Skip to main content
Log in

The use of subtractive hybridization to obtain a DNA probe specific for Pseudomonas solanacearum race 3

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

Pseudomonas solanacearum, the causal agent of bacterial wilt, has been classified into three races based on host range and into five biovars based on physiological properties. Strains of race 3 belong exclusively to biovar 2 and primarily affect potatoes. Although this race is thought to have originated in the Andean highlands, it has unusual physiological properties that make it a potential threat to potatoes grown at the cooler latitudes worldwide. Consequently, there is need for a rapid and sensitive method for detection of race 3 strains. We have used subtractive hybridization to enrich for race 3-specific DNA sequences in total race 3 genomic DNA, and thereby obtained a 2 kb clone homologous to DNA from all 28 race 3 strains tested, but with only five of 90 non-race 3 strains. In addition, two larger regions of the genome, containing a minimum of 23 kb of DNA, are also specific for race 3. Deletion of this DNA did not affect virulence. This race 3-specific DNA is a potentially useful diagnostic tool for the detection of race 3 strains.

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

  • Amasino RM (1986) Acceleration of nucleic acid hybridization rate by polyethylene glycol. Anal Biochem 152:304–345

    Google Scholar 

  • Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1988) Current Protocols in Molecular Biology. John Wiley and Sons, New York

    Google Scholar 

  • Barker RH, Suebsaeng L, Rooney W, Alicrim GC, Dourado HV, Wirth DF (1986) Specific DNA probe for the diagnosis of Plasmodium falciparum malaria. Science 231:1434–1436

    Google Scholar 

  • Boucher CA, Barberis PA, Arlat M (1988) Acridine orange selects for deletion of hrp genes in all races of Pseudomonas solanacearum. Mol Plant-Microbe Interact 1:282–288

    Google Scholar 

  • Buddenhagen I, Sequeira L, Kelman A (1962) Designation of races in Pseudomonas solanacearum. Phytopathology 52:726

    Google Scholar 

  • Buddenhagen I (1985) Bacterial wilt revisited. In: Persley GJ (ed) Bacterial Wilt Disease in Asian and the South Pacific, ACIAR Proceedings No 13. ACIAR, Canberra, pp 126–143

    Google Scholar 

  • Ciampi L, Sequeira L (1980) Influence of temperature on virulence of race 3 strains of Pseudomonas solanacearum. Am Potato J 57:307–317

    Google Scholar 

  • Cook D, Barlow L, Sequeira L (1989) Genetic diversity of Pseudomonas solanacearum: detection of restriction fragment length polymorphisms with DNA probes that specify virulence and the hypersensitive response. Mol Plant-Microbe Interact 2:113–121

    Google Scholar 

  • Coplin DL, Cook D (1990) Molecular genetics of extracellular polysaccharide biosynthesis in vascular phytopathogenic bacteria. Mol Plant-Microbe Interact, in press

  • Harris D (1972) Intra-specific variation in Pseudomonas solanacearum. In: Proceedings of the third international conference of plant pathogenic bacteria, Wageningen pp 289–292

  • Hayward A (1964) Characteristics of Pseudomonas solanacearum. J Appl Bact 27:265–277

    Google Scholar 

  • Hendrick CA, Sequeira L (1984) Lipopolysaccharide-defective mutants of the wilt pathogen Pseudomonas solanacearum. Appl Environ Microbiol 48:389–395

    Google Scholar 

  • Hyman HC, Yogev D, Razin S (1987) DNA probes for detection and identification of Mycoplasma pneumoniae and Mycoplasma genitalium. J Clin Microbiol 25:726–728

    Google Scholar 

  • Johansen IE, Rasmussen OF, Heide M (1989) Specific identification of Clavibacter michiganense subsp. sepedonicum by DNA-hybridization probes. Phytopathology 79:1019–1023

    Google Scholar 

  • Kelman A (1954) The relationship of pathogenicity of Pseudomonas solanacearum to colony appearance on a tetrazolium medium. Phytopathology 44:693–695

    Google Scholar 

  • Kelman A, Hruschka J (1973) The role of motility and aerotaxis in the selective increase of avirulent bacteria in still broth cultures of Pseudomonas solanacearum. J Gen Microbiol 76:177–188

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular Cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Martin C, French ER, Nydegger U (1981) Bacterial wilt of potatoes in the Amazon basin. Plant Disease 65:246–248

    Google Scholar 

  • Miller JH (1972) Experiments in Molecular Genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Morton D, Dukes P, Jenkins S (1966) Serological relationships of races 1, 2 and 3 of Pseudomonas solanacearum. Plant Disease Reporter 50:275–277

    Google Scholar 

  • Olson K (1976) Overwintering of Pseudomonas solanacearum in Sweden. In: Sequeira L, Kelman A (eds) Planning Conference and Workship on the Ecology and Control of Bacterial Wilt Caused by Pseudomonas solanacearum. North Carolina State University, Raleigh, pp 105–109

    Google Scholar 

  • Seneviratne A (1969) On the occurrence of Pseudomonas solanacearum in the hill country of Ceylon. J Hort Sci 44:393–402

    Google Scholar 

  • Thurston HD (1963) Bacterial wilt of potatoes in Colombia. Am Potato J 40:381–390

    Google Scholar 

  • Wood PK, Morrie JG, Small PLC, Sethabutr O, Regina M, Trabulsi L, Kaper JB (1986) Comparison of DNA probes and the Sereny test for identification of invasive Shigella and Escherichia coli strains. J Clin Microbiol 24:498–500

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by J. Schell

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cook, D., Sequeira, L. The use of subtractive hybridization to obtain a DNA probe specific for Pseudomonas solanacearum race 3. Molec. Gen. Genet. 227, 401–410 (1991). https://doi.org/10.1007/BF00273930

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00273930

Key words

Navigation