Identification of a maize (Zea mays) chitinase allele sequence suitable for a role in ear rot fungal resistance

Abstract

Chitinases are thought to play a role in plant resistance to pathogens, but the extent of this role is unknown. The gene for a maize (Zea mays) chitinase, “chitinase 2”, previously reported to be induced by two ear rot pathogens in one maize inbred, was cloned from mRNA isolated from milk stage kernels of several different inbreds reported to be susceptible or resistant to ear rot pathogens. The chitinase gene sequence of some of the susceptible inbreds had frame shifts which would result in a nonfunctional protein. Other susceptible inbreds, and a few resistant inbreds, were missing regions that would contribute to chitin binding. A putative functional clone from a resistant inbred was expressed in yeast, and produced a protein with chitinase activity against different oligomers of N-acetyl glucosamine and modified chitin. The yeast-produced chitinase was also resistant to degradation by proteases from maize ear rot fungi, and enhanced antifungal activity of miconazole towards Fusarium graminearum. When introduced in maize callus transgenically, the callus expressing the chitinase 2 gene had significantly less growth of the ear rot pathogen, F. graminearium, than callus which did not express the gene. This information suggests susceptibility and resistance to ear rot pathogens is influenced not only by expression levels, but also by the sequence of putative resistance genes. Previous reports of increased expression of putative resistance genes need to be tempered with the realization that they may play no role in resistance if the genes code for nonfunctional or reduced function proteins.