森林草莓Fv CAX基因家族的鉴定及生物学功能分析

Identification and biological function analysis of FvCAX gene family in Fragaria vesca

CAXs是存在于植物细胞质膜或液泡膜上的多基因家族,能调控Ca2+的平衡,是重要的阳离子转运蛋白。因此,从全基因组水平确定森林草莓中CAX基因家族成员并通过数据库分析CAX家族成员的进化关系和基因结构、分析表达模式和顺式作用元件,可为研究森林草莓中CAX基因家族的功能奠定理论基础。首先获得拟南芥CAX的保守序列作为参考,通过TBtools软件比对得到森林草莓FvCAXs基因家族成员。利用数据库分析家族成员的染色体定位并进行命名,分析得到FvCAXs的理化性质、亚细胞定位、基因结构及保守结构域。比较FvCAXs基因在不同组织及同一组织不同发育时期的表达量并利用R软件对FvCAXs家族的基因表达模式进行可视化分析,通过植物顺式调控数据库分析FvCAXs启动子元件。森林草莓FvCAXs家族包含7个成员,编码427~543个氨基酸、分子量介于47.02~58.82k Da,等电点介于4.90~5.81,且都为疏水性蛋白;大多数位于细胞质膜和液泡膜上,少部分位于叶绿体类囊体膜;进化上森林草莓与月季、苹果的CAX亲缘关系较近。森林草莓FvCAXs具有组织特异性表达特点:Fv CAX1~3和Fv CAX5在整个植株发育时期表达量较低,仅在花发育的部分时期有表达量;Fv CAX6和Fv CAX7在苗期、叶片、花的发育过程和种子的形成过程中表达量都有显著性增加。启动子分析表明,顺式作用元件主要有MYB、Dof(AAAG)、WRKY、W-box和MBS,表明FvCAXs基因家族成员的大多数基因能够参与外界的胁迫反应。

CAXs,a kind of gene family,located in the membrane of plasma or vacuole,regulate the balance of Ca2+concentration and are important for the transportation of position ions.Thus,identification the members,phylogenetic analysis,gene structures,expression profiles and cis-acting elements of FvCAXs in Fragaria vesca on the whole genome level will provide a theoretical basis for further analyzing of their functions.The conserved sequences of CAX in Arabidopsis thaliana were firstly obtained,and then the FvCAXs were identified using TBtools software.FvCAXs were named according to the chromosome location.The physical and chemical properties,subcellular localizations,gene structures and conserved domains were also analyzed.The expressional profiles in different tissues or the same tissue at various development stages were performed using R software.Besides,the cis-acting regulatory elements of FvCAXs were obtained based on the promoter analysis.In this study,a total of seven FvCAXs were identified based on the genome,containing 427-543 amino acids with the molecular weight of 47.02-58.82 kDa.The isoelectric points(pI)were from 4.90 to 5.81,and all proteins belonged to hydrophobic proteins.Most of them were in the membrane of plasma or vacuole,while a few were in the chloroplast of thylakoid membrane.Fv CAX showed a close relationship with Rosa cultivars and Malus domestica.The tissue-specific expression profiles proved that:the expression level of Fv CAX1-3 and FvCAX5 were low at the development stage,only a few expressed higher at the flower stage;FvCAX6 and FvCAX7 expressed higher in seedlings,leaves,flowers and seed.The major cis-acting regulatory elements were determined,including MYB,Dof(AAAG),WRKY,W-box and MBS,suggesting that most of FvCAXs are regulated by environmental stresses.