Raznolikost gena za molekularni šaperonin GroEL u genomima bakterija roda Candidatus Phytoplasma

master's thesis

• Title (english): Diversity of genes encoding molecular chaperonin GroEL in the genomes of bacteria from the genus Candidatus Phytoplasma
• Author: Anamarija Slović
• Mentor: Martina Šeruga Musić (mentor)
• Committee member: Martina Šeruga Musić (predsjednik povjerenstva)
• Committee member: Inga Marijanović (član povjerenstva)
• Committee member: Ana Galov (član povjerenstva)
• Committee member: Željka Vidaković-Cifrek (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2014-01-21, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Fitoplazme ('Candidatus Phytoplasma') su bakterije bez stanične stijenke iz razreda Mollicutes. Obligatni su simbionti biljaka i kukaca te uzrokuju brojne ekonomski značajne bolesti mnogih poljoprivrednih kultura u cijelome svijetu. Nemoguće ih je uzgajati u mediju bez stanica, čime je njihova detekcija i klasifikacija otežana. Molekularni šaperonin GroEL pomaže u smatanju drugih proteina, a prisustvo ovog gena detektirano je u svim potpuno sekvenciranim genomima fitoplazmi. Zbog mnogobrojnih uloga proteina GroEL, ovaj je gen istražen u novim izolatima fitoplazmi ribosomske podskupine 16SrXII-A, koje pripadaju vrsti 'Ca. P. solani' i uzrokuju ekonomski značajne štete vinove loze (Vitis vinifera L.). Gen za molekularni šaperonin GroEL iz nekoliko izolata uspješno je umnožen PCR-om, nakon čega je određen slijed nukleotida ovog gena, a metodom Southern blot detektirana je jedna kopija gena u genomu. Raznolikost gena novih izolata i gena iz poznatih genoma ispitana je filogenetskim analizama, koje su pokazale očekivano grupiranje fitoplazmi s obzirom na 16Sr skupine, što ovaj gen čini pogodnim markerom za analizu evolucijskih odnosa među fitoplazmatskim vrstama kao i varijabilnost izolata unutar pojedine vrste.

Abstract (english)

Phytoplasmas ('Candidatus Phytoplasma') are wall-less bacteria from the class Mollicutes. They are obligate symbionts of plants and insects and cause diseases in numerous agriculturally significant plant species. It is impossible to cultivate them in cell-free media, making their detection and classification difficult. Molecular chaperonin GroEL assists in the folding of other proteins, and its presence was detected in all phytoplasma genomes that have been sequenced to completion. Because of many various functions of GroEL protein, this gene was studied in new phytoplasma isolates from the 16SrXII-A ribosomal subgroup, belonging to the 'Ca. P. solani' species that cause significant economic damage of the grapevine (Vitis vinifera L.). Gene encoding molecular chaperonin GroEL was successfully amplified by PCR from several isolates, and subsequently nucleotide sequence of the gene was determined. The presence of a single copy of the gene in the genome was confirmed by Southern blot method. Phylogenetic analysis of groEL gene sequences belonging to new phytoplasma isolates and those available in GenBank was also performed. Considering the 16S rDNA gene based groups, the expected phytoplasma grouping was observed, making this gene a suitable marker for analysis of evolutionary relationships among different phytoplasma species as well as the studies of intraspecies variability.

• Keywords: Candidatus Phytoplasma solani; fitoplazme; stolbur skupina; Southern blot; filogenetska analiza; molekularni marker
• Keywords (english): Candidatus Phytoplasma solani; phytoplasma; stolbur group; Southern blot; phylogenetic analysis; molecular marker
• Language: croatian
• URN:NBN: urn:nbn:hr:217:781702
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-04-26 12:24:23