摘要
寒兰(Cymbidium kanran)是中国传统名花,其观赏与经济价值极高。简单重复序列(simple sequence repeats,SSR),又名微卫星标记,有多态性高,非共显性等优势。基于转录组数据对SSR标记的开发具有经济效能高等优点,通过高通量测序平台对寒兰EST-SSR标记进行开发,了解其在转录组序列中的分布类型及特性,为寒兰的SSR多样性的分析及遗传图谱构建奠定基础。通过MISA程序筛选寒兰转录组测序得到的68699条unigene(序列总长60.06 Mb),并分析了其SSR位点分布情况,获得了9837个SSR位点,在总unigene的比例为14.32%,平均每6.25kb出现1个SSR。SSR位点中二核苷酸重复频率达到最高,占55.22%,其中AG/CT高达38.83%;接下来为三、单核苷酸重复,频率依次为25.37%和14.49%。利用Primer3共设计出6017对SSR引物。随机选取141对引物进行PCR扩增,有79对引物扩增得到的条带清晰可重复,15对在15个不同地区寒兰中表现出多态性。结果表明,本研究中设计的SSR引物在寒兰遗传多样性分析中具有较高的适用性,可用于寒兰的遗传研究。
Cymbidium kanranis a traditional flower,with high ornamental and economic value.Simple sequence repeats（SSR）is also called microsatellite markers,with the advantages of high polymorphism and non-co-dominant.In this study,we used high-throughput sequencing platform to develop EST-SSR markers for C.kanran.Distribution patterns of the markers in the transcriptome sequences and their characteristics were analyzed,in order to provide a basis for analysis of SSR diversity analysis and genetic map construction.9837 SSR loci（14.32%）were retrieved from 68,699unigenes（60.06 Mb）,by using MISA program,and the unigene were obtained from transcriptome sequencing.1SSR appears every 6.25 kb,and repeat frequency of dinucleotide in SSR was the highest（55.22%）,among which AG/CT was up to 38.83%,followed by trinucleotide and mononucleotide at 25.37% and 14.49%,respectively.6017 SSR primers were designed using Primer3.Randomized selection of 141 primers was carried out by PCR amplification,of which 79 pairs were amplified by clearly repeating bands,and 15 pairs showed polymorphism in 15 different regions of C.kanran.The results showed that the SSR primers designed in this study had high availability and could be used for the genetic research.
出处
《南昌大学学报（理科版）》
CAS
北大核心
2017年第3期249-254,共6页
Journal of Nanchang University(Natural Science)
基金
国家自然科学基金资助项目(31360491)
江西省博士后基金(2014KY40)
关键词
寒兰
SSR
转录组
多态性
Cymbidium kanran
SSR
Transcriptome
Polymorphism
