基于色木枫转录组数据的核低拷贝基因引物开发  

Low-copy nuclear gene primers based on transcriptome data of Acer pictum

作　　者：邓雅文 陈荣樟 鲍蕾[1,2,3] 管宇 王红芳 DENG Yawen;CHEN Rongzhang;BAO Lei;GUAN Yu;WANG Hongfang(Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering,100875,Beijing,China;National Forestry and Grassland Administration Key Laboratory for Conservation Ecology in the Northeast Tiger and Leopard National Park,100875,Beijing,China;College of Life Sciences,Beijing Normal University,100875,Beijing,China)

机构地区：[1]生物多样性与生态工程教育部重点实验室,北京100875 [2]东北虎豹国家公园保护生态学国家林草局重点实验室,北京100875 [3]北京师范大学生命科学学院,北京100875

出　　处：《北京师范大学学报（自然科学版）》2022年第6期844-852,I0014,I0015,共11页Journal of Beijing Normal University(Natural Science)

基　　金：国家自然科学基金面上项目(32071494,32171654)。

摘　　要：基于色木枫的转录组数据初步设计166对引物,并根据琼脂糖凝胶电泳与Sanger测序结果筛选出其中的26对核低拷贝基因引物.在东亚63个种群的182个个体中大批量扩增所开发引物,单倍型多样性范围为0.45~0.97,核苷酸多样性为(1.90~18.03)×10^(-3);9~15对引物可在金钱槭、茶条槭、青榨槭和鸡爪槭中扩增;STRUCTURE聚类结果与之前核微卫星研究结果类似,且展示了更多的谱系地理结构,结果表明引物多态性高、拓展性强、具有可靠性.本研究的转录组数据为槭属的研究提供重要遗传信息,所用方法被证明可以有效开发大量核低拷贝基因引物,已开发出的26对引物将为色木枫的进化历史和槭属系统发育研究提供丰富有效的分子标记.Tertiary relic temperate tree species Acer pictum is excellent to study evolutionary history in East Asia.Abundant morphological variations exist in this species,contributing to controversial classification at the species and subspecies level.Therefore low-copy primers with clear mutation mechanism and high diversity are of great significance in the evolution and taxonomy of A.pictum.In this study,we designed 166 pairs of primers based on transcriptome data,and screened for low-copy genes by agarose gel electrophoresis and Sanger sequencing.A total of 26 pairs of low-copy primers were used.Effectiveness of primers was verified by amplifications in 182 individuals of 63 populations in East Asia.Nucleotide diversity was found to range from 1.90×10-3to 18.03×10^(-3),haplotype diversity from 0.45 to 0.97.Nine to fifteen pairs of primers were used for successful amplifications in Dipteronia sinensis,Acer ginnala,A.davidii and A.palmatum.STRUCTURE clustering data were found similar to previous nuclear microsatellite studies and showed more phylogenetic geography.Transcriptome data provided important genetic information for Acer,the method was effective.The 26 pairs of primers will provide rich and effective molecular markers for the study of evolutionary history of A.pictum and the phylogeny of Acer.

关 键 词：引物开发 转录组测序 核低拷贝基因 槭属 

分 类 号：Q37[生物学—遗传学]