星油藤转录组SSR序列特征分析及其分子标记开发  

Characteristic Analysis of SSR Sequences and Development of Molecular Markers Based on Plukenetia volubilis Transcriptome

作　　者：朱阅 郭亚静 冯程 潘帮珍[2] 何惠英[2] 徐增富[2] 杜光辉 付乾堂[2] Zhu Yue;Guo Yajing;Feng Cheng;Pan Bangzhen;He Huiying;Xu Zengfu;Du Guanghui;Fu Qiantang(College of Agriculture,Yunnan University,Kunming,650500;CAS Key Laboratory of Tropical Plant Resources and Sustainable Use,Innovative Academy of Seed Design,Xishuangbanna Tropical Botanical Garden,Chinese Academy of Sciences,Mengla,666303)

机构地区：[1]云南大学农学院,昆明650500 [2]中国科学院西双版纳热带植物园,种子创新研究院,中国科学院热带植物资源可持续利用重点实验室,勐腊666303

出　　处：《分子植物育种》2021年第9期2985-2992,共8页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31870291);云南省自然科学基金(2016FB051);中科院西双版纳热带植物园一三五项目(2017XTBG-T02)共同资助。

摘　　要：基于星油藤(Plukenetia volubilis L.)转录组信息,分析其表达序列标签-简单序列重复(expressed sequence tag-simple sequence repeat,EST-SSR)并设计引物进行验证,为星油藤的SSR分子标记开发提供数据支持。本研究利用MISA软件对星油藤雌雄花序芽转录组测序所获得的57664条Unigenes进行分析,共检测出10572个SSR位点;其中8825条Unigenes分布有SSR位点,占总Unigenes的15.30%。SSR位点的平均分布频率为每5.34 kb出现1个SSR位点。各类型的SSR位点共有154种,其中单核苷酸重复(40.48%)、二核苷酸重复(25.82%)和三核苷酸重复(28.89%)为主要的重复类型,A/T、AG/TC和AAG/CTT分别为单核苷酸、二核苷酸和三核苷酸类型中主导的重复基序。利用Primer 3.0设计EST-SSR引物,并随机挑选75对SSR引物,以星油藤及其2个近缘种(亚洲星油藤和大果星油藤)为材料进行筛选验证,其中31对引物可进行有效扩增,并最终获得7对在3个物种间具有多态性的引物。本研究结果表明,星油藤转录组测序产生的Unigenes序列信息作为开发SSR分子标记是可行的,开发的这些SSR分子标记可用于星油藤及其近缘种的遗传多样性评价、种质资源鉴定和分子标记辅助育种等研究。In order to develop simple sequence repeat(SSR)molecular markers in Plukenetia volubilis,the expressed sequence tag-SSR(EST-SSR)loci information was analyzed based on the transcriptome of this plant,and specific primers of SSRs were designed to confirm their validity.A total of 10572 SSR loci were screened from 57664 Unigenes obtained from the transcriptome of male and female inflorescence buds in P.volubilis using MISA software,and 8825(15.30%)unigenes were identified containing SSR loci.The average frequency of SSR in unigenes was one in every 5.34 kb of analyzed sequences.The dominant motifs in the 154 SSR types were mononucleotide repeat,trinucleotide repeat and dinucleotide repeat,accounting for 40.48%,28.89%and 25.82%,respectively.A/T,AG/TC,and AAG/CTT were the dominant repeat motifs in mononucleotide,dinucleotide and trinucleotide types,respectively.EST-SSR primers were designed using Primer 3.0,and 75 pairs of SSR primers were randomly selected and verified in P.volubilis and its two related species(Plukenetia corniculata and Plukenetia conophora).Clear bands were amplified by 31 pairs of primers and good polymorphism bands were produced by seven pairs of primers in these three species.These results indicated that it was feasible to develop SSR markers based on the transcriptome information of P.volubilis.These SSR markers could be used for the genetic diversity evaluation,germplasm identification,and molecular marker-assisted breeding in P.volubilis and its related species.

关 键 词：星油藤 亚洲星油藤 大果星油藤 转录组 EST-SSR 

分 类 号：S565.9[农业科学—作物学]