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


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.


Sweet Orange Optical Density Current Microbiology Alkaline Phosphatase Conjugate Xylella Fastidiosa 
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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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