摘要
【目的】WRKY转录因子在植物响应非生物胁迫过程中发挥着重要的调控作用,目前超积累植物伴矿景天(Sedum plumbizincicola)WRKY转录因子的研究较少。进行SpWRKY基因家族的全基因组鉴定及其在镉胁迫下的表达模式分析可为后续SpWRKYs基因克隆和耐镉功能分析提供参考。【方法】对伴矿景天WRKY基因家族进行全基因组鉴定和生物信息学分析,利用转录组数据和qRT-PCR技术检测WRKY基因在镉胁迫下的表达模式并进行转基因拟南芥耐镉性的分析。【结果】伴矿景天基因组中共鉴定出77个SpWRKYs,非均匀地分布在各条染色体上;系统进化树分析发现,伴矿景天WRKY成员被分成3大类群(Ⅰ—Ⅲ),第Ⅱ类群又被分成5个亚群(Ⅱa—e);共线性分析发现,伴矿景天WRKY基因家族和拟南芥(Arabidopsis thaliana)之间存在7个共线基因对,和玉吊钟(Kalanchoe fedtschenkoi)之间存在53个共线基因对;SpWRKYs基因之间存在19个片段复制基因对;启动子序列分析发现,SpWRKYs启动子有多种与激素和逆境响应等相关的顺式调控元件;镉胁迫表达分析发现,7个SpWRKYs受镉胁迫显著上调表达,整体表达趋势呈现先增后减的状态;在拟南芥中过表达SpWRKY69发现,SpWRKY69可以增强Cd离子向地上部的转运速率,对植株镉耐性起到负调控作用。【结论】伴矿景天WRKY基因家族与其他物种的WRKY家族结构相似,片段复制是其主要进化动力之一。镉胁迫条件下部分成员相对表达量显著变化,而SpWRKY69能增加Cd离子向地上部的转运速率,故其他WRKY成员可能也参与调控植株镉耐性。
【Objective】WRKY transcription factors play important regulatory roles for a plant abiotic stress response.However,little information is available about WRKY transcription factors in the hyperaccumulating plant Sedum plumbizincicola.The identification of SpWRKY gene family members and analysis of their expression patterns under cadmium stress can provide a reference for molecular cloning of SpWRKYs gene and functional characterization of cadmium tolerance.【Method】In this study,the genome⁃wide identification and bioinformatics analysis of WRKY gene family members were carried out.Expression patterns of WRKY genes under cadmium stress were derived from transcriptome data and qPCR,and the roles of SpWRKY69 in cadmium tolerance were assessed by the heterologous expression in Arabidopsis thaliana.【Result】There were 77 SpWRKYs identified in S.plumbizincicola,unevenly distributed on the chromosomes.Based on the phylogenetic analysis,the SpWRKY proteins were classified into three groups(Ⅰ-Ⅲ),among which the second group was further divided into five subgroups(Ⅱa-e).The synteny analysis showed that there were seven collinear gene pairs in the S.plumbizincicola and A.thaliana WRKY gene family and 53 collinear gene pairs for Kalanchoe fedtschenkoi;19 pairs of SpWRKYs were identified as the segmental duplication.Cis⁃regulatory elements related to stress and hormone responses were found in the promoters of SpWRKYs.The expression profiles showed that the expression levels of the seven SpWRKYs were significantly up⁃regulated under cadmium stress,exhibiting a pattern of increasing initially and then decreasing.Over⁃expression of SpWRKY69 in A.thaliana showed that SpWRKY69 can enhance the transport rate of Cd ions to the shoot and play a negative role in regulating plant cadmium tolerance.【Conclusion】Structural features of WRKY gene family members in S.plumbizincicola are similar to those of other species,and the fragment duplication is one of the main evolutionary forces.The relative expression level
作者
王剑超
邱文敏
金康鸣
陆铸畴
韩小娇
卓仁英
刘晓光
何正权
WANG Jianchao;QIU Wenmin;JIN Kangming;LU Zhuchou;HAN Xiaojiao;ZHUO Renying;LIU Xiaoguang;HE Zhengquan(Key Laboratory of Three Gorges Regional Plant Genetics&Germplasm Enhancement(CTGU),Biotechnology Research Center,China Three Gorges University,Yichang 443002,China;Key Laboratory of Tree Breeding of Zhejiang Province,The Research Institute of Subtropical Forestry,Chinese Academy of Forestry,Hangzhou 311400,China;Tangshan Academy of Agriculture Science,Tangshan 063001,China)
出处
《南京林业大学学报（自然科学版）》
CAS
CSCD
北大核心
2023年第2期49-60,共12页
Journal of Nanjing Forestry University：Natural Sciences Edition
基金
国家重点研发计划子课题(2016YFD080080104)。
关键词
伴矿景天
WRKY转录因子
镉胁迫
基因表达
拟南芥转基因
Sedum plumbizincicola
WRKY transcription factor
cadmium stress
gene expression
Arabidopsis transformation
