Functional analysis of a beta-1,6-glucanase gene from the grass endophytic fungus Epichloë festucae.
beta-1,6-glucanases degrade the polysaccharide beta-1,6-glucan, a cell wall component in some filamentous fungi. A single copy of a beta-1,6-glucanase gene, designated gcnA, was identified in each of the grass endophytic fungi Neotyphodium lolii and Epichloë festucae. Phylogenetic analysis indicates that the GcnA protein is a member of glycosyl hydrolase family 5, and is closely related to fungal beta-1,6-glucanases implicated in mycoparasitism. The E. festucae gcnA gene was expressed in mycelium grown in culture and in both vegetative and reproductive tissues of perennial ryegrass. A gcnA replacement mutant had reduced beta-1,6-glucanase activity when grown in media containing pustulan as the major carbon source. beta-1,6-glucanase activity was restored in the replacement mutant by introducing multiple copies of the gcnA gene. Growth of DeltagcnA and gcnA-overexpressing strains in vegetative grass tissues was indistinguishable from wild type strains.
Bryant
Michelle K
MK
Centre for Functional Genomics, Institute of Molecular Biosciences, Massey University, Private Bag 11222, Palmerston North, New Zealand.
May
Kimberley J
KJ
Bryan
Gregory T
GT
Scott
Barry
B
D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000502:physiology*; D017353:Gene Deletion; D006026:Glycoside Hydrolases / Q000235:genetics / Q000502:physiology*; D006999:Hypocreales / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development*; D008969:Molecular Sequence Data; D010802:Phylogeny; D006109:Poaceae / Q000382:microbiology*; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA
D004271:DNA, Fungal; D005656:Fungal Proteins; D006026:Glycoside Hydrolases; C020105:endo-1,6-beta-glucanase