magistrsko delo
Urban Česnik (Author), Lorena Butinar (Mentor)

Abstract

Opravili smo dve hitri molekularni tehniki za identifikacijo enoloških kvasovk, ki so bile izolirane iz vinogradov z ekološko in intergrirano pridelavo grozdja. Najprej smo določili 15 Saccharomyces kvasovk s pomočjo analize multipleks PCR med zbranimi 131 izolati, nato smo s pomočjo ribotipizacije ITS določili približno 98 kvasovk do nivoja vrste, ki so bile potrjene tudi s sekvenciranjem regije ITS. Določili smo 16 vrst vinskih kvasovk: Hanseniaspora uvarum, H. valbyensis, H. opuntiae / H. uvarum, Lachancea thermotolerans, Candida tropicalis, Candida sp. / Starmerella sp., Metschnikowia pulcherrima, Pichia kudriavzevii, P. terricola, P. kluyveri, Wickerhamomyces anomalus, Kodamaea ohmeri, S. bacillaris, Saccharomyces cerevisiae, Saturnispora diversa in Torulaspora delbrueckii. Ugotovili smo, da so najdene kvasovke običajne naseljevalke grozdne mikroflore ter da je način pridelave grozdja vplival na pojavljanje določenih vrst v obravnavanih vinogradih. Nadalje smo testirali kvasovke na tvorbo sideroforov, tvorbo protiglivnih hlapnih organskih spojin (HOS), hidrolitično aktivnost in toleranco kvasovk na fungicide, kot so baker, iprodion, fluazinam ter kombinacijo ciprodinil / fludioksonil. Za kvalitativno detekcijo hidrolitične aktivnosti smo uporabili presejalne teste na osnovi trdnega gojišča z dodanim hitinom ali β-D-glukozidi kot substratom in ugotovili, da je veliko testiranih kvasovk sposobnih proizvajati litične encime, ki bi lahko bili odgovorni za razgradnjo celičnih sten fitopatogenih gliv in potencialno sposobni proizvajati HOS s hidrolizo grozdnih glikozidov zaradi prisotnosti β-glukozidaz. Prav tako smo opazili, da so te encimske aktivnosti običajno odvisne od seva. Na splošno so kvasovke tudi tolerantne na testirane fungicide. Ugotovili smo tudi, da je odpornost biofungicidnih kvasovk na fungicide koristna lastnost za razvoj biofungicidov v prihodnosti pri aplikacijah v strategijah z zmanjšanim vnosom pesticidov.

Keywords

kvasovke;biološki nadzor;fitopatogeni;Botrytis cinerea;vinogradništvo;magistrske naloge;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UNG FZO - Faculty of Environmental Sciences
Publisher: [U. Česnik]
UDC: 54
COBISS: 5487867 Link will open in a new window
Views: 3517
Downloads: 115
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Other data

Secondary language: English
Secondary title: Yeasts as promising biological control agents (BCAs) against phytopathogenic fungi in viticulture
Secondary abstract: We performed two rapid molecular techniques for the identification of enological yeasts that were collected from vineyards with organic and integreated pest management. First we distinguised 15 Saccharomyces yeasts with multiplex PCR analysis from collected 131 isolates, further on we identified approx. 98 isolates to species level based on ITS ribotyping that were confirmed also with ITS sequencing. Overall we distinguished 16 wine yeast species: Hanseniaspora uvarum, H. valbyensis, H. opuntiae / H. uvarum, Lachancea thermotolerans, Candida tropicalis, Candida sp. / Starmerella sp., Metschnikowia pulcherrima, Pichia kudriavzevii, P. terricola, P. kluyveri, Wickerhamomyces anomalus, Kodamaea ohmeri, S. bacillaris, Saccharomyces cerevisiae, Saturnispora diversa and Torulaspora delbrueckii. We observed that obtained yeasts are common inhabitants of grape microflora, and that applied pest managament in studied vineyards had an influence on occurence of certain species. Thereafter, we tested yeasts for siderophore production, antifungal VOCs production, hydrolysis activity and yeast tolerance to fungicides like copper, iprodione, fluazinam and cyprodinil / fludioxonil combination. With a qualitative detection of the hydrolysis activity by using screening methods based on soild medium with chitin or β-D-glucosides as substrates, we recognized that many tested yeasts were able to produce lytic enzymes that could be responsible for cell wall degradation of phytopathogenic fungi, and potentially able to produce VOCs from the hydrolysis of grape glycosides due to the β-glucosidase presence. We also observed that these enzymatic activities are usually strain-dependent. In general yeasts were also tolerant to tested fungicides, and we concluded that fungicide resistance of biofungicide yeasts is beneficial trait for future biofungicide development for possible applications in low input pest management strategies.
Secondary keywords: Yeasts;biological control;phytopathogen;Botrytis cinerea;viticulture;
URN: URN:SI:UNG
Type (COBISS): Master's thesis/paper
Thesis comment: Univ. v Novi Gorici, Fak. za znanosti o okolju
Pages: X, 45 str.
ID: 11290942