基于松科树种EST序列的落叶松SSR引物开发  

Development of Larix leptolepis SSR Primers from EST Sequence of Pinaceae Family

作　　者：贯春雨[1] 张含国[1] 张磊[1] 朱航勇[1] 邓继峰[1] 

机构地区：[1]东北林业大学,哈尔滨150040

出　　处：《东北林业大学学报》2011年第1期20-23,共4页Journal of Northeast Forestry University

基　　金：国家自然科学基金项目(30640034);国家科技基础性工作专项(2007FY110400-3)资助

摘　　要：登陆NCBI的dbEST数据库搜索松科树种EST序列,应用SSRIT软件在线搜索EST-SSR,用Primer3设计EST-SSR引物。选用日本落叶松(L.leptolepis)×兴安落叶松(L.gmelinii)及其145个F1为检测植株,对所开发EST-SSR引物进行扩增,并以丙烯酰胺凝胶检测扩增产物,以获得适合落叶松的SSR引物,以期用于该家系的图谱构建及QTL定位的研究。NCBI数据库搜索松科EST序列40608条,使用SSRIT检测得到SSR位点1011个,频率为2.49%。使用primer 3设计EST-SSR引物132对。SSRs以二核苷酸重复最为常见,重复基元11种,重复数量5598个,占全部重复基元比例达到83.79%;其次为三核苷酸,重复基元40种,重复数量1067,占全部重复单元的比例为15.97%;四核苷酸为3种重复基元,重复数量16,所占比例为1.95%。经扩增检测125对EST-SSR引物在日×兴亲本及F中获得扩增产物,占94.7%。A search for EST sequence of Pinaeeae species were made by dbEST database of NCBI. SSRIT software was used to search EST-SSR online and primer3 to design EST-SSR primers. Larix leptolepis, L. gmelinii and their 145 full-sib F1 individuals were amplified by the EST-SSR primers, and then the amplified products were detected by acrylamide gel. Suit- able SSR primers were selected for larch to construct genetic map and QTL mapping for the family. Through the sequence analysis of 40608 ESTs in Pinaceae deposited in NCBI, a total of 1011 SSR loci were obtained using the SSRIT software, and the frequency was 2.49%. One hundred and thirty-two pairs of EST-SSR primers were designed by primer3. The di- nucleotide repeats were most common in SSRs, and the repeated primitives were 11, accounting for 83.79% of the total repeated primitives, and the repeat number was 5 598. The trinueleotide repeats were more common, and the repeated primitives and repeat number were 40 and 1 067, respectively, accounting for 15.97% of the total repeated primitives. Tetranucleotide repeats included three types of repeated primitives, and the repeat number was 16, accounting for 1.95% of the total. A total of 125 EST-SSR primer pairs were obtained which amplified products in the parents （L. leptolepisxL. gmelinii） and F1, accounting for 94.7% of the total.

关 键 词：松科 落叶松 EST—SSR 引物开发 

分 类 号：S722.7[农业科学—林木遗传育种] S791.22[农业科学—林学]