滇水金凤花距发育相关基因TIP的克隆及表达分析  

Cloning and expression analysis of TIP genes related to petal spur development in Impatiens uliginosa

作　　者：魏春梅 李洋[1] 孟丹晨 李泽凤 李新艺 黄美娟[1] 黄海泉[1] WEI Chunmei;LI Yang;MENG Danchen;LI Zefeng;LIXinyi;HUANG Meijuan;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工程研究中心,云南省功能性花卉资源及产业化技术工程研究中心,园林园艺花卉研发中心,昆明650224

出　　处：《植物生理学报》2022年第9期1757-1765,共9页Plant Physiology Journal

基　　金：云南省重大科技专项(202102AE090052);国家自然科学基金(32060364、32060366、31860230和31560228);云南省高校园林植物与观赏园艺科技创新团队(51700204);云南省中青年学术和技术带头人培养项目(2018HB024)。

摘　　要：本文通过对滇水金凤(Impatiens uliginosa)花距发育相关基因TIP的克隆与表达分析,探讨其对滇水金凤花距发育的分子机理。本研究运用RT-PCR技术成功克隆了滇水金凤花距发育相关基因TIP的3个片段,其cDNA全长为777、762和750 bp,分别编码258、253和249 aa,命名为IuTIP1-1、IuTIP1-2和IuTIP2-1,其蛋白均具有MIP(major intrinsic protein)超家族典型的2个NPA(Asn-Pro-A1a)保守结构域。同源性分析表明,滇水金凤TIP基因的氨基酸序列与雷蒙德氏棉(XP_012440576.1)、橡胶树(XP_021686336.1)同源性达77.95%左右;系统进化分析表明,IuTIP1-1与IuTIP1-2聚为一大支,IuTIP2-1单独成一支,三者为旁系同源关系。q RTPCR分析表明,3个基因在滇水金凤花距发育的3个时期及2个不同部位均有表达,其中IuTIP1-1在花苞期檐部的表达量最高,其次是IuTIP2-1在盛花期檐部表达量最高,随着花距的发育,两者在花距距部的表达逐渐下调;而IuTIP1-2在花距檐部的表达随花距发育逐渐下调,但在花距距部的表达逐渐上升,盛花期达到最高,可能与IuTIP1-2对花距距部发育起主要作用有关。根据以上结果,推测滇水金凤的3个TIP基因为旁系同源关系,IuTIP1-1和IuTIP2-1对花距檐部发育起主要作用,而IuTIP1-2对花距距部发育起主要作用。On the basis of cloning and expression analysis of TIP genes related to spur development of Impatiens uliginosa,the molecular mechanism of its spur development was discussed.In this study,TIP genes related to spur development of I.uliginosa were successfully cloned by using RT-PCR method,their full-length cDNA sequences were 777,762 and 750 bp.They were respectively encoded 258,253 and 249aa,and named IuTIP1-1,IuTIP1-2 and IuTIP2-1.All of them have two typical NPA(Asn-Pro-A1a)conserved domains of MIP(major intrinsic protein)superfamily.According to the result of homology analysis,it showed that the amino acid sequences of TIP genes reached 77.95%homology with Gossypium raimondii(XP_012440576.1)and Hevea brasiliensis(XP_021686336.1).Based on phylogenetic analysis,it found that IuTIP1-1and IuTIP1-2 clustered into one branch,while IuTIP2-1 formed a separate branch,so all of them were paralogous genes.In accordance with qRT-PCR analysis,it showed that three TIP genes were expressed in both three stages and two different parts of spur development of I.uliginosa.The expression level of IuTIP1-1 was the highest in the blade of bud stage,followed by the highest expression level of IuTIP2-1in the blade of blooming stage.With the development of spur,their expression levels decreased gradually in petal spur.While the expression level of IuTIP1-2 was decreased gradually in blade with development of spur,increased gradually in spur cup and reached the highest level at the full blooming stage,which conferred that IuTIP1-2 played major role in spur cup development.Based on above results,it was speculated that three TIP genes of I.uliginosa were belonged to paralogous genes,among which both IuTIP1-1 and IuTIP2-1 played major roles in the blade development,while IuTIP1-2 in the spur cup development.

关 键 词：滇水金凤 花距发育 TIP基因 基因克隆 表达分析 

分 类 号：S681.1[农业科学—观赏园艺]