杜鹃基因组微卫星富集文库构建.docVIP

杜鹃基因组微卫星富集文库构建 　　摘要：利用FIASCO方法（Fast Isolation by AFLP Sequences COntaining repeats）构建杜鹃（Rhododendron simsii Planch.）的（AG）n微卫星富集文库。通过菌液PCR检测，从（AG）n微卫星富集文库的356个阳性菌落中筛选了233个进行测序分析。结果表明，202个序列富含SSR位点，阳性比率达86.7%。去除杂峰、叠峰、信号弱的序列，最终筛选39个供后续引物开发使用，其中完美型SSR有22个，不完美型5个，混合型12个。磁珠富集法构建杜鹃基因组微卫星文库是一条高效可行的途径，为微卫星位点的分离、杜鹃种群遗传多样性和遗传结构的研究奠定了基础。 　　关键词：杜鹃（Rhododendron simsii Planch.）；微卫星；探针；富集文库 　　中图分类号：S685.21；Q343.1 文献标识码：A 文章编号：0439-8114（2015）03-0627-04 　　DOI：10.14088/ki.issn0439-8114.2015.03.030 　　Constructing Microsatellite-enriched Library of Rhododendron simsii Planch. 　　WANG Shu-zhen，JIN Wei-bin，XIANG Jun，ZHENG Yong-liang，FANG Yuan-ping， 　　GAN Jian-ping，CHENG Hua，DU Hang 　　（Hubei Key Laboratory of Economic Forest Germplasm Improvement and Resource Comprehensive Utilization/College of Life Science， Huanggang Normal University， Huanggang 438000， Hubei， China） 　　Abstract： （AG）n microsatellite-enriched library of Rhododendron simsii Planch. was constructed by FIASCO（Fast Isolation by AFLP Sequences COntaining repeats） method. Among 356 positive colonies verified by colony PCR of （AG）n microsatellite-enriched library， 233 colonies were chosen for further sequencing. Results showed that 202 colonies contained microsatellites， accounting for 86.7%. Discarding sequences with miscellaneous peak， overlapping peaks and weak signal， 39 sequences were selected for primer exploitation. The perfect， imperfect and compound microsatellite repeat types were 22， 5 and 12， respectively. The FIASCO method was efficient for constructing the enrichment library of R. simsii Planch. microsatellite. It will lay foundation for developing microsatellite markers and studying genetic diversity and genetic structure of Rhododendron species. 　　Key words： azalea（Rhododendron simsii Planch.）； microsatellite； probe； enriched library 　　杜鹃花（Rhododendron spp.）泛指杜鹃花科（Ericaceae）杜鹃花属（Rhododendron）植物，具有极高的观赏价值，并被誉为“花中西施”和“木本花卉之王”[1，2]。杜鹃花在亚洲、欧洲、北美洲等地广泛分布，并且因气候条件不同而垂直分布明显，从而形成了常绿大乔木、常绿小乔木、常绿灌木、落叶灌木等不同形态特征的地理居