文献类型：期刊文献

中文题名：超积累型东南景天Sa12F279基因的抗逆表达响应及功能关联分析

英文题名：Characterization of Expression Patterns of Sa12F279 from Sedum alfredii under Abiotic Stress and Its Functional Interaction Analysis

作者：冯童禹[1,2] 乔桂荣[1] 蒋晶[1] 邱文敏[1] 韩小娇[1] 卓仁英[1] 刘明英[1]

第一作者：冯童禹

机构：[1]中国林业科学研究院亚热带林业研究所,浙江杭州311400;[2]南京林业大学,江苏南京210037

年份：2020

卷号：33

期号：3

起止页码：12-21

中文期刊名：林业科学研究

外文期刊名：Forest Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2017】；CSCD:【CSCD2019_2020】；

基金：国家自然科学基金面上项目“SaREFl基因参与超积累型东南景天镉抗性的分子机理研究”(31870647)。

语种：中文

中文关键词：东南景天;ABC转运蛋白;非生物胁迫;表达模式

外文关键词：Sedum alfredii;ABC transporters;abiotic stress;expression pattern

分类号：S718.46

摘要：[目的]对超积累型东南景天ABC转运蛋白家族成员Sa12F279开展生物信息学分析及表达分析,为探究ABC类转运蛋白在镉、干旱、盐碱等非生物胁迫中的功能提供参考。[方法]通过对超积累型东南景天转录组数据库进行比对分析,获得1个ABC家族成员的基因。通过生物信息学方法分析Sa12F279基因的进化关系、蛋白结构域构成、核心结构域同源比对及在组学数据中相关的互作蛋白分类;借助实时定量PCR技术分析该基因在盐碱、干旱及ABA激素胁迫下根中的表达变化。[结果]通过比对分析获得ABC家族成员Sa12F279基因,该基因的开放阅读框长度为4 497 bp,编码蛋白长度为1 498个氨基酸,相对分子量为167.1 KD,等电点为6.92。进化分析显示:Sa12F279基因与C亚类成员聚为一簇,且在结构域构成上符合ABC家族的保守排布。共表达网络分析显示:Sa12F279与567个基因存在功能上的关联,对这些基因进行功能注释发现,其中39.8%的基因执行代谢相关功能,29.3%的基因与细胞内进程相关,10.2%的基因涉及生物调控,7.1%的基因参与转运活性。对镉胁迫前后转录组数据分析显示,Sa12F279基因在根、茎、叶3种组织中,取样点24 h和96 h的表达量均呈现对镉胁迫下调响应。实时定量结果显示:该基因对ABA激素、盐碱和干旱不同胁迫的应答模式存在差异,且应答响应较平缓。在ABA激素处理下,呈现先下调后上升的趋势;在盐胁迫下,呈现早期响应不显著而于胁迫后期出现上调应答;在干旱处理下,呈现先升后降又升的趋势。[结论]鉴定了超积累型东南景天ABC家族的1个Sa12F279基因,揭示了该基因在不同非生物胁迫下的表达模式。
[Objective]In order to elucidate the functions of the ATP-binding cassette(ABC)protein in Sedum alfredii,a Cd hyperaccumulator,the bioinformatics analysis and expression pattern study were performed on Sa12F279 belonging to ABC protein family.[Methods]A local blast was carried out in the transcriptomic datasets of S.alfredii and produced a transcript categorized to ABC protein family.Further bioinformatics analysis was conducted on the phylogenetic clustering,the protein domain structures and classification of possible interaction proteins.Quantitative real-time PCR(qRT-PCR)was applied to uncover the expression patterns of Sa12F279 in the root under drought,salt,abscisic acid stresses.[Result]In the present work,a gene belonging to C subfamily of ABCs was identified in S.alfredii using local blast and designated as Sa12F279.The length of Sa12F279 open reading frame was 4497 bp,coding a protein of 1498 amino acids with pI being 6.92.The phylogenetic analysis showed that Sa12F279 was clustered with the C-subfamily of ABC protein,exhibiting uniform domain structure of TMD-NBDTMD-NBD.Co-expression network analysis indicated that Sa12F279 served as hub genes mostly associated with metabolic process(39.8%),cellular process(29.3%),biological regulation(10.2%)and transporter activity(7.1%).The expression data from the transcriptomic datasets showed that Sa12F279 was suppressed by Cd stress in the three tissues(root,stem and leaf)at the sampling points of 24 h and 96 h.Transcriptional expression profiles responsive to drought,salt,abscisic acid(ABA)stress illustrated different response tendencies showing moderate induction.Under the treatment of ABA,the expression of Sa12F279 exhibited the tendency of decrease to elevation.Under the salt stress,Sa12F279 was up-regulated at the late stage of treatment while under the drought stress,Sa12F279 displayed the irregular tendency of elevation-decrease-elevation.[Conclusion]Based on the studies above,the authors characterize a gene named Sa12F279 belonging to the ABC protein family in S.alfredii and performs a comprehensive analysis including phylogenetic analysis,protein structure and co-expression network.Meanwhile,the expression profiles under different abiotic stress are also uncovered.The results could provide reference for the further studies on the ABC protein family.