Available online 4 May 2016
- a Plant Biotechnology Laboratory, Centre for Advanced Studies, Department of Botany, North-Eastern Hill University, Shillong 793022, Meghalaya, India
- b Research Centre for Plant Growth and Development, School of Life Sciences, University of KwaZulu-Natal, Pietermaritzburg, Private Bag X01, Scottsville 3209, South Africa
- Received 10 January 2016, Revised 12 March 2016, Accepted 11 April 2016, Available online 4 May 2016
Highlights
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- An efficient micropropagation protocol for Dendrobium crepidatum, a threatened, medicinal orchid with increased phytochemical yield and high genetic stability has been developed.
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- Genetic stability evaluated by SCoT and ISSR markers.
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- Higher antioxidant activity within the micropropagated plants over the mother plant.
Abstract
Orchids are one of the promising medicinal plant families of which Dendrobium crepidatum. Lindl & Paxton figures out prominently because of its multi directional medicinal attributes. In the present report, in vitro regeneration protocol has been developed from the nodal segments of D. crepidatum,
to cater sustainable commercial exploitation and conservation needs.
Thidiazuron (TDZ) at 3 mg/l singly resulted in the response frequency of
55% which could be increased to 97% by incorporating NAA at 0.5 mg/l
and TDZ at 2 mg/l in combination in the medium. Shoot induction rate was
further enhanced with the use of polyamines and at 0.8 mM putrescine
along with 2 mg/l TDZ and 0.5 mg/l NAA in the medium 11.8 shoots/explant
could be obtained. Highest rooting frequency of the shoots was achieved
in medium containing 2 mg/l IBA. Genetic stability of the acclimatized
plants was assessed using Start Codon Targeted (SCoT) polymorphism and
inters simple sequence repeats (ISSR). SCoT revealed a total variability
of 10% within the micropropagated plants whereas the cumulative ISSR
and SCoT data revealed 6.25% clonal variability indicating high genetic
fidelity amongst the regenerates. A comprehensive higher yield of the
secondary metabolites along with significant higher antioxidant
potentials as compared to the mother plant was revealed using DPPH and
FRAP assays. An increased regeneration frequency with a comparative
higher yield of secondary metabolite and genetic stability reported in
the present communication ensure the ingenuity of this clonal
propagation protocol developed for D. crepidatum which can be
further utilized in the sustainable commercial utilization and
conservation of other medicinally important orchid species.
Keywords
- Medicinal orchid;
- Traditional Chinese Medicine;
- Secondary metabolites;
- Natural antioxidants;
- Gene targeted markers
© 2016 Elsevier GmbH. All rights reserved.
