陆地棉GhUGT76C1基因克隆、生物信息学分析及功能初探  

Cloning,bioinformatics analysis and functional exploration of GhUGT76C1 gene in Gossypium hirsutum L.

作　　者：唐成芬 林文妮 Josee ornella musaniwabo 刘爱玉[1] 屠小菊 TANG Cheng-fen;LIN Wen-ni;Josee ornella musaniwabo;LIU Ai-yu;TU Xiao-ju(College of Agronomy of Hunan Agricultural University,Changsha 410128,China;College of Biological Science and Technology of Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]湖南农业大学农学院,长沙410128 [2]湖南农业大学生物科学技术学院,长沙410128

出　　处：《西南农业学报》2022年第9期1986-1991,共6页Southwest China Journal of Agricultural Sciences

基　　金：湖南省重点研发计划项目(2020NK2023);湖南省自然科学基金(2018JJ3233)。

摘　　要：【目的】研究GhUGT76C1基因调控陆地棉株高的生物学功能。【方法】以陆地棉矮化突变体LA-1为材料,克隆GhUGT76C1基因,并对克隆片段进行生物信息学及进化分析。构建过量表达载体,利用农杆菌介导法转化拟南芥获得过量表达转基因株系。通过不同光质光强处理条件下转基因株系表型鉴定分析GhUGT76C1基因在调控LA-1矮化中发挥的功能。【结果】成功克隆了陆地棉GhUGT76C1基因。GhUGT76C1基因全长1447 bp,cds序列长1365 bp,编码454个氨基酸,理论分子量为51.31 kD,等电点为5.40,编码不稳定疏水性蛋白。GhUGT76C1蛋白的二级结构含有10个α螺旋,17个β片层区,22个转角,22个无规则卷区。系统进化树分析发现,GhUGT76C1蛋白与拟南芥同源蛋白的亲缘关系最近。成功构建陆地棉GhUGT76C1基因过量表达载体,并转化拟南芥获得过量表达转基因株系。表型分析发现,在黑暗、低光强蓝光及远红光条件下过量表达株系均出现显著的下胚轴增长的表型。【结论】首次成功克隆了陆地棉GhUGT76C1基因,并初步解析了其调控株高的功能,为深入了解陆地棉高度的调控机理提供研究材料。【Objective】The present paper aimed to study the function of GhUGT76 C1 gene in regulating the hight of upland cotton.【Method】GhUGT76 C1 gene was cloned for bioinformatics and evolutionary analysis using upland cotton dwarf mutant LA-1 as material.Overexpression vector of GhUGT76 C1 were constructed and the overexpression transformed Arabidopsis plants were created using the transformation method mediated by Agrobacterium infection.The function of GhUGT76 C1 in regulating the dwarfnism of LA-1 were analyzed through phenotypic identification of transgenic lines under different light quality and intensity treatment.【Result】GhUGT76 C1 gene was cloned successfully.Bioinformatics analysis showed that the gene was 1447 bp in length and the CDS region was 1365 bp which encoded 454 amino acids,the theoretical relative molecular mass of the gene was 51.31 kD,the theoretical soelectric point was 5.40 and the protein was a unstable protein.In the secondary structure,there were 10 alpha helix regions,17 beta lamella regions,22 beta turn regions and 22 random coil regions in total.Phylogenetic tree analysis showed that this gene was most closely related to Arabidopsis AtUGT76 C1 gene.We successfully constructed GhUGT76 C1 gene overexpressed vector and got Arabidopsis overexpressed mutant through transformation.We analysed the phenotypic of GhUGT76 C1 gene overexpressed mutants under different light quality and intensity,the results showed that the mutants had significantly increased hypocotyl growth phenotype under dark,low light intensity blue and far red light conditions.【Conclusion】This study laid a foundation for the analysis of the function of GhUGT76 C1 gene in regulating plant height and the molecular mechanism of LA-1 dwarfism,and provided research materials for the in-depth understanding of the regulation mechanism of height in Gossypium hirsutum.

关 键 词：陆地棉 GhUGT76C1 基因克隆 生物信息学 过量表达转基因株系 

分 类 号：S562[农业科学—作物学]