夏橙果实离层中伤害诱导基因的克隆与表达分析  

Cloning of wound-induced protein genes from Citrus sinensis cv. Valencia and analysis of their expression patterns in fruit calyx abscission zone

作　　者：杨雪莲[1,2] 张凌云[1] 王志卫[1] 黄勇[1] 闫树堂[1,3] 钟广炎[1,3] 

机构地区：[1]西南大学,重庆北碚400715 [2]贵州大学,贵阳550025 [3]中国农业科学院柑桔研究所,重庆北碚400712

出　　处：《果树学报》2010年第6期864-870,共7页Journal of Fruit Science

基　　金：重庆市科技攻关计划项目(CSTC,2007AA1018);国家柑橘产业技术体系支持项目

摘　　要：从乙烯处理的伏令夏橙果实离层中克隆出4个伤害诱导基因(CsWIP1、CsWIP2、CsWIP3、CsWIP4)。通过RT-PCR、RACE等克隆技术分别获得了CsWIP1、CsWIP2、CsWIP3和CsWIP4的全长cDNA。测序结果表明CsWIP1、CsWIP2、CsWIP3和CsWIP4全长cDNA分别为588、549、412和629bp,开放阅读框分别编码98、95、83和88个氨基酸,结构模拟显示这些蛋白在C-端和N-端各有一个α-螺旋,中间是一个富丝氨酸域。与其他植物伤害诱导蛋白序列比对结果显示,CsWIP家族蛋白分为2类,CsWIP1、CsWIP2和CsWIP3与玉米WIP家族中NP_001148450.1、NP_001151919.1、NP_001151459.1、NP_001147041.1和拟南芥NP_192765.1、NP_849355.1、NP_179023.1聚成一类;CsWIP4与葡萄的AY159560.1聚成一类。通过实时定量PCR检测这些基因在乙烯处理下果实离层中的表达情况,结果显示CsWIP1和CsWIP2受乙烯强烈诱导,CsWIP3和CsWIP4几乎不受乙烯调控。Four wound-induced genes, Cs WIP1, Cs WIP2, Cs WIP3 and Cs WIP4, were cloned from ethylene-treated abscission zones （ AZ-C ） of citrus （Citrus sinensis cv. Olinda valencia ）. Their full-length cDNAs were obtained by using RTPCR and RACE. The full-length cDNAs of CsWIP1, CsWIP2, CsWIP3 and CsWIP4 were 588, 549, 412 and 629 bp and the corresponding ORFs would encode 98, 95, 83 and 88 amino-acid residues, respectively. Structural simulations showed that the deduced proteins possessed two -helices located separately at N- and C-terminals and were rich in serine residues in the middle. Alignments with other plant wound-induced proteins showed that the citrus wound-induced proteins could be grouped into two classes： CsWIP1, CsWIP2 and CsWIP3 were clustered together with corn NP_001148450.1, NP_001151919.1, NP_001151459.1, NP_001147041.1 and arabidopsis NP_192765.1, NP_849355.1, NP_179023.1 while Cs WIP4 was clustered together with grape AY159560.1. Quantitative real time PCR was used to study the expression patterns of these genes. In AZ-C, the expression of Cs WIP1 and Cs WIP2 was strongly induced by ethylene, whereas the expression of Cs WIP3 and Cs WIP4 was not significantly modulated by ethylene.

关 键 词：夏橙 伤害诱导蛋白 基因家族 克隆 乙烯诱导 

分 类 号：S666.4[农业科学—果树学]