苋菜生长素响应因子AtrARF7的克隆及表达  

Cloning and expression analysis of auxin response factor AtrARF7 in Amaranthus tricolor L.

作　　者：王乐 陈何 肖昉 郑友峰 安子贤 李宜轩 刘生财[1] WANG Le;CHEN He;XIAO Fang;ZHENG Youfeng;AN Zixian;LI Yixuan;LIU Shengcai(Institute of Horticultural Biotechnology,Fujian Agriculture and Forestry University,Fuzhou 350002,China)

机构地区：[1]福建农林大学园艺植物生物工程研究所,福州350002

出　　处：《应用与环境生物学报》2022年第3期623-630,共8页Chinese Journal of Applied and Environmental Biology

基　　金：福建省自然科学基金项目(2018J01700);福建农林大学科技创新专项基金项目(CXZX2017174,CXZX2016118,CXZX2017189)资助。

摘　　要：为了解苋菜生长素响应因子(ARF)的表达特性及功能,采用RT-PCR方法从全红苋菜(Amaranthus tricolor L.)中克隆获得1个生长素响应因子AtrARF7基因(GenBank登录号:MW036654)并进行生物信息学分析,同时利用qRT-PCR分析AtrARF7在不同浓度生长素处理下的苋菜幼苗及苋菜愈伤组织中的表达模式.结果表明:苋菜AtrARF7基因含有一个3324 bp的开放阅读框(ORF),编码1107个氨基酸,预测分子量为12.34×10^(3),等电点为6.07,是亲水不稳定蛋白.AtrARF7包括B3、Auxin-resp和AUX_IAA这3个ARF基因特有的结构域;多序列比对和系统进化树分析发现,苋菜AtrARF7蛋白与其他植物ARF7蛋白具有高度一致性,且与甜菜BvARF7蛋白亲缘关系最近.荧光定量结果表明,AtrARF7在不同浓度NAA处理下的苋菜幼苗中呈上调表达;在MS+6.0 mg/L 6-BA+0.5 mg/L 2,4-D培养基中诱导的苋菜愈伤组织处于诱导期时,AtrARF7的表达量达到最高;在添加了不同浓度NAA的继代培养基(MS+6.0 mg/L 6-BA+0.5 mg/L 2,4-D)中AtrARF7均受到调控.本研究表明AtrARF7可能具有ARF家族正调控因子表达特征,其转录水平受到生长素的诱导,且AtrARF7在苋菜愈伤组织形成的3个时期中均有响应,初步推测其参与生长素信号传导途径调控苋菜愈伤组织形成的过程.(图10表2参29)To investigate the expression characteristics and function of auxin response factor(ARF)in Amaranthus tricolor L.,an auxin response factor gene AtrARF7(GenBank Accession Number:MW036654)was cloned using reverse-transcription PCR(RT-PCR),and bioinformatics analysis was performed.In addition,quantitative RTPCR(qRT-PCR)analysis was performed to determine AtrARF7 expression patterns in amaranth seedlings and calli treated with different auxin concentrations.The results showed that AtrARF7 contains a 3324 bp open reading frame,which encodes a protein of 1107 amino acids.AtrARF7 is a hydrophilic unstable protein with a predicted molecular weight of 12.34×10^(3)and an isoelectric point at 6.07.AtrARF7 includes three ARF gene domains:B3,auxin-resp,and AUX_IAA.Multiple sequence alignment and phylogenetic tree analysis showed that AtrARF7 was highly consistent with ARF7 found in other plants and was closely related to BvARF7 of Beta vulgaris L.The qRT-PCR analysis showed that AtrARF7 expression was upregulated in amaranth seedlings treated with different concentrations of 1-naphthaleneacetic acid(NAA).In the induction stage of amaranth callus induced by Murashige and Skoog(MS)medium supplemented with 6.0 mg/L 6-benzylaminopurine(6-BA)and 0.5 mg/L 2,4-dichlorophenoxyacetic acid(2,4-D),the expression level of AtrARF7 was the highest.AtrARF7 was also regulated in the subculture medium(MS medium supplemented with 6.0 mg/L 6-BA and 0.5 mg/L 2,4-D)supplemented with different concentrations of NAA.We showed that AtrARF7 may possess the expression characteristics of positive regulators of the ARF family,and its transcription level was induced by auxin.In addition,AtrARF7 responded to the three stages of amaranth callus formation;thus,it was speculated that AtrARF7 is involved in the regulation of amaranth callus formation.

关 键 词：苋菜 生长素响应因子(ARF) 基因克隆 表达分析 

分 类 号：S636.4[农业科学—蔬菜学]