过表达烟草乙烯响应因子NtERF115促进烟草尼古丁的生物合成  

Overexpression of ethylene response factor NtERF115 increases nicotine biosynthesis in tobacco(Nicotiana tabacum L.)

作　　者：李梅云 隋学艺 宋中邦 高玉龙 李文正 赵璐 费玲玲 耿小晴 韩生成[2] 王丙武 LI Meiyun;SUI Xueyi;SONG Zhongbang;GAO Yulong;LI Wenzheng;ZHAO Lu;FEI Lingling;GENG Xiaoqing;HAN Shengcheng;WANG Bingwu(Tobacco Breeding and Biotechnology Center,Yunnan Academy of Tobacco Agricultural Science,650021,Kunming,Yunnan,China;Beijing Key Laboratory of Gene Resource and Molecular Development,College of Life Sciences, Beijing Normal University,100875,Beijing,China)

机构地区：[1]云南烟草农业科学研究院烟草育种与生物技术研究中心,云南昆明650021 [2]抗性基因资源与分子发育北京市重点实验室,北京师范大学生命科学学院,北京100875

出　　处：《北京师范大学学报（自然科学版）》2019年第3期341-348,共8页Journal of Beijing Normal University(Natural Science)

基　　金：中国烟草总公司云南省公司资助项目(2018530000241001)

摘　　要：乙烯响应因子ERF是一类与植物抗逆相关的转录因子,该因子通过激活下游一系列抗逆应答基因的表达而提高植株的综合抗性.本研究前期通过烟草转录组表达谱分析,从茉莉素(JA)处理后的烟草BY-2(Nicotiana tabacum Bright Yellow-2)细胞系中,筛选出一个受JA诱导的转录因子基因NtERF115,属于IX亚组.采用RT-PCR技术获得NtERF115基因cDNA全长,并对其进行序列分析、蛋白结构功能预测以及相同或不同物种间NtERF115同源基因的进化树构建.进一步在烟草云烟87(YN87)植株中,过表达NtERF115并检测其对尼古丁合成的影响.结果表明:NtERF115核苷酸序列ORF为690bp,编码229个氨基酸;在过表达NtERF115转基因株系中,NtPMT1a和NtQPT2表达水平显著增加;与空载体转基因对照相比,过表达NtERF115使烟叶中尼古丁质量分数显著增加,平均提高了54.8%.该结果暗示,NtERF115基因可作为利用植物基因工程技术提高烟叶中尼古丁含量的分子工具,也为进一步揭示尼古丁合成的调控机制奠定了基础.Ethylene response factor(ERF)is a plant stress-related transcription factor able to activate downstream responsive gene expression to increase plant stress resistance.Here,we have studied the function of JA-induced transcription factor NtERF115obtained from transcriptomic data of JA-induced BY2cells.Full length of NtERF115was cloned by RT-PCR.Sequence comparison and functional analysis of NtERF115were performed.Inter-/intra-homologs of NtERF115 were identified by homolog blast,followed by multiple sequence alignment to compare their phylogenetic relationship.NtERF115was over-expressed in YN87to detect its effect on nicotine content.CDS of NtERF115was found to be 690bp long,encoding a sequence of 229amino acid residues.NtERF115over-expressing lines showed significantly increased transcription of NtPMT1aand NtQPT2,with a 54.8% increase in nicotine content in the leaves,compared with vectortransfected control plants.

关 键 词：烟草 乙烯响应因子NtERF115 序列分析 尼古丁合成 

分 类 号：Q522[生物学—生物化学]