甘蔗ScHAK11基因启动子的克隆与初步分析  

Cloning and Preliminary Analysis of ScHAK11 Gene Promoter in Sugarcane

作　　者：罗海斌[1] 魏源文[2] 曹辉庆[1] 蒋胜理 吴兴剑 叶丽萍 黄诚梅[1] Luo Haibin;Wei Yuanwen;Cao Huiqing;Jiang Shengli;Wu Xingjian;Ye Liping;Huang Chengmei(Guangxi Crop Genetic Improvement and Biotechnology Laboratory,Guangxi Academy of Agricultural Sciences,Nanning,530007;Agro-products Quality Safety and Testing Technology Research Institute,Guangxi Academy of Agricultural Sciences,Nanning,530007)

机构地区：[1]广西农业科学院,广西作物遗传改良生物技术重点开放实验室,南宁530007 [2]广西农业科学院农产品质量安全与检测技术研究所,南宁530007

出　　处：《分子植物育种》2023年第15期4914-4922,共9页Molecular Plant Breeding

基　　金：广西自然科学基金项目(2018GXNSFBA281103;2020GXNSFAA259061);广西农业科学院基本科研业务专项(桂农科2020YM107;桂农科2021YT118);广西甘蔗遗传改良重点实验室运行补助项目(19-185-24-K-01-02)共同资助。

摘　　要：钾转运体ScHAK11基因是甘蔗钾转运体基因家族的重要成员。本研究以甘蔗为材料,通过染色体步移方法对ScHAK11上游启动子片段(pScHAK11)进行克隆,获得ScHAK11起始密码子ATG上游启动子序列,序列长度为2018 bp。序列分析表明,该序列包含多个真核生物启动子核心元件TATA-box、CAAT-box以及与逆境胁迫、光响应、激素诱导、分生组织和叶肉栅栏组织表达等顺式作用元件,推测pScHAK11启动子受到多种激素和逆境胁迫诱导表达,并通过分生组织和叶肉栅栏组织等顺式调控元件参与对甘蔗组织发育的调控。将p ScHAK11启动子序列与包含GUS基因的载体pBI121连接进行活性分析,发现pScHAK11启动子片段能驱动GUS基因在烟草茎和根中瞬时表达。荧光定量PCR结果表明,ScHAK11主要在甘蔗叶片和根系表达,且其表达受发育时期的影响,该结果与pScHAK11启动子驱动的GUS基因在烟草中的表达结果不一致,结果表明p Sc HAK11启动子是组织特异型启动子。本研究结果有助于深入了解ScHAK11基因表达调控的分子机制,为研究ScHAK11基因的转录调控机制奠定基础。Potassium transporter ScHAK11 gene was playing an important role in sugarcane potassium transporter gene family.In the present investigation,by using genomic walking methods,the 2018 bp promoter region of ScHAK11 gene from sugarcan was cloned(pScHAK11).Computational analysis affirmed the existence of abiotic stress responsive cis-elements and core cis-elements such as TATA box,CAAT box,stress response motifs,light response motifs,phytohormone responsive motifs,as well as regulatory motifs related to the meristem and mesophyll palisade tissue.It is speculated that ScHAK11 promoter induced by phytohormone and stress,and the promoter may participate in the regulation of sugarcane tiller.By constructing the target promoter into PBI121 plasmid that contains a GUS reporter,and transient expression in tobacco,the results showed that ScHAK11 promoter could result in transient expression of GUS gene in stems and roots of tobacco.The results of fluorescence quantitative PCR showed that ScHAK11 was mainly expressed in sugarcane leaves and roots,and its expression was affected by development stage,the results of fluorescence quantitative PCR were not consistent with the results of GUS expression driven by pScHAK11 in tobacco.The research results show that pScHAK11 promoter is a tissue-specific promoter,and there results are helpful to revealed the molecular mechanism of ScHAK11 expression regulation,and provide the foundation information for ScHAK11 on transcriptional regulation.

关 键 词：甘蔗 ScHAK11基因启动子 瞬时表达分析 

分 类 号：S566.1[农业科学—作物学]