| Abstract:Based on the latest grape genome database,members of the nucleobase-ascorbate transporters (NAT) family gene were identified on a genome-wide scale and subjected to bioinformatics and expression pattern analysis,laying a foundation for further functional investigation.The conserved sequence of the Xan_ur_permease domain (PF00860) was used as a seed sequence to screen NAT genes.Prot Prama,Clustalx,MEGA 7.0,PlantCare,GSDS 2.0,and MEME were used to analyze physicochemical properties,multiple sequence alignment,evolutionary characteristics,cis-acting elements,gene structure,and protein structure.The expression patterns of these genes in various tissues and in response to stress signals were analyzed using public database gene expression data and qRT-PCR.The results showed that there were 13 NAT family members in the grape genome,denoted as VvNAT1-VvNAT13,with the number of amino acids,molecular mass,and isoelectric point ranging from 304-755,33.01-81.49 ku,and 8.15-9.6,respectively.The phylogenetic analysis divided the NAT proteins from grape,apple,Arabidopsis thaliana,rice,and pineapple into four subgroups.Structural analysis showed that the number of VvNAT gene exons ranged from 1 to 14,and the corresponding protein sequences contained two conserved motifs,\[(Q/E/P)NXGXXXXT(R/K/G)\] and (QH).Moreover,the VvNAT gene promoter region contained elements responsive to various hormones and stress signals.The collinearity analysis showed that there were three NAT collinear gene pairs within the grape species,and 12 NAT collinear gene pairs between grape and A.thaliana.Gene expression analysis showed that VvNAT1, VvNAT3, VvNAT8, VvNAT11,andVvNAT12 had high expression levels in different tissues and developmental stages of grape.After 200 mmol/L NaCl salt stress and 4 ℃ low temperature treatment,multiple members were differentially expressed,among whichVvNAT8 was significantly up-regulated in both stems and roots.In summary,multiple NAT genes in grape are involved in tissue development and response to salt and low-temperature stress,while VvNAT8 may be involved in tissue development and plant response to low temperature and salt stress,and can be used as a candidate gene for subsequent functional analysis. |
