蓝莓VcILR1基因的克隆与表达分析  

Cloning and Expression of VcILR1 in Blueberry

作　　者：胡丽超 王小敏[2] 王鑫 吴雅琼 吴文龙[2] 闾连飞[2] 李维林 HU Lichao;WANG Xiaomin;WANG Xin;WU Yaqiong;WU Wenlong;LYU Lianfei;LI Weilin(Southern Modern Forestry Collaborative Innovation Center/College of Forestry,Nanjing Forestry University,Nanjing,Jiangsu 210037,China;Institute of Botany,Chinese Academy of Sciences in Jiangsu(Nanjing Mem.Sun Yat-Sen Botanical Garden)/Key Laboratory for Research and Utilization of Plant Resources,Nanjing,Jiangsu 210014,China;Xinghua Forestry Technical Guidance Station,Xnghua,Jiangsu 225700,China)

机构地区：[1]南京林业大学南方现代林业协同创新中心/南京林业大学林学院,江苏南京210037 [2]江苏省中国科学院植物研究所(南京中山植物园)/江苏省植物资源研究与利用重点实验室,江苏南京210014 [3]江苏省兴化市林业技术指导站,江苏兴化225700

出　　处：《福建农业学报》2023年第2期151-157,共7页Fujian Journal of Agricultural Sciences

基　　金：江苏省种业振兴揭榜挂帅项目(JBGS[2021]021);江苏省科技厅现代农业项目(BE2022372)。

摘　　要：[目的]克隆蓝莓(Vaccinium spp)生长素酰胺水解酶基因VcILR1,分析该基因在蓝莓花、叶、茎、果和根中的表达模式和经过赤霉素处理的青果和成熟果两个时期的表达情况,为进一步探究该基因功能和蓝莓胚珠败育的机制提供依据。[方法]以蓝莓果实的cDNA为模板克隆得到VcILR1基因,利用ProtParam等工具对VcILR1进行生物信息学分析,利用qRT-PCR方法对VcILR1基因在蓝莓不同组织及赤霉素处理的青果和成熟果中的表达量进行分析。[结果]成功克隆到蓝莓VcILR1基因,该基因含976 bp的开放阅读框(ORF),编码325个氨基酸,有两个保守结构域,分别是Peptidase_M20和M20_dimer;编码蛋白的分子质量35 364.20 kDa,理论等电点5.67,不稳定系数43.57,脂肪系数88.55,亲水性平均值-0.054,属于不稳定亲水性蛋白。系统进化树分析表明蓝莓VcILR1基因与山茶科植物的茶树(Camellia sinensis)生长素酰胺水解酶(IAA-Leucine Resistant 1-like,ILL)基因亲缘关系较近。qRTPCR分析结果显示,VcILR1在蓝莓花、叶、茎、果和根中均有表达,在叶片中表达量最高,具有组织特异性;赤霉素处理能上调果实中VcILR1的表达。[结论]赤霉素处理后蓝莓青果和成熟果中VcILR1的表达量均显著增高,推测赤霉素处理能够促进蓝莓果实中生长素含量的提高,从而抑制胚珠发育,导致果实无籽。【Objective】VcILR1 of blueberry was cloned to analyze the expressions in various plant organs as well as after being treated by gibberellin to decipher its function and association with ovule abortion.【Method】Using the cDNA of blueberry as the template,VcILR1 was cloned by RT-PCR.Bioinformatics on the amino acid sequence was performed utilizing ProtParam and other relevant tools.Expressions of the gene in the flowers,leaves,stems,fruits,and roots of the plant as well as those in green and ripe fruits after being treated by gibberellin were determined.【Result】VcILR1 was successfully cloned and found to contain 976 bp ORF encoding 325 amino acids with two conserved structural domains,peptidase_M20 and M20_dimer,a molecular mass of 35364.20 kDa,a theoretical isoelectric point of 5.67,an instability coefficient of 43.57,and a lipid coefficient of 88.55.It was an unstable protein with a hydrophilic mean value of−0.054.A phylogenetic tree analysis showed the gene to closely relate to Camellia sinensis IAA-Leucine Resistant 1-like(ILL)gene.The qRT-PCR analysis revealed that the gene was expressed tissue-specific with the highest in the leaves,and that the gibberellin treatment up-regulated the expression in the fruits.【Conclusion】The expressions of VcILR1 in the blueberries significantly increased after being treated by gibberellin.It suggested that the plant hormone could stimulate IAA generation in the fruits and might also affect the ovule development.

关 键 词：蓝莓 VcILR1基因 基因克隆 无籽 

分 类 号：Q785[生物学—分子生物学]