白花草木樨MaERF058基因耐旱功能验证  

Functional analysis of the MaERF058 gene in response to drought stress in Melilotus albus

作　　者：魏娜 敬文茂 许尔文 王荣新 赵晶忠 马雪娥 张吉宇[2] 刘文献[2] WEI Na;JING Wen-mao;XU Er-wen;WANG Rong-xin;ZHAO Jing-zhong;MA Xue-e;ZHANG Ji-yu;LIU Wen-xian(Academy of Water Resources Conservation Forests in Qilian Mountains of Gansu Province,Zhangye 734000,China;State Key Laboratory of Herbage Improvement and Grassland Agro-ecosystems,Lanzhou 730030,China;Gansu Qilian Mountain Forest Eco-System of the State Research Station,Zhangye 734000,China)

机构地区：[1]甘肃省祁连山水源涵养林研究院,甘肃张掖734000 [2]兰州大学草种创新与草地农业生态系统全国重点实验室,甘肃兰州730030 [3]甘肃祁连山森林生态系统国家定位观测研究站,甘肃张掖734000

出　　处：《草业学报》2024年第8期159-169,共11页Acta Prataculturae Sinica

基　　金：国家自然科学基金(32061143035,U21A20468);2022年张掖市市级科技计划(ZY2022RC03,ZY2022KY03);兰州大学中央高校基本科研业务费专项资金(lzujbky-2021-it05,lzujbky-2021-ct14)资助。

摘　　要：干旱是农业生产中最常见的逆境胁迫之一,对植物的生长发育、光合作用、气孔开闭、渗透功能和激素调节等方面都会产生不良影响。白花草木樨在我国北方干旱地区广泛种植,是具有耐旱、耐盐碱、耐贫瘠和粗蛋白含量高等特点的优质牧草。转录因子(TF)作为调控基因表达的重要蛋白分子,可作用于其下游启动子区的特定顺式元件以激活或抑制基因的转录,进而调控植物生长发育及响应逆境。植物ERF转录因子家族基因参与植物对非生物胁迫的抗性,是作物抗逆性改良的理想候选基因。本研究对白花草木樨关键耐旱基因MaERF058进行烟草亚细胞定位分析,发现该基因表达蛋白定位于细胞核。干旱胁迫下,过表达MaERF058的拟南芥长势明显优于野生型。干旱条件下转MaERF058基因白花草木樨毛状根脯氨酸含量较对照显著(P<0.01)增加;丙二醛含量显著(P<0.01)降低;过氧化氢酶活性显著(P<0.05)增加;单胺氧化酶染色(NBT法)结果显示转基因毛状根的活性氧含量低于对照。以上结果表明白花草木樨MaERF058基因编码的转录因子对调控白花草木樨响应干旱胁迫具有积极作用。研究结果为解析白花草木樨耐旱分子机制提供了新思路,同时也能够为加快耐旱牧草分子育种提供优异基因资源。Drought is one of the most common stresses in agricultural production.It has a deleterious effect on plant growth and many aspects of development,such as photosynthetic function,stomatal aperture,osmotic balance,and hormone regulation,thereby affecting crop yield and quality.Melilotus albus is a high-quality forage legume that is widely grown in the northern arid regions of China because of its high protein content and tolerance to drought,salinity,and barren environments.Transcription factors(TFs)are important proteins that regulate gene expression.They bind to specific cis-elements in the promoter regions of their target genes to activate or repress their transcription,thereby regulating plant growth and development in response to abiotic stress.Genes in the plant ethylene response factor(ERF)TF family respond to various abiotic stresses,and are important components of plant stress resistance mechanisms.MaERF058 is a key drought tolerance gene in M.albus.In this study,we analyzed the subcellular localization of MaERF058 in tobacco(Nicotiana tabacum),and found that the expressed protein was located in the nucleus.Overexpression of MaERF058 in Arabidopsis thaliana significantly increased its drought tolerance,compared with that of wild type.Compared with the wild type,the MaERF058-overexpressing lines showed significantly increased proline content(P<0.01)and catalase activity(P<0.05)and significantly decreased malondialdehyde content(P<0.01)under drought conditions.Monoamine oxidase staining using the nitroblue tetrazolium method showed that the reactive oxygen content in the roots was higher in the MaERF058-overexpressing lines than in the control.These results suggest that the TF encoded by MaERF058 in M.albus has a positive regulatory role in the drought stress response.

关 键 词：白花草木樨 ERF转录因子 干旱胁迫 遗传转化 

分 类 号：S541.9[农业科学—作物学]