JIN Fengmei,XUE Jun,SONG Jian,et al.Identification of Resistance Gene Sw-5 and Ty-3a by Multiplex PCR in Tomato[J].Northern Horticulture,2017,41(11):115-118.[doi:10.11937/bfyy.201711023]
番茄抗病基因Sw-5和Ty-3a的多重PCR检测体系的建立
- Title:
- Identification of Resistance Gene Sw-5 and Ty-3a by Multiplex PCR in Tomato
- 关键词:
- 番茄斑萎病毒(TSWV); 番茄黄化曲叶病毒; Sw-5; Ty-3a; 多重PCR
- Keywords:
- tomato spotted wilt virus(TSWV); tomato yellow leaf curl virus; Sw-5 gene; Ty-3a gene; multiplex PCR
- 文献标志码:
- A
- 摘要:
- 以含有斑萎病毒和黄化曲叶病毒抗病基因和感病基因的番茄为试材，采用多重PCR方法，建立番茄抗斑萎病毒Sw-5和抗番茄黄化曲叶病毒的Ty-3a基因同时扩增的体系，以期为番茄分子标记抗病育种提供更省时、省力、经济的方法。结果表明：多重PCR扩增的特异性片段与单引物扩增片段完全一致，与Sw-5基因紧密连锁的SCAR标记，在抗病基因型中可扩增出574 bp的条带，感病基因型中扩增出464 bp的条带；与Ty-3a基因紧密连锁的SCAR标记，在抗病基因型中可扩增出630 bp的条带，感病基因型中扩增出320 bp的条带，多重PCR体系可在同一PCR反应体系中对2个抗病基因进行筛选鉴定及苗期早期辅助选育。
- Abstract:
- Tomatoes with spotted virus resistant and susceptible genotypes and yellow leaf curl virus resistant and susceptible genotypes were used as test materials.The multiplex PCR system was developed to detect the Sw-5 and Ty-3a gene simultaneously in order to provide a more time-saving,labor-saving and cost-effective method for tomato resistance breeding.The results showed that the amplified fragments by multiplex PCR were identical with single-primer PCR.A 574 bp band was amplified in the resistant genotype,and a 464 bp band was amplified in the susceptible genotype by the SCAR marker of Sw-5 gene.A 630 bp band could be amplified in the resistant genotype and a 320 bp band was amplified in the susceptible genotype by the SCAR marker of Ty-3a gene.The multiplex PCR system could amplify two genes in one reaction.It could be useful marker assisted selection during seedling stage in tomato and efficiently speed up breeding procedure.
参考文献/References:
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备注/Memo
第一作者简介：金凤媚（1977-），女，天津宝坻人，硕士，副研究员，现主要从事番茄的分子育种等研究工作。E-mail:jinfm414@163.com.基金项目：天津市种业科技重大专项资助项目(16ZXZYNC00070)；国家星火计划资助项目（2015GA610）。