The flower bract of 28-day-old, greenhouse grown curcuma
plants was used for the in planta assay. Approximately
2 × 108 CFU/ml (were added with 0.001% Tween 20) of bacterial
cells and 107 cells/ml. of Colletotrichum sp. each were
prepared for the in planta test. The curcuma bracts were first
wounded using the prick technique described by May et al. [9].
The experiment was divided into two main components; (1) for
the single treatment experiment, a fungal cell suspension was
mixed at a ratio of 1:1 with the bacterial cells of interest and a
40-μl aliquot of this suspension was inoculated into the wound.
The two controls contained wounded bract with and without
inoculation by the fungus cell suspension. (2) In the second
component (the serial treatment), the fungal cell suspension was
first inoculated into the prick wound and then followed by
spraying the bacterial cells onto the wound separately every
3 days for 10 days. The development of spot lesions was
recorded daily during the 10 days of the experiment. In all
treatments, the bracts were wrapped with a transparent plastic