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杂交构树UGDH基因家族生物信息学分析及表达特性分析

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[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。
Bioinformatics analysis and expression characteristics analysis of UGDH gene family in hybrid Broussonetia kazinoki×Broussonetia papyifera
[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 1 895,927,1 658 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 hybrid B.papyrifera.It laid a theoretical foundation for the study of the regulation of hemicellulose and pectin content and the interaction between them.

Broussonetia papyriferaUGDH genebioinformatics analysisexpression analysis

高娜、吕昕霏、刘佳欣、白玉娥、吉仁花

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内蒙古农业大学 林学院,内蒙古 呼和浩特 010019

杂交构树 UGDH基因 生物信息学分析 表达分析

2024

中南林业科技大学学报
中南林业科技大学

中南林业科技大学学报

CSTPCD北大核心
影响因子:1.442
ISSN:1673-923X
年,卷(期):2024.44(11)