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.
Highlights
DNA barcoding is a standardized and automated biological specimen identification system (Casiraghi et al 2010)
The identification of fungal species was performed by combining the disease symptoms, the morphology of fruiting bodies, culture morphology, and DNA barcoding of the samples collected in the forest of the management unit East Psunj and Papuk Nature Park, (Tables 1 and 2, respectively)
On the site of management unit East Psunj, fungi were determined from three twig samples of Fagus sylvatica, two bark samples of Castanea sativa, and two fungal fruiting bodies on the branch of F. sylvatica
Summary
DNA barcoding is a standardized and automated biological specimen identification system (Casiraghi et al 2010). A DNA barcode is a relatively short gene sequence of the highly variable sequences of the genome, unique for the identification of species. The advantage of this technique is that a single specimen can provide complete information about the species, regardless of the morphology or lifestage characteristics. The procedure has been applied to flora and fungi. Further research into the genome of organisms revealed that for certain groups of organisms, different sequences of the genome are more acceptable for DNA barcoding. Common usage, and sufficient barcode distance of ITS sequence are characteristics that allow a wide application as a barcode for fungi (Raja 2017). Research by Yang et al (2018) proved that the operational taxonomic richness and details of the fungal community in the ITS2 sequence concerning the full-length ITS sequence produced similar results
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