摘要
利用松属5个树种的EST序列,分别树种查找SSR位点,采用Primer3.0软件进行引物设计。设计的5个树种的EST—SSR引物中:所占比例最高的均为三核苷酸重复,其次是六核苷酸重复,两者之和都达到了64%以上;四核苷酸重复所占比例均为最低。从设计的引物中分别不同树种各选取20对进行引物合成和通用性检测以及马尾松群体检测,结果表明:松属5个树种的EST序列开发的马尾松SSR引物,其在马尾松中的扩增成功率为60%～80%;最高的是辐射松,最低的是海岸松,平均为72%。通用性最好的为辐射松,属内的通用性为94%;属间和科间通用性也分别达到75%和44%。不同树种序列来源的EST—SSR引物,其扩增成功率在10%～30%;最高的是北美短叶松,其次是辐射松,最低的是扭叶松和火炬松;平均为17%。
The EST sequences from 5 species （P. banksiana, P. radiata, P. pinaster, P. contorta and P. taeda） of Pinus were ana- lyzed and the SSR locus were found respectively, and the primers of the SSR found above were designed using Primer 3.0 software. The types of EST-SSR primers designed from the 5 species of Pinus were compared and it showed that trinucleotide repeat had the highest proportion in all species, followed by hexanueleotide repeat, and the sum of them reached to 64%, the lowest was tetranucleotide re- peat. 20 pairs of SSR primers of each species were selected to detect the transferability and the rate of success amplification in P. mozsoniana. The results showed that the rate of success amplification was 60% ~ 80% for primers designed from the 5 species of Pi- nus, the highest rate of success amplification was P. radiata, the lowest was P. pinaster, and the average rate was 72%. The best trans- ferability in Pinus was P. radiata, with the transferability of 94%, and the transferability in intergeneric and interfamilial were 75% and 44% respectively. The EST-SSR primers designed by sequences from different species were tested in P. massoniana groups, it showed that the amplification rate was 10% - 30%, the highest was P. banksiana, followed by P. radiata, the lowest were P. contorta and P. taeda, and the average rate was 17%.
出处
《湖南林业科技》
2013年第1期14-17,64,共5页
Hunan Forestry Science & Technology
基金
林业公益性行业科研专项经费资助(201104010)
国家"十二五"科技支撑项目(2012BAD01B02)
江苏高校优势学科建设工程项目资助
