Articles
Xanthomonas euvesicatoria associated with pepper (Capsicum annuum L.) leaf spot in Montenegro
Article number
1391_53
Pages
389 – 394
Language
English
Abstract
During 2017/19 survey, pepper leaf spot symptoms were observed in seven localities in Montenegro.
From the leaf samples, yellow, convex and mucoid bacterial colonies were isolated on YDC medium.
All 17 strains were gram negative, HR positive, strictly aerobic, oxidase negative, catalase positive, hydrolyzed gelatin and esculin, did not reduce nitrate, and did not grow at 37°C or in presence of 0.1% triphenyl tetrazolium chloride.
Eight strains hydrolyzed starch and three showed pectolytic activity.
PCR analysis, with primer pair BSXeF/XeR, produced a single band of 173 bp in all tested strains.
Results of rep-PCR using the BOX A1R primer, showed 100% homology with Xanthomonas euvesicatoria (Xe) reference strain KFB 1. Amplification and sequencing of gyrB gene of nine selected strains was performed using primers described by Parkinson et al. (2007). Obtained partial DNA sequences showed that these strains (GenBank nos.
MZ569011, MZ574079, MZ574080, MZ574081, MZ574082, MZ574083, MZ574084, MZ574085, and MZ574086) share 99.86 to 100% identity with Xe type strain ICPM:109 and 98.71 to 100% identity with Xe strain LMG930 isolated from pepper in the United States.
Pathogenicity was confirmed by spraying young pepper plants with bacterial suspension (108 CFU mL-1 in SDW). According to the reaction of cultivar Early Calwonder (ECW) and its isogenic lines (ECW-10R, ECW-20R and ECW-30R) all tested strains belong to Xe race P8. All strains grew on SPA plates amended with 100 ppm of copper sulfate, indicating high level of resistance in Xe Montenegrin population, while 25 ppm of streptomycin sulfate inhibited their growth.
These results confirmed presence of copper tolerant race of Xanthomonas euvesicatoria in pepper production in Montenegro.
From the leaf samples, yellow, convex and mucoid bacterial colonies were isolated on YDC medium.
All 17 strains were gram negative, HR positive, strictly aerobic, oxidase negative, catalase positive, hydrolyzed gelatin and esculin, did not reduce nitrate, and did not grow at 37°C or in presence of 0.1% triphenyl tetrazolium chloride.
Eight strains hydrolyzed starch and three showed pectolytic activity.
PCR analysis, with primer pair BSXeF/XeR, produced a single band of 173 bp in all tested strains.
Results of rep-PCR using the BOX A1R primer, showed 100% homology with Xanthomonas euvesicatoria (Xe) reference strain KFB 1. Amplification and sequencing of gyrB gene of nine selected strains was performed using primers described by Parkinson et al. (2007). Obtained partial DNA sequences showed that these strains (GenBank nos.
MZ569011, MZ574079, MZ574080, MZ574081, MZ574082, MZ574083, MZ574084, MZ574085, and MZ574086) share 99.86 to 100% identity with Xe type strain ICPM:109 and 98.71 to 100% identity with Xe strain LMG930 isolated from pepper in the United States.
Pathogenicity was confirmed by spraying young pepper plants with bacterial suspension (108 CFU mL-1 in SDW). According to the reaction of cultivar Early Calwonder (ECW) and its isogenic lines (ECW-10R, ECW-20R and ECW-30R) all tested strains belong to Xe race P8. All strains grew on SPA plates amended with 100 ppm of copper sulfate, indicating high level of resistance in Xe Montenegrin population, while 25 ppm of streptomycin sulfate inhibited their growth.
These results confirmed presence of copper tolerant race of Xanthomonas euvesicatoria in pepper production in Montenegro.
Authors
J. Adamović, T. Popović, M. Pantelić, A. Obradović
Keywords
bacterial spot, identification, race, copper sulfate, streptomycin, resistance
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