First Report of Tumorigenic Agrobacterium radiobacter on Raspberry in Serbia
Prvi nalaz tumorogenih sojeva Agrobacterium radiobacter na malini u Srbiji
Author
Milijasevic, Svetlana
Gavrilovic, Veljko
Zivkovic, Svetlana
Trkulja, Nenad
Pulawska, Joanna
Keywords
Tumorigenic AgrobacteriumCrown gall
Raspberry
Identification
PCR
Tumorogeni sojevi
bakteriozni rak korena
malina
identifikacija
Metadata
Show full item recordAbstract
During the spring of 2003, gall symptoms on the roots and crowns of young raspberry
plants cv. Vilamette were observed near Valjevo. Phytopathogenic bacteria were isolated
from diseased plant samples. Based on the pathogenic, morphological, differential biochemical
and physiological characteristics, the isolated strains were identified as tumorigenic
Agrobacterium radiobacter (biovar 1 Agrobacterium). In order to confirm the identity of
isolated strains by polymerase chain reaction (PCR) primers complementary to tms2 gene
located on the Ti plasmid were used. In the first PCR protocol using a tms2F1 + tms2R2 primer
pair, 617 bp products specific for tumorigenic Agrobacterium strains were amplified.
The second PCR protocol, using a tms2F1 + tms2B primer pair, amplified the expected 458
bp products. On the basis of multiplex PCR with primers complementary to chromosomal
gene coding for 23S rRNA, the isolated strains were classified as biovar 1 Agrobacterium (A.
radiobacter). This is the first report of tumorigenic A. radiobacter on raspberry in Serbia. Tokom proleća 2003. godine, zapaženi su tumori na korenu i korenovom vratu mladih
biljaka maline sorte Vilamette u okolini Valjeva. Iz obolelih uzoraka izolovane su fitopatogene
bakterije. Proučavanjem patogenih, morfoloških i diferencijalnih biohemijsko-fizioloških
karakteristika izolovanih sojeva, utvrđeno je da proučavani sojevi pripadaju bakteriji
Agrobacterium radiobacter (biovar 1). U cilju potvrde identiteta izolovanih sojeva lančanom
reakcijom polimeraze (PCR), korišćeni su prajmeri komplementarni tms2 genu lociranom na
Ti plazmidu. U prvoj PCR reakciji izvedenoj korišćenjem tms2F1 + tms2R2 para prajmera, detektovani
su fragmenti nukleinske kiseline veličine 617 baznih parova (bp), specifièni za tumorogene
sojeve A. tumefaciens. U drugoj PCR reakciji korišćenjem para prajmera tms2F1 i
tms2B su amplifikovani PCR produkti očekivane veličine 458 bp, karakteristièni za ovu bakteriju.
Na osnovu multiplex PCR reakcije korišćenjem prajmera komplementarnih na hromozomalni
gen koji kodira 23S rRNA sojevi izolovani iz tumora na korenu maline svrstani su u
biovar 1 Agrobacterium (A. radiobacter). Ovo je prvi nalaz ove bakterije na malini u Srbiji.