番茄抗晚疫病基因ph-3的RAPD及CAPS标记

RAPD Marker Converting to CAPS Marker of ph-3 Gene Resistant to Phytophthora infestans

将番茄抗晚疫病ph-3基因已有的RAPD特异片段进行克隆、测序。根据该测序结果设计SCAR引物对抗病亲本、感病亲本、抗病池、感病池、F1个体进行扩增,均获得一条592bp的特异片段。感病基因型和杂合抗病基因型存在XbaⅠ酶切位点,酶切后分别产生了261bp、193bp和95bp以及592bp、261bp、193bp和95bp的特异性片段,纯合抗病基因型无此酶切位点,酶切结果仍为592bp的产物。这些片段能成功区分抗病材料、感病材料和F1个体,很有可能是与ph-3基因连锁的CAPS标记。

A RAPD fragment linking to ph-3 gene was cloned.According to the sequencing result,we designed SCAR primer.A 592bp fragment was amplified in resistant and susceptible parents,resistant and susceptible bulks,F1 individuals,and then the PCR products were digested with restriction enzyme XbaI.Susceptible genotypes and heterozygous resistant genotypes could produce respectively 261bp,193bp,95bp and 592bp,261bp,193bp,95bp bands.Homozygous resistant genotypes presented 592bp fragment.These DNA bands which could distinguish resistant material,susceptible material and F1 individuals successfully,were probably CAPS markers for ph-3 gene.