Abstract
Nothapodytes nimmoniana (Icacinaceae) yields camptothecin (isoquinoline alkaloid) which is a potent anti-cancer drug. The major objectives of the present study were to develop an efficient protocol for mass propagation of N. nimmoniana using liquid medium and to compare regeneration with semisolid cultures; as also to quantify the amount of camptothecin in regenerated plants. Adventitious shoots were induced from the callus derived from nodal explants on semisolid and liquid Murashige and Skoog (MS) medium supplemented with 1.0, 2.0, 5.0 and 10.0 μM 6-benzylaminopurine or kinetin or 2-isopentenyl adenine (2-iP). The highest number of adventitious shoots was regenerated on medium supplemented with 2.0 μM BAP. Compared to semisolid medium (41.9 shoots per explant), liquid medium (165.9 shoots per explant) was found suitable for shoot induction and shoot multiplication. Shoots were rooted on MS semisolid medium of one-fourth strength containing IBA (2.4 μM) and IAA (5.7 μM). The plantlets were acclimatized in a growth chamber at 25°C, 60% relative humidity, with 16-h photoperiod (40 μmol m−2 s−1). The camptothecin content was determined in ex vitro plants using HPLC. The analysis revealed that the leaves and stems of ex vitro plants had a considerable amount of camptothecin and these plants could be used as a raw material for camptothecin extraction.
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Acknowledgments
This work was partially financed by Special Assistance Program and Major Research Project [F. No. 33-188/2007 (SR)] of University Grants Commission, New Delhi, India. Authors are thankful to Dr. M.R. Davey, Plant Sciences, University of Nottingham, Sutton Bonington, LE12 5RD, UK for reviewing the manuscript.
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Communicated by J. Van Huylenbroeck.
V. S. Dandin and H. N. Murthy contributed equally for the manuscript.
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Dandin, V.S., Murthy, H.N. Enhanced in vitro multiplication of Nothapodytes nimmoniana Graham using semisolid and liquid cultures and estimation of camptothecin in the regenerated plants. Acta Physiol Plant 34, 1381–1386 (2012). https://doi.org/10.1007/s11738-012-0934-x
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DOI: https://doi.org/10.1007/s11738-012-0934-x