甘蔗线虫病抗性基因的PCR检测研究

PCR Detection of Nematode Resistant Gene in Sugarcane

以38份未经线虫病常规鉴定的甘蔗种质资源为供试材料,根据番茄抗根结线虫病基因和甜菜抗胞囊线虫病基因的保守序列区域,分别设计了2条上游引物2、条下游引物,对设计的引物进行组合后,应用PCR特异扩增从中分别筛选出各1对特异引物。用抗根结线虫基因设计的特异引物进行扩增最终获得了1条大约460 bp大小的特异片段;用抗胞囊线虫基因设计的特异引物进行扩增最终获得了1条大约690 bp大小的特异片段,进一步进行了PCR-Southern杂交,确认了该2条特异引物的真实性。

According to the sequence of root-knot nematode resistant gene in tomato and the sequence of nematode resistant gene in sugar beet, two forward primers and two reverse primers were separately designed, and a pair of primers each was selected among them by PCR. Thirty-eight sugarcane cultivars （without conventional identification of nematode resistance） was tested by the selected PCR primers. The results showed that two special fragments of about 460 bp and 690 bp were obtained, and PCR-Southem blotting about them was made further in order to affirm the homogeneous sequences of nematode resistant gene.