Marker recycling in Lentinula edodes via 5-fluoroorotic acid counter-selection.

Shiitake (Lentinula edodes) contains various beneficial compounds and possesses several notable properties. However, there are few reports on its molecular breeding due to delay in development of its gene-modifying technology. Therefore, here, strain UV30 (pyrG -) was bred from the UV-irradiated protoplasts of strain M2.

Development of a gene-targeting system using CRISPR/Cas9 and utilization of pyrG as a novel selectable marker in Lentinula edodes.

First, we attempted to recombine the Shiitake (Lentinula edodes) pyrG (ura3) gene homologously by introducing a donor vector containing a carboxin resistance gene (lecbxR) flanked by homologous sequences of pyrG into protoplasts of the fungus. However, all the carboxin-resistant transformants only contained ectopic insertions of the exogenous gene and no homologous insertions. Agaricomycetes are generally known for their low efficiency of homologous recombination, and a similar result was shown for L.

An exocyst component, Sec5, is essential for ascospore formation in .

In this study, we identified Sec5 in , a homologue of Sec5 in and a possible exocyst component of the fungus. To examine how Sec5 affects the life cycle of , we generated null mutant strains of the gene (). The strains showed a strong reduction in hyphal growth and a slight reduction in pathogenicity.

Cla4 PAK-like kinase is required for pathogenesis, asexual/sexual development and polarized growth in Bipolaris maydis.

PAK (p21-activated protein kinases)-like kinases are master regulators of development and morphogenesis, which were conserved among eukaryotes, including fungi. In budding yeast, two types of PAK-like kinases, Ste20 and Cla4 have distinct but shared roles in the regulation of pseudohyphal development, budding and mating. In this study, to examine the broad functions of PAK-like kinases in growth, pathogenicity and asexual/sexual reproduction in filamentous fungi, we identified and characterized two PAK-like kinases, Ste20 and Cla4 in Bipolaris maydis.

Characterization of the autophagy-related gene BmATG8 in Bipolaris maydis.

Autophagy is involved in cellular development and the maintenance of viability under nutrient deprivation in a wide range of eukaryotes. A filamentous ascomycete Bipolaris maydis, responsible for southern corn leaf blight, is also studied as a model fungus for sexual reproduction in filamentous ascomycetes that form filiform ascospores. In order to clarify the roles of autophagy in various stages of the life cycle of B.