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Endogenous cytokinins in shoots of Aloe polyphylla cultured in vitro in relation to hyperhydricity, exogenous cytokinins and gelling agents

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Abstract

The process of hyperhydricity in tissue cultured plants of Aloe polyphylla is affected by both applied cytokinins (CKs) and the type of gelling agent used to solidify the medium. Shoots were grown on media with agar or gelrite and supplemented with different concentrations of N6-benzyladenine (BA) or zeatin (0, 5 and 15 μM). Endogenous CKs were measured in in vitro regenerants after an 8-weeks cycle to examine whether the hyperhydricity-inducing effect of exogenous CKs and gelling agents is associated with changes in the endogenous CK content. On media with agar a reduction in hyperhydricity occurred, while the gelrite treatment produced both normal and hyperhydric shoots (HS). The content of endogenous CKs, determined by HPLC-mass spectrometry, in the shoots grown on CK-free media comprised isopentenyladenine-, trans-zeatin- and cis-zeatin-type CKs. The application of exogenous CKs resulted in an increase in the CK content of the shoots. Following application of zeatin, dihydrozeatin-type CKs were also detected in the newly-formed shoots. Application of BA to the media led to a transition from isoprenoid CKs to aromatic CKs in the shoots. Shoots grown on gelrite media contained higher levels of endogenous CKs compared to those on agar media. Total CK content of HS was higher than that of normal shoots grown on the same medium. We suggest that the ability of exogenous CKs and gelrite to induce hyperhydricity in shoots of Aloe polyphylla is at least partially due to up-regulation of endogenous CK levels. However, hyperhydricity is a multifactor process in which different factors intervene.

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Abbreviations

BA:

N6-benzyladenine

BA9G:

N6-benzyladenine 9-glucoside

BAR:

N6-benzyladenine 9-riboside

BARMP:

N6-benzyladenine 9-riboside- 5′-monophosphate

CK:

Cytokinin

DHZ:

Dihydrozeatin

DHZ9G:

Dihydrozeatin 9-glucoside

DHZOG:

Dihydrozeatin O-glucoside

DHZR:

Dihydrozeatin 9-riboside

DHZRMP:

Dihydrozeatin 9-riboside-5′-monophosphate

DHZROG:

Dihydrozeatin 9-riboside O-glucoside

DMAPP:

Dimethylallyl diphosphate

DW:

Dry weight

HS:

Hyperhydric shoots

IAC:

Immunoaffinity chromatography

IBA:

Indole-3-butyric acid

iP:

N6-(Δ2-Isopentenyl)adenine

iP9G:

N6-(Δ2-Isopentenyl)adenine 9-glucoside

iPR:

N6-(Δ2-Isopentenyl)adenine 9-riboside

iPRMP:

N6-(Δ2-Isopentenyl)adenine 9-riboside-5′-monophosphate

LC(+) ES–MS:

Liquid chromatography + electrospray ionization–mass spectrometry

NS:

Normal shoots

T:

Topolin

mT:

meta-Topolin

mT9G:

meta-Topolin 9-glucoside

mTOG:

meta-Topolin O-glucoside

mTR:

meta-Topolin 9-riboside

mTRMP:

meta-Topolin 9-riboside- 5′-monophosphate

mTROG:

meta-Topolin 9-riboside O-glucoside

oT:

ortho-Topolin

oT9G:

ortho-Topolin 9-glucoside

oTOG:

ortho-Topolin O-glucoside

oTR:

ortho-Topolin 9-riboside

oTRMP:

ortho-Topolin 9-riboside- 5′-monophosphate

oTROG:

ortho-Topolin 9-riboside O-glucoside

pT:

para-Topolin

pTOG:

para-Topolin O-glucoside

pTR:

para-Topolin 9-riboside

pTRMP:

para-Topolin 9-riboside- 5′-monophosphate

pTROG:

para-Topolin 9-riboside O-glucoside

Z:

Zeatin

Z9G:

Zeatin 9-glucoside

cZ:

cis-Zeatin

cZOG:

cis-Zeatin O-glucoside

cZR:

cis-Zeatin 9-riboside

cZRMP:

cis-Zeatin 9-riboside- 5′-monophosphate

cZROG:

cis-Zeatin 9-riboside O-glucoside

tZ:

trans-Zeatin

tZOG:

trans-Zeatin O-glucoside

tZR:

trans-Zeatin 9-riboside

tZRMP:

trans-Zeatin 9-riboside- 5′-monophosphate

tZROG:

trans-Zeatin 9-riboside O-glucoside

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Acknowledgements

The National Research Foundation, Pretoria, Republic of South Africa, the Grant Agency of the Czech Republic (grant No. 301/05/0418) and the Czech Ministry of Education (grant No. MSM 6198959216) are thanked for financial assistance. M.I. thanks Dr W. Stirk and Prof R. Beckett for their helpful criticism and careful reading of the manuscript and A. Nsabimana for help with statistical analysis. M.S. thanks H. Martinková and P.␣Amarkorová for excellent technical assistance.

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Authors and Affiliations

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

    Mariyana Ivanova & Johannes Van Staden

  2. Laboratory of Growth Regulators, Palacký University and Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Šlechtitelů 11, 783 71, Olomouc, Czech Republic

    Ondřej Novák & Miroslav Strnad

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  1. Mariyana Ivanova
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Correspondence to Johannes Van Staden.

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Ivanova, M., Novák, O., Strnad, M. et al. Endogenous cytokinins in shoots of Aloe polyphylla cultured in vitro in relation to hyperhydricity, exogenous cytokinins and gelling agents. Plant Growth Regul 50, 219–230 (2006). https://doi.org/10.1007/s10725-006-9139-x

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