Cloning and expression analysis of maize fucosyltransferase gene SPINDLY

SPINDLY(SPY)is a novel nucleocytoplasmic protein O-fucosyltransferase that regulates target protein activity or stability via O-fucosylation. Previous studies have indicated that the SPY protein regulates plant growth and development by modulating various intracellular processes in Arabidopsis thaliana. Its mediated O-fucosylation plays an important role in maintaining cell homeostasis and regulating plant growth and development, however protein O-fucosylation regulated by SPY in other plants largely remain unknown. Maize(Zea mays )is one of the most important cereals crops for supplying foods, fibers, and fuels to humans. In order to explore the function of maize fucosyltransferase gene(ZmSPY), this study first analyzed the conserved domain, amino acid sequence and physicochemical properties of ZmSPY protein by bioinformatics means, and cloned the gene from maize root tissue to construct the GFP fusion protein expression vector. The subcellular localization of ZmSPY was analyzed, and its response to different hormone treatments(GA, IAA, 6BA, ABA)was determined by exogenous hormone application. And the results were as follows:(1)ZmSPY proteins belong to the TPR and SPY superfamilies, and structural analysis demonstrated that ZmSPY had TPR(Tetratricopeptide repeat)and catalytic domains.(2)Phylogenetic analysis shows that SPYs are highly conserved, and ZmSPY exhibits strong homology to SPY in Sorghum bicolor.(3)Sequence analysis shows that the CDS region of ZmSPY is 2736 bp. Physicochemical analysis indicates that ZmSPY, which contains 911 amino acids and 33 glycosylation sites, is hydrophilic and non-secretory. Its secondary and tertiary structure is largely composed of alpha helix and random coil.(4)The subcellular localization of ZmSPY is predominantly observed in the nucleus.(5)The expression of ZmSPY is induced by phytohormones including GA, IAA, 6BA and ABA, and exhibits various expression patterns. This study provides foundational information on SPY in maize, which could contribute to further investigation of SPY and its effect on O-fucosylation in cereal plants.