摘要
为探究生长素诱导相关基因NtIAA13在烟草钾吸收与利用中的作用,以烟草K326的cDNA为模板,克隆了一个全长879 bp,编码292个氨基酸的基因,并将该基因命名为NtIAA13b。采用qRT-PCR和亚细胞定位等生物技术对NtIAA13b基因在不同组织和不同非生物胁迫条件下的表达特性进行分析,通过转基因技术获得NtIAA13b基因过量表达株系并验证其生理功能。结果表明,NtIAA13b基因所编码的蛋白质相对分子质量为31.71 kD,理论等电点(pI)为6.48。NtIAA13b基因的启动子区包含生长素响应元件TGA-element、赤霉素响应元件P-box和参与厌氧诱导的调控元件ARE等。亚细胞定位的结果显示,该蛋白定位于细胞核。系统进化树结果表明,NtIAA13b和番茄SlIAA13亲缘关系最近。实时荧光定量PCR结果表明,NtIAA13b基因在根中的表达水平最高,其次为茎,叶中表达水平最低。在正常供钾条件下,过量表达株系叶片的钾含量极显著增加。NtIAA13b基因与烟草响应低钾胁迫有关,可能参与了烟草钾的吸收与利用。
To investigate the role of auxin-induced gene NtIAA13 in potassium uptake and utilization in tobacco,a gene designated NtIAA13b with a total length of 879 bp and encoding 292 amino acids was cloned using the cDNA of tobacco K326 as a template.The expression characteristics of NtIAA13b in different tissues under different abiotic stress conditions were systematically analyzed by qRT-PCR and subcellular localization,and the overexpression strain of NtIAA13b was obtained by transgenic technology to verify its physiological function.The results showed that the protein encoded by NtIAA13b had a relative molecular mass of 31.71 kD and a theoretical isoelectric point(pI)of 6.48.The promoter region of NtIAA13b contained auxin-responsive element TGA-element and gibberellin-responsive element P-box,as well as regulatory element ARE that was involved in anaerobic induction.The subcellular localization results showed that the protein was localized in the nucleus.The phylogenetic tree results showed that NtIAA13b and tomato SlIAA13 were closely related.The real-time PCR results showed that NtIAA13b had the highest expression level in roots,followed by stalks and leaves with the lowest expression levels.Under normal potassium supply conditions,overexpressed strains significantly increased leaf potassium content.The NtIAA13b gene is thus associated with tobacco response to hypokalemic stress and may be involved in the uptake and utilization of potassium in tobacco.
作者
喻奇伟
朱紫童
何轶
曹廷茂
康俊
罗贞宝
李磊磊
戴彬
李鹏志
高冬冬
吴学巧
贾宏昉
YU Qiwei;ZHU Zitong;HE Yi;CAO Tingmao;KANG Jun;LUO Zhenbao;LI Leilei;DAI Bin;LI Pengzhi;GAO Dongdong;WU Xueqiao;JIA Hongfang(Bijie Branch of Guizhou Provincial Tobacco Company,Bijie 551700,Guizhou,China;College of Tobacco,Henan Agricultural University,Zhengzhou 450002,China)
出处
《烟草科技》
CAS
CSCD
北大核心
2023年第8期12-19,共8页
Tobacco Science & Technology
基金
中国烟草总公司贵州省公司重点研发项目“烟草钾高效吸收利用功能基因挖掘及新品系创制”(2022XM06)。
