菠萝AcNINV家族全基因组分离及表达分析  

Genome-wide identification and expression analysis of AcNINV family in pineapple

作　　者：吴建阳 陈妹[3,4] 姚艳丽 张秀梅[3,4] WU Jianyang;CHEN Mei;YAO Yanli;ZHANG Xiumei(Zhanjiang Preschool Education College,Zhanjiang 524037,Guangdong,China;Basic Education College of Lingnan Normal University,Zhanjiang 524037,Guangdong,China;South Subtropical Crop Research Institute,Chinese Academy of Tropical Agricultural Science(CATAS)/Key Laboratory of Tropical Fruit Biology,Ministry of Agriculture and Rural Affairs,Zhanjiang 524091,Guangdong,China;National Key Laboratory for Tropical Crop Breeding,Sanya 572024,Hainan,China)

机构地区：[1]湛江幼儿师范专科学校,广东湛江524037 [2]岭南师范学院基础教育学院,广东湛江524037 [3]中国热带农业科学院南亚热带作物研究所·农业农村部热带果树生物学重点实验室,广东湛江524091 [4]热带作物生物育种全国重点实验室,海南三亚572024

出　　处：《果树学报》2024年第4期598-610,共13页Journal of Fruit Science

基　　金：国家重点研发计划项目(2023YFD2300804);广东省现代农业技术体系创新团队建设专项(2023KJ109);广东省普通高校特色创新项目(2023KTSCX377)。

摘　　要：【目的】鉴定出菠萝NINV家族全基因组成员,初步阐明NINV基因与蔗糖代谢的关系。【方法】采用生物信息学方法鉴定并分析菠萝NINV家族全基因组成员,通过实时荧光定量PCR分析其表达特性,利用HPLC对蔗糖含量进行测定。【结果】在菠萝中共鉴定出6个NINV基因,分布于5条不同染色体上,其二级结构主要由α-螺旋和无规则卷曲组成,该基因家族启动子区域光反应元件数量最多。AcNINV外显子数量介于4~6个之间,其中AcNINV3、6外显子的数量为6个,亚细胞定位预测发现这2个基因都分布于叶绿体,属于α亚族;AcNINV1、2、4、5外显子的数量为4个,亚细胞定位预测发现这4个基因都分布于质膜,属于β亚族。在果柄、果皮、果心中表达量最高的是AcNINV2基因,在果肉中表达量最高的是AcNINV6基因。蔗糖含量随着菠萝果实成熟呈先升高后降低的变化趋势,而AcNINV4基因在菠萝果实成熟过程中呈下调表达。【结论】AcNINV4可能是催化果实蔗糖降解的水解酶基因。【Objective】Sucrose plays a crucial role in plant growth and development.The alkaline/neu-tral invertase(NINV)proteins irreversibly cleave sucrose into fructose and glucose.To date,the ge-nome-wide identification,characterization,and expression profile analysis of the NINV gene families have been reported in many species but not in pineapple.【Methods】Genome sequence and annotation data were acquired from the pineapple genome database through the blastp against protein database and the tblastn against genome databases using the query sequences of Arabidopsis thaliana NINV proteins to identify the potential members of the NINV gene families in pineapple.The physicochemical charac-teristics,including molecular weight(MW),theoretical isoelectric points(pI),and instability index were obtained using the ExPASy tool.The CDS sequence and protein length were acquired from the pineapple genome database.The synteny analysis between different species and exon/intron structures were visualized using the TBtools.The conserved motifs of the AcNINV gene families were analyzed us-ing the online MEME tool.The phylogenetic tree was constructed using the MEGA 5.0 through the neighbor-joining approach.The total RNA was isolated in accordance with the method described by Wu et al.RT-qPCR was performed using the LightCycler480ⅡSystem(Roche,Switzerland)and the Dy-NAmo Flash SYBR Green qPCR kit(Thermo,USA).RT-qPCR was performed in a total volume of 10μL,which contained 1μL diluted cDNAs and 5µL SYBR Green PCR Master Mix,and the primer final concentration was 250 nmol·L-1.The sucrose content was determined by HPLC.【Results】In this study,six NINV genes(AcNINV1-6)were identified.The six AcNINV genes were distributed on five dif-ferent chromosomes(LG1,LG3,LG14,LG17,LG21).There were two genes on LG21:AcNINV5 and AcNINV6,while another AcNINV gene on the other chromosomes.AcNINV proteins had a length of 556-673 aa and MW between 63.03 and 74.80 ku.Only one collinear NINV gene pair between pineap-ple and A.thaliana,and eight col

关 键 词：菠萝(Ananas comosus) 蔗糖含量 碱性/中性转化酶 基因家族 基因表达 

分 类 号：S668.3[农业科学—果树学]