柠檬蔗糖代谢关键酶基因家族鉴定及表达分析  

Identification and expression analysis of key enzyme gene family in lemon sucrose metabolism

作　　者：王亚娜 卢闯 杨云龙 唐腾[1] 严晓云 黄桂香[1] WANG Ya-na;LU Chuang;YANG Yun-long;TANG Teng;YAN Xiao-yun;HUANG Gui-xiang(College of Agriculture,Guangxi University,Nanning,Guangxi 530004,China)

机构地区：[1]广西大学农学院,广西南宁530004

出　　处：《南方农业学报》2023年第5期1327-1340,共14页Journal of Southern Agriculture

基　　金：广西科技重大专项(桂科AA22068092)。

摘　　要：【目的】对柠檬蔗糖代谢关键酶基因家族成员进行鉴定,并分析其在蔗糖积累有明显差异的2个柠檬品种果实发育过程中的表达情况,为揭示柠檬蔗糖代谢关键酶在果实发育中的作用机制及柠檬与其他柑橘类果实蔗糖代谢分子机制提供理论依据。【方法】运用生物信息学方法鉴定柠檬蔗糖代谢关键酶[蔗糖合成酶(SUS)、蔗糖磷酸合成酶(SPS)、蔗糖转化酶(INV)]基因家族成员,对其基本理化性质、保守基序、基因结构、系统进化、蛋白质保守功能域、顺式作用元件和进化模式等进行全面分析。基于转录组数据分析柠檬蔗糖代谢关键酶基因家族成员在不同组织中的表达模式,并利用实时荧光定量PCR检测其在果皮和果肉发育过程的表达情况。【结果】从柠檬基因组共鉴定获得6个ClSUS基因家族成员(ClSUS1~ClSUS6)、4个ClSPS基因家族成员(ClSPS1~ClSPS4)和15个ClINV基因家族成员(ClINV1~ClINV15),其中ClSPSs基因编码的氨基酸数量最多,ClSUSs、ClSPSs和ClINVs基因编码的保守基序数量和种类大致相同,ClSUSs基因内含子数目为12~14个,ClSPSs基因的内含子数目为12~13个,ClINVs基因的内含子数量为1~11个。ClSUS基因家族成员和ClINV基因家族成员各分为3个亚家族,ClSPS基因家族分为2个亚家族。ClSUS6、ClINV1和ClINV8蛋白均扩张出新的结构域,依据结构域不同,ClINV蛋白家族成员分为2种不同水解酶类型。ClSUSs、ClSPSs和ClINVs基因启动子区域均含有激素响应、光响应、胁迫响应和生长发育调控元件。共25个基因不均匀地分布在染色体上,出现基因串联和片段复制现象。基于转录组数据分析,发现仅有21个基因表达,其中7个基因在柠檬不同组织中特异性表达。21个基因的相对表达量与葡萄糖、果糖和蔗糖含量呈不同程度相关性,其中有9个基因与柠檬葡萄糖、果糖和蔗糖含量呈明显正相关或负相关。实时荧光定量PCR�【Objective】The purpose of the study was to identify the members of the key enzyme gene family of lemon sucrose metabolism and analyze their expression during fruit development in two lemon cultivars with obvious differences in sucrose accumulation,so as to provide reference for revealing the role of lemon sucrose metabolic key enzyme in fruit development and the molecular mechanism of sucrose metabolism in lemon and other citrus fruits.【Method】Bioinforma-tics methodwas used to identify the gene family members of the key enzymes of lemon sucrose metabolism[sucrose syn-thetase(SUS),sucrose phosphatesynthase(SPS),sucrose invertase(INV)],and their basic physicochemical proper-ties,conserved motifs,gene structures,phylogeny,protein-conserved functional domains,cisacting elements and evolu-tionary mode were comprehensively analyzed.The expression patterns of the gene family members of key enzymes of lemon sucrose metabolism in different tissues were analyzed based on transcriptomic data.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect its expression during peel and pulp development.【Result】A total of 6 ClSUS gene family members(ClSUS1-ClSUS6),4 ClSPS gene family members(ClSPS1-ClSPS4)and 15 ClINV gene family mem-bers(ClINV1-ClINV15)were identified from the lemon genome,all of which were hydrophilic proteins,among which ClSPSs gene encoded the largest number of amino acids.The numbers and types of conserved motifs encoded by CISUSs,ClSPSs and ClINVs genes were roughly the same,with the number of introns in ClSUSs gene ranging from 12 to 14,the number of introns in ClSPSs gene ranging from 12 to 13,and the number of introns in ClINVs gene ranging from 1 to11.ClSUS gene family members and ClINV gene family members were each divided into 3 subfamilies,and ClSPS gene family was divided into 2 subfamilies.ClSUS6,ClINV1 and ClINV8 proteins all expanded into new structural domains,and ClINV protein family members were divided into two different hydrolase types according to different structural dom

关 键 词：柠檬 蔗糖合成酶(SUS) 蔗糖磷酸合成酶(SPS) 蔗糖转化酶(INV) 基因家族鉴定 表达分析 

分 类 号：S666.503.6[农业科学—果树学]