滇水金凤花距发育相关基因Aux/IAA的克隆及表达分析  

Cloning and expression analysis of Aux/IAA genes related to spur development in Impatiens uliginosa

作　　者：杨建园 童桢开 闻永慧 汪琼[1] 黄美娟[1] 鄢波[1] 黄海泉[1] YANG Jianyuan;TONG Zhenkai;WEN Yonghui;WANG Qiong;HUANG Meijuan;YAN Bo;HUANG Haiquan(College of Landscape Architecture and Horticulture Sciences,Southwest Research Center for Engineering Technology of Landscape Architecture(State Forestry and Grassland Administration),Yunnan Engineering Research Center for Functional Flower Resources and Industrialization,Research and Development Center of Landscape Plants and Horticulture Flowers,Southwest Forestry University,Kunming 650224,China)

机构地区：[1]西南林业大学园林园艺学院,国家林业和草原局西南风景园林工程技术研究中心,云南省功能性花卉资源及产业化技术工程研究中心,西南林业大学园林园艺花卉研发中心,昆明650224

出　　处：《植物生理学报》2023年第5期899-908,共10页Plant Physiology Journal

基　　金：国家自然科学基金(32060364、32060366、31860230和31560228);国家重点研发计划(2019YFD1001002);云南省重大科技专项(202102AE090052);云南省高校园林植物与观赏园艺科技创新团队(51700204);云南省园林植物遗传改良与高效繁育博士生导师团队、云南省教育厅科学研究基金(2022Y612);云南省中青年学术和技术带头人培养项目(2018HB-024)。

摘　　要：Aux/IAA(Auxin/Indole-3-Acetic Acid)作为诱导植物主要生长素的关键基因,在植物生长发育、细胞分裂及伸长等方面发挥重要调控作用。为研究Aux/IAA基因在滇水金凤(Impatiens uliginosa)花距发育中的功能,本试验从滇水金凤中克隆出Aux/IAA基因的2个拷贝,分别命名IuIAA1和IuIAA2,其cDNA全长序列分别为648和564 bp,分别编码215和187个氨基酸;分析发现, IuIAA1和IuIAA2分别含2个和1个内含子,其长度分别为84和108 bp;IuIAA1和IuIAA2蛋白都是不稳定蛋白。氨基酸序列分析表明, IuIAA1蛋白具有Domain Ⅰ、Domain Ⅱ、Domain Ⅲ和Domain Ⅳ 4个完整的保守结构域,而IuIAA2蛋白则缺乏完整的Domain Ⅳ,但是2个Aux/IAA蛋白中均发现核定位信号(NLS)。系统进化分析表明, IuIAA1和IuIAA2均与拟南芥的IAA1/AXR5、IAA2、IAA3/SHY2和IAA4聚为1支,其亲缘关系较近。qRT-PCR分析发现, 2个Aux/IAA基因在花发育的檐部和距部均呈现先升后降的趋势,且在始花期表达量最高,花苞期表达量最低,推测2个Aux/IAA基因在距部和檐部的始花期促进了细胞的分裂和伸长,而在盛花期Aux/IAA基因表达减弱。在始花期和盛花期, 2个Aux/IAA基因在尖部、弯部、基部和檐部相对表达量呈逐渐上升趋势,且均以檐部相对表达量最高,尖部相对表达量最低,推测2个Aux/IAA基因在檐部的始花期和盛花期均促进了细胞的分裂和伸长;上述结果为进一步研究滇水金凤花距发育提供一定的基础理论。Aux/IAA(Auxin/Indole-3-acetic Acid)a key auxin-inducing gene plays an important role in plant growth and development,cell division and elongation.To studye the function of Aux/IAA gene in spur development of I.uliginosa,two copies of Aux/IAA gene were cloned from I.uliginosa,and named IuIAA1 and IuIAA2 respectively.Their full-length of cDNA sequences were 648 and 564 bp,encoding 215 and 187 aa respectively.The results showed that IuIAA1 and IuIAA2 contained two introns and one intron,with length of 84 and 108 bp respectively.Both IuIAA1 and IuIAA2 proteins were unstable.Amino acid sequence alignment analysis showed that the IuIAA1 protein has four complete conserved structural domains of DomainⅠ,DomainⅡ,DomainⅢand DomainⅣ,while the IuIAA2 protein lacks the complete DomainⅣ,but nuclear localization signals(NLS)were found in both Aux/IAA proteins.sequence(NLS)was found in both Aux/IAA proteins.IuIAA1 and IuIAA2 are clustered with IAA1/AXR5,IAA2,IAA3/SHY2 and IAA4 of Arabidopsis thaliana into one clade,and were closely related to each other.On the basis of their qRT-PCR results,it showed that the expression of two Aux/IAA genes increased first and then decreased in both spur and blade of flower development,the expression were the highest at the initial flowering stage,and the lowest at the bud stage,which suggested that two Aux/IAA genes promoted cell division and elongation in the initial flowering stage of spur and blade.The expression of Aux/IAA genes decreased dramatically during the full flowering stage.Based on the relative expression of the two Aux/IAA genes in the tip,camber,base and blade of spur,it showed a gradual increasing trend appeared during both initial flowering and full flowering stages,their relative expression reached the highest in the blade of spur and the lowest in the tip.It was speculated that both Aux/IAA genes in the blade promoted cell division and elongation during the initial flowering and full flowering stages.The above-mentioned results provided some basic theory for furthe

关 键 词：滇水金凤 花距发育 Aux/IAA基因 基因克隆 表达分析 

分 类 号：S681.1[农业科学—观赏园艺]