Search

Gibberella zeae causes Fusarium head blight of cereal crops, and sexual spores of the fungus play an important role as primary inocula. We isolated a restriction enzyme-mediated integration (REMI) transformant, ZH431, of G. zeae with defects in perithecia formation and virulence. Integration of the REMI vector resulted in disruption of GzCHS7 gene, which encodes a putative class VII chitin synthase with high similarity to Fusarium oxysporum ChsVb. A second chitin synthase, GzCHS5, is adjacently located in a head-to-head configuration with GzCHS7, and its deduced protein sequence showed similarity with a class V chitin synthase in F. oxysporum. Neither Δ GzChs5 nor Δ GzChs7 mutants produced perithecia or caused disease on barley heads. Microscopic observation revealed that both mutants formed balloon-shaped hyphae and intrahyphal hyphae and that cell wall rigidity of the mutants was weaker than that of the wild-type strain. Transcription profiles of GzCHS5 and GzCHS7 were not altered in Δ GzChs7 and Δ GzChs5, respectively, suggesting that transcription regulations of the genes are independent of each other. Our results demonstrate that GzCHS5 and GzCHS7 are indispensable for perithecia formation and pathogenicity as well as normal septa formation and hyphal growth in G. zeae. [ABSTRACT FROM AUTHOR]

Gibberella zeae is a self-fertile ascomycetous fungus that causes important diseases of cereal crops. A comprehensive understanding of sexual reproduction in G. zeae is needed for disease control. To identify fungal proteins involved in this process, we compared the protein profiles of a wild-type strain and its self-sterile strain deleted for MAT1-2, a master regulator of sexual reproduction in G. zeae. Using 2-DE and either MALDI-TOF or ESI-Q-TOF MS, we identified 13 protein spots that showed statistically significant differences in expression levels between the two strains; 11 were reduced and two were increased in abundance in the Δ MAT1-2 strain. Six of the 13 proteins were similar to those related to cell wall structure and the others were orthologs of proteins involved in metabolism or environmental stress responses. We confirmed that all the genes of the proteins examined were down-regulated during the sexual development stage in the Δ MAT1-2, Δ MAT1-1, and other strains deleted for a MAP kinase or a G-protein gene. These data suggest that differences in the protein expression levels are mostly affected by down-regulation of the corresponding genes in the Δ MAT1-2 strain. To date, this is the first proteomics approach successfully identifying proteins differentially regulated by MAT1-2 in G. zeae. [ABSTRACT FROM AUTHOR]

Gibberella zeae (anamorph, Fusarium graminearum) is an important pathogen of cereal crops found in many regions of the world. In this study, we have characterized two auxotrophic strains, designated S4B1279 and S4B3008, which were discovered from a collection of insertional mutants of G. zeae generated by restriction enzyme-mediated integration (REMI). Both mutant strains exhibited pleiotropic phenotypic changes that include reduction of mycelial growth and virulence and abolished sexual reproduction. Molecular analysis of the REMI mutants has shown that the auxotrophy of S4B1279 is due to a mutation of the ARG2 gene encoding an acetylglutamate synthase, and the auxotrophy of S4B3008 is due to a mutation of the ADE5 gene encoding a phosphoribosylamine-glycine ligase. Subsequent gene disruption and complementation studies have confirmed the functions for ARG2 and ADE5, respectively, in G. zeae. Our study has demonstrated the feasibility of using the REMI technique in studying G. zeae virulence mechanisms, in addition to providing two new selectable markers allowing genetic transformation of the fungus. [ABSTRACT FROM AUTHOR]