MoTlg2, a t-SNARE component is important for formation of the Spitzenkörper and polar deposition of chitin in Magnaporthe oryzae

Abstract

T-SNAREs are a family of conserved proteins involved in intracellular transport of membrane-coated cargo among subcellular compartments. In this study, we identified a putative t-SNARE gene, MoTLG2, in Magnaporthe oryzae via insertion mutagenesis. Deletion of MoTLG2 resulted in slower vegetative growth and less conidiation relative to the wild-type strain, but the ΔMotlg2 null mutant was as virulent as the wild-type strain. MoTlg2 has 30% overall amino acid identity with Saccharomyces cerevisiae Tlg2, and rescued the defect of monensin de-sensitivity in the yeast strain where TLG2 had been deleted. More importantly, apical regions of the hyphae of the ΔMotlg2 null mutant were only weakly stained by FM4-64, which was reported as an excellent vesicle tracer, suggesting that the Spitzenkörper was not well formed in the ΔMotlg2 null mutant. In addition, more uneven lateral deposition of chitin was observed in the cell wall of vegetative hyphae of the ΔMotlg2 null mutant. Taken together, this study shows that the t-SNARE Tlg2 is important for both vegetative hyphal growth and conidiation, but dispensable for plant infection in filamentous fungi, and suggests that Tlg2 is important for formation of the Spitzenkörper and polar distribution of chitin.