Use of response surface methodology to examine chitinase regulation in the basidiomycete Moniliophthora perniciosa.
We report here the first analysis of chitinase regulation in Moniliophthora perniciosa, the causal agent of the witches' broom disease of cacao. A multivariate statistical approach was employed to evaluate the effect of several variables, including carbon and nitrogen sources and cultivation time, on M. perniciosa non-secreted (detected in mycelium, i.e. in symplasm and cell wall) and secreted (detected in the culture medium) chitinase activities. Non-secreted chitinase activity was enhanced by peptone and chitin and repressed by glucose. Chitinase secretion was increased by yeast extract alone or in combination with other nitrogen sources, and by N-acetylglucosamine, and repressed in presence of chitin. The best cultivation times for non-secreted and secreted chitinase activities were 30 and 20 d, respectively. However, chitinase activity was always higher in the mycelium than in the culture medium, suggesting a relatively poor chitinase secretion activity. Conversely, higher mycelial growth was observed when the activity of the non-secreted chitinase was at its lowest, i.e. when the fungus was grown on glucose and yeast extract as sources of carbon and nitrogen, respectively. Conversely, the induction of non-secreted chitinase activity by chitin decreased the mycelium growth. These results suggest that the culture medium, by the induction or repression of chitinases, affected the hyphal growth. Thus, as an essential component of M. perniciosa growth, chitinases may be a potential target for strategies to control disease.
Lopes
Maíza Alves
MA
Laboratório de Genômica e Expressão Gênica, DCB, Universidade Estadual de Santa Cruz, Rodovia Ilhéus-Itabuna, Km 16, 45650-000 Ilhéus-BA, Brazil.
Gomes
Dayane Santos
DS
Koblitz
Maria Gabriela Bello
MG
Pirovani
Carlos Priminho
CP
Cascardo
Júlio Cézar de Mattos
JC
Góes-Neto
Aristóteles
A
Micheli
Fabienne
F
D000363:Agaricales / Q000201:enzymology* / Q000502:physiology; D018533:Biomass; D002244:Carbon / Q000378:metabolism*; D002688:Chitinases / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry; D003593:Cytoplasm / Q000201:enzymology; D005110:Extracellular Space / Q000201:enzymology; D005656:Fungal Proteins / Q000378:metabolism; D015999:Multivariate Analysis; D025282:Mycelium / Q000201:enzymology / Q000254:growth & development; D009584:Nitrogen / Q000378:metabolism*; D021381:Protein Transport; D013997:Time Factors
D003470:Culture Media; D005656:Fungal Proteins; D002244:Carbon; D002688:Chitinases; D009584:Nitrogen