Asian Journal of Microbiology, Biotechnology & Environmental Sciences Paper

Vol 08, Issue 2, 2006; Page No.(373-379)

ISOLATION AND DETECTION OF SPIROPLASMA CITRI THE CAUSAL AGENT OF CITRUS STUBBORN DISEASE IN EGYPT

AYMAN F. OMKAR, KAMARUZAMAN SIJAM, HABIBUDDIN H AND INON SULAIMAN

Abstract

Spiroplasma citri was isolated from common orange (Citrus sinensis) trees showing typical symptoms of citrus stubborn disease in Egypt. The organism was isolated from young leaves. Fried-egg shape colonies were obtained when S. citri was subcultured on C-3G solid medium. S. citri was detected using dark-field microscope, electron microscope, ELISA, and Dienes' stain. Small helical filamentous and small round bodies were observed by dark field microscope, whereas electron microscope inspection detected helical structure of the filamentous organism and vesicular blebs on the filaments. Moreover, electron microscope examination of ultra-thin sections of leaf midribs of stubborn infected-citrus plants revealed the presence of numerous polymorphic bodies of S. citri in the phloem tissue. The isolated organism was confirmed by a positive reaction in ELISA test. Dienes' stain was used successfully to detect plant disease infected by S. citri.

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