Razvoj metode z zanko posredovanega izotermalnega pomnoževanja za določanje gliv rodu Verticillium.

Grgič, Zarja (author), Štajner, Nataša (mentor), Radišek, Sebastjan (comentor)

Glive rodu Verticillium predstavljajo ene izmed najbolj škodljivih rastlinskih patogenov. Predstavniki rodu lahko parazitirajo širok spekter gostiteljskih rastlin, med katerimi je tudi veliko gospodarsko in kmetijsko pomembnih kultur. V Sloveniji največ škode povzročata vrsti V. nonalfalfae, ki povzroča blago in letalno obliko verticilijske uvelosti hmelja in V. dahliae. Za lažje nadzorovanje in preprečevanje širjenja bolezni, je ključnega pomena razvoj ustreznih diagnostičnih metod, ki bi omogočile hitro in zanesljivo identifikacijo ter razločevanje med posameznimi vrstami in patotipi. Cilj naše raziskave je vključeval razvoj metode LAMP za hitro in učinkovito določanje vrst rodu Verticillium ter letalnega patotipa vrste V. nonalfalfae (PG2). Ker je za obravnavan rod fitopatogenih gliv značilna velika heterogenost, smo izvedli tudi molekularno analizo variabilnosti. Preko pomnoževanja DNA z začetnimi oligonukleotidi, specifičnimi za letalno regijo patotip PG2, smo želeli pokazati veliko heterogenost znotraj posameznih vrst gliv rodu Verticillium in težavnost uporabe ustrezne diagnostične metode ter načrtovanja začetnih oligonukleotidov, ki bi omogočili razločevanje med vrstami in patotipi tega rodu.

• Language: Slovenian
• Keywords: Verticillium, identifikacija, LAMP, molekularna genetika, začetni oligonukleotidi, PCR, patogene glive
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [Z. Grgič]
• Year: 2019
• PID: 20.500.12556/RUL-111070
• UDC: 601.4:577.21:602:632.4(043.2)
• COBISS.SI-ID: 9362041
• Publication date in RUL: 22.09.2019

GRGIČ, Zarja, 2019, Razvoj metode z zanko posredovanega izotermalnega pomnoževanja za določanje gliv rodu Verticillium. [online]. Master’s thesis. Z. Grgič. [Accessed 2 April 2025]. Retrieved from: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=eng&id=111070

Secondary language

• Language: English
• Title: Development of loop-mediated isothermal amplification method for detection of fungi from the genus Verticillium.

Abstract

Fungi from the genus Verticillium are known to be one of the most harmful plant pathogenic organisms. The genus consists of species that can infect a broad spectrum of plants, many of which are agriculturally relevant crops. In Slovenia a great amout of harm is caused by two species, V. nonalfalfae and V. dahliae. By infecting the hop plant, they can cause a mild or lethal form of a disease, called verticillium wilt. In order to control and potentially even prevent the disease from spreading, the development of suitable diagnostic methods for rapid detection and identification is crucial. The aim of our study was to develop the LAMP method, which can enable a rapid and reliable detection of species of the genus Verticillium and the pathotype PG2. Due to the genus' heterogeneity, we analised a series of isolates via PCR primers specific for the amplicifation of different regions from lethal region of PG2 isolate, by which we wanted to emphasize the difficulties of development of the suitable method and designe of specific primers for molecular diagnostics of the genus Verticillium.

• Keywords: Verticillium, identification, LAMP, molecular genetics, primers, PCR, pathogenic fungi