根据番茄叶霉病高抗基因Cf-9的基因序列设计3对特异扩增引物,以近等基因系和本组的多重PCR检测材料为试材,扩增Cf-9基因1~2867 bp之间的单拷贝片段,3对引物分别扩增出560 bp,1 000 bp,1 080 bp的特异片段.分别用限制性内切酶TaqⅠ,Hind Ⅲ,HinfⅠ,EcorⅠ酶切,限制性内切酶TaqⅠ酶切特异引物SCAR1扩增的560特异片段具有酶切多态性,抗病品种酶切出450 bp,330 bp,290 bp的特异片段,感病品种酶切出450 bp,290 bp的特异片段.初步建成番茄叶霉病高抗基因Cf-9的CAPS标记,经近等基因系及其杂交种检验,结果稳定可靠,可用于番茄Cf-9抗病基因辅助选育.
Leaf mould is one of the serious diseases on tomato under protective cultivation (greenhouses and plastic houses), and is also a worldwide disease, which had caused heavy losses in tomato production. It is important to breed the resistant varieties to prevent the diseases. Disease-resistant genes are the basis for molecular screening of resistant varieties. With advances in molecular biology and molecular techniques, many resistant genes in tomato against Cladosporiurn fulvum have been identified and cloned, some of which have already been used in molecular breeding, providing a new approach to efficiently controlling the disease. In this study, three pairs of PCR primers were designed based on the sequence of Cf-9 gene, which produced 560 bp, 1 000 bp and 1 080 bp, respectively. PCR products were then digested with the restriction enzymes Taq Ⅰ , HindⅢ, Hinf Ⅰ and Ecor Ⅰ. Only the product amplified by SCAR1 showed polymorphism, with resistant varieties giving 3 restriction fragments (450 bp, 330 bp and 290 bp) and susceptible varieties producing 2 fragments (450 bp and 290 bp). The CAPS marker was developed based on the polymorphism of restriction sites. This result provides a useful tool for molecular-assisted selection of high resistance gene Cf 9 to leaf mould in tomato breeding.