| Title |
Characterization of Cultivated Fungi Isolated from Grape Marc Wastes Through the Use of Amplified rDNA Restriction Analysis and Sequencing |
| Author |
Spyridon Ntougias1,2, Nektarios Kavroulakis3, Kalliope K. Papadopoulou4, Constantinos Ehaliotis5, and Georgios I. Zervakis1,6* |
| Address |
1National Agricultural Research Foundation, Institute of Kalamata, Lakonikis 87, 24100 Kalamata, Greece, 2Democritus University of Thrace, Department of Environmental Engineering, Laboratory of Wastewater Management and Treatment, Vas. Sofias 12, 67100 Xanthi, Greece, 3National Agricultural Research Foundation, Institute of Chania, Agrokipion, 73100 Chania, Crete, Greece, 4University of Thessaly, Department of Biochemistry & Biotechnology, Ploutonos 26 & Aiolou, 41221 Larissa, Greece, 5Agricultural University of Athens, Department of Natural Resources and Agricultural Engineering, Soils and Agricultural Chemistry Laboratory, Iera Odos 75, Athens 118 55, Greece, 6Agricultural University of Athens, Department of Agricultural Biotechnology, Laboratory of General and Agricultural Microbiology, Iera Odos 75, Athens 118 55, Greece |
| Bibliography |
Journal of Microbiology, 48(3),297-306, 2010,
|
| DOI |
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| Key Words |
fungal diversity, agricultural residues, environmental quality, ITS, 18S rRNA gene, winery by-products |
| 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. |
