Improved PCR for identification of members of the genus Xanthomonas

Adriko, John; Mbega, Ernest R.; Mortensen, Christian; Wulff, E. G.; Tushemereirwe, Wilberforce; Kubiriba, Jerome; Lund, Ole

doi:10.1007/s10658-013-0329-x

Cited by 26 publications

(14 citation statements)

References 37 publications

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“…phaseoli (ICMP 22099). Bacterial isolates, as those originally inoculated, were consistently reisolated from inoculated plants with symptoms on YPGA medium and their identity was confirmed by using Xanthomonas ‐specific PCR primers (Table ; Adriko et al ., ) as described above.…”

Section: Resultsmentioning

confidence: 99%

“…Koch's postulates were accomplished by reisolating the inoculated isolates on YPGA from the inoculated plants showing symptoms. Confirmation of the reisolated bacteria was made using Xanthomonas ‐specific PCR primers X‐ITS‐F3j/X‐ITS‐F3k/X‐ITS‐F3c/X‐ITS‐F3d/X‐ITS‐R2 (Table ; Adriko et al ., ). The pathogenicity tests were repeated twice.…”

Section: Methodsmentioning

confidence: 97%

“…All isolates were tested using the generic primers for Xanthomonas spp. (Table ; Adriko et al ., ) and the specific ones for X. euvesicatoria , X. vesicatoria , X. perforans , X. gardneri (Table ; Koenraadt et al ., ), and X. axonopodis pv . phaseoli (Table ; Audy et al ., ).…”

Section: Methodsmentioning

confidence: 99%

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Monitoring the occurrence of tomato bacterial spot and range of the causal agent Xanthomonas perforans in Iran

et al. 2016

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A 2-year comprehensive field survey was conducted across major tomato-growing areas of Iran. Two hundred and thirty-four tomato fields and six tomato-producing greenhouses were surveyed for the potential presence of bacterial spot disease. Five hundred and ninety-six tomato samples with and without symptoms were analysed. While Xanthomonas spp. were found in association with tomato plants both with and without symptoms from five surveyed counties, the bacterial spot disease was observed only in plants from three of them. Only strains isolated from plants with symptoms induced disease symptoms on tomato, while those isolated from symptomless plants caused symptoms only on cabbage and common bean. None of the isolates caused disease symptoms on pepper and eggplant. Phylogenetic analysis showed that X. perforans is the causal agent of tomato bacterial spot in Iran, although X. campestris and X. axonopodis were also associated with symptomless tomato plants. All X. perforans isolates in this study were sensitive to streptomycin, copper sulphate and copper oxychloride at concentrations of 50 mg L À1 , 200 mg L À1 and 0.8 g L À1 , respectively. Unlike the type strain of X. perforans, isolates in this study did not produce bacteriocin against other Xanthomonas spp., nor were they detected using the usual species-specific primer pair Bs-XpF/Bs-XpR. This suggests an atypical nature of X. perforans strains in Iran, which leads to the hypothesis that X. perforans strains in Iran may have a separate origin to those causing disease epidemics elsewhere. The aggregated dispersal pattern of the diseased tomato fields signifies the seedborne introduction of the pathogen into the country.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 97%

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Monitoring the occurrence of tomato bacterial spot and range of the causal agent Xanthomonas perforans in Iran

et al. 2016

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“…Cycling conditions were modified from the original publication to simplify the original two-step scheme: 95°C 5 min, 359 (95°C 30 s, 68°C 90 s, 72°C 30 s), 68°C 10 min. PCR runs involving plant extracts were verified in parallel reactions with an additional internal positive control primer pair (IPC), targeting plant 26S mitochondrial rDNA (primers M26SF4/M26SR3 of Adriko et al, 2014; at 0.1 lM each) and amplifying a 754-bp DNA fragment. Validation of the different PCR tests was performed according to the requirements specified in EPPO Standard PM 7/ 98 (2) Specific requirements for laboratories preparing accreditation for a plant pest diagnostic activity (EPPO, 2014).…”

Section: Pcr Tests and Validationmentioning

confidence: 99%

Validation of a specific PCR screening test for Pantoea stewartii subsp. stewartii in maize (Zea mays) samples

et al. 2018

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Validation data is presented for a conventional PCR test that specifically detects the quarantine pathogen Pantoea stewartii subsp. stewartii in maize leaf and seed samples and does not cross‐react with the non‐pathogenic P. stewartii subsp. indologenes. The PCR tests currently recommended by the EPPO Diagnostic Protocol PM 7/60 (2) for initial screening bear the risk of false positives as they detect both subspecies of P. stewartii. The test presented here has high analytical sensitivity (102 cells mL–1 in leaf extracts, at least 103 cells mL−1 in seed extracts), high repeatability and high reproducibility and performed well on the two matrices tested: maize seed and leaf extract. Its improved analytical specificity and analytical sensitivity compared to the currently recommended tests lead the authors to suggest that this test should be included as a first screening test in the diagnostic protocol for P. stewartii subsp. stewartii when it is next revised.

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“…were identified using generic and specific primers for xanthomonads ( Table 2). The generic primers X-ITS-F3j, X-ITS-F3k, X-ITS-F3c, X-ITS-F3d and X-ITS-R2 (Table 2)capable to amplify all Xanthomonas species (Adriko et al 2014)were used for the identification of the gram-negative yellow-pigmented bacterial isolates to the genus level. Subsequently, we used species-specific primer pairs -Bs-XeF/Bs-XeR, Bs-XvF/ Bs-XvR, Bs-XgF/Bs-XgR and Bs-XpF/Bs-XpR (Table 2) for X. euvesicatoria, X. vesicatoria, X. gardneri and X. perforans, respectivelyto determine the identity of the causal agent to the species level (Koenraadt et al 2009).…”

Section: Genus-and Species-specific Polymerase Chain Reactionmentioning

confidence: 99%

Occurrence and Characterization of the Bacterial Spot Pathogen Xanthomonas euvesicatoria on Pepper in Iran

Osdaghi

Taghavi

Hamzehzarghani

et al. 2016

Journal of Phytopathology

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We report in this study for the first time the occurrence of bacterial spot of pepper in Iran and both phenotypic and genetic characterization of its causal agent, Xanthomonas euvesicatoria. Pepper plants grown in 15 of 30 surveyed private gardens and commercial fields were infected by the pathogen in Marand County, East Azerbaijan Province, north‐western Iran. The obtained strains of X. euvesicatoria had different amylolytic and pectolytic activities compared with those reported for this species elsewhere. Pathogenicity tests showed that strains isolated from diseased pepper are able to infect tomato, in addition to pepper. Host range of the pathogen was assessed on eight annual plant species including crops and weeds by measuring the population dynamics. The host range assessment showed that in addition to pepper and tomato, known hosts of X. euvesicatoria, the Iranian strains were able to colonize a number of new hosts such as nightshade and common bean. In contrast, none of them were able to build up their population on cowpea, eggplant, bindweed and zucchini. All X. euvesicatoria strains obtained in this study were sensitive to copper sulphate and streptomycin at concentrations higher than 20 and 50 mg/l, respectively. Phylogenetic analyses of the strains using the sequences of gyrB and hrpB genes confirmed their species as X. euvesicatoria. Given a direct commercial trade of fresh solanaceous vegetables between Iran and Turkey, it is hypothesized that the pathogen entered north‐western Iran from eastern parts of Turkey through infected plant materials. Finally, the role of prevention – based on the use of healthy planting materials and resistant and/or tolerant plant varieties – to contain the potential disease epidemics is discussed.

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Improved PCR for identification of members of the genus Xanthomonas

Cited by 26 publications

References 37 publications

Monitoring the occurrence of tomato bacterial spot and range of the causal agent Xanthomonas perforans in Iran

Monitoring the occurrence of tomato bacterial spot and range of the causal agent Xanthomonas perforans in Iran

Validation of a specific PCR screening test for Pantoea stewartii subsp. stewartii in maize (Zea mays) samples

Occurrence and Characterization of the Bacterial Spot Pathogen Xanthomonas euvesicatoria on Pepper in Iran

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