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Optimized in vitro micro-tuber production for colchicine biosynthesis in Gloriosa superba L. and its anti-microbial activity against Candida albicans

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Abstract

Gloriosa superba L. tubers are a rich source of commercially important colchicine and due to overexploitation, the species has become vulnerable. In the present investigation, in vitro tuber productions were carried out for its propagation and conservation. The in vitro and field-grown tubers were assessed for their colchicine content and antimicrobial activities. Maximum callusing was obtained when the medium was supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-furfurylaminopurine (kinetin). Among different auxins tested for in vitro tuberous root production, IAA (1.5 mg/L) induced 78.2% tuberous root per callus. In vitro micro-tuber raised in media supplemented with 1.5 mg/L thidiazuron (TDZ) recorded the highest response (72.9%) with 28.4 tubers per explant. Sucrose (6%) with TDZ (1.5 mg/L) produced significantly more micro-tubers per callus. Elicitor treatment with AlCl3 at 125 µM and 150 µM resulted in a significant increase in the micro-tuber and tuberous root production respectively. N6-(2-Isopentenyl) adenine (2ip) (1.0 mg/L) induced the highest frequency of in vitro micro-tuber sprouting and tuber formation compared to 6-benzylaminopurine (BAP) and 1-naphthalene acetic acid (NAA). The elicitor-treatments with AlCl3 significantly increased the colchicine content of in vitro tuberous root and tubers than that of the field grown tubers. The anti-microbial activity of in vitro raised tubers, tuberous roots and AlCl3 treated samples were significantly higher compared to the field grown samples. An optimized tissue culture system for mass propagation of G. superba with conservation aspects and the production of high-value colchicine is presented here, which can be used in various medicinal systems.

Key message

Manipulation of PGR’S significantly enhanced in vitro tuberization from noncorm bud explants. Elicitor treatment with AlCl3 enhanced the production of colchicine. HPLC analysis revealed significantly higher colchicine content in in vitro raised plants compared to field grown tubers. The study will help in mass propagation, conservation, and commercialization of Gloriosa superba L. for large scale production of colchicine.

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Acknowledgements

Dr. S. Sivakumar would like to acknowledge the Department of Plant Science, Bharathidasan University, Tiruchirappalli for providing HPLC facility (UGC-SAP) for this research. Mr. S. Sathish acknowledges ICMR, New Delhi (No. 3/1/2/102/2018-Nut.) for fellowship support.

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Author notes

  1. Sivakumar Subiramani and Sathish Sundararajan contributed equally to this work.

Authors and Affiliations

  1. Department of Plant Science, School of Life Sciences, Bharathidasan University, Tiruchirappalli, Tamil Nadu, 620 024, India

    Sivakumar Subiramani, Siva Govindarajan, Prem Kumar Ganesan, Vigneswaran Manickam, Senthil Kumar Thiruppathi & Jayabalan Narayanasamy

  2. Department of Biotechnology, AarupadaiVeedu Institute of Technology, Vinayaka Missions University, Chennai, Tamil Nadu, 603 104, India

    Vinoth Sadasivam

  3. Plant Genetic Engineering Laboratory, Department of Biotechnology, Bharathiar University, Coimbatore, Tamil Nadu, 641 046, India

    Sathish Sundararajan & Sathishkumar Ramalingam

  4. Department of Botany, Bharathiar University, Coimbatore, Tamil Nadu, 641 046, India

    Gurusaravanan Packiaraj

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  1. Sivakumar Subiramani
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Contributions

SS designed and executed all the experiments. SS prepared the manuscript. GS, SV, GPK, PG, and MV contributed substantially in experimental analysis and discussion. TSK, RS, and NJ mobilized funds and critically evaluated the manuscript.

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Correspondence to Sivakumar Subiramani.

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Subiramani, S., Sundararajan, S., Govindarajan, S. et al. Optimized in vitro micro-tuber production for colchicine biosynthesis in Gloriosa superba L. and its anti-microbial activity against Candida albicans. Plant Cell Tiss Organ Cult 139, 177–190 (2019). https://doi.org/10.1007/s11240-019-01675-7

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