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In vitro propagation, proscillaridin A production and antibacterial activity in Drimia robusta

Abstract

Drimia robusta is a threatened traditional medicinal plant extensively used in South Africa. Rapid in vitro mass propagation of the species was developed for commercial cultivation from leaf explants using various concentrations and combinations of plant growth regulators and organic elicitors. The highest number of regenerated shoots per explant (14.6 ± 0.54) was obtained on Murashige and Skoog (MS) medium supplemented with a combination of 2.27 μM thidiazuron (TDZ), 2.22 μM benzyladenine (BA) and 20 μM glutamine. Adventitious shoots were rooted and the plantlets were successfully acclimatized (100 %) in a vermiculite-soil mixture (1:1 v/v) in the greenhouse. Proscillaridin A (PsA) content and the antibacterial activity of in vitro and ex vitro regenerated plants were evaluated in different tissues in comparison to naturally-grown plants. The highest content of PsA (19.68 μg mg−1 DW) was recorded in roots of ex vitro plants which were grown on MS medium containing 2.27 μM TDZ, 2.22 μM BA and 100 mg l−1 casein hydrolysate. In vitro regenerated plants grown on MS medium containing 2.27 μM TDZ, 2.22 μM BA and 50.8 μM MBZ gave high antibacterial activity (MIC of 0.156 mg ml−1) against both Gram-positive and Gram-negative bacteria. Using this protocol the regenerated plants can be used in traditional medicine as an alternative to naturally collected plants.

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Abbreviations

BA:

6-Benzyladenine

CH:

Casein hydrolysate

GM:

Glutamine

HB:

Haemoglobin

HPLC:

High-performance liquid chromatography

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

MBZ:

Mebendazole

MS:

Murashige and Skoog medium

mT:

meta-Topolin

mTR:

meta-Topolin Riboside

NAA:

α-Naphthaleneacetic acid

OEs:

Organic elicitors

PGRs:

Plant growth regulators

PsA:

Proscillaridin A

PPF:

Photosynthetic photon flux

TDZ:

Thidiazuron

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Acknowledgments

The financial support by the National Research Foundation (NRF), Pretoria and the University of KwaZulu-Natal, Pietermaritzburg are gratefully acknowledged.

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Correspondence to Johannes Van Staden.

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Baskaran, P., Singh, S. & Van Staden, J. In vitro propagation, proscillaridin A production and antibacterial activity in Drimia robusta . Plant Cell Tiss Organ Cult 114, 259–267 (2013). https://doi.org/10.1007/s11240-013-0322-2

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Keywords

  • Antibacterial activity
  • Drimia robusta
  • Organic elicitors
  • Plant growth regulators
  • Proscillaridin A