For establishing aseptic cultures of pepper growing in vitro,
dry mature seeds were surface sterilized according to method
described by El Nagar, 2012. Sodium hypochlorite a common
disinfectant surface sterilize plant tissues was used. Seeds of
the pepper genotypes were immersed in a 2.5% sodium hypochlorite
for 10 min which is present in commercial bleach
solutions (Clorix). Then they were rinsed five times with sterile
distilled water for 10 min each. During immersion and rinsing
the solution was stirred on a shaker at 200 rpm under the
laminar air flow hood. The sterilized seeds were placed into
sterile tissue culture jars (30 seeds/jar) containing a half concentrated
basal MS medium (Murashige and Skoog 1962)
supplemented with B5 vitamins (Gamborg et al.,1968), 3.0%
sucrose and solidified with 0.7% Oxoid-Agar. The medium
was adjusted to pH 5.8 before autoclaving at 121°C and 1.2
kg/cm2 to 1.3 kg/cm2 pressure for 20 min. All cultures were
incubated at 25°C ±1°C under florescent light (2000 LUX) and
a 16 h photoperiod.