Detection of the plant pathogenic bacterium Xanthomonas campestris pv. Campestris in seed extracts of Brassica sp. Applying fluorescent antibodies and flow cytometry

Chitarra, L. G.; Langerak, C.J.; Bergervoet, J.H.W.; Bulk, R.W. van den

doi:10.1002/cyto.10058

Cited by 35 publications

(21 citation statements)

References 33 publications

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“…Examples are the development of easy-to-handle, field-compatible tests such as lateralflow (Delmulle et al 2005), but also more complex test set-ups like flow-cytometry (Chitarra et al 2002), and multiplex detection using Luminex (Rao et al 2004;Peters et al 2007) or protein chips (Taitt et al 2004). Use of recombinant antibodies, fused to specific tags, may facilitate multiplex design (Wingren and Borrebaeck 2006).…”

Section: Discussionmentioning

confidence: 99%

Stable recombinant alpaca antibodies for detection of Tulip virus X

Beekwilder¹,

Houwelingen²,

Beckhoven

et al. 2008

Eur J Plant Pathol

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For detection of the plant pathogenic Tulip virus X (TuVX), a panel of six recombinant antibodies was identified. To this end, a repertoire of variable domains from heavy-chain immunoglobulins (VHH) was cloned from an alpaca, which had been immunized with TuVX. Binding domains were selected by phage display and panning on immobilized TuVX particles. Recombinant VHH antibodies were tested for sensitivity in a sandwich ELISA, and were demonstrated to be readily able to distinguish TuVX-infected tulip leaf material from uninfected leafs. No cross-reactivity of the VHH antibodies to related flexiviridae was observed. Recombinant VHHs maintained their reactivity upon storage at −20°C for over a year. The effect of incubation at higher temperatures for prolonged time was studied. Two out of three VHH proteins retained activity after several weeks of storage at 37°C.

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Section: Discussionmentioning

confidence: 99%

Stable recombinant alpaca antibodies for detection of Tulip virus X

Beekwilder¹,

Houwelingen²,

Beckhoven

et al. 2008

Eur J Plant Pathol

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“…Therefore, planting seed free of bacterial contamination is an important disease management strategy. Several molecular and immunological methods, such as PCR (3,4,5,13,17,30), enzymelinked immunosorbent assay (1,35), and flow cytometry (7), were developed to test seeds for the presence of these pathogens. However, these methods cannot directly distinguish dead and viable cells and may vary in specificity and sensitivity according to seed and other sample types (plant tissue, soil, and water).…”

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confidence: 99%

Combination of Chromogenic Differential Medium and estA -Specific PCR for Isolation and Detection of Phytopathogenic Xanthomonas spp

Lee

Sung

Liu

et al. 2009

Appl Environ Microbiol

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A xanthomonad differential medium (designated Xan-D medium) was developed, on which streaks and colonies of xanthomonads, including 13 species of the genus Xanthomonas, turned wet-shining yellow-green and were surrounded with a smaller milky zone and a bigger clear zone in 3 to 4 days. The characteristics could easily be differentiated from those of yellow nonxanthomonads and other bacteria. The mechanism of color change and formation of a milky zone on the medium are mainly due to the Tween 80 hydrolytic capacity of xanthomonads. The gene, estA, responsible for Tween 80 hydrolysis was cloned and expressed in Escherichia coli, which acquired a capacity to hydrolyze Tween 80 and could turn green and form a milky zone on the Xan-D medium. The nucleotide sequence of estA is highly conserved in the xanthomonads, and the sequence was used to design a specific PCR primer set. The PCR amplification using the primer set amplified a 777-bp specific DNA fragment for all xanthomonad strains tested. The Xan-D medium was used to isolate and differentiate Xanthomonas campestris pv. campestris from naturally infected cabbages with black rot symptoms for a rapid diagnosis. All isolated X. campestris pv. campestris strains developed characteristic colonies and were positive in the PCR with the estA primer set. The Xan-D medium was further amended with antibiotics and successfully used for the detection of viable X. campestris pv. campestris cells from plant seeds. Although some yellow nonxanthomonads and other saprophytic bacteria from plant seeds could still grow on the medium, they did not interfere with the color development of X. campestris pv. campestris. However, Stenotrophomonas maltophilia, which is closely related to xanthomonads, existing in a seed lot could also develop yellow-green color but had different colony morphology and was negative in the PCR with the estA primer set. Accordingly, the combination of the Xan-D medium with the estA-specific PCR is a useful and reliable method for the isolation and detection of viable xanthomonad cells from plant materials.

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“…The isolate 3811 is the reference strain for Xcc race 1 and is often used in studies of Xcc identification, pathogenicity or diversity (Guy et al, 2013;Jensen et al, 2010;Vicente et al 2001). The isolate 1279A represents the reference strain for Xcc race 4 and is also used in scientific researches (Bila et al, 2013;Chitarra et al, 2002;Taylor et al, 2002). The mixture of all isolates showed lower symptoms than individual isolates in case of resistant 'Cerox' but similar values on susceptible 'Sonja' where reached mostly 4 th and 5 th degree.…”

Section: Discussionmentioning

confidence: 99%

Testing of Inoculation Methods and Susceptibility Testing of Perspective Cabbage Breeding Lines (Brassica Oleracea convar. Capitata) to the Black Rot Disease Caused by Xanthomonas Campestris pv. Campestris

Peňázová¹,

Kopta²,

Jurica³

et al. 2018

Acta Univ. Agric. Silvic. Mendelianae Brun.

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The susceptibility of twenty-four cabbage breeding lines to Xanthomonas campestris pv. campestris was evaluated. The selection of appropriate inoculation method was done on 4 cabbage cultivars ('Cerox', 'Sintex', 'Sonja' and 'Avak'). One month old plants were infected by 5 inoculation methods (spraying, injection by syringe, multiple pricking, carborundum abrasion and scissor clipping method). Four different bacterial isolates of Xcc (WHRI 3811, 3971A, 1279A; SU) and their mixture were evaluated for the aggressiveness on 'Cerox' and 'Sonja' cultivars. On the basis of obtained results, breeding lines of head cabbage were inoculated by mixture of all tested isolates using multiple pricking method. The disease severity of inoculated seedlings proved high susceptibility of young plants to the Xcc infection. The disease incidence determined 75 and 105 days after sowing showed changes for 16 of tested lines and indicated that resistance testing should be observed until mature stage. The study revealed five breeding lines (DP25, T1, IT10, Kalibos and Avak1) with disease incidence lower than 20 % as perspective sources of resistance for further breeding.

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Detection of the plant pathogenic bacterium Xanthomonas campestris pv. Campestris in seed extracts of Brassica sp. Applying fluorescent antibodies and flow cytometry

Cited by 35 publications

References 33 publications

Stable recombinant alpaca antibodies for detection of Tulip virus X

Stable recombinant alpaca antibodies for detection of Tulip virus X

Combination of Chromogenic Differential Medium and estA -Specific PCR for Isolation and Detection of Phytopathogenic Xanthomonas spp

Testing of Inoculation Methods and Susceptibility Testing of Perspective Cabbage Breeding Lines (Brassica Oleracea convar. Capitata) to the Black Rot Disease Caused by Xanthomonas Campestris pv. Campestris

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