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
郑秋玲 1王建萍 2肖慧琳 2史圣朋 3宁友峥 3杜远鹏 4管雪强 5唐美玲 6宋志忠7
作者信息
- 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;鲁东大学农林工程研究院,山东 烟台 264025
- 2. 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
- 3. 剑桥大学植物系,英国 剑桥 CB2 3EA
- 4. 山东农业大学园艺科学与工程学院,山东 泰安 271018;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
- 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
- 6. 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;鲁东大学农林工程研究院,山东 烟台 264025;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
- 7. 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;鲁东大学农林工程研究院,山东 烟台 264025;剑桥大学植物系,英国 剑桥 CB2 3EA
- 折叠
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Fe2+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.
Abstract
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 mate-rial in this study.The IRT encoding iron regulatory transporter was isolated and cloned,and the distribution and struc-ture 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 Fe2+transporters or zinc/iron transporters(PF02535),which were typical iron regulatory proteins.The proteins enco-ded by VvIRT1,VvIRT2,VvIRT4,VvIRT5 and VvIRT8 were alkaline proteins,while the other five proteins were acid-ic proteins.VvIRT5 protein and VvIRT8 protein were unstable proteins,and the other eight proteins were stable pro-teins.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 de-creased significantly from 85 d after flowering(second expansion stage)to 115 d after flowering(mature stage).Therefore,the response characteristics of VvIRT gene to foliar application of iron fertilizer are closely related to fruit development stage and organ type.The proteins encoded by VvIRT7 and VvIRT9 genes are two important iron regulatory transporters during grape fruit development.
关键词
葡萄/铁调节转运蛋白/铁肥/VvIRT基因Key words
grape/iron-regulated transporter/iron fertilizer/VvIRT gene引用本文复制引用
基金项目
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)
出版年
2024
国家科技期刊平台
NSTL
万方数据