title: Culture and serological detection of the xylem-limited bacterium causing citrus variegated chlorosis and its identification as a strain ofXylella fastidiosa.
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.
authors.affiliation: Laboratory of Cellular and Molecular Biology, INRA and University of Bordeaux II, Villenave d'Ornon Cedex, France.
authors.forename: C J
authors.initials: CJ
authors.lastname: Chang
authors.forename: M
authors.initials: M
authors.lastname: Garnier
authors.forename: L
authors.initials: L
authors.lastname: Zreik
authors.forename: V
authors.initials: V
authors.lastname: Rossetti
authors.forename: J M
authors.initials: JM
authors.lastname: Bové