Molecular and General Genetics MGG

, Volume 218, Issue 1, pp 127–136 | Cite as

Genetic and structural characterization of the avirulence gene avrBs3 from Xanthomonas campestris pv. vesicatoria

  • Ulla Bonas
  • Robert E. Stall
  • Brian Staskawicz


The avirulence gene avrBs3 from Xanthomonas campestris pv. vesicatoria was cloned and found to be localized on a self-transmissable plasmid. Genetic analysis of an avrBs3 insertion mutation revealed that avrBs3 constitutes a single locus, specifying the resistant phenotype on pepper plants. Southern blot experiments showed that no DNA sequences homologous to avrBs3 were present in other races of X. c. pv. vesicatoria, which are unable to induce a hypersensitive reaction on ECW-30R. However, the DNA of several different pathovars of X. campestris hybridized to the avrBs3 probe. A deletion analysis defined a region of 3.6–3.7 kb essential for avrBs3 activity. The nucleotide sequence of this region was determined. A 3561 nucleotide open reading frame (ORF1), encoding a 125000 dalton protein, was found in the 3.7 kb region that was sufficient for avrBs3 activity. A second long ORF (2351 nucleotides) was identified on the other strand. A remarkable feature of both ORFs is the presence of 17 direct repeats of 102 bp which share 91%–100% homology with each other.

Key words

Xanthomonas campestris Avirulence gene Disease resistance DNA sequence Deletion analysis 


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  1. Anderson S (1981) Shotgun DNA sequencing using cloned DNa-se I-generated fragments. Nucleic Acids Res 9:3015–3027Google Scholar
  2. Aota S, Gojobori T, Ishibashi F, Maruyama T, Ikemura T (1988) Codon usage tabulated from the GenBank Genetic Sequence Data. Nucleic Acids Res 16 Suppl:r315–402Google Scholar
  3. Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523Google Scholar
  4. Boyer HW, Roulland-Dussoix D (1969) A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol 41:459–472Google Scholar
  5. Chen J, Varner JE (1985) An extracellular matrix protein in plants: characterization of a genomic clone for carrot extensin. EMBO J 4:2145–2151Google Scholar
  6. Chou PY, Fasman GD (1974) Prediction of protein conformation. Biochemistry 13:222–245Google Scholar
  7. Cook AA, Stall RE (1963) Inheritance of resistance in pepper to bacterial spot. Phytopathology 53:1060–1062Google Scholar
  8. Daniels MJ, Barber CE, Turner DC, Cleary SG, Sawzyc MK (1984) Isolation of mutants of Xanthomonas campestris pv. campestris showing altered pathogenicity. J Gen Microbiol 130:2447–2455Google Scholar
  9. Day PR (1974) Genetics of host-parasite interaction. WH Freeman and Co, San Francisco, pp 1–238Google Scholar
  10. Ditta G, Stanfield S, Corbin D, Helinski D (1980) Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 77:7347–7351Google Scholar
  11. Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13Google Scholar
  12. Figurski D, Helinski DR (1979) Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci USA 76:1648–1652Google Scholar
  13. Gabriel DW, Burges A, Lazlo GR (1986) Gene-for-gene interactions of five cloned avirulence genes from Xanthomonas campestris pv. malvacearum with specific resistance genes in cotton. Proc Natl Acad Sci USA 83:6415–6419Google Scholar
  14. Green RL, Warren GJ (1985) Physical and functional repetition in a bacterial ice nucleation gene. Nature 317:645–648Google Scholar
  15. Hibberd AM, Stall RE, Bassett MJ (1987) Allelism tests of three dominant genes for hypersensitive resistance to bacterial spot of pepper. Phytopathology 77:1304–1307Google Scholar
  16. Kim BS, Hartmann RW (1985) Inheritance of a gene (Bs3) conferring hypersensitive resistance to Xanthomonas campestris pv. vesicatoria in pepper (Capsicum annuum). Plant Disease 69:233–235Google Scholar
  17. Ludwig A, Jarchau T, Benz R, Goebel W (1988) The repeat domain of Escherichia coli haemolysin (HlyA) is responsible for its Ca2+-dependent binding to erythrocytes. Mol Gen Genet 214:553–561Google Scholar
  18. Maclan DJ, Sargent JA, Tommerup IC Ingram DS (1974) Hypersensitivity as a primary event in resistance to fungal parasites. Nature 249:186–187Google Scholar
  19. Martin R (1987) Overcoming DNA sequencing artifacts: stops and compressions. BRL Focus 9:8–10Google Scholar
  20. Maxam AM, Gilbert W (1980) Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol 65:499–560Google Scholar
  21. Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New YorkGoogle Scholar
  22. Napoli C, Staskawicz BJ (1987) Molecular characterization of an avirulence gene from Race 6 of Pseudomonas syringae pv. glycinea. J Bacteriol 169:572–578Google Scholar
  23. Prentki P, Krisch HM (1984) In vitro insertional mutagenesis with a selectable DNA fragment. Gene 29:303–313Google Scholar
  24. Ronald PC, Staskawicz BJ (1988) The avirulence gene avrBs1 from Xanthomonas campestris pv. vesicatoria encodes a 50-kD protein. Mol Plant-Microbe Interact 1:191–198Google Scholar
  25. Sanger, F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467Google Scholar
  26. Schneider A, Hemphill A, Wyler T, Seebeck T (1988) Large microtubule-associated protein of T. brucei has tandemly repeated, near identical sequences. Science 241:459–462Google Scholar
  27. Sequeira L (1984) Plant bacterial interactions. In: Linskens AF, Heslop-Harrison J (eds) Encyclopedia of plant physiology, New Ser, vol. 7. Springer, Berlin, pp 187–211Google Scholar
  28. Shine J, Dalgarno L (1974) The 3′-terminal sequence of Escherichia coli, 16S ribosomal RNA: complementary to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci USA 71:1342–1346Google Scholar
  29. Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517Google Scholar
  30. Speicher DW, Marchesi VT (1984) Erythrocyte spectrin is comprised of many homologous triple helical segments. Nature 311:177–180Google Scholar
  31. Stachel SE, An G, Flores C, Nester EW (1985) A Tn3 lacZ transposon for the random generation of β-galactosidase gene fusion: application to the analysis of gene expression in Agrobacterium. EMBO J 4:891–898Google Scholar
  32. Stall RE, Cook AA (1966) Multiplication of Xanthomonas vesicatoria and lesion development in resistant and susceptible pepper. Phytopathology 56:1152–1154Google Scholar
  33. Staskawicz BJ, Dahlbeck D, Keen N (1984) Cloned avirulence gene of Pseudomonas syringae pv. glycinea determines race-specific incompatibility on Glycine max (L.) Merr. Proc. Natl. Acad Sci USA 81:6024–6028Google Scholar
  34. Staskawicz B, Dahlbeck D, Keen N, Napoli C (1987) Molecular characterization of cloned avirulence genes from race 0 and race 1 of Pseudomonas syringae pv glycinea. J Bacteriol 169:5789–5794Google Scholar
  35. Swanson J, Kearney B, Dahlbeck D, Staskawicz B (1988) Cloned avirulence gene of Xanthomonas campestris pv. vesicatoria complements spontaneous race-change mutants. Mol Plant-Microbe Interact 1:5–9Google Scholar
  36. Tamaki S, Dahlbeck D, Staskawicz B Keen NT (1988) Characterization and expression of two avirulence genes cloned from Pseudomonas syringae pv. glycinea. J Bacteriol 170:4846–4854Google Scholar
  37. Vieira J, Messing J (1987) Production of single-stranded plasmid DNA. Methods Enzymol 153:3–11Google Scholar
  38. Wahl GM, Stern M, Stark GR (1979) Efficient transfer of large DNA fragments from agarose to diazobenzoyloxymethyl paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci USA 76:3683–3687Google Scholar
  39. Whalen MC, Stall RE, Staskawicz BJ (1988) Characterization of a gene from a tomato pathogen determining hypersensitive resistance in non-host species and genetic analysis of this resistance in bean. Proc Natl Acad Sci USA 85:6743–6747Google Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • Ulla Bonas
    • 1
  • Robert E. Stall
    • 2
  • Brian Staskawicz
    • 1
  1. 1.Department of Plant PathologyUniversity of CaliforniaBerkeleyUSA
  2. 2.Department of Plant PathologyUniversity of FloridaGainesvilleUSA

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