In Vitro Seedlings as Dynamic Explants for Establishment of Root Cultures of Pyrenacantha volubilis Hook. for Camptothec
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RESEARCH ARTICLE
In Vitro Seedlings as Dynamic Explants for Establishment of Root Cultures of Pyrenacantha volubilis Hook. for Camptothecin Production S. Hima1 • C. K. Midhu1 • G. Krishnakumar2 • K. Satheeshkumar1
Received: 31 October 2018 / Revised: 22 May 2019 / Accepted: 1 June 2019 The National Academy of Sciences, India 2019
Abstract Pyrenacantha volubilis, an important medicinal plant, is reported to contain camptothecin (CPT), a high value anticancer compound with a global demand of USD 2.2 billion in 2008. Isolated root cultures of P. volubilis could be established using radicle-developed roots from the seeds cultured in 0.3 mg L-1 kinetin. The tap roots isolated from the germinated seeds were transferred to half-strength MS liquid media supplemented with 0.3 mg L-1 indole-3acetic acid and 0.2 mg L-1 indole-3-butyric acid to establish isolated root cultures and were further subcultured into the same media to study the growth and CPT production over a period of 60 days at an interval of 10 days. Maximum biomass (3.34 ± 0.19 g) was obtained from roots cultured in MS liquid media augmented with 0.3 mg L-1 IAA and 0.2 mg L-1 IBA with alternating salt strength over a period of 45 (17 ? 28) days. The concentration of CPT in the in vitro root-derived culture sample was estimated through HPLC and recorded as 0.135% (on g DW basis, 30th day) which is comparable to that of field grown roots (0.13%). The overall results obtained with the present experiment demonstrated the use of in vitro
Significance Statement In vitro production of camptothecin (CPT) from root cultures of Pyrenacantha volubilis using in vitro-derived seedling explants was established for the first time which yielded more camptothecin than the field grown roots. Root culture can be used as an alternative source for CPT production instead of the conventional sources. & K. Satheeshkumar [email protected] 1
Biotechnology and Bioinformatics Division, Jawaharlal Nehru Tropical Botanic Garden and Research Institute, Palode, India
2
Department of Biotechnology, University of Kerala, Thiruvananthapuram, India
seedlings as a dynamic explant source for root culture establishment and the feasibility of the system for further use in the production of CPT. Keywords Kinetin Root culture Camptothecin estimation In vitro seedlings Pyrenacantha volubilis
Introduction Plant secondary metabolites are a diverse group of organic compounds which facilitate the interaction of plants with the environment and also in the establishment of a defence mechanism. Among the important plant-derived compounds prescribed for cancer therapy, camptothecin (CPT), a water insoluble cytotoxic monoterpene-derived indole alkaloid, has been recognized as a very potent anticancer compound for its amazing inhibitory action against tumour cells by blocking the eukaryotic topoisomerase-I. Over a dozen derivatives of CPT are currently under clinical trials. Derivatives such as Hycamtin (topotecan) and Camptosar (irinotecan or CPT 11) are currently used widely for the tr
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