Since disinfection with hypochlorite was
found to be inadequate, antimicrobial substances
(antibiotic and fungicide) were
incorporated into the culture medium. After
standard decontamination, the explants
were inoculated on WPM [6] medium solidified
with 0.7% (w/v) agar (Sigma) and supplemented
with 3% sucrose and the combinations
of different concentrations of the
commercial fungicide Benlate 500 (50% ofbenomyl) at (0.0, 1.0, 2.0 or 4.0) g·L–1 and
ampicillin antibiotic at (0.0, 1.0, 2.0 or
4.0) mg·L–1. The benomyl was added to the
medium prior to autoclavation. The ampicillin
was filter-sterilized (Millipore 0.22 µm)
and introduced to the medium after autoclaving.