利用多重PCR反应同时鉴定番茄Ty-3a基因和抗溃疡病QTL  

Identification of the tomato Ty-3a gene and a QTL site of tomato bacterial canker by multiplex PCR markers

作　　者：张建盈 严景日 杨石奇[1] 王平勇 刘辰[1] 沈火林[1] 

机构地区：[1]设施蔬菜生长发育调控北京市重点实验室.中国农业大学农学与生物技术学院,北京100193

出　　处：《中国瓜菜》2015年第3期5-8,共4页China Cucurbits And Vegetables

基　　金：现代农业产业技术体系北京市果类蔬菜创新团队项目

摘　　要：番茄黄化曲叶病毒病和番茄溃疡病是两种分布广泛、危害严重的世界性病害,严重影响了番茄的产量,是国内外番茄抗病育种的重要目标。为了提高抗病育种的效率,研究建立了抗番茄黄化曲叶病毒病Ty-3a基因和抗番茄溃疡病QTL的多重PCR反应体系。该体系利用同一PCR反应,对分别与番茄抗黄化曲叶病毒病的Ty-3a基因和抗番茄溃疡病的一个QTL位点Rcm5.1紧密连锁的标记进行了扩增、筛选,扩增的特异性片段与单引物扩增片段吻合。其中与Ty-3a基因紧密连锁的标记Ty-3F/Ty-3R为共显性标记,基因型为Ty-3a/Ty-3a的材料均产生630 bp的特异性片段,基因型为ty-3/ty-3的材料均产生320 bp的特异性条带,基因型为Ty-3a/ty-3的材料产生630 bp和320 bp的特异性片段;与番茄溃疡病QTL位点Rcm5.1连锁的标记TG318F/TG318R为显性标记,只有抗病材料产生580 bp的特异性片段,感病材料不产生特异性条带。经反复验证结果准确、稳定,可用于同一PCR反应体系中对番茄抗黄化曲叶病毒病Ty-3a基因和番茄溃疡病QTL的同时筛选鉴定。Yellow leaf curl virus and bacterial canker of tomato are worldwide diseases. They are widely distributed, which reduced yields of tomato seriously, are important goals of tomato breeding at home and abroad. The study establishes PCR reaction system on Ty-3 gene of yellow leaf curl virus and a QTL site Rcm5.1 about bacterial canker of tomato, in order to improve the efficiency of breeding for disease resistance. The two markers tightly linked respectively with the Ty-3 gene resistant to yellow leaf curl virus and a QTL site Rcm5.1 to bacterial canker of tomato were amplified and screened by the same PCR system.The specific fragment amplified was identical with one amplified by single primer. Among them, co-dominant primer Ty-3F/Ty-3R tightly linked with Ty-3 gene produced the specific fragments of 630 bp, 320 bp, 630 bp and 320 bp in the materials with the genotype Ty-3/Ty-3, ty-3/ty-3, Ty-3/ty-3. The dominant primer TG318F/TG318R tightly linked with a QTL site Rcm5. 1 produced a specific fragment of 580bp. The replicated stable results proved that using corresponding PCR primer under adaptable condition could identifiy the Ty-3 gene resistant to yellow leaf curl virus and a QTL site Rcm5. 1 to bacterial canker of tomato simultaneously.

关 键 词：番茄 黄化曲叶病毒病 番茄溃疡病 多重PCR 

分 类 号：S641.2[农业科学—蔬菜学]