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Characterization of cultivated fungi isolated from grape marc wastes through the use of amplified rDNA restriction analysis and sequencing

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Abstract

Microbial assessment of grape marc wastes, the residual solid by-product of the wine-industry, was performed by identifying phylogenetically the fungal culturable diversity in order to evaluate environmental and disposal safety issues and to discuss ecological considerations of applications on agricultural land. Fungal spores in grape marc were estimated to 4.7×106 per g dry weight. Fifty six fungal isolates were classified into eight operational taxonomic units (OTUs) following amplified ribosomal DNA restriction analysis (ARDRA) and colony morphology. Based on 18S rRNA gene and 5.8S rRNA gene-ITS sequencing, the isolates representing OTUs #1, #2, #3, and #4, which comprised 44.6%, 26.8%, 12.5%, and 5.3%, respectively, of the number of the total isolates, were identified as Aspergillus fumigatus, Bionectria ochroleuca, Haematonectria haematococca, and Trichosporon mycotoxinivorans. The isolates of OTU#5 demonstrated high phylogenetic affinity with Penicillium spp., while members of OTUs #6 and #7 were closer linked with Geotrichum candidum var. citri-aurantii and Mycocladus corymbifer, respectively (95.4 and 97.9% similarities in respect to their 5.8S rRNA gene-ITS sequences). The OTU#8 with a single isolate was related with Aspergillus strains. It appears that most of the fungal isolates are associated with the initial raw material. Despite the fact that some of the species identified may potentially act as pathogens, measures such as the avoidance of maintaining large and unprocessed quantities of grape marc wastes in premises without adequate aeration, together with its suitable biological treatment (e.g., composting) prior to any agriculture-related application, could eliminate any pertinent health risks.

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Authors and Affiliations

  1. National Agricultural Research Foundation, Institute of Kalamata, Lakonikis 87, 24100, Kalamata, Greece

    Spyridon Ntougias & Georgios I. Zervakis

  2. Department of Environmental Engineering, Laboratory of Wastewater Management and Treatment, Democritus University of Thrace, Vas. Sofias 12, 67100, Xanthi, Greece

    Spyridon Ntougias

  3. National Agricultural Research Foundation, Institute of Chania, Agrokipion, 73100, Chania, Crete, Greece

    Nektarios Kavroulakis

  4. Department of Biochemistry & Biotechnology, University of Thessaly, Ploutonos 26 & Aiolou, 41221, Larissa, Greece

    Kalliope K. Papadopoulou

  5. Department of Natural Resources and Agricultural Engineering, Soils and Agricultural Chemistry Laboratory, Agricultural University of Athens, Iera Odos 75, Athens, 118 55, Greece

    Constantinos Ehaliotis

  6. Department of Agricultural Biotechnology, Laboratory of General and Agricultural Microbiology, Agricultural University of Athens, Iera Odos 75, Athens, 118 55, Greece

    Georgios I. Zervakis

Authors
  1. Spyridon Ntougias
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  2. Nektarios Kavroulakis
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  3. Kalliope K. Papadopoulou
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  4. Constantinos Ehaliotis
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  5. Georgios I. Zervakis
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Correspondence to Georgios I. Zervakis.

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Ntougias, S., Kavroulakis, N., Papadopoulou, K.K. et al. Characterization of cultivated fungi isolated from grape marc wastes through the use of amplified rDNA restriction analysis and sequencing. J Microbiol. 48, 297–306 (2010). https://doi.org/10.1007/s12275-010-9193-y

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  • DOI: https://doi.org/10.1007/s12275-010-9193-y

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