杂交构树UGDH基因家族生物信息学分析及表达特性分析  

Bioinformatics analysis and expression characteristics analysis of UGDH gene family in hybrid Broussonetia kazinoki×Broussonetia papyifera

作　　者：高娜 吕昕霏 刘佳欣 白玉娥[1] 吉仁花[1] GAO Na;LYU Xinfei;LIU Jiaxin;BAI Yu’e;JI Renhua(College of Forestry,Inner Mongolia Agricultural University,Hohhot 010019,Inner Mongolia,China)

机构地区：[1]内蒙古农业大学林学院,内蒙古呼和浩特010019

出　　处：《中南林业科技大学学报》2024年第11期120-132,共13页Journal of Central South University of Forestry & Technology

基　　金：内蒙古自治区地区科学基金项目(32360402);内蒙古自治区教育厅高等学校科学研究项目(NJZY22501);内蒙古农业大学人才引进科研启动项目(NDYB2020-14)。

摘　　要：【目的】对杂交构树UGDH基因家族成员进行克隆、生物信息学及表达特性分析。【方法】利用杂交构树(Broussonetia kazinoki×Broussonetia papyifera)的转录组数据获取UGDH基因序列,分别命名为BpUGDH1、BpUGDH2和BpUGDH4,设计引物克隆杂交构树的UGDH基因家族成员(其中BpUGDH3在前期研究中已克隆),采用ExPASy、TMHMMServerv2.0、Swiss-ModeL、SOPMA、MEGA11.0等软件进行了生物信息学分析;采用qRT-PCR对不同器官组织进行基因表达差异分析。【结果】通过克隆获得了杂交构树UGDH1、UGDH2、UGDH3和UGDH4基因全长序列分别为1 895、927、1 658和481 bp,编码480、294、480和131个氨基酸;其中BpUGDH2为疏水性蛋白,其余为亲水性蛋白。BpUGDH基因均不含信号肽;均无跨膜结构域,蛋白均存在PLNO2353 superfamily一个超家族保守结构域。多序列比对得出BpUGDH1、BpUGDH2、BpUGDH3、BpUGDH4蛋白序列之间相似性为68.39%;系统进化树BpUGDH1同大豆UGDH2聚为一类;BpUGDH2和大豆UGDH4亲缘关系较近,同时BpUGDH2和BpUGDH4聚为一类,BpUGDH4和水稻UGDH1、水稻UGDH2以及玉米UGDH2亲缘关系近;BpUGDH3和拟南芥UGDH3有较近的亲缘关系。蛋白二级结构分析表明BpUGDH1、BpUGDH2、BpUGD4均以α螺旋和无规则卷曲为主,BpUGDH3则以无规则卷曲为主,占39.79%。qRT-PCR结果显示BpUGDH1在叶的表达量最高;BpUGDH2在根的表达量最高;BpUGDH3在茎中的表达量最高,BpUGDH4在茎的表达量最高,该基因家族均在茎中表达。【结论】本研究克隆得到了杂交构树UGDH1、UGDH2、UGDH4基因并与吉仁花克隆所得的杂交构树UGDH3基因共同进行生物信息学分析,为研究探讨各基因之间在调控半纤维素和果胶含量及二者相互作用奠定了一定理论基础。【Objective】Cloning,bioinformatics and expression analysis of UGDH gene family members in Broussonetia kazinoki×Broussonetia papyifera.【Method】The UGDH gene sequences were obtained by using the transcriptome data of B.kazinoki×B.papyifera.,named BpUGDH1,BpUGDH2 and BpUGDH4,respectively.The primers were designed to clone the UGDH gene family members of the hybrid B.papyrifera(BpUGDH3 has been cloned in the previous study).Bioinformatics analysis was performed using ExPASy,MHMMServerv2.0,Swiss-ModelL,SOPMA,MEGA11.0 and other software.The gene expression differences of different organs and tissues were analyzed by qRT-PCR.【Result】The full-length sequences of BpUGDH1,BpUGDH2,BpUGDH3 and BpUGDH4 were 1895,927,1658 and 481 bp,respectively,encoding 480,249,480 and 131 amino acids.BpUGDH2 was a hydrophobic protein, and the rest was a hydrophilic protein. BpUGDH gene did not contain signal peptide. There is no transmembrane domain, and the proteins all had a conserved domain of the PLNO2353 superfamily. Multiple sequence alignment showed that the similarity between BpUGDH1, BpUGDH2, BpUGDH3 and BpUGDH4 protein sequences was 68.39 %. The phylogenetic tree BpUGDH1 was clustered with soybean UGDH2. BpUGDH2 was closely related to soybean UGDH4, and BpUGDH2 and BpUGDH4 were clustered together. BpUGDH4 was closely related to rice UGDH1, rice UGDH2 and maize UGDH2. BpUGDH3 had a close relationship with Arabidopsis. Protein secondary structure analysis showed that BpUGDH1, BpUGDH2 and BpUGD4 were dominated by alpha helix and random coil, while BpUGDH3 was dominated by random coil, accounting for 39.79%. The results of qRT-PCR showed that the expression of BpUGDH1 was the highest in leaves. The expression of BpUGDH2 was the highest in root. The expression level of BpUGDH3 was the highest in the stem, and the expression level of BpUGDH4 was the highest in the stem.【Conclusion】In this study, the UGDH1, UGDH2 and UGDH4 genes of hybrid B. papyrifera were cloned and analyzed by bioinformatics with the UGDH3 gene of hyb

关 键 词：杂交构树 UGDH基因 生物信息学分析 表达分析 

分 类 号：S794.9[农业科学—林木遗传育种]