Skip to main content
Log in

Rapd Pcr-based differentiation of Xanthomonas campestris pv. phaseoli and Xanthomonas campestris pv. phaseoli var. fuscans

  • Published:
European Journal of Plant Pathology Aims and scope Submit manuscript

Abstract

A RAPD PCR-based method was used to differentiate between isolates of Xanthomonas campestris pv. phaseoli and Xanthomonas campestris pv. phaseoli var. fuscans. Using random primer OP-G11, a single, high intensity band of 820 bp was amplified from DNAs of all X. c. pv. phaseoli var. fuscans isolates, while multiple amplification products of varying sizes were generated from X. c. pv. phaseoli DNAs. Whereas RAPD PCR differentiation gave an unambiguous result in under 4 h, standard differentiation by recording the production of a brown pigment by X. c. pv. phaseoli var. fuscans isolates took up to 7 days and showed variation both between isolates and between media. The unequivocal nature of the RAPD PCR method was demonstrated when isolate 408, originally classified as X. c. pv. phaseoli var. fuscans, failed to produce the 820 bp band typical of X. c. pv. phaseoli var. fuscans isolates, and after also failing to produce a brown pigment, was re-classified as X. c. pv. phaseoli.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Allen DJ (1983) The Pathology of Tropical Food Legumes. In: John Wiley (ed) Disease Resistance in Crop Improvement. Chichester, UK

    Google Scholar 

  • Audy P, Laroche A, Saindon G, Huang HC and Gilbertson L (1994) Detection of the bean common blight bacteria, Xanthomonas campestris pv. phaseoli and X. c. phaseoli var. fuscans, using the polymerase chain reaction. Phytopathology 84: 1185-1192

    Google Scholar 

  • Audy P, Braat CE, Saindon G, Huang HC and Laroche A (1996) A rapid and sensitive assay for concurrent detection of bacteria causing common and halo blights in bean seed. Phytopathology 86: 361-366

    Google Scholar 

  • Basu PK and Wallen VR (1967) Factors affecting virulence and pigment production of Xanthomonas phaseoli var. fuscans. Canadian Journal of Botany. 45: 2367-2374

    Google Scholar 

  • Burkholder WH and Bullard ET (1946) Varietal susceptibility of beans. Xanthomonas phaseoli var. fuscans. Plant Disease Reporter 30: 446-448

    Google Scholar 

  • Dye DW(1962) The inadequacy of the usual determination tests for the identification of Xanthomonas spp. New Zealand Journal of Science 5: 393-416

    Google Scholar 

  • Ekpo EJA and Saettler AW (1976) Pathogenic variation in Xanthomonas phaseoli and X. phaseoli var. fuscans. Plant Disease Reporter 60: 80-83

    Google Scholar 

  • El-Sharkawy TA and Huisingh D(1971) Differentiation among Xanthomonas species by polyacrylamide gel electrophoresis of soluble proteins. Journal of General Microbiology 68: 155-165

    Google Scholar 

  • Goodwin PH and Sopher CR (1994) Brown pigmentation of Xanthomonas campestris pv. phaseoli associated with homogentisic acid. Canadian Journal of Microbiology 40: 28-34

    Google Scholar 

  • Hildebrand DC, Palleroni NJ and Scheath MN (1990) Deoxyribonucleic acid relatedness of 24 xanthomonad isolates representing 23 Xanthomonas campestris pathovars and Xanthomonas fragariae. Journal of Applied Bacteriology 68: 263-269

    Google Scholar 

  • King EO, Ward MK and Raney DE (1954) Two simple media for the demonstration of pyocyanin and fluorescein. Journal of Laboratory and Clinical Medicine44: 301-307

    Google Scholar 

  • Lazo GR and Gabriel DW (1987) Conservation of plasmid DNA sequences and pathovar identification of isolates of Xanthomonas campestris. Phytopathology 77: 448-453

    Google Scholar 

  • Lazo GR, Roffey R and Gabriel DW (1987) Pathovars of Xanthomonas campestris are distinguishable by restriction fragmentlength polymorphism. International Journal of Systematic Bacteriology 37: 214-221

    Google Scholar 

  • Leakey CLA(1973) A note on Xanthomonas blight of beans (Phaseolus vulgaris (L.) Savi) and prospects for its control by breeding for tolerance. Euphytica 22: 132-140

    Google Scholar 

  • Maraite H (1989) Resistance genetique aux bacterioses des cultivures vivrieres en Afrique centrale (Genetic resistance of food crops to bacterial diseases inCentralAfrica).Abstracts field beans (Phaseolus vulgaris L.) 14: 40-41

    Google Scholar 

  • Opio AF, Allen DJ and Teri JM (1996) Pathogenic variation in Xanthomonas campestris pv. phaseoli, the causal agent of common bacterial blight in Phaseolus beans. Plant Pathology 45: 1126-1133

    Google Scholar 

  • Rudolph K (1990) Toxins as taxonomic features. Pages 251-267. In: Klement Z, Rudolph K and Sands DC (eds) Methods in Phytobacteriology. Akademia Kiado, Budapest, Hungary

    Google Scholar 

  • Sambrook J, Fritsch EF and ManiatisT (1989) In: Molecular cloning: a laboratory manual 2nd Ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. </del>

    Google Scholar 

  • Schuster ML and Coyne DP (1981) Biology, epidemiology, genetics and breeding for resistance to bacterial pathogens of Phaseolus vulgaris L. Horticultural Reviews 3: 28-58

    Google Scholar 

  • Vidaver AK (1967) Synthetic and complexmedia for the rapid detection of fluorescence of phytopathogenic pseudomonads: effects of the carbon source. Applied Microbiology 15: 1523-1524

    Google Scholar 

  • Vidaver AK (1993) Xanthomonas campestris pv. phaseoli: Cause of common bacterial blight of bean. Pages 40-44 In: Swings JG and Civerolo EL (eds) Xanthomonas. Chapman and Hall, London

    Google Scholar 

  • Wallen VR and Jackson HR (1975) Model for yield loss determination of bacterial blight of field beans utilizing aerial infrared photography combined with field plot studies. Phytopathology 65: 942-948

    Google Scholar 

  • Xue BG and Goodwin PH (1993) Amplified DNA polymorphisms of putitive two-component regulatory genes of several Xanthomonas campestris pathovars. Canadian Journal of Plant Pathology 16: 1-7

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Birch, P.R., Hyman, L.J., Taylor, R. et al. Rapd Pcr-based differentiation of Xanthomonas campestris pv. phaseoli and Xanthomonas campestris pv. phaseoli var. fuscans. European Journal of Plant Pathology 103, 809–814 (1997). https://doi.org/10.1023/A:1008698726790

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1008698726790

Navigation