花生NBS-LRR类基因P9的克隆与序列分析  

Cloning and Sequence Analysis of NBS-LRR Gene P9 in Peanut

作　　者：朱晓峰[1] 姜涛[1] 赵西拥 王嵩[1] 汪清[1,2] 倪皖莉 Zhu Xiaofeng;Jiang Tao;Zhao Xiyong;Wang Song;Wang Qing;Ni Wanli(Institute of Crops,Anhui Academy of Agricultural Sciences,Hefei,230031;Collge of Agronomy,Anhui Agnicultural University,Hefei,230036)

机构地区：[1]安徽省农业科学院作物研究所,合肥230031 [2]安徽农业大学农学院,合肥230036

出　　处：《分子植物育种》2020年第3期737-743,共7页Molecular Plant Breeding

基　　金：国家重点研发项目子课题(2018YFD0201009-4);安徽省农业科学院国家产业技术体系岗站团队项目(18T0201)和安徽省农业科学院院长青年基金项目(17B0205)共同资助。

摘　　要：为探索花生NBS-LRR类基因在花生抗病中的分子作用机制,本研究以花生品种‘Z525’为试验材料,采用电子克隆与RT-PCR相结合的方法,克隆了花生叶片中的P9基因。结果表明,获得一个长度为3557 bp的序列,该序列包含一个完整的开放阅读框(ORF),ORF的长度为3195 bp(93 bp^3287 bp),编码1064个氨基酸(120.4 kD),等电点pI为5.92。序列分析表明,该基因编码的蛋白与花生中假定的抗性蛋白RPP13-like及花生二倍体野生种Arachis duranensis和Arachis ipaensis推测的抗病蛋白At3g14460、RPP13-like高度相似;P9蛋白属于NBS-LRR家族,具有典型的NB-ARC和LRR-3两个保守结构域,推测该基因参与花生抗病调控过程。蛋白质亚细胞定位预测表明该基因编码的蛋白主要位于叶绿体中,可能少量分布于细胞核中,推测该基因可能主要作为叶绿体蛋白参与细胞的抗氧化、抗衰老等抗逆过程,其次作为转录因子参与转录调控作用。本研究克隆了P9基因的全长cDNA序列,并对该基因序列、结构和功能等方面进行了分析,为进一步研究花生抗病分子机制提供理论基础,同时为花生抗病品种的选育提供理论支持。In order to explore the molecular mechanism of NBS-LRR genes in peanut disease resistance,In silico cloning and RT-PCR were used to isolate c DNA sequence of P9 from peanut leaves by using’Z525’as experimental material in this study.The results showed that it obtained a sequence of length 3557 bp.The obtained sequence contained a 3195 bp complete open reading frame(ORF),from 93 bp to 3287 bp,and encoded 1064 amino acids residues with a predicted molecular weight of 120.4 kD,bioelectric point value of 5.92.Sequence analysis showed that the amino acids sequence encoded by P9 gene was highly conserved with the putative resistant protein RPP13-like in peanut and the disease-resistant protein At3 g14460 and RPP13-like in Arachis duranensis and Arachis ipaensis.The P9 protein belongs to NBS-LRR family and contained two conserved domains named NB-ARC and LRR-3.It is inferred that this NBS-LRR gene may be involved in the resistance regulation in peanut.Subcellular localization indicated that this protein is mainly located in the chloroplast,and a little bit in the nucleus.The result speculated that this gene may be mainly involved in the cell anti-oxidant and anti-aging process as chloroplast protein,and then participated in the transcriptional regulation as a transcription factor.In this study,the full-length cDNA sequence of P9 gene was cloned,and the sequence,structure and function of this gene were analyzed and predicted using bioinformatics methods,providing a theoretical basis for further research on the molecular mechanism of disease resistance and laying a foundation for the breeding of disease resistance varieties in peanut.

关 键 词：花生 P9基因 克隆 序列分析 

分 类 号：S565.2[农业科学—作物学] Q943.2[生物学—植物学]