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A high efficiency in vitro regeneration protocol and clonal uniformity analysis in Hypericum hookerianum Wight & Arn., a lesser known plant of ethnomedicinal and economic importance

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Abstract

Hypericum hookerianum (Hypericaceae), a critically endangered plant of Western Ghats, India, has acquired significant importance due to its medicinal implications and ornamental flowers. This species is under severe anthropogenic pressure due to urbanization, tourism, and plantation activities taking place in their natural habitat. A clonal propagation strategy is standardized in this species which offers an opportunity for a stable production of active metabolites from this species as well as their conservation. The study deals with the optimization of axillary bud proliferation using nodal explants followed by genetic stability analysis of regenerants. Maximum number of shoots (3.66) was observed on the Murashige and Skoog (MS) medium supplemented with Kinetin (2.325 µM) with 85 % shoot multiplication frequency. In vitro grown shoots were rooted best in 1/2 MS medium supplemented with indole-3-butyric acid (2.45 µM) with an average of 6.8 ± 0.79 roots/shoot and 95.5 % rooting frequency. Plantlets were acclimatized best (90 %) in a mixture of sterile sand and farmyard manure (3:1). Micropropagated plants were subjected to random amplified polymorphic DNA analysis and phytochemical analysis to confirm their clonal stability. In RAPD analysis, 1032 amplicons were collectively generated which were monomorphic and similar to the mother plant. The comparable major phytochemical constituents in regenerants and mother plants together with genetic uniformity data obtained from RAPD analysis confirmed clonal fidelity of the regenerants. Findings in this study are the first report on micropropagation and assessment of genetic stability of micropropagated plantlets in H. hookerianum which suggests that in vitro axillary shoot proliferation can effectively be used as a tool for propagation and conservation of H. hookerianum.

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Acknowledgments

The authors thank Jeppiaar Educational Trust, Sathyabama University, Chennai for the financial support.

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  1. Department of Biotechnology, Centre for Bioresource Research and Development, Sathyabama University, Chennai, 600 119, India

    Reji Joseph Varghese, Anusha Bayyapureddy, Selvadoss Pradeep Pushparaj & Sooriamuthu Seeni

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  1. Reji Joseph Varghese
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Correspondence to Reji Joseph Varghese.

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Joseph Varghese, R., Bayyapureddy, A., Pushparaj, S.P. et al. A high efficiency in vitro regeneration protocol and clonal uniformity analysis in Hypericum hookerianum Wight & Arn., a lesser known plant of ethnomedicinal and economic importance. Braz. J. Bot 39, 377–386 (2016). https://doi.org/10.1007/s40415-015-0201-7

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