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High frequency multiple shoot induction from nodal segments and rhinacanthin production in the medicinal shrub Rhinacanthus nasutus (L.) Kurz

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Abstract

High frequency multiple shoots have been induced from nodal segments of Rhinacanthus nasutus (L.) Kurz., a potent anticancerous ethnomedicinal plant. For initiation of cultures, nodal segments were cultured on MS medium supplemented with various concentrations (1.0–5.0 μM) of 6-benzyladenine or thidiazuron (TDZ) alone or in combination with α-naphthalene acetic acid (NAA 0.5–1.0 μM). The optimum frequency of response (85 %) and shoot number (3.3) was observed on MS medium supplemented with 4.0 μM TDZ and 0.8 μM NAA. The shoots developed on initiation media were excised and nodal segments were subcultured on MS medium supplemented with TDZ (4.0 μM) and NAA (0.5–1.0 μM). This subculturing process was repeated thrice, each with 45 days of duration and the multiple shoot formation was recorded at the end of every subculture stage. The highest frequency of response (100 %) and number of multiple shoots (24.1) per explant were recorded at the end of the third subculture passage on MS medium supplemented with 4.0 μM TDZ and 0.8 μM NAA. The optimum rooting of shoots was observed on ½ MS medium fortified with 3.0 μM indole-3-butyric acid. The rooted plants were successfully transplanted to soil. The estimation of rhinacanthin (RC) content in shoots and roots was carried out in 6-month-old ex vitro plants (i.e., plants regenerated via in vitro culture) and field grown natural plants by high performance liquid chromatography. Both shoots and roots of naturally grown plants showed slightly higher RC content than ex vitro grown plants. The highest RC content (4.6 mg/g DW RC-C, 0.14 mg/g DW RC-D and 0.10 mg/g DW RC-N) was recorded in roots of naturally grown plants.

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Abbreviations

BA:

6-Benzyladenine

HPLC:

High performance liquid chromatography

IBA:

Indole-3-butyric acid

NAA:

α-Naphthalene acetic acid

RC:

Rhinacanthin

TDZ:

Thidiazuron (N-phenyl-N’-1,2,3-thiadiazol-5-yl urea)

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Acknowledgments

M.K.C is thankful to M.G. University, Kottayam, Kerala for the financial assistance in the form of University Junior Research Fellowship and Principal, St. Thomas College, Pala for providing lab facilities. T.D.T acknowledges the financial support from Indian National Science Academy (INSA, New Delhi) and Polish Academy of Sciences (PAS), Poland in the form of bilateral exchange programme.

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  1. T. Dennis Thomas

    Present address: Department of Pharmaceutical Biology and Botany, Medical University in Wroclaw, Al. Jana Kochanowskiego 10, 51-601, Wroclaw, Poland

Authors and Affiliations

  1. Post Graduate and Research Department of Botany, St. Thomas College, Pala, Arunapuram (P.O), Kottayam, 686574, Kerala, India

    Meena K. Cheruvathur & T. Dennis Thomas

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Correspondence to T. Dennis Thomas.

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Cheruvathur, M.K., Thomas, T.D. High frequency multiple shoot induction from nodal segments and rhinacanthin production in the medicinal shrub Rhinacanthus nasutus (L.) Kurz. Plant Growth Regul 74, 47–54 (2014). https://doi.org/10.1007/s10725-014-9895-y

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