葡萄IRT基因的克隆、鉴定及其对氨基酸-铁复合肥的响应特征  

Cloning and identification of IRT genes and their response to amino acid-iron compound fertilizer in grape

作　　者：郑秋玲[1,2] 王建萍 肖慧琳[1] 史圣朋[3] 宁友峥 杜远鹏 管雪强[5] 唐美玲 宋志忠 ZHENG Qiuling;WANG Jianping;XIAO Huilin;SHI Shengpeng;NING Youzheng;DU Yuanpeng;GUAN Xueqiang;TANG Meiling;SONG Zhizhong(Shandong Engineering Research Center for Grape Breeding and Cultivation,Yantai Academy of Agricultural Sciences,Yantai 265500,China;The Engineering Research Institute of Agriculture and Forestry,Ludong University,Yantai 264025,China;Department of Plant Science,University of Cambridge,Cambridge CB23EA,United Kingdom;College of Horticulture Science and Engineering,Shandong Agriculture University,Tai’an 271018,China;Shandong Technology Innovation Center of Wine Grape and Wine/COFCO Great Wall Wine(Penglai)Co.,Ltd.,Yantai 264000,China)

机构地区：[1]山东省烟台市农业科学研究院葡萄良种选育与良法栽培山东省工程研究中心,山东烟台265500 [2]鲁东大学农林工程研究院,山东烟台264025 [3]剑桥大学植物系,英国剑桥CB23EA [4]山东农业大学园艺科学与工程学院,山东泰安271018 [5]山东省酿酒葡萄与葡萄酒技术创新中心/中粮长城葡萄酒(蓬莱)有限公司,山东烟台264000

出　　处：《江苏农业学报》2024年第5期913-921,共9页Jiangsu Journal of Agricultural Sciences

基　　金：山东省重点研发计划项目(重大科技创新工程)(2022CXGC010605);国家现代农业产业技术体系建设专项(CARS-29-17);国家留学基金项目(202208370080);烟台市科技计划项目(2020XCZX026)。

摘　　要：为揭示葡萄铁素吸收与转运的分子机制,明确铁调节转运蛋白编码基因VvIRT在葡萄果实不同发育时期的表达差异及其对叶面喷施氨基酸-铁复合肥的响应特征,本研究以烟酿1号为材料,分离并克隆葡萄铁调节转运蛋白编码基因IRT,分析IRT基因分布、结构及其编码蛋白质的保守基序等特征;设置叶面喷施氨基酸-铁复合肥处理,利用实时荧光定量PCR分析烟酿1号IRT基因在果实不同发育时期的表达差异及其对叶面喷施处理的响应特征。结果表明:在葡萄基因组中克隆得到10个VvIRT基因(VvIRT1~VvIRT10),分布在7条染色体上。其编码蛋白质均含有Fe^(2+)转运蛋白或锌/铁转运体(PF02535),属于典型的铁调节蛋白。VvIRT1、VvIRT2、VvIRT4、VvIRT5、VvIRT8等基因编码的蛋白质属于碱性蛋白质,而其他5个蛋白质属于酸性蛋白质。VvIRT5蛋白和VvIRT8蛋白属于不稳定蛋白,其他8个蛋白为稳定蛋白。VvIRT7蛋白主要定位于液泡膜,其他蛋白质均主要定位于细胞质膜。VvIRT7在葡萄果实不同发育时期各组织中的整体表达水平都是最高的,其次为VvIRT9和VvIRT6,而VvIRT1、VvIRT3、VvIRT5、VvIRT8和VvIRT10在葡萄果实发育过程中均无表达。叶面喷施铁肥处理下,韧皮部中VvIRT7表达量和果实及韧皮部中VvIRT9表达量在花后35 d(幼果期)至70 d(转色期)明显增加,而在花后85 d(第2次膨大期)至115 d(成熟期),韧皮部和叶片中VvIRT7和VvIRT9表达量明显减少。因此,葡萄VvIRT基因对叶面喷施铁肥的响应特征与果实发育时期及器官类型密切相关,VvIRT7、VvIRT9基因编码蛋白质是葡萄果实发育过程中2个重要的铁调节转运蛋白。In order to reveal the molecular mechanism of iron absorption and transport in grape,and to clarify the expression difference of iron regulatory transporter gene VvIRT in different development stages of grape fruit and its response characteristics to foliar application of amino acid-iron compound fertilizer,Yanniang 1 was used as the material in this study.The IRT encoding iron regulatory transporter was isolated and cloned,and the distribution and structure of IRT gene and conserved motifs of proteins encoded by IRT gene were analyzed.The expression difference of IRT gene in different development stages of Yanniang 1 fruit and its response characteristics to foliar spraying treatment were analyzed by real-time fluorescence quantitative PCR.The results showed that 10 VvIRT genes(VvIRT1-VvIRT10)were cloned from the grape genome and distributed on seven chromosomes.The encoded proteins contained Fe 2+transporters or zinc/iron transporters(PF02535),which were typical iron regulatory proteins.The proteins encoded by VvIRT1,VvIRT2,VvIRT4,VvIRT5 and VvIRT8 were alkaline proteins,while the other five proteins were acidic proteins.VvIRT5 protein and VvIRT8 protein were unstable proteins,and the other eight proteins were stable proteins.The VvIRT7 protein was mainly located in the vacuolar membrane,and the other proteins were mainly located in the plasma membrane.The overall expression level of VvIRT7 was the highest in all tissues at different developmental stages of grape fruit,followed by VvIRT9 and VvIRT6,while VvIRT1,VvIRT3,VvIRT5,VvIRT8 and VvIRT10 were not expressed during the development of grape fruit.Under foliar application of iron fertilizer,the expression levels of VvIRT7 in phloem and VvIRT9 in fruit and phloem increased significantly from 35 d after flowering(young fruit stage)to 70 d after flowering(veraison stage),while the expression levels of VvIRT7 and VvIRT9 in phloem and leaves decreased significantly from 85 d after flowering(second expansion stage)to 115 d after flowering(mature stage).Therefore,th

关 键 词：葡萄 铁调节转运蛋白 铁肥 VvIRT基因 

分 类 号：Q786[生物学—分子生物学] S663.1[农业科学—果树学]