Somaclonal variation was induced using in vitro micropropagated
plants. Microplantlets were regenerated through different
tissue culture techniques including micropropagation, direct shoot
organogenesis from leaf, indirect organogenesis via callus culture
and somatic embryogenesis (Fig. 2). Seventy five explants were
used of each regeneration treatments. Our results showed that
maximum number of plants regenerated from meristem culture.
Direct organogenesis using cut leaf segments produced the lowest
number of regenerants (Fig. 3). However, there was no statistical
difference in regeneration of plants between the meristem culture
and micropropagation. These treatments were however significantly
different from others (leaf organogenesis, callus culture and
somatic embryogenesis). There was also no difference in regeneration
from callus culture or via somatic embryogenesis. The
regenerated plantlets were subsequently evaluated for somaclonal
variation under field conditions. Qualitative and quantitative traits
observed in the putative somaclones and quantitative data are
summarized in Table 2. In most cases somaclones were more
vigorous than control. However, different morphological characters
differed in the different populations and there was no set
trend. We observed several phenotypic differences in the in vitro
derived plants such as leaf shape in most plants regenerated from
the twelfth subculture, direct regeneration from leaves, callus
culture or somatic embryogenesis (Fig. 4H–N). Leaf petiole was
shorter and thicker and lamina was comparatively bigger than
control. Most of the leaves were lighter green and number of leaves
was less than control. Plants regenerated via somatic embryogenesis
had a significantly bigger canopy size while numbers of
runners were more in plants derived from meristem culture
(Fig. 4A–G). All the in vitro derived plants were late to flower when
compared to control. However, most of the in vitro plants had
significantly more flowers per plant and number of fruits. In