番茄SlTIP-1基因的克隆、表达及启动子活性分析  

Cloning, expression and promoter activity analysis of Sl TIP-1 from Solanum lycopersicum

作　　者：李文静 李甜饴 乔洁[1] 朱姝 程兴茹 王聪艳[1,2] 张新业 LI Wenjing;LI Tianyi;QIAO Jie;ZHU Shu;CHENG Xingru;WANG Congyan;ZHANG Xinye(College of Life Sciences,Langfang Normal University,Langfimg,Hebei 065000,China;Hebei Key Laboratory of Animal Diversity,Langfang,Hebei 065000,China;Langfang Key Laboratory of Cell Engineering and Application,Langfang,Hebei 065000,China)

机构地区：[1]廊坊师范学院生命科学学院,河北廊坊065000 [2]河北省动物多样性重点实验室,河北廊坊065000 [3]廊坊市细胞工程与应用研究重点实验室,河北廊坊065000

出　　处：《植物生理学报》2020年第6期1221-1230,共10页Plant Physiology Journal

基　　金：河北省高等学校自然科学研究重点项目(ZD2020122);河北省高等学校科学技术研究项目(QN2018017)。

摘　　要：为研究水孔蛋白分子功能,本研究以番茄(Solanum lycopersicum)矮化品种‘Micro-Tom’为材料,采用RTPCR技术克隆得到1个液泡膜内在蛋白(tonoplast intrinsic proteins,TIPs)类的水孔蛋白基因,命名为SlTIP-1,其开放阅读框(ORF)长753 bp,编码250个氨基酸。基因结构分析发现SlTIP-1有3个外显子和2个内含子。生物信息学分析表明SlTIP-1蛋白含有6个跨膜区及2个NPA(天冬酰胺-脯氨酸-丙氨酸)基序,具有MIP超家族典型的蛋白保守区序列特征。系统进化分析表明,SlTIP-1蛋白与野生种潘那利番茄TIP共聚于一支。实时定量PCR结果表明,SlTIP-1在番茄根中特异性表达。将SlTIP-1基因起始密码子上游1085 bp的序列构建到植物表达载体pBI121中,转化拟南芥(Arabidopsis thaliana)并进行GUS组织化学染色分析。结果表明,SlTIP-1基因启动子驱动GUS基因在拟南芥根部特异性表达,与实时定量PCR结果一致。本研究拟为进一步探索该基因的功能及番茄分子辅助育种提供理论依据。To investigate the molecular function of aquaporin,a tonoplast intrinsic proteins(TIPs)gene TIP-1 was isolated from Solanum lycopersicum‘Micro-Tom’by RT-PCR,and named as SlTIP-1.The SlTIP-1 gene had an open reading frame of 753 bp,encoding 250 amino acid residues.Gene structure analysis showed that it consisted of 3 exons and 2 introns.Bioinformatics analysis revealed that SlTIP-1 included 6 transmembrane domains and 2 typical NPA(Asn-Pro-Ala)motif of MIP superfamily.Phylogenetic analysis displayed that SlTIP-1 was grouped with wild tomato variety pannali TIP protein.Quantitative RT-PCR results demonstrated that SlTIP-1 was specifically expressed in tomato roots.We isolated the 1085 bp upstream sequence of the initial codon of SlTIP-1,constructed it into plant expression vector pBI121 infusing with GUS reporter gene,and transfered the vectors into Arabidopsis thaliana.Histochemical staining showed that SlTIP-1 promoter drove GUS gene to accumulate specifically in Arabidopsis thaliana roots,which was accordance with the results of quantitative RT-PCR.These results will provide a foundation for further functional research of SlTIP-1 protein and molecular marker assisted breeding in tomato.

关 键 词：番茄 水孔蛋白 基因克隆 根特异性启动子 GUS组织化学染色 

分 类 号：S641.2[农业科学—蔬菜学]