野生茄托鲁巴姆抗根结线虫相关基因的克隆与表达分析

Cloning and Expression Analysis of Genes Related to Root Knot Nematode Resistance from Solanum torvum Swartz

克隆茄子抗根结线虫相关基因并利用荧光定量PCR技术分析其表达特性。根据在NCBI上找到的抗性基因NBS-LRR保守结构域,设计简并引物,以野生茄托鲁巴姆(Solanum torvum Swartz)的c DNA为模板采用c DNA末端快速扩增(RACE)技术克隆抗根结线虫基因,利用荧光定量PCR技术分析其表达特性。获得15条抗性基因片段,且E1、E3、E6、E7、E12、E13、E15为茄子抗根结线虫基因片段。通过聚类分析和氨基酸比对确定E7为Mi基因的一部分。利用RACE技术获得到了野生茄托鲁巴姆抗根结线虫Mi同源基因序列。本研究在野生茄托鲁巴姆中获得了抗病基因同源序列(resistance gene analogs,RGAs)基因,为茄子抗根结线虫分子育种奠定基础。

The aim of the study was to clone gene related to root knot nematode(Meloidogyne incognita) resistance from Solanum torvum Swartz, and real-time PCR was used to analyze its expression. The degenerated primers were designed based on the conserved regions of nucleotide binding site(NBS)-leucine rich repeat(LRR) domain from plant resistance genes reported in NCBI. RACE was used to clone the resistance genes from Solanum torvum Swartz. The expression characteristics of resistance genes were analyzed by fluorescence quantitative PCR technique. Fifteen resistance gene segments were obtained and E1, E3, E6, E7, E12, E15 were determined as root knot nematode resistance gene segments. Cluster analysis and alignment of amino acid showed that E7 was one part of the root knot nematode resistance gene Mi. Resistance gene analogs( RGAs) of root knot nematode resistance gene Mi were obtained from Solanum torvum Swartz by RACE. The acquisition of RGAs from Solanum torvum Swartz laid a promising foundation for unravelling molecular mechanism of eggplant disease resistance and molecular breeding.