石蒜属种间杂交种的鉴定和分子身份证构建  

Construction of molecular identification card of Lycoris interspecific hybrids

作　　者：王黎[1] 周琪 高燕会[1] WANG Li;ZHOU Qi;GAO Yanhui(State Key Laboratory of Subtropical Silviculture,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China)

机构地区：[1]浙江农林大学省部共建亚热带森林培育国家重点实验室,浙江杭州311300

出　　处：《浙江农林大学学报》2022年第3期562-570,共9页Journal of Zhejiang A&F University

基　　金：浙江省大学生科技创新活动计划暨新苗人才计划项目(2019R412008);“十四五”浙江省科技计划项目(2021C02071-6);“十三五”浙江省科技计划项目(2016C02056-13);国家自然科学基金资助项目(31670696)。

摘　　要：【目的】鉴定石蒜属Lycoris种间杂交种F_(1)的真实性。【方法】利用荧光标记表达序列标签简单序列重复(ESTSSR)方法,构建亲本石蒜L.radiata、中国石蒜L.chinensis、换锦花L.sprengeri和石蒜-中国石蒜、石蒜-换锦花杂交种F_(1)共78个样品的分子身份证,并分析遗传关系。【结果】15对多态性强的荧光标记EST-SSR引物得到扩增条带92条,每对引物平均扩增6.13条;多态性信息量(PIC)为0.6292~1.0000,平均为0.9137。引物SSR203+SSR115对石蒜-换锦花、石蒜-中国石蒜杂交种F_(1)代的真实性鉴定率分别为96.30%和96.15%。非加权组平均法(UPGMA)聚类分析表明:石蒜、中国石蒜、换锦花及其种间杂交种的遗传相似系数为0.76~0.98,在相似系数为0.77处25个亲本和53个杂交种聚为Ⅰ和Ⅱ两大类,Ⅰ类包括亲本换锦花、石蒜-换锦花杂交种,Ⅱ类包括石蒜、中国石蒜以及中国石蒜-石蒜杂交种,Ⅱ类又可分为Ⅱa、Ⅱb和Ⅱc等3个亚类。利用荧光标记EST-SSR基因型编码,构建了78个石蒜属亲本及杂交种的分子身份证。【结论】荧光标记EST-SSR可用于石蒜属种间杂交种的早期鉴定。[Objective]This study is aimed to identify the authenticity of F_(1),an interspecific hybrids among Lycoris.[Method]With 78 samples collected of F_(1),an interspecific hybrids among L.radiata,L.chinensis and L.sprengeri,expressed sequence tag simple sequence repeat(EST-SSR)was employed to construct their molecular identification cards before an analysis was conducted of their genetic relationships.[Result]fluorescence labeled EST-SSR primers yielded 92 amplified bands that averaged 6.13 per pair;polymorphism information(PIC)was 0.6292 to 1.0000 and averaged 0.9137.The authenticity identification rate of primers SSR203+SSR115 for the F_(1) generation of L.radiata-L.sprengeri and L.chinesis-L.radiata was 96.30%and 96.15%,respectively.UPGMA cluster analysis showed that the genetic similarity coefficients of parents(L.radiata,L.chinensis and L.sprengeri)and their hybrids ranged from 0.76 to 0.98,and with the similarity coefficient being 0.77,the 25 parents and 53 hybrids could be clusted into 2 groups,namely groupⅠand groupⅡ.GroupⅠincluded L.sprengeri parents and hybrids between L.radiata and L.sprengeri whereas groupⅡ,including L.radiata,L.chinensis and the hybrids between L.radiata and L.chinensis,could be divided into three subclades:Ⅱa,Ⅱb andⅡc.The unique molecular identity cards of 78 Lycoris parents and hybrids were constructed based on the fluorescent EST-SSR genotypes coding.[Conclusion]Fluorescence labelled EST-SSR can be used for early identification of interspecific hybrids in Lycoris,which can provide important reference for variety identification and cross breeding of Lycoris.

关 键 词：石蒜属 杂种鉴定 简单序列重复(SSR) 分子身份证 

分 类 号：S682[农业科学—观赏园艺]