Results
The medium successfully supported growth of all explants.
Phytoplasma-diseased plants showed significantly reduced shoot height, little leaf and witches’broom symptoms when compared to the healthy control
(figure 1). The best time for subculture was every six weeks. Jar
explants grew faster than the tube explants.
Amplification of two pathogen-specific DNA fragments
320 bp and 1,830 bp with two primer pairs P3/P7
and P1/P7, respectively by polymerase chain reaction
(PCR) confirm the presence of WBDL phytoplasma in
the diseased plants after 10 months of in vitro propagation
(figure 2).
healthy control. The shoot height and leaf width were
reduced by increasing interval subculture until 8 weeks,
however the witches’ broom symptom seems not to be
correlated with phytoplasma concentration.
The maintainance of phytoplasma through tissue culture
of diseased key lime can present a useful way to
maintain living phytoplasma for studies of phytoplasmaplant
interactions toward the understanding of phytoplasma
pathogenicity and also to study possibility of
their elimination by cryotherapy, or other in vitro techniques.