马蔺IlWRKY28基因的克隆与表达分析  

Cloning and Expression Analysis of the IlWRKY28 Gene in Iris lactea var.chinensis

作　　者：刘清泉[1] 张永侠[1] 王银杰 顾春笋[1] 佟海英[1] 黄苏珍[1] 原海燕[1] LIU Qingquan;ZHANG Yongxia;WANG Yinjie;GU Chunsun;TONG Haiying;HUANG Suzhen;YUAN Haiyan(Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing 210014, China)

机构地区：[1]江苏省中国科学院植物研究所,南京210014

出　　处：《西北植物学报》2020年第9期1490-1497,共8页Acta Botanica Boreali-Occidentalia Sinica

基　　金：江苏省自然科学基金(BK20180317);国家自然科学基金(41907135);江苏省海洋科技创新专项(HY2018-5)。

摘　　要：马蔺(Iris lactea var.chinensis)是鸢尾属多年生草本盐生植物,具有很高的耐盐性和观赏价值。为研究马蔺耐盐的分子机制,通过cDNA末端快速扩增技术(RACE)从马蔺中克隆到一个WRKY转录因子基因IlWRKY28,获得了1302 bp的全长cDNA序列,其包含一个108 bp 5′末端非翻译区(UTR),一个174 bp 3′末端UTR和一个1020 bp开放阅读框。IlWRKY28编码339个氨基酸,预测的蛋白质分子量为37.22 kD,等电点为7.04。氨基酸序列分析显示,IlWRKY28包含一个保守的WRKY基序和一个C2H2型锌指结构域。系统发育分析表明,马蔺IlWRKY28与菠萝(Ananas comosus)AcWRKY28和藏北嵩草(Kobresia littledalei)ClWRKY28亲缘关系最近。荧光定量PCR分析显示,盐处理后,IlWRKY28基因在马蔺地上部显著上调表达。该研究结果为进一步研究IlWRKY28在马蔺适应高盐胁迫中的功能和作用机制奠定了重要的分子基础。Iris lactea var.chinensis is a perennial herbaceous halophyte of the genus Iris and shows high salt tolerance and ornamental value.In order to study the molecular mechanism of salt tolerance in I.lactea,we cloned a WRKY transcription factor gene IlWRKY28 from I.lactea through rapid amplification of cDNA ends technology(RACE),and obtained a 1302 bp full-length cDNA sequence,which contains a 5′untranslated region(UTR)of 108 bp,a 3′UTR of 174 bp and an open reading frame of 1020 bp.IlWRKY28 encodes 339 amino acids,the predicted protein molecular weight and isoelectric point is 37.22 kD and 7.04,respectively.Amino acid sequence analysis showed that IlWRKY28 contains a conserved WRKY motif and a C2H2-type zinc finger domain.Phylogenetic analysis showed that I.lactea IlWRKY28 was much closer to Ananas comosus AcWRKY28 and Kobresia littledalei ClWRKY28.Quantitative PCR analysis showed that after salt treatment,the IlWRKY28 gene was significantly up regulated in the I.lactea shoots.The results of this study laid an important molecular foundation for the further study of the function and mechanism of IlWRKY28 in the tolerance of I.lactea to high salt stress.

关 键 词：马蔺 IlWRKY28 盐胁迫 RACE 基因表达 

分 类 号：Q785[生物学—分子生物学] Q786