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A high-frequency in vitro multiplication, micromorphological studies and ex vitro rooting of Cadaba fruticosa (L.) Druce (Bahuguni): a multipurpose endangered medicinal shrub

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Abstract

An efficient and reproducible in vitro propagation protocol has been established for Cadaba fruticosa (L.) Druce. Surface-sterilized nodal stem segments of mature plant were used as explants for culture establishment. Multiple shoots were optimally differentiated from the nodal stem explants through bud breaking on Murashige and Skoog (1962) medium containing 3.0 mg l−1 benzyladenine (BA). The effect of different plant growth regulators and minerals were studied on different stages of micropropagation procedure (i.e., explant establishment, shoot multiplication/growth and ex vitro rooting). Additionally, for enhancing shoot multiplication during subculture, MS medium was modified (MMS) with higher levels of magnesium, potassium and sulphate ions. Out of these, MMS3 medium containing 0.25 mg l−1 each of BA and Kin (N6-furfuryladenine), with 0.1 mg l−1 NAA (α-naphthalene acetic acid) was found the best for shoot multiplication (42.45 ± 3.82 per culture vessel). The in vitro regenerated shoots were rooted under ex vitro conditions on treating the shoot base with 500 mg l−1 of IBA (indole-3 butyric acid) for 3 min on sterile Soilrite®. The ex vitro rooted plants were hardened in the greenhouse and transferred to the field with ≈85 % survival rate. There were not any visual differences between wild and micropropagated plants in the field, although the later underwent significant changes during acclimatization. Micromorphological changes on leaf surface characters from in vitro to acclimatized plantlets were studied in terms of development of glandular trichomes, changes in vein spacing and vein structure in order to understand the nature of plant responses towards environmental conditions. The method developed and defined can be applied for commercial cultivation, which may be important for extraction of bioactive compounds and may facilitate conservation of this multipurpose endangered medicinal shrub.

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Abbreviations

BA:

6-benzyladenine

IBA:

Indole-3-butyric acid

2-iP:

N6–(2-isopentenyl) adenine

Kin:

Kinetin (N6-furfuryladenine)

MS:

Murashige and Skoog (1962) medium

MMS:

Modified MS medium

NAA:

α-naphthalene acetic acid

NaOH:

Sodium hydroxide

NOA:

Naphthoxyacetic acid

PFD:

Photon Flux Density

PGRs:

Plant Growth Regulators

RH:

Relative Humidity

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Acknowledgments

DL gratefully acknowledges the financial support from Council of Scientific and Industrial Research (CSIR), New Delhi in the form of Junior and Senior Research Fellowship (JRF-SRF). AKP is thankful to the University Grant Commission (UGC), New Delhi, for providing Special Assistance Program (SAP) in the form of Centre for Advanced Study (CAS) to the Department of Botany, Jai Narain Vyas University, Jodhpur (INDIA).

Author’s contribution

D. Lodha designed and performed the experiments and wrote the first draft of the manuscript. A. K. Patel helped in data analysis and organized it in figures and tables. N. S. Shekhawat guided the research and edited the final version of the manuscript.

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Authors and Affiliations

  1. Biotechnology Unit, Department of Botany (UGC – Centre for Advanced Study), Jai Narain Vyas University, New Campus, Prof. V. V. John Road, Jodhpur, 342 001, Rajasthan, India

    Deepika Lodha, Ashok Kumar Patel & N. S. Shekhawat

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  1. Deepika Lodha
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  2. Ashok Kumar Patel
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  3. N. S. Shekhawat
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Correspondence to N. S. Shekhawat.

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Lodha, D., Patel, A.K. & Shekhawat, N.S. A high-frequency in vitro multiplication, micromorphological studies and ex vitro rooting of Cadaba fruticosa (L.) Druce (Bahuguni): a multipurpose endangered medicinal shrub. Physiol Mol Biol Plants 21, 407–415 (2015). https://doi.org/10.1007/s12298-015-0310-6

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