Detection of Antagonistic activities in planta: The
flower bract of 28 day-old, greenhouse grown curcuma
plants was used for the in planta assay. Initially, each
leaf was cleaned with 70% ethyl alcohol to form a
surface sterile flower for the microorganisms. On the
order of 2x108
CFU/ml (were added with 0.001% tween
20) of bacterial cells and 107
cells/ml. of C. musae each
were prepared for the test. The curcuma bracts were
each wounded in three regions with a total of ten
pinpricks using the technique described by[25]. Using a
fungal cell suspension at a ratio of 1:1 with the bacterial
species of interest, a 40µl aliquot of this suspension was
inoculated into the wound. The two controls contained
wounded bract with and without inoculation by the
fungus cell suspension. The development of spot
lesions was recorded daily during the 10 days of the
experiment where the total number of infected leaves
was recorded to determine the disease incidence as the
percentage of leaves infected. In all treatments, the
bracts were wrapped with a transparent plastic bag after
inoculation and secured within the greenhouse to
minimize coinfection by unknown bacteria. Each
treatment was run with 3 replicates in which twenty
curcuma bracts were used for each replicate.