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Spatial and temporal changes in endogenous cytokinins in develo** pea roots

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Abstract

Germination and seedling establishment follows a distinct pattern which is partly controlled by hormones. Roots have high levels of cytokinins. By quantifying the fluctuations in endogenous cytokinins over time, further insight may be gained into the role of cytokinins during germination and seedling establishment. Radicles were excised from sterile Pisum sativum L. seeds after 30 min and 5 h imbibition. Seedlings germinated on agar were harvested after 1, 3, 6 and 9 days. The roots were divided into the root tip, root free zone, secondary root zone and from day 6, the secondary roots. Samples were purified by various chromatographic methods and endogenous cytokinins detected by LC(+)ES-MS. Benzyladenine levels doubled after 5 h imbibition and then gradually decreased over time. Low concentrations of cis-Zeatin (cZ) type cytokinins were detected in the radicle after 30 min imbibition. After 5 h imbibition, cis-zeatin riboside-5′-monophosphate had greatly increased. The total cytokinin content of the roots increased over time with the ribotides being the predominant conjugates. From day 3 onwards, there was a gradual increase in the free bases, O-glucosides and their ribosylated forms. Mainly N 6-(2-isopentenyl)adenine (iP)-type cytokinins were detected in the root tip, whereas trans-zeatin- (tZ), dihyrozeatin- (DHZ) and iP-type cytokinins were found in the secondary roots and root zone. Cytokinin biosynthesis was only detected after day 6. Biosynthesis of iP and tZ derivatives was quite rapid, whereas biosynthesis of cZ derivatives remained at a low basal level. These fluctuations in cytokinin types and concentrations suggest the cytokinins may be synthesized from various pathways in pea roots.

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Abbreviations

BA:

N 6-benzyladenine

CBP:

Cytokinin binding proteins

cZ:

cis-Zeatin

cZOG:

cis-Zeatin-O-glucoside

cZR:

cis-Zeatin riboside

cZROG:

cis-Zeatin riboside-O-glucoside

cZRMP:

cis-Zeatin riboside-5′-monophosphate

DEAE:

Diethylaminoethyl

DHZ:

Dihydrozeatin

DHZRMP:

Dihydrozeatin riboside-5′-monophosphate

IAC:

Immunoaffinity chromatography

iP:

N 6-(2-isopentenyl)adenine

iPRMP:

N 6-(2-isopentenyl)adenosine-5′-monophosphate

IPT:

Isopentenyltransferase

LC(+)ES-MS:

Liquid chromatography with electrospray interface mass spectrometry

MS:

Mass spectrometry

mT:

meta-Topolin

tRNA:

Transfer ribonucleic acid

tZ:

trans-Zeatin

tZR:

trans-Zeatin riboside

tZRMP:

trans-Zeatin riboside-5′-monophosphate

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Acknowledgments

Georgina Arthur, Marnie Light and Nicky Taylor, University of KwaZulu-Natal are thanked for their assistance in harvesting the seedlings and Hana Martinková and Petra Amarkorová, Palacký Univeristy and Institute of Experimental Botany AS CR for their help with cytokinin analyses. The National Research Foundation, South Africa and the Czech Ministry of Education (grant No. MSM 6198959216) are thanked for financial 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, South Africa

    W. A. Stirk & J. van Staden

  2. Laboratory of Growth Regulators, Palacký University & Institute of Experimental Botany AS CR, Šlechtitelů 11, 783 71, Olomouc, Czech Republic

    O. Novák, K. Václavíková, P. Tarkowski & M. Strnad

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  1. W. A. Stirk
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Correspondence to W. A. Stirk.

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Stirk, W.A., Novák, O., Václavíková, K. et al. Spatial and temporal changes in endogenous cytokinins in develo** pea roots. Planta 227, 1279–1289 (2008). https://doi.org/10.1007/s00425-008-0699-z

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