Identifikacija, karakterizacija i genetički diverzitet sojeva Agrobacterium spp., prouzrokovača bakterioznog raka vinove loze
Identification, characterization and genetic diversity of strains of Agrobacterium spp., grapevine crown gall causal agent
Author
Kuzmanović, Nemanja S.Mentor
Obradović, Aleksa![](/themes/MirageNardus/images/orcid.png)
Committee members
Balaž, JelicaGavrilović, Veljko
Marković, Nebojša
Ivanović, Milan
![](/themes/MirageNardus/images/orcid.png)
Metadata
Show full item recordAbstract
Bakteriozni rak smatra se jednim od najznačajnijih i najrasprostranjenijih
bakterioznih oboljenja vinove loze u svetu. Glavni prouzrokovač ovog oboljenja je vrsta
Agrobacterium vitis. Tokom poslednjih nekoliko godina primećena je izražena pojava
bolesti u većini vinogradarskih rejona u Srbiji. Stoga je u ovom radu proučena etiologija
bakterioznog raka vinove loze u našoj zemlji. Takođe, izvršena je karakterizacija i
proučen genetički diverzitet prouzrokovača bolesti. Primenjene su kako klasične
bakteriološke, tako i molekularne metode. U radu su takođe korišćeni i brojni referentni
sojevi A. vitis poreklom iz međunarodnih kolekcija.
Simptomatični uzorci vinove loze prikupljeni su u 22 lokaliteta, raspoređena u
gotovo svim vinogradarskim rejonima u našoj zemlji. Izolacija bakterija vršena je na
neselektivne podloge sa manitolom i kvaščevim ekstraktom (YMA). Preliminarna
identifikacija izolovanih sojeva, izvedena je primenom PCR metode, korišćenjem
prajmera specifičnih za plazmidne (virC..., virF, virD2, ipt, tms2) i hromozomske (pehA,
23S rRNK) gene. Među izolovanim sojevima identifikovan je tumorogeni A. vitis, a za
dalji rad izdvojeno je ukupno 36 reprezentativnih sojeva. Rezultati dobijeni primenom
PCR metode potkrepljeni su filogenetskom analizom zasnovanoj na parcijalnoj sekvenci
16S rRNK gena.
Taksonomska pripadnost proučavanih sojeva dalje je potvrđena primenom
klasičnih bakterioloških metoda. Svi proučavani sojevi ispoljili su morfološke,
odgajivačke i biohemijsko-fiziološke odlike karakteristične za vrstu A. vitis. Jedini
izuzetak bio je soj KFB 243, koji je i pri ponavljanju testa stvaranja kiseline iz eritritola
kao izvora ugljenika ispoljio odlike svojstvene vrsti A. rhizogenes/biovar 2.
U cilju provere patogenosti proučavanih sojeva, vršena je inokulacija biljaka
vinove loze, suncokreta i paradajza. Od ukupno 36 proučavanih sojeva, 34 su
prouzrokovala razvoj tipičnih tumora na inokulisanim biljkama vinove loze. U testu
patogenosti na biljkama suncokreta i paradajza, zabeležene su značajne varijacije u
intenzitetu simptoma, kako unutar pojedinačnog ogleda, tako i među različitim ponavljanjima...
Crown gall is one of the most important and widespread bacterial diseases of
grapevine throughout the world. It is predominantly caused by Agrobacterium vitis.
During the last few years, serious outbreaks of the disease were observed in major
viticultural regions in Serbia. Therefore, the objective of this research was to study
etiology of the grapevine crown gall in this country. Characterization and study of
genetic diversity of the pathogen were also performed, using conventional
bacteriological and molecular methods. Various reference strains of A. vitis originating
from international collections were also included.
Symptomatic samples of grapevine were collected from 22 localities distributed
in almost all viticultural regions in the country. Isolation of bacteria was performed on
non-selective media with mannitol and yeast extract (YMA ). Preliminary identification
of the isolated strains was performed using PCR method with primers specific for the
plasmid (virC, virF, virD2, ipt..., tms2) and chromosomal (pehA, 23S rRNA) genes.
Among isolated strains, 36 representative tumorigenic A. vitis strains were selected for
further study. Results obtained by PCR method were supported by phylogenetic
analysis based on partial sequence of 16S rRNA gene.
Taxonomic position of studied strains was further confirmed using conventional
bacteriological methods. All the strains showed morphological, biochemical and
physiological characteristics of A. vitis. The only exception was strain KFB 243, which
showed characteristic of A. rhizogenes/biovar 2 in test of acid production from
erythritol.
Pathogenicity of the strains was studied by inoculation of grapevine, sunflower,
and tomato plants. Out of 36 strains studied, 34 caused development of typical tumors
on inoculated grapevine. In pathogenicity assay on tomato and sunflower plants,
considerable variations in the intensity of symptoms were recorded, both within a single
experiment, and among different repetitions. Therefore, use of tomato and sunflower as test plants, despite the rapid symptom development, cannot be recommended as a
reliable test...
Faculty:
Универзитет у Београду, Пољопривредни факултетDate:
07-05-2014Projects:
- Development of integrated management of harmful organisms in plant production in order to overcome resistance and to improve food quality and safety (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-46008)