magistrsko delo
Sara Fišer (Author), Martina Turk (Reviewer), Cene Gostinčar (Mentor), Polona Kogovšek (Co-mentor)

Abstract

Karantenska patogena gliva Phyllosticta citricarpa je povzročiteljica črne pegavosti agrumov. Zaradi nevarnosti njene razširitve v evropski prostor v ogroženih nasadih potekajo vzorčenja zraka in dežja. Analiza vzorcev je potekala v sklopu magistrske naloge, kjer smo z uporabo optimiziranega postopka izolacije DNA in testov qPCR preverjali prisotnost glive. Poleg kontrolnih testov 18S (komercialni test, Applied Biosystems), Yeast (Hierro in sod., 2006) in FungiQuant (Liu in sod., 2012), za specifično določevanje glive uporabljamo testa PC (van Gent-Pelzer in sod., 2007) in PC-2 (Schirmacher in sod., 2019). Testa na podlagi zaporedja ITS ločujeta P. citricarpa od P. capitalensis in P. citriasiana, ne pa tudi od P. paracitricarpa, zato je bil naš namen vzpostaviti in validirati nov qPCR test TEF1, ki bi to omogočal. Z zastavljenimi poskusi smo dokazali njegovo strogo specifičnost za P. citricarpa, visoko občutljivost in robustnost. Pokazali smo linearnost testa v širokem območju in z večkratnim izvajanjem poskusov dokazali njegovo ponovljivost. Ocenili smo ga kot primernega za nadaljnjo uporabo v diagnostične namene. Analizirali smo realne vzorce zraka in ugotovili, da iskani patogen v njih ni prisoten, kar pomeni, da se še ni razširil v evropski prostor.

Keywords

Phyllosticta citricarpa;črna pegavost agrumov;patogena gliva;rastlinski patogen;izolacija DNA;PCR v realnem času;vzpostavitev metode;ITS;TEF1;magistrska dela;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [S. Fišer]
UDC: 577
COBISS: 115677443 Link will open in a new window
Views: 82
Downloads: 17
Average score: 0 (0 votes)
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Other data

Secondary language: English
Secondary title: Establishing a method for detection of a pathogenic fungus Phyllosticta citricarpa
Secondary abstract: The quarantine pathogenic fungus Phyllosticta citricarpa is the causative agent of citrus black spot disease. Due to the risk of its spread to the European region, air and rain sampling is taking place in endangered orchards. The samples were analysed as part of this master's thesis, in which the presence of the fungus is monitored using an optimized DNA extraction protocol and qPCR tests. In addition to the control tests – 18S (Applied Biosystems), Yeast (Hierro et al., 2006), and FungiQuant (Liu et al., 2012) – the tests PC (van Gent-Pelzer et al., 2007) and PC-2 (Schirmacher et al., 2019) are used for specific fungal identification. Tests based on the ITS sequence are used to distinguish P. citricarpa from P. capitalensis and P. citriasiana, but not from P. paracitricarpa. Therefore, our goal was to establish and validate a new qPCR assay TEF1 that would allow this. With the set experiments, we demonstrated its strict specificity for P. citricarpa, its high sensitivity and robustness. We demonstrated the linearity of the test over a wide range and proved its repeatability by repeated experiments. It was evaluated as suitable for further use in diagnostics. We have analysed real air samples and found that the target pathogen is not present in them, which means that it has not yet spread to the European region.
Secondary keywords: citrus black spot;pathogenic fungi;plant pathogen;DNA isolation;real-time PCR;method establishing;master thesis;Molekularna biologija;Glive;Univerzitetna in visokošolska dela;
Type (COBISS): Master's thesis/paper
Study programme: 0
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Oddelek za biologijo
Pages: XII, 52 f., [17] f. pril.
ID: 15907818