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Plant Growth Regulation

, Volume 63, Issue 2, pp 105–114 | Cite as

Changes in endogenous cytokinin profiles in micropropagated Harpagophytum procumbens in relation to shoot-tip necrosis and cytokinin treatments

  • Michael W. Bairu
  • Ondřej Novák
  • Karel Doležal
  • Johannes Van Staden
SI : Tissue Culture

Abstract

Changes in cytokinin (CK) profiles and their physiological implications in micropropagated Harpagophytum procumbens [(Burch.) DC. ex Meisn.] tissues in relation to shoot-tip necrosis (STN) and CK treatments were studied. Total CK content was quantified in benzyladenine (BA)-treated necrotic and normal plantlets and in plantlets treated with the CKs BA, meta-topolin (mT) and meta-topolin riboside (mTR) with and without the auxin indole-3-acetic acid (IAA). Generally necrotic shoots yielded more total CK compared to normal shoots. Cytokinin accumulation was higher at the basal section (basal > middle > top). Further analysis of the CKs based on structural and functional forms revealed excessive accumulation of 9-glucosides (deactivation products—toxic metabolites) and limited amounts of O-glucosides (storage forms—re-utilizable) in necrotic and BA-treated shoots compared to normal and topolin-treated cultures. The addition of IAA enhanced the formation of 9-glucosides in BA-treated cultures but reduced it in topolin-treated cultures. The symptom of STN could therefore be attributed to conversion of active cytokinins to other forms such as 9-glucosides which are neither active nor reversibly sequestrated to active forms. Literature shows that metabolites like 9-glucosides of BA have a detrimental effect in plant tissue culture.

Keywords

Auxin cytokinin interaction Cytokinin purification and analysis Devil’s claw Glucosylation Harpagophytum procumbens Micropropagation Pedaliaceae Shoot-tip necrosis 

Abbreviations

BA

N6-benzyladenine

BA9G

N6-benzyladenine-9-glucoside

BAR

N6-benzyladenosine

BAR5′MP

N6-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

N6-isopentenyladenine

iP9G

N6-isopentenyladenine-9-glucoside

iPR

N6-isopentenyladenosine

iPR5′MP

N6-isopentenyladenosine-5′-monophosphate

IAC

Immunoaffinity chromatography

MRM

Multiple reaction monitoring

MS medium

Murashige and Skoog (1962) basal medium

mT

meta-topolin

mT9G

meta-topolin-9-glucoside

mTOG

meta-topolin-O-glucoside

mTR

meta-topolin riboside

mTR5′MP

meta-topolin-5′-monophosphate

mTROG

meta-topolin-O-glucoside riboside

oT

ortho-topolin

oT9G

ortho-topolin-9-glucoside

oTOG

ortho-topolin-O-glucoside

oTR

ortho-topolin riboside

oTR5′MP

ortho-topolin-5′-monophosphate

oTROG

ortho-topolin-O-glucoside riboside

pT

para-topolin

pTOG

para-topolin-O-glucoside

pTR

para-topolin riboside

pTR5′MP

para-topolin-5′-monophosphate

pTROG

para-topolin-O-glucoside riboside

STN

Shoot-tip necrosis

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

Notes

Acknowledgments

The financial support of the National Research Foundation-South Africa, the Grant Agency of the Czech Republic (GA 206/07/0570) and Czech Ministry of Education (MSM 6198959216, 1M06030). We also wish to thank Hana Martínková for technical assistance.

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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Michael W. Bairu
    • 1
  • Ondřej Novák
    • 2
  • Karel Doležal
    • 2
  • Johannes Van Staden
    • 1
  1. 1.Research Centre for Plant Growth and Development, School of Biological and Conservation SciencesUniversity of KwaZulu-Natal PietermaritzburgScottsvilleSouth Africa
  2. 2.Laboratory of Growth RegulatorsPalacký University & Institute of Experimental Botany AS CROlomoucCzech Republic

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