西藏野生垂穗披碱草EnCBL10基因的克隆与功能研究  

Cloning and Function Identification of EnCBL10 Gene from Tibetan Wild ElymusnutansGriseb.

作　　者：张慎彤 高浩[1] 张金鹏[1] 任智慧[1] 姜惠娜 其美拉姆 普布卓玛 付娟娟[1] ZHANG Shen-tong;GAO Hao;ZHANG Jin-peng;REN Zhi-hui;JIANG Hui-na;QI MEI La-mu;PU BU Zhuo-ma;FU Juan-juan(Department of Grassland Science,College of Grassland Agriculture,Northwest A&F University,Yangling,Shaanxi Province 712100,China;Natural Resources Bureau of Qushui County,Lhasa,Tibet 850000,China;College of Animal Science,Agricultural and Animal Husbandry College of Tibet University,Lhasa,Tibet 850000,China)

机构地区：[1]西北农林科技大学草业与草原学院,陕西杨凌712100 [2]西藏自治区拉萨市曲水县自然资源局,西藏拉萨850000 [3]西藏自治区农科院草业科学研究所,西藏拉萨85000

出　　处：《草地学报》2022年第9期2315-2324,共10页Acta Agrestia Sinica

基　　金：国家自然科学基金项目(31901382,U21A20240);国家自然科学面上项目(31872411);国家自然科学基金-区域联合发展重点基金(U20A2050);大学生创新创业训练计划创新训练项目(202110712056)共同资助。

摘　　要：钙调磷酸酶B类蛋白(Calcineurin B-Like Proteins,CBLs)是钙信号通路中重要成员,在植物应答多种非生物胁迫中具有重要的作用。本研究基于前期转录组数据,在西藏野生垂穗披碱草(Elymus nutans Griseb.)中筛选EnCBL10基因,将其异源表达于烟草(Nicotiana tabacum)中,分析转基因植株EnCBL10在低温和干旱胁迫下的生长及生理响应。结果表明:EnCBL10开放阅读框为792bp,编码263个氨基酸。EnCBL10蛋白的理论等电点为5.17,为疏水蛋白和非分泌蛋白。低温和干旱胁迫下,对比于野生型烟草,转基因烟草叶片的细胞膜稳定性显著增加,过氧化物酶(Peroxidase,POD),抗坏血酸过氧化物酶(Aseorbateperoxidase,APX)和谷胱甘肽还原酶(Gluathione reductase,GR)活性显著升高。低温胁迫下,两个转基因株系的脯氨酸含量分别提高了79.7%和70.8%(P<0.05);在干旱胁迫下,野生型植株(WT)和过表达植株中脯氨酸含量均有所降低,但转基因植株比WT的下降幅度小。因此,EnCBL10基因可能在垂穗披碱草的耐寒和耐旱调控中发挥重要功能。Calcineurin B-Like Proteins(CBLs)are important members of the calcium signaling pathway and play an important role in plant response to various abiotic stresses.Based on the previous transcriptome data,EnCBL10 gene was screened from wild Elymus nutans Griseb.in Tibet,and its heterologous expression in tobacco was performed to analyze the growth and physiological response of transgenic plants EnCBL10in low temperature and drought.The results showed that the open reading frame of EnCBL10 gene was 792bp,encoding 263amino acids.The theoretical isoelectric point of EnCBL10protein was 5.17,which was a hydrophobic and nonsecretory protein.Under cold and drought stress,the cell membrane stability of transgenic tobacco leaves was significantly higher than that of wild-type tobacco leaves.Furthermore,the activities of peroxidase(POD),ascorbate peroxidase(APX),and glutathione reductase(GR)in EnCBL10 transgenic plants increased significantly under cold and drought stress.Under cold stress,the proline content in the leaves of the two transgenic lines increased by 79.7%and 70.8%(P<0.05)compared with their respective controls.Under drought stress,the proline content decreased in WT and overexpressed plants,but the decrease of transgenic plants was less than in WT.Therefore,EnCBL10 may play an important role in low temperature and drought tolerance regulation in Elymus nutans.

关 键 词：垂穗披碱草 EnCBL10 基因克隆 低温 干旱 基因调控 生理机制 

分 类 号：S543.9[农业科学—作物学] Q945.78[生物学—植物学]