Molecular characteristics and expression pattern analysis of grapevine VvWRKY53
To explore the biological function of VvWRKY53,the VvWRKY53 gene containing a coding sequence(CDS)of 453 bp was cloned by RT-PCR from the cDNA of'Thompson seedless'.The molecular characteristics of the VvWRKY53 gene and its encoded protein were analyzed using bioinformatics tools.The subcellular localization of candidate protein was determined by transient expression in Nicotiana benthamiana.The expression patterns of the VvWRKY53 gene after spraying with four hormones:salicylic acid(SA),abscisic acid(ABA),methyl jasmonate(MeJA),and 1-aminocyclopropane-1-carboxylic acid(ACC);two elicitors:chitin and flg22,and after inoculation with Lasiodiplodia theobromae were detected using quantitative real-time PCR(qRT-PCR).The transcriptional activation or inhibitory activity of VvWRKY53 was determined by yeast system.The results showed that:The molecular weight of the VvWRKY53 protein is 18.1 ku,which has a theoretical isoelectric point(pl)of 9.67.The VvWRKY53 protein is located in the nucleus of Nicotiana benthamiana.The phylogenetic tree analysis indicates that VvWRKY53 has high similarity to AtWRKY45 and AtWRKY75 proteins.The results of qRT-PCR reveal that the relative expression level of VvWRKY53 changed after spraying SA,MeJA and ACC on'seedless white'grape leaves.The relative expression level of VvWRKY53 gene is gradually increased after ABA treatment.Both chitin and flg22 can rapidly induce the expression of the gene.The relative expression of VvWRKY53 gene continued to be up-regulated after inoculation of'summer black'semi-woody branches with Lasiodiplodia theobromae.It is confirmed that VvWRKY53 has transcriptional activation activity in yeast.The results indicate that the transcription factor VvWRKY53 may play an important role in the process of Lasiodiplodia theobromae infection.
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