No abstract

No abstract

No abstract

No abstract

In a previous study, we reported a transient transformation system using repeated screening for hygromycin B (Hyg) resistance in the basidiomycete Ceriporiopsis subvermispora. In the present study, by combining this technique with CRISPR/Cas9, we demonstrated successful marker-free genome editing in Pleurotus ostreatus, which is one of the most economically important cultivated mushrooms as well as a model white-rot fungus. At first, transformant selection mediated by the transient expression of marker genes was demonstrated using a plasmid harboring the Hyg resistance gene (hph) in P. ostreatus. Then, genome editing of fcy1, which confers 5-fluorocytosine (5-FC) resistance to the host cell, was performed by the transient expression of Cas9, gRNA, and hph and strains with 5-FC resistance and Hyg sensitivity were isolated. Additionally, genome editing of fcy1 in these strains was confirmed by Sanger sequencing. To our knowledge, this is the first report of marker-free genome editing through the transient expression of Cas9, gRNA, and hph in agaricomycetes, which opens the door for repeated genome editing in these fungi.

Cochliobolus heterostrophusMitogen-activated protein kinase cascade Signal transduction Southern corn leaf blight a b s t r a c tIn this study, we identified a gene, Ste7, in the genome of Bipolaris maydis encoding a mitogen-activated protein kinase (MAPK) kinase homologous to yeast Ste7. A Dste7 strain was defective in conidiation and conidial germination, and was severely impaired in lesion formation, mainly due to the loss of appressoria, whereas the effect on infectious hyphal development was not significant. The Dste7 strain did not have a maternal ability for sexual reproduction, mainly in protothecium formation. These phenotypes of Dste7 were also observed in Dchk1 and Dste11, null mutant strains of MAPK and MAPK kinase kinase, respectively. These observations strongly suggested that Ste7 is a component of the Chk1 MAPK cascade, and is involved in the regulation of various developmental and morphogenetic processes in the asexual and sexual lifecycles of B. maydis.

Dicyma pulvinata strain 414-3, isolated from the surface of a tomato leaf, is a mycoparasitic fungus of Cladosporium fulvum, which causes leaf mold of tomato. We report here the draft genome sequence of strain 414-3, which will contribute to elucidating the molecular mechanisms involved in the mycoparasitism.

No abstract