Molecular characterization of 'Candidatus Phytoplasma solani' isolates from periwinkle (Catharanthus roseus (L.) G. Don) of the Zagreb area

Zovkić, Tanja (2014) Molecular characterization of 'Candidatus Phytoplasma solani' isolates from periwinkle (Catharanthus roseus (L.) G. Don) of the Zagreb area. Diploma thesis, Faculty of Science > Department of Biology.

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Abstract

Phytoplasmas (genus "Candidatus Phytoplasma") are bacteria from the class Mollicutes without cell wall found in hosts from two kingdoms, in plant phloem and insect hemolymph. They are intracellular organisms that have undergone reduction of genome with a loss of many metabolically important genes. Therefore they cannot be cultured in vitro in cell-free media. Phytoplasma infect plants all over the world causing significant economic damage. Species 'Ca. Phytoplasma solani' is phytoplasma with widest natural host species range which is characteristic for the European continent. The aim of this thesis was to characterize 'Ca. Phytoplasma solani' isolates from periwinkle (Catharanthus roseus (L.) G. Don) collected at the Zagreb downtown area. In multigene typization of isolates following genes were analyzed: 16S rRNA, secY, tufB, vmp1 and stamp. Fragments of 16S rRNA, tufB and vmp1 genes were amplified by polymerase chain reaction using phytoplasma specific primers and further analyzed by RFLP (restriction fragment length polymorphism) analyses. Amplified fragments of genes secY and stamp were sequenced and analyzed by using bioinformatics tools. Phylogenetic analysis of secY and stamp nucleotide sequences together with RFLP analysis of other genes confirmed the affiliation of all isolates to the 'Ca. Phytoplasma solani' species and their intraspecies variability. Analyses of housekeeping genes secY and tufB showed that these sequences were highly conserved, while the analyses of stamp i vmp1 species specific genes, revealed more genotype variability.

Item Type: Thesis (Diploma thesis)
Keywords: 16S rDNA, phytoplasma, secY, tufB, vmp1, stamp, PCR, RFLP, phylogenetic analysis, genetic variability
Supervisor: Šeruga Musić, Martina
Date: 2014
Number of Pages: 52
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Silvana Šehić
Date Deposited: 12 Nov 2014 13:23
Last Modified: 12 Nov 2014 13:23
URI: http://digre.pmf.unizg.hr/id/eprint/3243

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