基于松科树种EST序列的落叶松SSR引物开发

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F1中获得扩增产物,占94.7%.%A search for EST sequence of Pinaceae species were made hy dhEST datahase 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. Suitable 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 dinucleotide repeats were most c:ommon 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 trinucleotide repeats were more common, and the repeated primitives and repeat numher 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. leptolepi.sxL.gmelinii ) and F1 , accounting for 94. 7％ of the total.