Naslov
Molecular variability of phytoplasmas from 16SrXII-A ribosomal subgroup (stolbur/’ Candidatus Phytoplasma solani’ ) in Croatian grapevines

Autori
Šeruga Musić, Martina ; Krajačić, Mladen ; Škorić, Dijana

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Abstract book of the Central European Symposium on Industrial and Microbial Ecology: Power of Microbes in industry and environment / Kosalec, Ivan ; Pigac, Jasenka ; Vujaklija, Dušica - Zagreb : Pressum, 2007

ISBN
978-953-96567-5-9

Skup
Central European Symposium on Industrial and Microbial Ecology: Power of Microbes in industry and environment

Mjesto i datum
Zadar, Hrvatska, 19.09.2007. - 22.09.2007

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
method; phytoplasma; polymorphism; SSCP; tuf

Sažetak
Phytoplasmas (genus ‘ Candidatus Phytoplasma’ ) are wall-less prokaryotes with reduced genomes that reside endocellularly in the plant phloem and insects. Since they are not cultivable in vitro, the detection and classification rely mainly on the 16S rRNA gene sequence and its polymorphism. The aim of this research was to examine the molecular variability of phytoplasma isolates from the Croatian grapevines previously assigned to the 16SrXII-A ribosomal subgroup (stolbur/’ Ca. P. solani’ ). Almost 60 isolates from different geographic regions were included. Four gene/pseudogene regions were investigated: a part of the 16S rDNA, tuf gene, dnaB pseudogene and plsC gene. In order to find polymorphism, RFLP and SSCP analyses were performed. Fragments amplified by PCR were subjected to the RFLP analysis with a set of routinely used restriction enzymes. Conditions for the SSCP analysis had to be optimized for each set of PCR products, because of their different lengths. SSCP analyses revealed the presence of polymorphisms in the phytoplasma 16S rRNA gene, tuf gene and dnaB pseudogene that was undetectable by RFLP analyses. To confirm the results, cloning and sequencing of selected fragments showing different SSCP profiles were also performed, followed by phylogenetic analyses using various bioinformatics’ tools. Thus, SSCP analysis can be used as a new approach in search for phytoplasma molecular variability, particularly as a fast method for screening a large number of field samples.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

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