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Current Microbiology

, Volume 27, Issue 3, pp 137–142 | Cite as

Culture and serological detection of the xylem-limited bacterium causing citrus variegated chlorosis and its identification as a strain ofXylella fastidiosa

  • Chung J. Chang
  • Monique Garnier
  • Leyla Zreik
  • Victoria Rossetti
  • Joseph M. Bové
Article

Abstract

A xylem-limited bacterium resemblingXylella fastidiosa has been shown previously by electron mmcroscopy to be associated with citrus variegated chlorosis (CVC), a new disease of sweet organe tress in Brazil. A bacterium was consistently cultured from plant tissues from CVC twigs of sweet orange trees but not from tissues of healthy trees on several cell-free media known to support the growth ofXylella fastidiosa. Bacterial colonies typical ofX. fastidiosa became visible on PW, CS20, and PD2 agar media after 5 and 7–10 days of incubation, respectively. The cells of the CVC bacterium were rod-shaped, 1.4–3 μm in length, and 0.2–0.4 μm in diameter, with rippled walls. An antiserum against an isolate (8.1.b) of the bacterium gave strong positive reactions to double-antibody-sandwich (DAS), enzyme-linked immunosorbent assay (ELISA) with other cultured isolates from CVC citrus, as well as with several type strains ofX. fastidiosa. This result indicates that the CVC bacterium is a strain ofX. fastidiosa. ELISA was also highly positive with all leaves tested from CVC-affected shoots. Leaves from symptomless tress reacted negatively. Sweet organe seedlings inoculated with a pure culture of the CVC bacterium supported multiplication of the bacterium, which became systemic with 6 months after inoculation and could be reisolated from the inoculated seedlings. Symptoms characteristic of CVC developed 9 months post inoculation.

Keywords

Sweet Orange Optical Density Current Microbiology Alkaline Phosphatase Conjugate Xylella Fastidiosa 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Literature Cited

  1. 1.
    Ball EM, Hampton RO, De Boer SH, Schaad NW (1990) Polyclonal antibodies. In: Hampton R, Ball E., de Boer S. (eds) Serological methods for detection and identification of viral and bacterial plant pathogens. St. Paul, Minnesota, APS Press, pp. 33–54Google Scholar
  2. 2.
    Beretta MJ, Lee RF, Derrik KS, Davis CL, Branlsky RH (1992) Culture and serology of aXylella fastidiosa associated with citrus variegated chlorosis in Brazil. Abstract of the VII International Citrus Congress, Acireale (Italy) march 8–13, 1992Google Scholar
  3. 3.
    Chagas CM, Rossetti V, Beretta MJG (1992) Electron microscopy studies of a xylem-limited bacterium in sweet orange affected with citrus variegated chlorosis disease in Brazil. J Phytopathol 134:306–312CrossRefGoogle Scholar
  4. 4.
    Chang CJ, Walker TJ (1988) Bacterial leaf scorch of northern red oak: isolation, cultivation and pathogenicity of xylemlimited bacterium. Plant Dis 72:730–733CrossRefGoogle Scholar
  5. 5.
    Chen J, Chang CJ, Jarret RL, Gawel N (1992) Genetic variation amongXylella fastidiosa strains. Phytopathology 82:973–977CrossRefGoogle Scholar
  6. 6.
    Clark MF, Adams AN (1977) Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. J Gen Virol 34:475–483PubMedCrossRefGoogle Scholar
  7. 7.
    Davis MJ, Purcell AH, Thomson SV (1980) Isolation media for the Pierce's disease bacterium. Phytopathology 70:425–429CrossRefGoogle Scholar
  8. 8.
    Davis MJ, French WJ, Schaad NW (1981) Axenic culture of the bacteria associated with phony disease of peach and plum leaf scald. Curr Microbiol 6:309–314CrossRefGoogle Scholar
  9. 9.
    Hopkins DL (1989)Xylella fastidiosa: xylem-limited bacterial pathogen of plants. Annu Rev Phytopathol 27:271–290CrossRefGoogle Scholar
  10. 10.
    Kamper SM, French WJ, de Kloet SR (1985) Genetic relationships of some fastidious xylem-limited bacteria. Int J Syst Bacteriol 35:185–188CrossRefGoogle Scholar
  11. 11.
    Leite RP. RMVBC Leite (1991) Associaçao deXylella fastidiosa com clorose variegada dos citrus. Abstract of the XIV Congresso Paulista de Fitopatologia. Summa Fitopathol, 18:1Google Scholar
  12. 12.
    Lowry RH, Rosebrough NJ, Farr AL, Randall NJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265PubMedGoogle Scholar
  13. 13.
    Mackenzie DJ (1990) Preparation of antibody enzyme conjugates. In: Hampton R, Ball E, de Boer S (eds) Serological methods for detection and identification of viral and bacterial plant pathogens. St. Paul, Minnesota, APS Press, pp 87–92Google Scholar
  14. 14.
    Neto R, Beriam LOS, Komori N (1991) Caracterizaçao de isolados deXylella fastidiosa de citros por electroforese em gel de poliacrilamida/SDS. Abstract of the XIV Congresso Paulista de Fitopatologia. Summa Phytopathol 18:72Google Scholar
  15. 15.
    Purcell AH (1989) Homopteran transmission of xylem-inhabiting bacteria. In: Advances in disease vector research, vol 6. New York: Springer Verlag, pp 243–263Google Scholar
  16. 16.
    Rossetti V, Garnier M, Bové JM, Beretta MJG, Teixeira ARR, Quaggio JA, De Negri JD (1990) Présence de bactéries dans le xylème d'orangers atteints de chlorose variégée, une nouvelle maladie des agrumes au Brésil, C R Séances Acad Sci [III] 310:345–349Google Scholar
  17. 17.
    Wells JM, Raju BC, Hung HY, Weisburg WG, Mandelco-Paul L, Brenner DJ (1987)Xylella fastidiosa gen. nov. sp. nov.: Gram-negative, xylem-limited, fastidious plant bacteria related to Xanthomonas spp. Int J Syst Microbiol 36:136–143Google Scholar

Copyright information

© Springer-Verlag New York Inc. 1993

Authors and Affiliations

  • Chung J. Chang
    • 1
  • Monique Garnier
    • 1
  • Leyla Zreik
    • 1
  • Victoria Rossetti
    • 1
  • Joseph M. Bové
    • 1
  1. 1.Laboratory of Cellular and Molecular BiologyINRA and University of Bordeaux IIVillenave d'Ornon CedexFrance

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