Gljive iz agroindustrijskog otpada kao antagonisti fitopatogenim gljivama

Abstract

Kompostiranjem agroindustrijskog otpada prevazilaze se ekološki problemi i dobija se proizvod znaĉajne upotrebne vrednosti. Dodavanje kompostiranog agroindustrijskog otpada poljoprivrednom zemljištu ima mnoge korisne efekte po useve, a meĊu njima je i smanjenje uĉestalosti oboljenja biljaka. Smatra se da je za efekat na prouzrokovaĉe bolesti biljaka odgovoran diverzitet mikroorganizama koji stupaju u složene interakcije sa fitopatogenima. Predmet sprovedenih istraživanja su dve vrste agroindustrijskog otpada: komina grožĊa i duvanski otpad i kompost od džibre, sporednog proizvoda koji nastaje pri proizvodnji šljivovice. Cilj istražvanja bio je ispitivanje antagonizma filamentoznih gljiva izolovanih iz agroindustrijskog otpada prema fitopatogenim vrstama. Izvršena je mikrobiološka karakterizacija otpada i izolacija filamentoznih gljiva, a zatim i selekcija izolata koji pokazuju antagonizam prema tri fitopatogene vrste: Botrytis cinerea, Fusarium oxysporum i Pythium aphanidermatum. Izolacija je izvršena standardnim mikrobiološkim metodama, a u cilju selekcije je primenjen konfrontacijski test. Odabrano je šest izolata koji su okarakterisani i identifikovani na osnovu morfoloških, odgajivaĉkih i ekoloških svojstava, proizvodnje hidrolitiĉkih enzima i molekularnim metodama. Molekularna identifikacija izvršena je umnožavanjem i sekvencioniranjem ITS regiona rDNK i proteinskih gena koji kodiraju sintezu β-tubilina i Tef-1α i poreĊenjem dobijenih sekvenci sa sekvencama dostupnim u GenBank bazi podataka. Selektovani izolati su identifikovani kao: Penicillium paneum, Penicillium chrysogenum, Aspergillus fumigatus, Trichoderma longibrachiatum, Rhyzopus oryzae i Aspergillus sp. sekcija Nigri. Ispitan je uticaj ekstracelularnih metabolita šest odabranih izolata sadržanih u filtratu teĉne kulture na rast fitopatogena. Primenjene su razliĉite koncentracije filtrata i razliĉiti naĉini sterilizacije (autoklaviranje i mikrobiološka filtracija). Indukcija proizvodnje antifungalnih metabolita usled prisustva fitopatogene vrste ispitana je putem efekta filtrata dvojne kulture (izolata i fitopatogena) na rast istog biljnog patogena...

Composting of agro-industrial waste is the way to overcome ecological problems and to get a product of the significant utility value. Soil amendment with the composted agroindustrial waste possesses many beneficial effects, including the decrease of plant diseases incidence. It is considered that diversity of microorganisms that interact with plant pathogens is responsible for the disease suppression effect. The subject of the conducted research were two types of agro-industrial waste: grape marc and tobacco waste, and compost made of plum stillage, by product in plum brandy production. The aim of the research was to study the antagonism of filamentous fungi from agroindustrial waste towards phytopathogenic species. Microbiological characterization of wastes and isolation and characterization of filamentous fungi was performed, followed by selection of the isolates that exhibits antagonistic effect on three phytopathogenic species: Botrytis cinerea, Fusarium oxysporum and Pythium aphanidermatum. The isolation was carried out by standard microbiological methods, and confrontation test allowed selection of six isolates which were identified and characterized on the bases on morphological, cultural and ecological properties, production of hydrolytic enzymes and by molecular methods. Molecular identification was done by amplification and sequencing of ITS rDNA, and protein coding genes β-tubulin and Tef-1α, and comparisons of the obtained sequences with sequences available in GenBank. Selected isolates are identified as Penicillium paneum, Penicillium chrysogenum, Aspergillus fumigatus, Trichoderma longibrachiatum, Rhyzopus oryzae and Aspergillus sp. section Nigri. The effect of extracellular metabolites, contained in the liquid culture filtrate, on the growth of phytopathogens was tested. Different concentrations of filtrates were applied as well as different sterilization methods (autoclaving and microbiological filtration). The induction of antifungal metabolites production by the presence of phitophatogenic species was examined by the effect of dual culture filtrates (of the isolate and plant pathogen) on the growth of the respective plant pathogen...