Automated image analysis to observe pellet morphology in liquid cultures of filamentous fungi such as the basidiomycete Coprinopsis cinerea

In this study, an image analysis system for fungal pellets was developed using the commercial software system analySIS® and the protocol was evaluated in morphological studies of pellet formation in submerged cultures of the basidiomycete Coprinopsis cinerea. Pellets were analysed on large scale (60 to 130 pellets per image, 225 to 400 pellets per culture). Morphologies of pellets were characterised by the parameters grey value, pellet area, convexity, shape factor, sphericity and pellet diameter. Threshold values were defined for all parameters for object filtering. By application of the parameter filter, aggregated hyphal fragments present in larger amounts particularly in cultures grown at higher temperature (37°C) could be clearly distinguished in image analysis from compact pellets. At a lower growth temperature (25°C), there was little background of loose hyphal material, fungal pellets were regularly shaped and the pellets remained constant in shape and size over a longer cultivation period.

[1]  S. Kajiwara,et al.  Secretional expression of a Bacillus subtilis xylanase gene in the Basidiomycete Coprinus cinereus , 1999 .

[2]  Maria Papagianni,et al.  Fungal morphology and metabolite production in submerged mycelial processes. , 2004, Biotechnology advances.

[3]  M. Aebi,et al.  Restriction enzyme-mediated DNA integration in Coprinus cinereus , 1997, Molecular and General Genetics MGG.

[4]  G. Foster,et al.  Expression of laccase gene lcc1 in Coprinopsis cinerea under control of various basidiomycetous promoters , 2006, Applied Microbiology and Biotechnology.

[5]  G. C. Paul,et al.  Characterisation of mycelial morphology using image analysis. , 1998, Advances in biochemical engineering/biotechnology.

[6]  K. Sang-woo,et al.  Influence of Agitation Intensity and Aeration Rate on Production of Antioxidative Exopolysaccharides from Submerged Mycelial Culture of Ganoderma resinaceum , 2006 .

[7]  U. Kües Life History and Developmental Processes in the Basidiomycete Coprinus cinereus , 2000, Microbiology and Molecular Biology Reviews.

[8]  H. Hwang,et al.  Morphological and rheological properties of the three different species of basidiomycetes Phellinus in submerged cultures , 2004, Journal of applied microbiology.

[9]  M. Schülein,et al.  Comparison of structure and activities of peroxidases from Coprinus cinereus, Coprinus macrorhizus and Arthromyces ramosus. , 1992, Biochimica et biophysica acta.

[10]  S. Kajiwara,et al.  Molecular breeding of the basidiomycete Coprinus cinereus strains with high lignin-decolorization and -degradation activities using novel heterologous protein expression vectors , 1998, Applied Microbiology and Biotechnology.

[11]  E. Grulke,et al.  Development of a stirred tank reactor system for the production of lignin peroxidases (ligninases) byPhanerochaete chrysosporium BKM-F-1767 , 1990, Journal of Industrial Microbiology.

[12]  P. Cox,et al.  Classification and measurement of fungal pellets by automated image analysis , 1992, Biotechnology and bioengineering.

[13]  L. H. Grimm,et al.  Investigations of the Morphogenesis of Filamentous Microorganisms , 2006 .

[14]  Maria Papagianni,et al.  Quantification of the fractal nature of mycelial aggregation in Aspergillus niger submerged cultures , 2006, Microbial cell factories.

[15]  T. Anke,et al.  Influence of morphology and rheology on the production characteristics of the basidiomycete Cyathus striatus. , 1998, Biotechnology and bioengineering.

[16]  S. Braun,et al.  Mycelial morphology and metabolite production , 1991 .

[17]  J. Nielsen,et al.  Pellet Formation and Fragmentation in Submerged Cultures of Penicillium chrysogenum and Its Relation to Penicillin Production , 1995, Biotechnology progress.

[18]  L. Harvey,et al.  Submerged culture fermentation of "higher fungi": the macrofungi. , 2008, Advances in applied microbiology.

[19]  P. Pukkila,et al.  DNA‐mediated transformation of the basidiomycete Coprinus cinereus. , 1987, The EMBO journal.