园艺学报 ›› 2003, Vol. 30 ›› Issue (6): 678-682.

• 研究论文 • 上一篇    下一篇


李君明 宋 燕 徐和金 周永健 Carole CaraJ1ta2 冯兰香   

  1. ( 中国农业科学院蔬菜花卉研究所,北京100081; Institute National Breeding of Fruits and Vegetables,Monfavet Avignon 84143,France)
  • 收稿日期:2003-04-19 修回日期:2003-07-17 出版日期:2003-12-25 发布日期:2003-12-25

Simultaneous Identification of Multi-genes、 th Resistance Respectively to Rootknot Nematode and TS、VV by PCR M arkers in Toma to

lJ Jurlnling ,Song yan ,Xu Hejin ,Zhou Yongjian ,Carole Caran~ ,and Feng 1.anxiang   

  1. ( Institute of and Flowers,Chinese Academy of Agricultural Sciences,Bejing 100081, China; Institute National Breeding of Fruits and Vegetables,Montfavet Avignon 84143,France)
  • Received:2003-04-19 Revised:2003-07-17 Online:2003-12-25 Published:2003-12-25

摘要: 利用同一PCR反应体系,对分别与番茄抗根结线虫的 基因和抗斑萎病毒(1w V)的Sw一5
基因紧密连锁的SCAR标记进行了同时扩增筛选,扩增的特异性片段与单引物扩增片段完全吻合,其中与基因紧密连锁的SCAR1标记为共显性标记,抗感试材均产生750 bp的特异片段,纯合和杂合抗病基因型试材存在 I酶切位点,酶切后分别产生了570 bp、160 bp和750 bp、570 bp、160 bp的不同特异性片段,而感病基因型试材无 I酶切位点;与Sw一5基因紧密连锁的SCAR2标记为显性标记,只有抗病试材扩增出400 bD的特异性片段。经反复验证,结果稳定准确可靠,可用于在同一PCR反应体系中对两个抗病基因进行同时筛选鉴定。

关键词: 番茄, 分子标记, PCR, Mi基因, 一5基因

Abstract: Abstract:Single PCR reaction with two SCAR markers respectively tightly linkedwith M/an d Sw一5 genes in tomato,which resistant to root—knot nema tode an d toma to spotted wi lt virus,has been used to screen the multiplex
ban ds.The PCR products with multiplex primers were completely correspond to the amplified bands produced by single SCAR primer.Among them,codominant SCAR 1 Balker tightly linked with Mi gene produce 750 bp flag—
ment in both resistant an d susceptible toma to lines.Th e am plified ban ds from susceptible an d resistant lines were distinguishable after cleavage wi th the restriction enzyme I. Genotype wi th homozygous an d heterozygous M /
gene could produce respectively 570 bp, 160 bp an d 750, 570 bp, 160 bp bands . Susceptible genotypes still present 750 bp fragment.The dominant SCAR2 marker tightly linked with Sw一5 gene would produce only 400 bp PCR product in resistant genotype. Th e replicated stable results pmved that tw o resistant genes could be identified simultaneously by using corresponding SCAR primer under adaptable condition.This system compared with single
primerPCR would be time saving, less labor an d low cost. It could be very useful for marker—as sisted selection during early stage in tommo and fficiently speed up breed ing procedure.

Key words: Tomato, PCR, M/gene, Sw一5 gene