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Optimization of factors affecting in vitro regeneration, flowering, ex vitro rooting and foliar micromorphological studies of Oldenlandia corymbosa L.: a multipotent herb

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Abstract

The conditions were optimized for efficient in vitro regeneration of shoots and roots of Oldenlandia corymbosa L. using nodal shoot explants. Murashige and Skoog’s (MS) medium augmented with additives and 2.0 mg L−1 BAP was recorded optimum for shoot bud induction from the nodal meristems. The shoots were proliferated by subsequent subcultures on half strength MS medium fortified with 1.0 mg L−1 BAP and 0.5 mg L−1 kinetin + additives. This media combination yielded maximum number of shoots (223 ± 4.12 shoots/culture bottle) with 13.4 cm average length. Flower buds were induced (4.2 ± 0.28 flowers) from the in vitro multiplied shoots on MS medium contained 1.0 mg L−1 BAP and 0.5 mg L−1 of kinetin and IAA under 50 µmol m−2 s−1 SFPD light intensity for 12 h/day photoperiod. In vitro regenerated shoots were rooted on half strength MS medium conjunct with IAA, IBA and NAA singly at different concentrations. The best rooting response was observed on half strength MS medium containing IBA at 2.0 mg L−1 with activated charcoal. Roots were also induced from the cut ends of the shoots using ex vitro rooting techniques in O. corymbosa by pulse treating the shoots with 300 mg L−1 IBA for 4 min. Better roots were achieved in this method than the in vitro roots in terms of numbers (14.7 ± 0.21) and firmness. The foliar micromorphological studies could help to understand the structural adaptations of micropropagated O. corymbosa plantlets towards field environments. The in vitro induced anomalies in stomatal apparatus were repaired during hardening of plantlets in the greenhouse and after field transfer. Decrease in stomatal density (from 70.78 ± 0.55 to 55.6 ± 0.10), and increase in veins, trichomes and crystals/raphides densities revealed the developments of structural changes in the leaves to withstand in the harsh field conditions. The acclimatized plantlets with well developed root systems were successfully shifted to the natural soils with 98% survival rate.

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Abbreviations

BAP:

6-Benzylaminopurine

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

Kin:

Kinetin

MS:

Murashige and Skoog’s basal medium

NAA:

α-Naphthalene acetic acid

RH:

Relative humidity

SPFD:

Spectral photon flux density

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Acknowledgements

Authors are grateful to the Department of Science, Technology and Environment, Government of Puducherry, India for providing financial support to their laboratory under the Grant-In-Aid Scheme.

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Authors and Affiliations

  1. Department of Botany, Kanchi Mamunivar Center for Postgraduate Studies, Puducherry, 605 008, India

    J. Revathi & M. Manokari

  2. Department of Plant Science, M.G.G.A.C., Mahe, Puducherry, 673 311, India

    Mahipal S. Shekhawat

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  1. J. Revathi
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  2. M. Manokari
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  3. Mahipal S. Shekhawat
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Contributions

JR performed the experiments and analyzed data. MM designed the experiment and contributed to writing the manuscript. MSS conceived the idea, and contributed to writing and editing the manuscript.

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Correspondence to Mahipal S. Shekhawat.

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The authors declare that they have no conflict of interest.

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The authors declare that the study was carried out following accepted professional conduct. However no ethical approval was needed for the study as it did not involve the use of animals or human subjects.

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Communicated by Sergio J. Ochatt.

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Revathi, J., Manokari, M. & Shekhawat, M.S. Optimization of factors affecting in vitro regeneration, flowering, ex vitro rooting and foliar micromorphological studies of Oldenlandia corymbosa L.: a multipotent herb. Plant Cell Tiss Organ Cult 134, 1–13 (2018). https://doi.org/10.1007/s11240-018-1395-8

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