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In vitro and ex vivo vegetative propagation and cytokinin profiles of Sceletium tortuosum (L.) N. E. Br.: a South African medicinal plant

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Abstract

Sceletium tortuosum is a South African protected species with tremendous value in traditional and modern medicine. The plants’ mesembrine-type alkaloids are potential therapeutics for a plethora of psychological, neurological and inflammatory disorders. In our in vitro and ex vivo studies, vegetative propagation and growth of this species were investigated. Cytokinin (CK) profiles were also explored. Shoot multiplication was induced on Murashige and Skoog (MS) medium supplemented with 2.5 µM indole-3-butyric acid (IBA). In vitro-generated shoots were inoculated on MS medium supplemented with 0, 2.5, 5.0 and 10.0 µM IBA or indole-3-acetic acid (IAA). Optimal rooting (55%), mean number of roots (3.80 ± 0.83) and new leaf pairs (4.65 ± 0.67) were achieved by 10.0 µM IBA. After greenhouse acclimatization, 45–90% of plantlets survived. All ex vivo shoot cuttings rooted well (90–100%). The highest mean number of roots (11.20 ± 1.37) and root length (57.18 ± 3.85 mm) were obtained by 5.0 µM IBA. Although spontaneous rooting was observed in both experiments, auxins enhanced multiple growth parameters. Cytokinin analyses of tissue-cultured (auxin-treated) and greenhouse (untreated) plants revealed higher cytokinin levels in vitro. These investigations provide rapid and efficient propagation techniques for Sceletium tortuosum which will be valuable to conservationists and pharmaceutical companies.

Key message

Plant tissue culture and cuttings enabled rapid propagation of Sceletium tortuosum. Exogenous plant growth regulators were not essential for shoot multiplication, flowering and rooting. Auxins effectively improved growth parameters.

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Data availability

All data are available from the corresponding author on request.

Code availability

Software applications used to analyse data and generate figures are disclosed.

Abbreviations

BAP :

N6-Benzylaminopurine

BAP7G :

N6-Benzylaminopurine-7-glucoside

BAP9G :

N6-Benzylaminopurine-9-glucoside

BAPR :

N6-Benzylaminopurine riboside

BAPR5´MP :

N6-benzylaminopurine-5´-monophosphate

CK:

Cytokinin

cZ :

cis-zeatin

cZ7G :

cis-zeatin-7-glucoside

cZ9G :

cis-zeatin-9-glucoside

cZOG :

cis-zeatin-O-glucoside

cZR :

cis-zeatin riboside

cZR5´MP :

cis-zeatin riboside-5´-monophosphate

cZROG :

cis-zeatin-O-glucoside riboside

DHZ:

Dihydrozeatin

DHZ7G:

Dihydrozeatin-7-glucoside

DHZ9G:

Dihydrozeatin-9-glucoside

DHZOG:

Dihydrozeatin-O-glucoside

DHZR:

Dihydrozeatin riboside

DHZR5´MP:

Dihydrozeatin riboside-5´-monophosphate

DHZROG:

Dihydrozeatin-O-glucoside riboside

GMP:

Greenhouse mother plant

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

iP:

N6-isopentenyladenine

iP7G:

N6-isopentenyladenine-7-glucoside

iP9G:

N6-isopentenyladenine-9-glucoside

iPR:

N6-isopentenyladenosine

iPR5´MP:

N6-isopentenyladenosine-5´-monophosphate

K:

Kinetin

K9G:

Kinetin-9-glucoside

KR:

Kinetin riboside

MNLP:

Mean number of new leaf pairs

MNR:

Mean number of roots

MRL:

Mean root length

MS medium:

Murashige and Skoog (1962) medium

mT:

meta-topolin

mT7G:

meta-topolin-7-glucoside

mT9G:

meta-topolin-9-glucoside

mTR :

meta-Topolin riboside

oT:

ortho-topolin

oT7G:

ortho-topolin-7-glucoside

oT9G:

ortho-topolin-9-glucoside

oTR:

ortho-topolin riboside

PAR:

Photosynthetic active radiation

PGR:

Plant growth regulator

pT:

para-topolin

pT7G :

para-topolin-7-glucoside

pT9G :

para-topolin-9-glucoside

pTR :

para-topolin riboside

tZ:

trans-zeatin

tZ7G:

trans-zeatin-7-glucoside

tZ9G:

trans-zeatin-9-glucoside

tZOG:

trans-zeatin-O-glucoside

tZR:

trans-zeatin riboside

tZR5´MP:

trans-zeatin riboside-5´-monophosphate

tZROG:

trans-zeatin-O-glucoside riboside

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Acknowledgements

We are indebted to Dr J. H. de Lange for generously donating plant material for this study. This research was supported by an ERDF project entitled “Development of Pre-Applied Research in Nanotechnology and Biotechnology” (No. CZ.02.1.01/0.0/0.0/17_048/0007323).

Funding

This research was supported by a European Regional Development Fund project entitled “Development of Pre-Applied Research in Nanotechnology and Biotechnology” (No. CZ.02.1.01/0.0/0.0/17_048/0007323).

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Authors and Affiliations

  1. Research Centre for Plant Growth and Development, School of Life Sciences, University of KwaZulu-Natal Pietermaritzburg, Private Bag X01, 3209, Scottsville, South Africa

    Amrisha Sreekissoon, Jeffrey F. Finnie & Johannes Van Staden

  2. Laboratory of Growth Regulators, Faculty of Science, Institute of Experimental Botany, Czech Academy of Sciences and Palacký University, Šlechtitelů 27, 78371, Olomouc, Czech Republic

    Lenka Plačková & Karel Doležal

  3. Department of Chemical Biology, Faculty of Science, Palacký University, Šlechtitelů 27, 78371, Olomouc, Czech Republic

    Karel Doležal

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  5. Johannes Van Staden
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Contributions

A.S, J.F.F and J.V.S conceived the research idea and design. J.V.S was responsible for the distribution of the materials necessary for the study. A.S conducted the vegetative propagation experiments. L.P and K.D performed the UHPLC-MS/MS analysis. A.S performed the subsequent data analysis and writing of the manuscript. J.F.F and J.V.S supervised the research. All authors reviewed and approved the final manuscript.

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Correspondence to Johannes Van Staden.

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Communicated by Ali R. Alan.

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Sreekissoon, A., Plačková, L., Doležal, K. et al. In vitro and ex vivo vegetative propagation and cytokinin profiles of Sceletium tortuosum (L.) N. E. Br.: a South African medicinal plant. Plant Cell Tiss Organ Cult 145, 191–202 (2021). https://doi.org/10.1007/s11240-020-02001-2

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