滇水金凤花距发育相关基因GIF的克隆及表达分析  

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

作　　者：蔡斌 魏春梅 刘婕 李新艺 李洋[1] 黄海泉[1] 黄美娟[1] CAI Bin;WEI Chunmei;LIU Jie;LI Xinyi;LI Yang;HUANG Haiquan;HUANG Meijuan(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,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6期1117-1124,共8页Plant Physiology Journal

基　　金：国家自然科学基金(32060364、32060366、31860230和31560228);云南省重大科技专项计划(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;云南省中青年学术和技术带头人培养项目(2018HB024)。

摘　　要：GIF(GRF-interacting factor)是植物体内特有的转录共激活因子之一,对植物生长发育调控具有重要作用。本研究以滇水金凤(Impatiens uliginosa)花距为材料,采用RT-PCR等技术克隆得到GIF基因的2个拷贝,命名为IuGIF1和IuGIF2,其cDNA序列全长分别为591和540 bp,分别编码196和179 aa。对其编码蛋白进行生物信息学分析,发现2个GIF蛋白均属于亲水性不稳定蛋白,且均具有SSXT超家族典型的保守结构域。多序列比对发现,IuGIF1和IuGIF2所编码的氨基酸序列与烟草、松蒿以及茶树等物种的同源性达54%左右。系统进化分析表明,IuGIF1基因和IuGIF2基因单独分为两支,其中IuGIF1与林烟草亲缘关系最近,而IuGIF2与松蒿亲缘关系最近,推测两者为旁系同源关系。qRT-PCR分析表明,随着花距发育,IuGIF1和IuGIF2基因在花距距部的表达量均呈下降趋势,且IuGIF1在两个部位的表达均高于IuGIF2,推测IuGIF1对促进花距檐部和距部细胞生长作用强于IuGIF2基因。此外,IuGIF1在始花期尖部的表达量最高,随着花距的发育逐渐降低,可能与IuGIF1在花距尖部的发育中发挥重要作用有关;而IuGIF2在始花期花距弯部中表达量最高,随着花距的发育逐渐降低,可能与IuGIF2在花距弯部的发育中发挥重要作用有关。综上表明,IuGIF1基因对花距发育的作用强于IuGIF2基因;IuGIF1主要在花距尖部发育中发挥着重要作用,而IuGIF2基因主要对花距弯部发育起重要作用。GIF is one of the unique transcription co-activators in the plants,which plays an important role in plant growth and development.In this study,the spur of Impatiens uliginosa was used as the materials,two GIF genes related to spur development of I.uliginosa were cloned by RT-PCR method and other techniques and named luGIF1and luGIF2 respectively.Their cDNA full-length were 591 and 540 bp,encoding 196 and 179 aa respectively.On the basis of their bioinformatics analysis,it showed that both luGIF1 and luGIF2 were hydrophilic and unstable proteins,and had typical conserved domains of ssxT superfamily.According to the phylogenetic analysis,it showed that the homology of their amino acid sequences reached 54%with those of Nicotiana sylvestris,Phtheirospermum japonicum and Camellia sinensis.Based on the phylogenetic analysis,it showed that luGIF1 and luGIF2 genes were separated into two branches,among which luGIF1 was more closely with that of N.sylvestris,while luGIF2 was more closely with that of P.japonicum.It was inferred that the two genes were paralogous genes.According the analysis of qRTPCR results,it showed that the expression levels of luGIF1 and luGIF2 genes in the cup decreased with the development of spur.Meanwhile,the expression levels of luGIF1 in both parts were higher than luGIF2.It was speculated that luGIF1 had a stronger effect on promoting the growth of blade and cup cells than lu-GIF2 gene.In addition,the expression level of luGIF1 was the highest in the spur tip of beginning flowering stage and gradually decreased with the development of spur,which inferred that luGIF1 played an important role in spur tip development.The expression level of luGIF2 was the highest in the spur camber of beginning flowering stage,and gradually decreased with the spur development,which inferred that luGIF2 played an important role in the spur camber development.In conclusion,luGIF1 played a stronger role in promoting the growth of spur cells than luGIF2.luGIF1 mainly played a key role in the development of spur tip,w

关 键 词：滇水金凤 花距发育 GIF基因 基因克隆 表达分析 

分 类 号：S681.1[农业科学—观赏园艺]