泰山海棠抗旱基因MhDREB2A的克隆与功能鉴定  

Cloning and Functional Idenfication of Drought-Resistant Related Gene in Malus hupehensis

作　　者：党明青 王京平 冉昆 刘加芬 李慧峰[3] DANG Ming-qing;WANG Jing-ping;RAN Kun;LIU Jia-fen;LI Hui-feng(College of Plant Protection,Nanjing Agricultural University,Nanjing 210095,China;Integrated Service Station of Sikou County in Qixia City,Yantai Shandong 265300,China;Shandong Institute of Pomology,Taian Shandong 271000,China)

机构地区：[1]南京农业大学植物保护学院,南京210095 [2]栖霞市寺口镇综合服务站,山东烟台265300 [3]山东省果树研究所,山东泰安271000

出　　处：《沈阳农业大学学报》2022年第4期462-468,共7页Journal of Shenyang Agricultural University

基　　金：山东省自然科学基金项目(ZR2019MC071)。

摘　　要：泰山海棠(Malus hupehensis)是我国特有的苹果属种质资源,具有抗旱等优良性状。DREB转录因子(dehydration responsive element binding protein)是AP2/EREBP(APETALA 2/an ethylene responsive element binding protein)转录因子家族的一个亚家族,含有保守的AP2结构域,能够特异性地结合到抗逆基因启动子区域的DRE顺式作用元件上。以泰山海棠为材料,克隆得到抗旱相关基因MhDREB2A。该基因有1个AP2(APETALA 2)结构域,cDNA全长1197bp,编码398个氨基酸。实时荧光定量PCR分析发现MhDREB2A明显受到干旱或ABA的诱导表达,并且MhDREB2A在泰山海棠花和根中表达水平高于茎、叶和果中。亚细胞定位结果表明MhDREB2A定位于细胞核中。为分析其功能,构建35S::MhDREB2A过量表达载体,遗传转化了烟草,并获得转基因过表达株系。表型分析和相关生理指标测定表明,在干旱处理下转基因株系抗旱性明显高于对照野生型,转基因株系中的叶绿素总含量明显高于野生型植株,电导率(EL)和丙二醛(MDA)含量则明显低于野生型植株,同时抗干旱胁迫相关基因NtDREB3、NtERD10C、NtERD10D、NtNCED1和NtLEA5在转基因株系表达水平比野生型显著升高,而转基因烟草株系和野生型在对照条件下,抗干旱胁迫相关基因的表达水平没有显著差异。结果表明MhDREB2A具备干旱胁迫响应和抗旱功能,揭示其在植物抵抗干旱胁迫中具有重要作用。Malus hupehensis is a specific apple variety of China, with many good characteristics such as drought resistance.Transcription factor DREB(dehydration responsive element binding protein) belongs to subfamily of APETALA 2/an ethylene responsive element binding protein transcription factors family. The DREB transcription factor contains a conserved AP2(APETALA2)domain which can bind to DRE cis-acting element in promoter regions of resilience genes. In this study, we cloned a drought stress responsive gene named as MhDREB2A. MhDREB2A was 1197bp in length and the encoding products containing 398 AA. The real time PCR analysis showed that the expression of MhDREB2A could be induced by drought or ABA. In M. hupehensis,MhDREB2A was expressed in all organs tested, but with higher expression level in flower and root than that in stem, leaf or fruit.Subcellular localization results showed that MhDREB2A was located in nucleus. To gain insight into the function of MhDREB2A, a35S::MhDREB2A cassette was transformed into tobacco plants. In our results, the phenotypes and physiological indexes associated with drought response were higher in transformed plants compared with wild-type plants. In the condition of drought, the chlorophyll content of the transgenic plant was higher than that of the wild-type plant, while the electrolyte leakage and MDA content were lower.The expression levels of the five selected drought stress induction marker genes NtDREB3, NtERD10C, NtERD10D, NtNCED1 and NtLEA5 in transgenic plants were significantly higher than that in wild-type plants. In the condition of control,there were no significant differences between transgenic tobacco strains and wild type in the expression levels of drought-resistant genes. In brief, we found the MhDREB2A can respond to drought stress and resist drought, and it plays an important role in plant resistance to drought stress.

关 键 词：泰山海棠 MhDREB2A 抗旱 基因表达 

分 类 号：S661.1[农业科学—果树学]