Multigenska tipizacija izolata fitoplazme 'Candidatus Phytoplasma solani' s područja Istre

master's thesis

• Title (english): Multilocus sequence typing of 'Candidatus Phytoplasma solani' phytoplasma isolates from Istria
• Author: Magdalena Delač
• Mentor: Martina Šeruga Musić (mentor)
• Committee member: Martina Šeruga Musić (predsjednik povjerenstva)
• Committee member: Sunčica Bosak (član povjerenstva)
• Committee member: Sven Jelaska (član povjerenstva)
• Committee member: Željka Vidaković-Cifrek (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2023-11-02, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Fitoplazme (rod 'Candidatus Phytoplasma') su unutarstanične biljne patogene bakterije čiji su domaćini i vektori kukci koji se hrane floemom biljaka. Nemoguće ih je kultivirati u čistoj kulturi in vitro te su stoga istraživanja otežana. Imaju veliku ekonomsku važnost te su povezane s velikim gubicima u proizvodnji mnogih usjeva diljem svijeta. Uzročnici su bolesti žutica vinove loze (eng. grapevine yellows; GY) i ozbiljno ugrožavaju europske vinograde. Povezane su sa zarazom fitoplazmama bois noir (BN; ribosomska podskupina 16SrXII-A; 'Ca. P. solani') i flavescence dorée (ribosomska skupina 16SrV). Cilj ovog diplomskog rada bila je multigenska tipizacija izolata fitoplazme 'Ca. P. solani' dobivenih iz uzoraka vinove loze, koprive i brijesta s dva odabrana lokaliteta na području Istre. Prisutnost fitoplazmi sam potvrdila testom triplex real-time PCR, nakon čega sam filogenetski analizirala sekvence gena secY, stamp, vmp1 i tufB koji su prethodno umnoženi PCR-om. Veća raznolikost genotipova uočena je kod gena secY i stamp, a manje varijabilni bili su vmp1 i tufB. Po prvi puta je napravljena multigenska analiza nekog izolata BN fitoplazme iz brijesta. Navedene analize omogućile su detaljniji uvid u epidemiologiju fitoplazmi na području Istre te dale podatke o prisutnosti do sada nezabilježenih skupnih genotipova fitoplazme 'Ca. P. solani' u Hrvatskoj.

Abstract (english)

Phytoplasmas (genus 'Candidatus Phytoplasma') are intracellular plant pathogenic bacteria whose hosts and vectors are insects that feed on the phloem of plants. It is not possible to cultivate them in pure culture in vitro, and therefore research is difficult. They are of great economic importance and are associated with major losses in the production of many crops worldwide. They are the causative agents of grapevine yellows (GY) and seriously threaten European vineyards. They are associated with infection with phytoplasmas bois noir (ribosomal subgroup 16SrXII-A; 'Ca. P. solani') and flavescence dorée (ribosomal group 16SrV). The aim of this thesis was the multigene typing of phytoplasma isolates 'Ca. P. solani' obtained from grapevine, nettle, and elm samples from two selected locations in Istria. I confirmed the presence of phytoplasmas by using triplex real-time PCR test, followed by phylogenetic analyses secY, stamp, vmp1 and tufB gene sequences which were previously amplified by PCR. A greater diversity of genotypes was assessed for the secY and stamp genes, while vmp1 and tufB were less variable. A multigene analysis of an isolate of BN phytoplasma from elm was done for the first time. These analyses enabled a more detailed insight into the epidemiology of phytoplasma in Istria and further data on the presence of yet unrecorded collective genotypes of phytoplasma 'Ca. P. solani' in Croatia.

• Keywords: fitoplazma; filogenetska analiza; genotipizacija; PCR; MLST
• Keywords (english): MLST; phytoplasma; PCR; phylogenetic analysis
• Language: croatian
• URN:NBN: urn:nbn:hr:217:880446
• Study programme: Title: Experimental Biology-University graduate programme; Study programme type: university; Study level: graduate; Academic / professional title: sveučilišni/a magistar/magistra eksperimentalne biologije (sveučilišni/a magistar/magistra eksperimentalne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2024-11-02
• Created on: 2023-11-29 12:42:12