RGA法克隆NBS-LRR类抗病基因同源序列及其在葫芦科作物上应用的研究进展

RGA克隆法是利用抗病基因产物的保守结构域人工设计简并引物,以植物gDNA或cDNA为模板进行PCR扩增而克隆植物抗病基因同源序列的方法。随着各种植物基因组测序计划的完成及计算机和生物信息学的发展,植物抗病基因的克隆取得了很大进展,目前人们已经从植物中克隆得到100多个抗病基因。研究表明,大多数抗病基因都存在NBS-LRR、STK、LZ、TIR等功能保守的结构域,其中大部分都为NBS-LRR类型,利用NBS-LRR保守结构域设计PCR引物已经从植物中扩增出大量的RGA。简要综述了已克隆NBS-LRR类抗病基因的结构特点和功能、RGA法克隆NBS-LRR类抗病基因同源序列及其在葫芦科作物上应用的研究进展,并探讨其未来的发展与应用。

Cloning of NBS-LRR-encoding Resistance Gene Analogues Using RGA Approach and the Advance in Cucurbit Crops

As majority resistance genes encode a highly conserved protein domain,RGA approach is a method of using the conserved protein domain design degenerate primers to clone resistance gene analogs from plant gDNA or cDNA.With the assembling of genome sequence in various plant species and the development of bioinformatics,the cloning of plant disease resistance genes have made great progress,more than 100 resistance genes have been cloned.Related studies showed that majority of plant disease resistance genes encode a conserved NBS-LRR,STK,LZ,TIR functional structure and majority of them belonging to NBS-LRR class.Using conserved NBS-LRR-encoding domain to design degenerate primer has amplified a number of RGA.In this article,the structure characteristics and function of NBS-LRR-encoding resistance genes and the advance in cloning NBS-LRR-encoding resistance genes using RGA approach and application in cucurbit crops has been reviewed and its future development and application has also been discussed.