术中检测到48对SSR引物具有多态性,占设计引物的38.09%。结论姜黄EST资源中含有高频率的SSR位点,且EST-SSR标记开发效率较高。

Objective Based on available sequence information of Curcuma longa L.in NCBI website, and develop the EST-SSR primers from the closely related species-Curcama kwangsinesis. Methods Download the ESTs from Curcuma longa L.which were published in NCBI database, use SSR-FINDER to search the SSR loci and Primer 5. 0 to design primers; chooses 30 phenotype differences germplasm resources of Curcama kwangsinesis to detect the effectiveness and polymorphic of primers. Results A total of 1 243 microsatellite repeats were detected from 12 678 EST sequences of Curcuma longa L. After redundancy elimination, the dinucleotide frequency is 50.36%, and the trinucleotide and the tetranucleotide, 31.54% . AT/TA and CT/GA have the highest frequency.About 325 primers were designed, the PCR detection showed that 104 primers can amplify the clear zone type; Detected in 30 different germplasm in Curcama kwangsinesis. About 48 primers had polymorphism, accounting for 38.09% of all the designed primers. Conclusion The EST resources of Curcuma longa L. has the high frequency of SSR loci, and the EST - SSR markers development is efficiency.