STUDIES ON IN VITRO MORPHOGENESIS IN Gentiana kurroo Royle - AN ENDANGERED MEDICINAL PLANT
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Date
2014
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Abstract
ABSTRACT
Gentiana kurroo Royle is an endangered medicinal plant and hence an urgent need arises to propagate
and conserve this species. Protocol was developed for in vitro morphogenesis through direct and indirect
somatic embryogenesis and organogenesis by providing different growth regulators and culture conditions. The
morphogenic response of different explants (leaf, petiole and root) varied for callus induction according to
concentration of NAA and BA. The petiole explants were best responding for callus induction on MS
basal+1.00 mg/l BA and 3.00 mg/l NAA. Petiole, leaf and petiole derived callus was cultured on MS medium
supplemented with different growth regulators for induction of somatic embryogenesis directly and indirectly.
Somatic embryos were observed from both direct and indirect method. In direct pathway somatic embryos were
in contact with maternal tissue in a suspensor like structure. In indirect pathway, the explants first proliferated to
give rise to callus before embryoids were induced. The best performance was observed on MS basal medium
supplemented with 1.00 mg/l dicamba. Maturation of somatic embryos was observed on MS basal+0.50 mg/l
GA3 and the maximum plantlets formation was achieved when cotyledonary-stage somatic embryos were
shifted to half strength MS basal medium. MS medium supplemented with 0.10 mg/l NAA+0.75mg/l TDZ was
recorded as the best medium for direct regeneration. However, for indirect regeneration the maximum number
of shoot emergence was observed on MS basal fortified with 0.10 mg/l NAA and 1.00 mg/l TDZ. Half strength
MS basal supplemented with 1.00 mg/l IBA gave best response for root induction. Subsequently, the plantlets
were transferred to different potting mixtures and 100% survival rate was recorded on autoclaved cocopeat
which has been recorded to be the best of all other potting mixtures. For encapsulation shoot tips and somatic
embryos were excised from in vitro stock cultures, were encapsulated into beads. The beads were dehydrated by
air drying in laminar-flow chamber to reduce the water content, followed by direct immersion in liquid nitrogen
in cryovials for one month. Frozen beads were quickly thawed in water bath at 38oC for 3 minutes and cultured
on MS medium fortified with 0.50 mg/l BA and 0.50 mg/l Kn for shoot retrieval. A maximum of 53.30 % shoot
retrieval was recorded. For checking the genetic fidelity of the in vitro raised plants a total of four RAPD and
four ISSR markers were used and no morphological variations were recorded in the in vitro raised plants.
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Keywords
yields, heterosis, hybrids, planting, crossing over, genotypes, grain, diseases, biological phenomena, plant oils, Gentiana kurroo Royle ,in vitro morphogenesis,embryogenesis and organogenesis