Genome-Wide Identification and Analysis of Chitinase GH18 Gene Family in Valsa mali.

Chitinases are enzymes that catalyze the hydrolysis of chitin and play a significant biophysiological role in fungal growth, development, and pathogenesis. Valsa mali is a necrotrophic fungus that is a primary contributor to apple Valsa canker. Our study focused on the identification of chitinase gene families from V. mali and the analysis of their expression profiles during infection and nutritional growth. A phylogenetic analysis and conservation of catalytic domains were used to classify these genes into three classes, and their chromosome distribution was random. The qRT-PCR analysis identified five differentially expressed VmGH18 genes during infection and nutritional growth. GH18 chitinases use glutamate, whereas VmGH18-4 (VM1G_05900) and VmGH18-10 (VM1G_03597) use glutamine as the catalytic motif. To further test whether it can induce cell death in apple, the recombinant protein was produced in E. coli. It showed that the purified VmGH18-4 recombinant protein retained cell-death inducing activity, and it could also induce cell death in apple. But the enzyme activity shows that neither VmGH18-4 nor VmGH18-10 have chitinases enzyme activity. These results suggest that VmGH18-4 can elicit cell death in multiple plant species, while VmGH18-10 cannot.