向日葵盐诱导HD-Zip类转录因子HB-12的克隆及表达分析  

Cloning and Expression Analysis of a Salt-Stress-Induced HD-Zip Transcription Factor HB-12 from Sunflower(Helianthus annuus L.)

作　　者：孙瑞芬[1] 张艳芳[1] 郭树春[1] 于海峰[1] 李素萍[1] 聂惠[1] 安玉麟[1] 

机构地区：[1]内蒙古农牧业科学院,呼和浩特010031

出　　处：《分子植物育种》2017年第6期2077-2087,共11页Molecular Plant Breeding

基　　金：内蒙古农牧业创新基金项目(2013CXJJN14)资助

摘　　要：HB转录因子家族基因在植物生长、发育、生物和非生物胁迫中发挥着重要作用。根据已知序列Unigene551_All设计引物,利用SMARTer RACE技术从向日葵中克隆了一个HD-Zip类转录因子HB-12,该基因全长cDNA序列821 bp,包括长度为573 bp的开放阅读框,编码190个氨基酸。预测该蛋白的分子量和等电点分别为22.55 kD和5.58,具有一个同源异型框结构域(homeobox domain,HD)和一个同源异型框结合类亮氨酸拉链结构域(homeobox associated leucine zipper domain,HALZ),属于向日葵HD-ZipⅠ类蛋白,基因登录号为KU315052。HB-12蛋白不存在跨膜域,亚细胞定位预测其可能位于细胞核。聚类分析发现向日葵HB-12与茄科作物马铃薯和番茄HB-12基因亲缘关系较近。利用PCR技术扩增了HB-12全长cDNA对应的g DNA序列,该序列自起始密码子ATG至终止密码子TAA长度为652 bp,由2个外显子和1个内含子组成,基因登录号为KU315053。Real-time PCR结果表明,HB-12在向日葵根、下胚轴和叶中的表达存在器官特异性,且受盐、ABA及PEG的诱导表达。HB-12基因的克隆及表达分析为向日葵抗逆分子育种和功能研究奠定了基础。The HB transcription factor genes play an important role in plant growth, development, biotic and abiotic stress. A HD-Zip transcription factor HB-12 was cloned from sunflower by using SMARTer RACE technology based on the known sequence of Unigene551_All. The full length cDNA sequence of HB-12 was821 bp, including the open reading frame of 573 bp and encoding 190 amino acids. The predicted protein molecular weight and isoelectric point were 22.55 kD and 5.58 respectively, with a homeobox domain（HD）and a homeobox associated leucine zipper domain（HALZ）. HB-12 belongs to the sunflower subfamilyⅠof HD-Zip proteins, the sequence accession number was KU315052 in Gen Bank. HB-12 protein did not exist in the transmembrane domain, and its subcellular localization predicted that it might be located in the nucleus. The cluster analysis revealed that the HB-12 of sunflower was closely related to the HB-12 of potato and tomato crops. The g DNA sequence corresponding to the full-length cDNA of HB-12 was amplified by PCR technique.The full length of the coding region of g DNA was 652 bp, and two exons were separated by one intron. The sequence had been submitted to Gen Bank（Accession No. KU315053）. The results of real-time PCR showed that the expression levels of HB-12 had specific expression differences in different organs, such as roots, hypocotyls and leaves,and its transcript levels were strongly induced when sunflower plants were treated with salt, ABA or PEG. The cloning and expression analysis of HB-12 have laid a foundation for the research of molecular breeding and function of sunflower.

关 键 词：向日葵 HB-12 HD-ZIP 胁迫 表达分析 

分 类 号：Q943.2[生物学—植物学] S565.5[农业科学—作物学]