利用RAPD和ISSR 2种分子标记鉴定西瓜杂交种的遗传纯度  

Evaluation of Genetic Purity of Watermelon Hybrid(Citrullus lanatus) Using RAPD and ISSR Molecular Markers

作　　者：羊杏平[1,2] 刘广[2] 侯喜林[1] 徐锦华[2] 高长洲[2] 

机构地区：[1]南京农业大学园艺学院作物遗传与种质创新国家重点实验室,江苏南京210095 [2]江苏省农业科学院蔬菜研究所,江苏南京210014

出　　处：《江苏农业学报》2010年第6期1313-1318,共6页Jiangsu Journal of Agricultural Sciences

基　　金：江苏省科技基础设施建设计划项目(BM2008008);国家科技支撑计划项目(2006BAD01A7);国家西甜瓜产业技术体系项目(nycytx-36-01-01-03);江苏省科技支撑计划项目(BE2009307);江苏省农业科技自主创新基金项目[CX(09)601]

摘　　要：为了鉴定西瓜杂交种抗病苏蜜和苏蜜5号的遗传纯度,利用RAPD和ISSR 2种分子标记技术对F1杂种及其相应亲本的基因组DNA进行了分析。结果显示:在抗病苏蜜杂交种的分析中,对450个随机RAPD引物进行了筛选,其中139个引物产生了235个多态性条带,平均每个引物扩增出了3.12个条带,多态率为16.7%。139个多态性引物中,3个引物产生了母本特异标记,4个引物产生了父本特异标记,2个引物产生了共显性标记。引物JAASRP388的扩增图谱显示产生了1个母本特异标记JAASRP3881200及2个父本特异标记JAASRP3882010、JAAS-RP388500。引物JAASRP410的扩增图谱显示也产生了1个母本特异标记JAASRP410625及2个父本特异标记JAAS-RP4102000、JAASRP4101000。在杂交种苏蜜5号及其亲本的分析中,100个RAPD引物产生了325条扩增带,其中27个RAPD引物成功地扩增出了37个多态性条带,平均多态率为11.38%。在27个多态性RAPD引物中,4个引物产生了母本特异标记,1个引物产生了父本特异标记。此外对62个ISSR引物进行了检测,45个引物成功地产生了188个扩增条带,每个引物平均产生了4.18个条带,15个为多态性引物产生了23条多态性带,平均多态率为12.23%。在ISSR检测中,只发现了2个引物能产生母本特异条带。研究结果说明,不管是1个母本特异引物和1个父本特异引物组合还是利用1个共显性引物都是杂交种种子质量检测过程中一个非常有效的工具。Two molecular marker systems,RAPD（random amplified polymorphic DNA） and ISSR（inter-simple sequence repeat）,which are based on polymerase chain reaction,were used to evaluate seed genetic purity of two watermelon hybrid Kangbingsumi and Sumi No.5.Genomic DNA from the F1 hybrids and their corresponding parental lines was analyzed with RAPD primers and ISSR primers.In Kangbingsumi,of 450 random RAPD primers screened,139 primers generated 235 polymorphic bands.On average each primer amplified 3.12 scorable bands,with polymorphic ratio of 16.7%.Out of 139 polymorphic primers,three primers produced female parent-specific markers,four primers produced male parent-specific markers,and two primers produced male and female parent-specific markers simultaneously in the hybrid.The codominant primer JAASRP388 produced one female parent-specific marker JAASRP3881200 and two male parent-specific markers JAASRP3882010 and JAASRP388500 simultaneously.The codominant primer JAASRP410 produced one female parent-specific marker JAASRP410625 and two male parent-specific markers JAASRP4102000 and JAASRP4101000.In the hybrid Sumi No.5 and its parental lines,27 RAPD primers successfully amplified 37 polymorphic bands out of 100 RAPD primers screened which generated 325 bands,and the average polymorphism rate was 11.38%.Of the 27 primers,four primers generated the female parent-specific bands,and one primer generated the male parent-specific bands.Out of the 62 ISSR primers detected,45 primers successfully generated 188 bands with the average of 4.18 bands per primer.Fifteen of the forty-five primers generated 23 polymorphic bands.The average polymorphism rate was 12.23%.There were only two primers which produced the female parent-specific bands useful to test the hybrid Sumi No.5 purity.The results indicated that either the combination of one female parent-specific primer and one male parent-specific primer or one codominant primer could be an efficient implement in the process of quality testing of hybrid watermelon seeds.

关 键 词：西瓜 分子标记 杂交种 遗传纯度 

分 类 号：S651[农业科学—果树学]