As. Pac. J. Mol. Biol. & Biotech., Jan 2015 Vol. 1, 10-19
A Reliable Protocol for Micropropagation of Gloriosa superba L. (Colchicaceae)
Kuldeep Yadav, Ashok Aggarwal and Narender Singh*
Department of Botany, Kurukshetra University, Haryana 136119, India.
* Author for correspondence: Dr. Narender Singh
Professor and Head, Department of Botany, Kurukshetra University, Kurukshetra-136119, Haryana, India.
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Abstract.
An efficient, rapid and improved in vitro plant regeneration protocol has been established for large scale multiplication of Gloriosa superba, an endangered ornamental and medicinal plant with limited reproductive capacity. Shoot tip explants from mature plants were sterilized using different concentrations (0.5- 1.0 w/v) of sodium hypochlorite (NaOCl) and then cultured on BAP or Kn (0.5-2.0 mg/l) alone or in combination with NAA (0.5 mg/l) at 25±2°C under a 16/8 hour light/dark photoperiod cycle. The most effective sterilization was achieved using 1.0% NaOCl treatment for 8 minutes. The highest regeneration frequency (76.6%) and average number of shoots (1.2) were obtained on MS medium fortified with BAP (2.0 mg l-1) + NAA (0.5 mg l-1). A high frequency of rooting (66.6%) with early root initiation (20.2 days) and maximum growth response was obtained when in vitro shoots were sub-cultured onto half strength MS medium supplemented with NAA (1.0 mg l-1) + IBA (0.5 mg l-1) at 3% (w/v) sucrose. Simultaneously, a successful attempt was made to acclimatize the tissue culture-raised plants of G. superba using different organic manures as hardening media. Among all the combinations tried, sand: soil: vermicompost (1:2:1) produced the highest percentage survival (93.3%) upon transplantation of plants to the field conditions, followed by sand: soil: farm yard manure (1:2:1) which produced 86.7% survival. The hardened plantlets were successfully acclimatized in the greenhouse and transferred to open field conditions. Parameters including plant height, number of leaves per plant, leaf area, root length and tuber length were also monitored periodically. The protocol can be successfully applied for the mass multiplication of this valuable threatened taxon as well as to facilitate experiments involving its genetic modification.
