Bioinformatics analysis,gene cloning and function study of poplar UGT198 gene
[Objective]The present paper aimed to study whether UGT198 gene was involved in the regulation of insect resistance of(Nicoti-ana tabacum)tobacco,and lay a foundation for the future study of UGT198 gene in the regulation of poplar insect resistance.[Method]By analyzing the public transcriptome data,it was found that PtrUGT198 was highly expressed in the transcriptome of poplar responding to insect infestation,and the bioinformatics and evolutionary analysis were conducted.Using 107 poplar as material,UGT198 gene was cloned,over-expression vector was constructed,and overexpression transgenic strain of tobacco was obtained by Agrobacterium mediated transformation.The feeding experiments preliminarily verified whether PtrUGT198 gene participated in the regulation of insect resistance,and analyzed the function of UGT198 gene in the regulation of insect resistance.[Result]The complete ORF region of PtrUGT198 gene was 1440 bp,encoding 479 amino acids,theoretical molecular weight was 53.14 kDa,and isoelectric point was 5.92,encoding unstable hydrophobic protein.The secondary structure of PtrUGT198 protein consisted of α-helix(41.75％),β-fold(14.61％),β-comer(7.10％)and random curl(36.53％).PtrUGT198 did not have a transmembrane region,which was predicted to be a non-secreted protein,and the phosphorylation of this protein was mainly serine.The results of homologous sequence comparison showed that highly conserved plant secondary product glyco-syltransferase(PSPG)motif existed in the C-terminal of PtrUGT198 protein.Phylogenetic tree analysis showed that PtrUGT198 protein was the most closely related to Populus tomentosa,P.alba and P.euphratica in the same genus,followed by a close relationship with tobacco model plants.UGT198 gene of poplar was cloned,pNC-UGT198 gene overexpression vector was constructed,and overexpression transgenic strain of tobacco was obtained.[Conclusion]UGT198 gene was successfully cloned for the first time,and its function of regulating insect re-sistance was initially analyzed,which provided reference value for further understanding of the mechanism of regulating poplar insect resist-ance by UGT198.
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