To develop an efficient micropropagation protocol for Stevia rebaudiana
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Date
2010
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Publisher
CCSHAU
Abstract
The present investigation has been undertaken to: (i) study the effect of various
concentrations of different growth regulators on in vitro shoot multiplication and growth in Stevia
rebaudiana and (ii) to develop the micropropagation protocol for efficient and quick multiplication of
Stevia rebaudiana. The shoot-tip and nodal segments were used as explants. Different sterilization
protocols were tried for surface sterilization of the explants in Stevia rebaudiana. The nodal and shoottip
explants were surface sterilized with 0.2% bavistin for 20 min and 70% alcohol treatment for 50 sec
followed by 0.1% HgCl2 for 3 min which resulted in 100% sterilization of the explants. The nodal and
shoot-tip explants were best regenerated on MS medium+BAP 2.0mg/l. The per cent explants
regenerating shoots on nodal and shoot –tip explants were 90% and 50% respectively. The average no.
of shoots was 2.00 and 3.2 from nodal and shoot-tip explants respectively. The primary proliferated
cultures were transferred on to various MS media supplemented with different concentration and
combination of growth regulators for shoot multiplication. The best shoot multiplication medium was
MS medium supplemented with BAP (0.3mg/l) + Kinetin (0.3 mg/l), on which an average 50.20
number of shoots developed from nodal and shoot-tip explants after 40 days on culture medium. The
other growth regulators viz. zeatin, TDZ and GA3 were also tried to check the effect on in vitro shoot
multiplication at different interval of time but the response was not satisfactory. The multiple shoots
were surgically separated and transferred on to various rooting media. The frequency of root
regeneration was 100% on half MS medium containing IBA (0.5 mg/l) while the rooting frequency was
50.00% on half MS medium containing IAA (2.0 mg/l) and 42.86% on ½ MS+IAA (0.1 mg/l).
Regenerated plantlets were shifted to pots containing sterile soil and sand mixture (3:1) for
propagation. The survival percentage of plants after hardening and transfer to potted soil was 80%.
Description
Keywords
Planting, Stevia, Stevia rebaudiana, Regeneration, Cytokinins, Biological development, Vegetative propagation, Biological phenomena, Tissue culture, Concentrates