Diverzitet populacije Xanthomonas spp. patogena paprike u Srbiji
Diversity of Xanthomonas spp. population - pepper pathogens in Serbia
Author
Ignjatov, Maja V.
Mentor
Obradović, Aleksa
Committee members
Balaž, JelicaMilošević, Mirjana
Gvozdenović-Varga, Jelica

Ivanović, Milan

Metadata
Show full item recordAbstract
Tokom 2008, 2009 i 2010. godine prikupljeni su uzorci obolelog lišća paprike sa
simptomima bakteriozne pegavosti iz različitih lokaliteta Republike Srbije. Izolacijom iz
zaraženih listova dobijeno je 116 sojeva bakterija.
Proučavanja patogenih, biohemijsko-fizioloških odlika ukazuju da proučavani
sojevi pripadaju vrsti Xanthomonas euvesicatoria.
Primenom dva serološka testa (DAS - ELISA i test aglutinacije) potvrđena je
antigena uniformnost proučavanih sojeva, kao i serološka sličnost sa bakterijom X. c. pv.
vesicatoria.
Diferencijacija Xanthomonas spp. prouzrokovača bakteriozne pegavosti paprike
izvršena je primenom dve molekularne metode. Prvi metod zasnivao se na restrikcionoj
analizi - RFLP (Restriction Fragment Lenght Polymorphism) fragmenta hrp gena (HrpB
regiona) nakon digestije (sečenja) dobijenih proizvoda restrikcionim enzimima (CfoI, TaqI
i HaeIII). Poređenjem restrikcionih profila proučavanih sojeva sa kontrolnim sojevima,
potvrđena je sličnost sa vrstom X. euvesicatoria.
D...rugi molekularni metod je lančana reakcija polimeraze primenom specifičnih
prajmera koji amplifikuju proizvode različite veličine specifične za: X. euvesicatoria – 173
bp; X. vesicatoria – 138 bp; X. perforans – 197 bp i X. gardneri – 154 bp. Kod svih
proučavanih sojeva, uključujući i kontrolni soj X. euvesicatoria KFB (189) detektovani su
fragmenti DNK veličine 173 bp, koji odgovaraju vrsti X. euvesicatoria. Utvrđeno je da je
prag detekcije ove metode 0,4 x 103 CFU/ml.
Proučeno je dejstvo bakar-sulfata (CuSO4), antibiotika streptomicina i
kasugamicina, pri različitim koncentracijama, na razvoj bakterijskih ćelija u in vitro
uslovima. U podlogu su dodati filterom sterilisani rastvori bakar-sulfata do konačne
koncentracije od 100 i 200 ppm tj. aktivne komponente navedenih antibiotika do konačne
koncentracije od 50, 100 i 200 ppm. Sojevi rezistentni na streptomicin nisu detektovani
ovim istraživanjima. Rezistentnost prema 50 ppm kasugamicina utvrđena je kod 6 sojeva, a
19 sojeva je bilo rezistentno na 200 ppm bakar-sulfata. Dobijeni rezultati ukazuju na
opasnost od razvoja rezistentnosti bakterija prema ovim jedinjenjima...
During 2008, 2009 and 2010 samples of diseased pepper leaves with bacterial
spot symptoms were collected from different localities in Republic of Serbia. Total of 116
strains of bacteria were obtained by isolation from infected leaves.
Studies of pathogenic, biochemical and physiological traits showed that tested
strains belong to species Xanthomonas euvesicatoria.
Two serological tests (DAS - ELISA and agglutination test) confirmed antigenic
identity of tested bacterial isolates and serological similarity to bacteria X. c. pv.
vesicatoria.
Differentiation of Xantomonas spp. causal agent of bacterial spot of pepper was
performed using two molecular methods. The first method was based on restriction analysis
– RFLP (Restriction Fragment Length Polymorphism) of the hrp gene fragment (HrpB
region) after digestion of obtained products by restriction enzymes (CfoI, TaqI and HaeIII).
Comparison of restriction profiles of studied strains to control strains confirmed the
similarity with the X.... euvesicatoria. The other molecular method is polymerase chain
reaction of specific primers which allow amplification of different size products sprecific
for: X. euvesicatoria – 173 bp; X. vesicatoria – 138 bp; X. perforans – 197 bp and X.
gardneri – 154 bp. As a result, DNA fragments of 173 bp which belong to species X.
euvesicatoria were detected at all studied strains, including test strain Xe KFB (189).
Threshold of detection was confirmed to be 0.4x103 CFU/ml.
The effects of copper-sulphate (CuSO4), antibiotics streptomycin and kasugamycin
on the development of bacterial cells were studied. The sensitivity of strains to bactericides
was studied in vitro by culturing bacteria on sucrose pepton agar (SPA) plates, amended
with filter-sterilized aqueous solution of streptomycin and kasugamycin (50, 100, 200 ppm)
or copper-sulphate (100, 200 ppm). Streptomycin resistant strains were not detected, but 6
strains were resistant to kasugamycin (50 ppm) and 19 strains to copper-sulphate (200
ppm), indicating bacterial resistance development...
Faculty:
Универзитет у Београду, Пољопривредни факултетDate:
22-11-2013Projects:
- Development of vegetable cultivars and hybrids intended for outdoor and indoor production (RS-31030)