Abstract:
For half a century, the limitations of obtaining cross-combinations in lilies because of
the incompatibility and incongruity between different varieties have been known. Somatic
hybridization is one of the most powerful tools for achieving distant interspecific hybrids.
For this purpose, protoplast preparation is a first and important step in efficient system
for the regeneration of plants from protoplasts. Protoplast isolation method was
previously developed in Lilium ledebourii (Baker) Boiss. In this study, several valuable
experiments were done based on completely randomized design with 3 replications and
also each experiment was repeated twice. The results revealed that cell wall and colony
formation were better in a liquid medium than those on a semi-solid medium. The highest
plating efficiency (1.34×106
per gr FW) and callus formation was obtained by using a
medium containing 1 mg L-1 2,4-D, 0.2 mg L-1 Kin and 2 g L-1 Yeast extract. Micro calli
were formed after one month of culture. Many plantlets were formed on the calli after
transfer of the proliferated calli to regeneration medium. The highest plantlet
regeneration (91.66%) was obtained by using a medium containing 0.5 mg L-1 NAA, 1.5
mg L-1 BA. Means comparison revealed that the semi- solid MS medium containing 0.5
mg L-1 NAA and 1.5 mg L-1 BA had the highest percentage of regeneration (91.66%), bulb
number (8.83), and length (0.7366 cm), root length (0.421cm) and leaf number (13.66) and
length (0.5052cm).
Keywords: Callus formation, In-vitro culture, Medium, Proliferated calli.
