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How does exogenously applied cytokinin type affect growth and endogenous cytokinins in micropropagated Merwilla plumbea?

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Abstract

Merwilla plumbea (Lindl.) Speta is a popular and highly sought after South African medicinal plant with diverse therapeutic values. Using Ultra performance liquid chromatography (UPLC), the effect of five cytokinins (CKs) [either isoprenoid = N 6-isopentenyladenine (iP) or aromatic = benzyladenine, meta-topolin (mT), meta-topolin riboside (mTR), and 6-(3-methoxybenzylamino)-9-tetrahydropyran-2-ylpurine] MemTTHP on growth and level of endogenous CKs during micropropagation and acclimatization stages was evaluated. Aromatic CK (mT/mTR) elicited the highest shoot proliferation (7–8 shoots per explant) during in vitro culture. Following acclimatization, iP-treated and the control plants were healthier with longer leaves, roots and higher fresh weight when compared to aromatic CKs. A total of 37 (22 isoprenoid and 15 aromatic) CK variants were quantified in both in vitro and acclimatized plants. Based on their metabolic function, these were grouped into five types including free bases, ribosides, ribotides, O- and 9-glucosides. In addition to enhancing our understanding of the hormone physiology in M. plumbea, the current findings are discussed in line with the effect of the exogenously applied CK on the observed differences in growth before and after the important stage of acclimatization. The observed dynamics in endogenous CK provide an avenue to manipulate in vitro growth and development of investigated species.

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Abbreviations

ANOVA:

Analysis of variance

BA:

N 6-Benzyladenine

BA9G:

N 6-Benzyladenine-9-glucoside

BAR:

N 6-Benzyladenine riboside

BAR5′MP:

N 6-Benzyladenosine-5′-monophosphate

CK:

Cytokinin

cZ:

cis-Zeatin

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

DHZ9G:

Dihydrozeatin-9-glucoside

DHZOG:

Dihydrozeatin-O-glucoside

DHZR:

Dihydrozeatin riboside

DHZR5′MP:

Dihydrozeatin riboside-5′-monophosphate

DHZROG:

Dihydrozeatin-O-glucoside riboside

iP:

N 6-Isopentenyladenine

iP9G:

N 6-Isopentenyladenine-9-glucoside

iPR:

N 6-Isopentenyladenosine

iPR5′MP:

N 6-Isopentenyladenosine-5′-monophosphate

KIN:

Kinetin

KIN9G:

Kinetin-9-glucoside

KINR:

Kinetin riboside

MemT:

meta-Methoxy topolin

MemTTHP:

6-(3-Methoxybenzylamino)-9-tetrahydropyran-2-ylpurine

MS:

Murashige and Skoog medium

mT:

meta-Topolin

mT9G:

meta-Topolin-9-glucoside

mTR:

meta-Topolin riboside

oT:

ortho-Topolin

oTOG:

ortho-Topolin-O-glucoside

oTR:

ortho-Topolin riboside

PGR:

Plant growth regulator

PPF:

Photosynthetic photon flux density

pT:

para-Topolin

pTOG:

para-Topolin-O-glucoside

tZ:

trans-Zeatin

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

UPLC:

Ultra performance liquid chromatography

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Acknowledgments

A.O.A. was supported by the Claude Leon Foundation, South Africa. L.P., O.N., L.S. and K.D. were supported by the Ministry of Education Youth and Sports, Czech Republic (grant LO1204 from the National Program of Sustainability and Agricultural Research) as well as by IGA of Palacký University (IGA_PrF_2014006). We thank Dr. W.A. Stirk for providing valuable suggestions towards improving the manuscript.

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  1. Michael W. Bairu

    Present address: Institute for Commercial Forestry Research, P. O. Box 100281, Pietermaritzburg, Scottsville, 3209, South Africa

Authors and Affiliations

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

    Adeyemi O. Aremu, Michael W. Bairu, Jeffrey F. Finnie & Johannes Van Staden

  2. Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University and Institute of Experimental Botany, Academy of Sciences of Czech Republic, 78371, Olomouc-Holice, Czech Republic

    Lenka Plačková, Ondřej Novák, Lucie Plíhalová & Karel Doležal

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  1. Adeyemi O. Aremu
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Correspondence to Johannes Van Staden.

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Aremu, A.O., Plačková, L., Bairu, M.W. et al. How does exogenously applied cytokinin type affect growth and endogenous cytokinins in micropropagated Merwilla plumbea?. Plant Cell Tiss Organ Cult 118, 245–256 (2014). https://doi.org/10.1007/s11240-014-0477-5

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