摘要
该研究主要开发筛选适用于杂交兰的EST-SSR引物,为杂交兰种质资源评价和遗传变异研究等提供可靠的分子标记。该研究对杂交兰进行转录组高通量测序,挖掘SSR位点和开发EST-SSR标记,并对不同种质的遗传多样性进行分析。结果表明,从31724条杂交兰Unigene中检测出18603个SSR位点,SSR出现频率为58.64%;SSR位点中的主导类型是单核苷酸重复,占总SSR的65.10%,其次是二核苷酸(23.56%)和三核苷酸(10.76%)重复;优势重复基元为A/T、AG/CT、AT/AT和AAG/CTT,分别占总位点的64.72%、13.74%、8.19%和2.51%。利用Primer Premier 5.0共设计了565对SSR引物,从筛选出的64对有效扩增引物中随机选择28对引物,对40份杂交兰种质进行多态性验证与遗传关系分析,其中16对(占57.14%)引物表现出可重复的高多态性,平均多态信息量(PIC)达0.789。基于扩增的多态性SSR信息,40份种质资源可聚为4类,聚类结果与其遗传背景基本一致。该研究印证了转录组测序获得的Unigene是SSR标记开发的有效来源,开发的EST-SSR引物可为杂交兰及近缘种的良种鉴别、遗传图谱构建、分子标记辅助育种及功能基因挖掘等提供有价值的候选标记。
EST-SSR primers suitable for hybrid Cymbidium were developed and screened to provide reliable molecular markers for germplasm resource evaluation and study on genetic variation.In the present study,transcriptome data of hybrid Cymbidium was obtained by high-throughput sequencing technology.The SSR loci were screened and the EST-SSR markers were developed,and then the genetic diversity of different germplasm were analyzed.The results showed that a total of 18603 SSR loci were mined from 31724 Unigenes with a frequency of 58.64%.Mononucleotide repeat was the main type,accounted for as much as 65.10% of all SSRs,followed by dinucleotide repeat(23.56%)and trinucleotide repeat(10.76%).The dominant repeat elements were A/T,AG/CT,AT/AT and AG/CTT,accounting for 64.72%,13.74%,8.19% and 2.51% of the total loci,respectively.565 pairs of SSR primers were found by Primer Premier 5.0 and 28 efficient pairs of primers were randomly selected from 64 valid pairs for polymorphism analysis.Thereinto,16 pairs of primers showed clear and reproducible results indicating the high poly morphism among 40 different germplasms,with the average PIC was 0.789.Based on the amplified polymorphic SSR data,the 40 materials were divided into 4 major groups by UPGMA.The dendrogram results were accordance with the genetic backgrounds.In conclusion,the Unigenes generated from transcriptome data of hybrid Cymbidium can be used as an effective source to development EST-SSR markers.The SSR markers obtained in the study could be valuable candidate markers for improved varieties identification,genetic map construction,molecular marker-assisted breeding and functional gene mining of hybrid Cymbidium and other Cymbidium species.
作者
钟淮钦
林榕燕
林兵
吴建设
樊荣辉
叶秀仙
ZHONG Huaiqin;LIN Rongyan;LIN Bing;WU Jianshe;FAN Ronghui;YE Xiuxian(Crop Research Institute,Fujian Academy of Agricultural Sciences,Flower Research Center,Fujian Academy of Agricultural Sciences,Fujian Engineering Research Center for Characteristic Floriculture,Fuzhou 350013,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2020年第2期286-295,共10页
Chinese Journal of Cell Biology
基金
福建省省属公益类科研院所专项(批准号:2018R1026-3、2017R1026-8)
福建省农业科学院科技创新团队(批准号:STIT2017-2-9)资助的课题。
