Preparation and maintenance of multiple meristem
cultures. Banana (Musa acuminata AAA cv. ‘Grand Naine’)
meristem cultures were initiated from suckers collected from
the farmer's field. In the laboratory, suckers were trimmed to
3–4-cm3 pieces containing the shoot meristem. They were
surface sterilized with 0.1% mercuric chloride (v/v) for 8 min
and washed several times with sterile distilled water. Leaf
sheaths were removed and trimmed into 0.5–1-cm3 pieces
containing meristems in a laminar flow hood. Sterile meristems
were then inoculated on to Murashige and Skoog (MS)
medium (Murashige and Skoog 1962) supplemented with
13.31 μM 6-benzylaminopurine (BA), 30 g/L sucrose and
7 g/L agar. The pH of the media was adjusted to 5.7 using
1 M NaOH or HCl, autoclaved at 121°C for 20 min and
poured into 25×150-mm culture tubes. Cultures were incubated
at 27°C under 50–60 μE m−2 s
−1 light intensity at 16-h
light/8-h dark photoperiod. Shoot multiplication was
performed by splitting the shoot clumps into two to three
pieces and inoculating onto fresh medium. Multiple meristem
cultures were regularly sub-cultured at 4-wk intervals.