利用多重PCR反应同时筛选番茄Cf-9和Tm-1基因  

Identification of the Genes with Resistance to Leaf Mold and Tomato Mosaic Virus by PCR Reaction in Tomato

作　　者：宋燕[1] 陈丽静[1] 李君明[1] 张智[1] 李天来[2] 徐和金[1] 周永健[1] 

机构地区：[1]中国农业科学院蔬菜花卉研究所,北京100081 [2]沈阳农业大学园艺学院,沈阳110161

出　　处：《植物遗传资源学报》2006年第2期165-169,共5页Journal of Plant Genetic Resources

基　　金：国家自然科学基金项目(30100124);国家"863"计划项目(2002AA244011)

摘　　要：利用同一PCR反应体系,对分别与番茄抗叶霉病的Cf-9基因和抗番茄烟草花叶病毒病的Tm-1基因紧密连锁的PCR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吻合。其中与Cf-9基因紧密连锁的CAPs标记在抗感试材均可扩增出560bp的特异片段,且都存在TaqI酶切位点,抗病基因型酶切后分别产生了450bp、330bp和290bp的不同特异性片段,而感病基因型试材酶切后产生450bp和290bp的特异性片段;与Tm-1基因紧密连锁的SCAR标记为显性标记,只有抗病试材产生750bp的特异片段,不能被TaqI酶切。经反复验证,结果稳定准确,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。该体系的建立不仅省时、省工、节省费用,而且可用于苗期辅助选育,加快番茄抗病育种进程。Single PCR reaction with two pairs of PCR primers designed on the markers respectively tightly linked with Cf-9 and Tm-1 genes in tomato, which be resistant to leaf mold and tomato mosaic virus. For the genes the PCR products were almost completely correspond to the amplified bands produced by single PCR primer. Among them, 560bp fragment linked with Cf-9 gene was amplified in both resistant and susceptible tomato lines. The amplified bands from susceptible and resistant lines were distinguishable after cleavage with the restriction enzyme TaqⅠ. Genotype with Cf-9 gene could produce respectively 450bp, 330bp and 290bp bands. Susceptible genotypes could produce respectively 450bp and 290bp fragments. 750bp fragment linked with Tm-1 gene was amplified in resistant tomato line. The amplified bands could not be cleavaged with the restriction enzyme TaqⅠ. The replicated stable results proved that two resistant genes could be identified simultaneously by using corresponding PCR primer under adaptable condition. This system compared with single primer PCR would be time saving, less labor and low cost. It could bevery useful for marker-assisted selection during early stage in tomato and efficiently speed up breeding procedure.

关 键 词：番茄 多重PCR反应 叶霉病 番茄烟草花叶病毒病 

分 类 号：S641.2[农业科学—蔬菜学]