海稻86转录因子HD-ZIP的克隆及表达分析  

Cloning and Expression Analysis of HD-ZIP Transcription Factor in Haidao 86

作　　者：陶相如 颜彦 肖勇[1] 夏薇[1] 胡远艺[4,5] 胡伟[2,3] Tao Xiangru;Yan Yan;Xiao Yong;Xia Wei;Hu Yuanyi;Hu Wei(College of Tropical Crops,Hainan University,Haikou 570228,China;Sanya Research Institute,Chinese Academy of Tropical Agricultural Sciences,Sanya 572025,China;Institute of Tropical Biotechnology,Chinese Academy of Tropical Agricultural Sciences,Haikou 571101,China;Hunan Hybrid Rice Research Center,Changsha 410125,China;National Technology Innovation Center for Saline-Alkali Tolerant Rice in Sanya,Sanya 572025,China)

机构地区：[1]海南大学热带作物学院,海南海口570228 [2]中国热带农业科学院三亚研究院,海南三亚572025 [3]中国热带农业科学院热带生物技术研究所,海南海口571101 [4]湖南杂交水稻研究中心,湖南长沙410125 [5]三亚市国家耐盐碱水稻技术创新中心,海南三亚572025

出　　处：《山东农业科学》2024年第11期21-27,共7页Shandong Agricultural Sciences

基　　金：海南省重大科技计划项目(ZDKJ202001)。

摘　　要：同源结构域亮氨酸拉链(HD-ZIP)转录因子是植物独有的转录因子,分为四个不同的亚家族(HDZIPⅠ~Ⅳ),其中HD-ZIPⅠ在植物发育过程的调控、信号网络和对环境胁迫的反应中发挥关键作用。海稻86是一种耐盐碱能力较高的特异水稻种质资源。为解析HD-ZIPⅠ在盐胁迫应答过程中的功能,本研究以海稻86为材料,利用RT-PCR技术克隆了一个HD-ZIPⅠ基因,命名为OsHDZ1,并对其进行生物信息学分析、亚细胞定位及盐胁迫下表达分析。结果表明,OsHDZ1基因的CDS序列全长831 bp,编码276个氨基酸,蛋白分子量为30.65 kDa,理论等电点为5.19,为亲水性不稳定蛋白;OsHDZ1蛋白不含跨膜结构,含有25个磷酸化位点,二级结构主要由α-螺旋和无规则卷曲组成;同源性分析发现该蛋白序列与禾本科植物弯叶画眉草和小麦具有较高的亲缘关系;对OsHDZ1启动子元件预测发现其具有脱落酸、茉莉酸等激素响应元件,同时具有环境胁迫等响应元件;细胞定位分析显示该蛋白定位于细胞核。荧光定量PCR结果发现,在盐胁迫处理下,Os-HDZ1表达量整体呈上升,在60 h之前逐渐上升,在60 h达到最高,在72 h表达量下降,表明该基因可能在海稻耐盐性中具有调控功能。本研究结果可为后续深入解析OsHDZ1基因调控海稻耐盐的功能奠定基础,为培育耐盐作物提供基因资源。Homologous domain leucine zipper(HD-ZIP)transcription factors are unique in the plant,which are divided into four different subfamilies(HD-ZIPⅠtoⅣ).HD-ZIPⅠplay key roles in regulation of plant development processes,signal networks and response to environmental stresses.Haidao 86 is a specific rice germplasm with high saline-alkali tolerance.In order to analyze the function of HD-ZipⅠin response to salt stress,an HD-ZIPⅠgene named OsHDZ1 was cloned by RT-PCR technology using Haidao 86 as materi-al,and its bioinformatics analysis,subcellular localization and expression analysis under salt stress were con-ducted.The results showed that the CDS sequence of OsHDZ1 gene was 831 bp in length,encoding 276 amino acids,with the molecular weight of 30.65 kDa and the theoretical isoelectric point of 5.19,which was a hydro-philic unstable protein.OsHDZ1 protein contained no transmembrane structure,contained 25 phosphorylation sites,and the secondary structure mainly consisted ofα-helix and random coil.Homology analysis showed that its protein sequence was highly related to Eragrostis curvula and Triticum aestivum.The prediction of the pro-moter element of OsHDZ1 showed that it had hormone(such as abscisic acid and jasmonic acid)response ele-ments,as well as response elements to environmental stresses.Subcellular localization analysis showed that the protein was localized in the nucleus.According to the results of fluorescence quantitative PCR,the expression of OsHDZ1 was significantly up-regulated and gradually increased in the former 60 hours of salt stress,and de-creased at 72 hours,but the overall trend was still on the rise,indicating that this gene might have the regula-tory function in salt tolerance of Haidao 86.The results of this study laied the foundation for subsequent in-depth analysis of the function of OsHDZ1 gene in regulating salt tolerance of Haidao 86,and could provide gene resources for breeding salt-tolerant crops.

关 键 词：海稻86 HD-ZIPⅠ 基因克隆 生物信息学 亚细胞定位 表达分析 

分 类 号：S511[农业科学—作物学] Q78[生物学—分子生物学]