Identifikacija Penicillium expansum prouzrokovača truleži plodova jabuke u skladištu
Sažetak
Penicillium expansum (Link) Thom. je jedan od najznačajnijih prouzrokovača truleži plodova jabučastog voća u toku čuvanja koji se odlikuje proizvodnjom mikotoksina patulina. U ovom radu je, na osnovu morfoloških i molekularnih osobina, identifikovana vrsta P. expansum kao prouzrokovač truleži plodova jabuke u skladištu. Plodovi jabuke sorti Ajdared, Zlatni delišes i Breburn sa simptomima truleži prikupljene su u skladištima u lokalitetima Pudarci, Suvodol, Čelarevo, Bela Crkva i Svilajnac. Na prikupljenim plodovima uočavale su se svetlosmeđe, meke i udubljene pege u okviru kojih je bila prisutna zelenoplava sporulacija gljive. Izolacija je obavljena standardnim postupkom na podlogu od krompir-dekstroznog agara (KDA), a patogenost izolata proverena je inokulacijom zdravih plodova jabuke. Morfološke odlike kolonija i reproduktivnih struktura izolata praćene su na dijagnostičkim hranljivim podlogama (MEA, CYA i YES). Molekularna identifikacija izolata obavljena je korišćenjem specifičnih prajmera za vrstu P. expansum zasnovanih na genu za poligalakturonazu (Pepg1), univerzalnih prajmera za ITS region i prajmera zasnovanih na genu za β-tubulin. Iz jabuke sa simptomima plave truleži dobijeno je devet izolata čija je patogenost potvrdjena veštačkim inokulacijama zdravih plodova jabuke. Ispitivani izolati su formirali kompaktne zelenoplave kolonije karakterističnog mirisa. Svi izolati su rasli na 5oC, a porasta nije bilo na 37oC. Konidije prosečne veličine 3,38 x 3 μm, bile su okruglaste do ovalne, formirane u dugim nizovima. Konidiofore su bile terverticillata, asimetričnog grananja sa glatkim stipama. Kod svih ispitivanih izolata amplikoni očekivane veličine oko 404 bp dobijeni su korišćenjem specifičnih prajmera za P. expansum PEF/PER, dok su primenom ITS1-ITS4 prajmera amplifikovani produkti veličine oko 600 bp, a primenom prajmera za deo β-tubulin gena, produkti veličine oko 800 bp. MegaBlast analizom dobijenih sekvenci ITS regiona i dela β-tubulin gena izabranog izolata JP1 utvrđena je 100%, odnosno 99-100% sličnost sa sekvencama odgovarajućih regiona vrste P. expansum deponovanim u NCBI bazi podataka. Na osnovu morfoloških i molekularnih osobina dobijenih izolata utvrđeno je da prouzrokovači truleži plodova jabuke u nekoliko skladišta u Srbiji pripadaju vrsti P. expansum.
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