葡萄铁蛋白基因Ferritin的鉴定、克隆及其在果实发育不同时期对氨基酸铁复合肥喷施的响应  

Identification and Cloning of Ferritin Family Genes in Grape and Response to Compound Amino Acid-Iron Spraying During Different Fruit Developmental Stages

作　　者：宋志忠 王建萍 史圣朋 曹晶雯 刘万好 徐维华 肖慧琳 唐美玲 SONG ZhiZhong;WANG JianPing;SHI ShengPeng;CAO JingWen;LIU WanHao;XU WeiHua;XIAO HuiLin;TANG MeiLing(Institute of Grape,Yantai Academy of Agricultural Science,Yantai 264000,Shandong,China;The Engineering Research Institute of Agriculture and Forestry,Universities of Shandong Ludong University/Key Laboratory of Molecular Module-Based Breeding of High Yield and Abiotic Resistant Plants,Yantai 264025,Shandong,China;Department of Plant Science,University of Cambridge,Cambridge CB23EA,United Kingdom;Wolfson College,University of Cambridge,Cambridge CB39BB,United Kingdom)

机构地区：[1]烟台农业科学研究院葡萄研究所,中国山东烟台264000 [2]鲁东大学农林工程研究院/山东省高等学校重点实验室(作物高产抗逆分子模块育种实验室),中国山东烟台264025 [3]剑桥大学植物系,英国剑桥CB23EA [4]剑桥大学沃尔森学院,英国剑桥CB39BB

出　　处：《中国农业科学》2023年第18期3629-3641,共13页Scientia Agricultura Sinica

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

摘　　要：【目的】铁蛋白Ferritin在植物生长发育过程中发挥重要作用,其在果树中的生物学功能尚未见报道。克隆葡萄Ferritin家族基因并揭示其在果实发育不同时期的表达模式及其对叶面喷施氨基酸-铁(Fe)复合肥处理的响应差异,可为研究果树铁素营养与代谢的分子机制提供理论依据。【方法】通过同源克隆法从‘马瑟兰’中克隆Ferritin家族基因,利用生物信息学手段分析葡萄Ferritin及其编码蛋白的详细特征;设置叶面喷施氨基酸-Fe复合肥处理,利用荧光实时定量PCR技术分析Ferritin在葡萄果实发育不同时期的表达特征及其对叶面喷施处理的差异响应。【结果】在葡萄基因组中检索并克隆获得4个Ferritin家族基因,命名为VvFer1—VvFer4,分布于第6、8和13号三条染色体上,均含有7个长度不一的内含子,且主要定位于叶绿体和细胞核。本文所选16种植物Ferritin蛋白序列的一致性高达61.48%,系统发育树分析表明同一属的Ferritin同源蛋白如十字花科的拟南芥和芜菁,茄科的烟草和马铃薯,豆科的大豆、落花生和鹰嘴豆,大戟科的橡胶树、木薯和蓖麻,蔷薇科的苹果、桃和草莓,遗传进化关系较近,葡萄VvFer3和茄科同源蛋白遗传关系最近。葡萄VvFer基因在五年生‘马瑟兰’果实发育不同时期、不同组织中的表达水平差异较大,其中,VvFer3在不同组织中的表达量均最高,最大值出现在硬核期至成熟期的果实中,其次是VvFer2和VvFer4。‘马瑟兰’葡萄发育不同时期果实中的Fe含量略有差异,从幼果期开始逐渐增加,并在转色期果实中达到最高值,从转色期到成熟期略有下降,但仍高于幼果期和硬核期果实中的Fe含量,叶面喷施处理显著提高了成熟期果实中的Fe含量、乌头酸酶ACO(aconitase)、硝酸还原酶NIR(nitrite reductase)和琥珀酸脱氢酶SDH(succinate dehydrogenase)的酶活性。VvFer2—4在转录水平易受叶面喷施铁肥的�【Objective】Ferritin plays an important role in plant growth and development,and its biological function in fruit trees are essentially unknown.Cloning of grape Ferritin family genes and revealing their expression patterns at different fruit developmental stages and their response differences to foliar spraying of amino acid iron(Fe)compound fertilizers could provide a theoretical basis for studying the molecular mechanisms of Fe nutrition and metabolism in fruit trees.【Method】The Ferritin family genes were screened and identified in grape genome by homologous cloning.The detailed characteristics of Ferritin genes and their encoded proteins were analyzed by using bioinformatical methods.The tissue-specific expression patterns of Ferritin family genes during distinct fruit developmental stages and differential response to foliar spraying of amino acid-iron compound fertilizer were determined by carrying out real-time fluorescent quantitative PCR.【Result】In total,4 Ferritin family genes were retrieved and cloned from grape,entitled with VvFer1-VvFer4,which were distributed on No.6,8 and 13 chromosomes,containing 7 introns with different lengths.VvFer proteins were mainly located in chloroplast and nucleus.The amino acid sequence identity of Ferritin homologs from 16 plant species was as high as 61.48%.Phylogenetic tree analysis indicated that Ferritin homologs belonging to the same genus,such as Arabidopsis and turnip of Cruciferae,tobacco and potato of Solanaceae,soybeans,peanuts and chickpeas of Leguminosae,rubber trees,cassava and castor of Euphorbiaceae,and apples,peaches and strawberries of Rosaceae,possessed a closer genetic distance during evolution.Grape VvFer3 was closely clustered with Solanaceae homologs.The expression levels of VvFer genes were different among distinct tissues or organs of 5-year-old Mathelan grape trees during different fruit developmental stages.In particular,the expression level of VvFer3 was the most abundant,and the maximum expression was observed in fruits from hard cor

关 键 词：葡萄 Fe的储藏与封存 Fe蛋白 氨基酸-Fe复合肥 叶面喷施 

分 类 号：S663.1[农业科学—果树学]