番杏LIM基因家族鉴定及TtWLIM1b的克隆与功能分析  

Characterization of LIM gene family from Tetragonia tetragonoides(Pall.)Kuntze and cloning and functional analysis of TtWLIM1b

作　　者：刘浩[1,2] 丁倩倩 王峥峰 简曙光[1] 张美 LIU Hao;DING Qian-qian;WANG Zheng-feng;JIAN Shu-guang;ZHANG Mei(CAS Engineering Laboratory for Vegetation Ecosystem Restoration on Islands and Coastal Zones/Guangdong Provincial Key Laboratory of Applied Botany/South China Botanical Garden,Chinese Academy of Sciences,Guangzhou 510650,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]中国科学院海岛与海岸带生态修复工程实验室/广东省应用植物学重点实验室/中国科学院华南植物园,广州510650 [2]中国科学院大学,北京100049

出　　处：《西南农业学报》2024年第5期944-954,共11页Southwest China Journal of Agricultural Sciences

基　　金：国家重点研发计划项目(2022YFC3103700);国家自然科学基金项目(32270380);广东省科技计划项目(2019B121201005)。

摘　　要：【目的】分析番杏[Tetragonia tetragonoides(Pall.) Kuntze]中TtLIM基因家族的序列组成、RNA-seq基因表达和异源表达生物学功能,探究其可能参与调控番杏植株发育和胁迫应答反应的分子机理,为发掘番杏抗逆功能基因及其在作物育种中的应用提供理论基础。【方法】通过前期对番杏[Tetragonia tetragonoides(Pall.) Kuntze]cDNA文库的筛选,获得一个编码番杏LIM基因的全长cDNA,命名为TtWLIM1b,对TtWLIM1b在酵母中进行功能分析。结合番杏全基因组测序信息,从番杏基因组中鉴定出16个TtLIM基因,对这些家族成员的染色体定位、所编码蛋白的理化性质、系统进化以及蛋白的保守模式进行综合的生物信息学分析;构建番杏与拟南芥、水稻和紫花苜蓿的LIM蛋白家族成员的进化树。最后,通过RNA-seq分析TtLIM基因家族成员在番杏不同组织以及在不同胁迫处理后的相对表达量。【结果】异源表达TtWLIM1b能够提高转基因酵母的重金属耐受性及耐热能力;番杏TtLIM家族蛋白成员均含有LIM结构域,而该结构域富含半胱氨酸。因此,TtLIMs也是富含半胱氨酸蛋白(Cysteine rich protein, CRP)的一种。序列分析表明,番杏LIM蛋白成员具有高度的保守性,且其成员的复制扩增较为明显;TtLIM基因家族成员在不同的番杏器官中广泛而有差异地表达。【结论】番杏TtLIMs可能参与调控番杏植株发育和胁迫应答反应。[Objective]The study aimed to explore the possible biological roles for development and abiotic stress responses of TtLIM gene family from Tetragonia tetragonoides(Pall.)Kuntze,mainly based on the RNA-seq expression patterns and the phenotype analysis of heter-ologous expression in yeast.[Method]In the study,we screened the cDNA library of T.tetragonoides(Pall.)Kuntze,and cloned a novel LIM gene from this species,named TtWLIMlb.The ectopic expression of TtWLIMIb in yeast was performed.Combined with the whole ge-nome sequencing data,a total of 16 TtLIM genes were identified from T.tetragonoides genome.The chromosomal localization,physicochemi-cal properties of the encoded proteins,phylogenetic evolution,and conserved motifs of these family members were comprehensively analyzed.A phylogenetic tree was constructed to compare the LIM protein family members between T.tetragonoides and Arabidopsis thaliana(L.)Heynh,Oryza sativa L,and Medicago sativa.Finally,RNA-seq analysis was performed to investigate the relative expression levels of TtLIM gene family members in different tissues and under various stress treatments.[Result]The heterologous expression of some TtLIMlb members could improve the heavy metals and heat tolerance of yeast.TtLIM proteins consisted of one or two LIM domains,and LIM domain was cyste-ine-rich zinc finger sequence,therefore TtLIM proteins also belonged to cysteine rich protein(CRP).The sequence analysis indicated that TtLIMs were highly conserved and their genes'reduplicating amplification was prominent during evolution.The RNA-seq results showed that TtLIMs were widely expressed with specific patterns in different organs in T.tetragonoides plant.[Conclusion]TtLIMs might play important roles in regulation of T.tetragonoides development and stress responses.

关 键 词：番杏 LIM基因 逆境胁迫 金属耐受 

分 类 号：S184[农业科学—农业基础科学]