These data indicate that the hydrophobic surface properties of conidia are a prerequisite for appropriate surface sensing under nutrient-limiting conditions. In order to test the role of hydrophobins in conidial and hyphal hydrophobicity, and therefore possibly in hydrophobic surface sensing, we performed a systematic search for the presence of hydrophobin genes in the B. cinerea genome, analysed IWR-1 chemical structure their expression, and performed a functional analysis of three hydrophobin genes and a hydrophobin-like gene. Surprisingly, Milciclib cost mutants lacking all these genes were found to be phenotypically

indistinguishable from the wild type in all parameters tested. Our results challenge the concept that hydrophobins are generally required for the formation of hydrophobic surface layers in conidia and hyphae of higher fungi. Results Cloning and sequence analysis of Botrytis cinerea hydrophobin genes In the B. cinerea strain B05.10 genome sequence, three hydrophobin encoding genes were identified. Using Magnaporthe

oryzae class I hydrophobin Mpg1 [4] as a query in a blastp search, a protein (BC1G_15273) with weak homology was detected. Its size, arrangement of the eight conserved cysteines, and overall hydropathicity was similar to M. oryzae Mpg1 and other class I hydrophobins, and it was called Bhp1 (for ‘ B otrytis h ydro p hobin’). Using Liothyronine Sodium M. oryzae class II Oligomycin A solubility dmso hydrophobin Mhp1 [6] in another blastp query, the B. cinerea proteins BC1G_03994 (called Bhp2) and BC1G_01012 (called Bhp3) were found to show significant

homologies (E values < e-10). With blastp and tblastn searches using known hydrophobin proteins, no further hydrophobin genes were identified in the B. cinerea genome. The identification of hydrophobin encoding genes in fungal genomes is sometimes difficult due to their small size, the variable spacing between the cysteine encoding codons, and their low sequence homologies, in particular among class I hydrophobin genes. In order to identify further candidates for B. cinerea hydrophobins, a systematic search was performed in the published genome sequences of B. cinerea strains B05.10 and T4. The following search parameters were used: a) Total size of the protein smaller than 250 amino acids; b) Presence of at least 6 cysteines, four of them in a tandem arrangement separated by two further cysteine residues (full cysteine motive of hydrophobins: C-(Xn)-CC-(Xn)-C-(Xn)-C-(Xn)-CC-(Xn)-C); c) Prediction of a signal peptide. The search resulted in the identification of six further hydrophobin-like B. cinerea proteins, which all had a small size (98-234 aa), and a similar pattern of eight cysteines after manual correction of annotations (Table 1; additional file 1 : Table S1).