South-East European Forestry (Jan 2020)
DNA Barcoding of Fungi in the Forest Ecosystem of the Psunj and Papuk Mountains in Croatia
Abstract
The saprotrophic, endophytic, and parasitic fungi were detected from the samples collected in the forest of the management unit East Psunj and Papuk Nature Park in Croatia. The disease symptoms, the morphology of fruiting bodies and fungal culture, and DNA barcoding were combined for determining the fungi at the genus or species level. DNA barcoding is a standardized and automated identification of species based on recognition of highly variable DNA sequences. DNA barcoding has a wide application in the diagnostic purpose of fungi in biological specimens. DNA samples for DNA barcoding were isolated from infected tree tissues, fungal fruiting bodies or fungal cultures. The ITS or ITS2 sequences of the fungal DNA were sequenced and aligned with the reference sequences in GenBank (NCBI) using BLAST. The size of ITS and ITS2 sequences were 512-584 bp and 248-326 bp, respectively. The sequences showed a high identity of 97.21%-100% at 98%-100% coverage with reference sequences in GenBank (NCBI). The exception was the species Amphilogia gyrosa that showed 95.65% identity at 100% coverage. Two fungi were determined at genus level: Cladosporium sp., and Cytospora sp., while 11 fungi were determined at species level: Alternaria alternata, Aureobasidium pullulans, Amphilogia gyrosa, Capronia pilosella, Cryphonectria parasitica, Exidia glandulosa, Epicoccum nigrum, Penicillium glabrum, Pezicula carpinea, Rosellinia corticium, and Stereum hirsutum.