Abstract
Witches’ broom disease of small-fruited lime (WBDL) is a severe disease associated with phytoplasmas in the south of Iran. In
this study maintenance of WBDL phytoplasma in key lime (Citrus aurantifolia Christm) in micropropagation is reported. After
the maintenance of tissue cultures in a growth chamber, symptoms of little leaves and shortening of internodes were observed in
shoots from infected plants. . Micropropagated shoots that developed in tissue culture were tested for the presence of phytoplasmas
using the two phytoplasma-specific primers pair P1/P7 and P3/P7. Amplification by polymerase chain reaction confirms the
presence of WBDL phytoplasmas in the diseased plants 10 months after the beginning of the in vitro propagation. To our knowledge,
this is the first report on maintenance of lime witches’ broom phytoplasma in key lime shoots through tissue culture in Iran.