Naslov
SSCP analysis as an approach for comparison of stolbur phytoplasma isolates

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

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

Izvornik
Book of Abstracts of the 15th Congress of the International Organization for Mycoplasmology (IOM) / Blanchard, Alain ; Browning, Glenn ; Firrao, Giuseppe ; Jacobs, Enno ; Miyata, Makoto ; Simecka, Jerry ; Waites, Kenn ; Citti, Christine ; Henrich, Birgit ; March, John ; Saillard, Colette ; Thiaucourt, Francois ; Yogev, David - Athens (GA) : International Organization for Mycoplasmology (IOM), 2004, 56-56

Skup
15th Congress of the International Organization for Mycoplasmology (IOM)

Mjesto i datum
Athens (GA), Sjedinjene Američke Države; Gruzija, 11.07.2004. - 16.07.2004

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
SSCP (single-strand conformation polymorphism); sequence changes; phytoplsama; stolbur; molecular differentiation

Sažetak
SSCP (single-strand conformation polymorphism) analysis is a rapid and sensitive method for detection of sequence changes, including single-base substitutions, as shifts in electrophoretic mobility. It is widely used for analyses of different genes and nucleic acid sequences including those of humans, plants, fungi, viruses and viroids. So far, there is no published data on the application of this method in phytoplasma sequence investigations. A phytoplasma from the ribosomal group 16SrXII-A, also known as stolbur phytoplasma, is an important pathogen of grapevine, vegetables and other plant species. It is distributed worldwide. One of the most utilized protocols for its characterization encompasses PCR and RFLP analyses of phytoplasma conserved gene regions such as 16S rDNA, a gene for the elongation factor EF-Tu and two stolbur-specific non-ribosomal regions amplified by primer pairs G35pm and STOL11f2/r1. More detailed characterization and classification of phytoplasmas from the same group often includes sequencing followed by phylogenetic analyses using bioinformatics tools. Although this is probably the most reliable way of completing the task, it is time consuming and relatively expensive. The aim of our study is to investigate the applicability of SSCP analysis for the finer differentiation of phytoplasmas within the same group using stolbur isolates as an experimental model group. The results of the tuf gene SSCP analyses showed two groups of roughly similar patterns for the samples that had identical RFLP patterns indicating nucleotide sequence differences among the isolates in the regions out of the restriction sites for the used enzymes. The cases of different SSCP patterns for tuf gene even for the stolbur phytoplasmas coming from the same vineyard have been found. RFLP of non-ribosomal genome portion amplified with primers G35pm showed different profiles indicating greater variability among the isolates in this region compared to the tuf gene portion and 16S rDNA. SSCP analyses of this region revealed the corresponding increase of variability of the patterns in the sample groups with the same RFLP profiles. In spite of very strong relatedness among stolbur isolates from different grapevine cultivars and regions in the 16S rDNA gene demonstrated by the RFLP analyses, SSCP analyses showed differences especially among geographically distant isolates. Therefore, our results suggest that SSCP analysis should be considered as a potential tool in phytoplasma research, especially in epidemiological studies with large number of samples. more detail study is needed to confirm its full applicability.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA