Plant Growth Regulation

, Volume 65, Issue 3, pp 427–437 | Cite as

Changes in cytokinin levels and metabolism in tobacco (Nicotiana tabacum L.) explants during in vitro shoot organogenesis induced by trans-zeatin and dihydrozeatin

  • Marek Klemš
  • Zdeňka Slámová
  • Václav Motyka
  • Jiří Malbeck
  • Alena Trávníčková
  • Ivana Macháčková
  • Josef Holík
  • Stanislav Procházka
Original paper


The uptake and metabolism of trans-zeatin and/or dihydrozeatin, in correlation with cytokinin oxidase/dehydrogenase (CKX) and β-glucosidase activity, were studied in leaf segments derived from wild-type (WT) and transgenic (T) tobacco (Nicotiana tabacum L. cv. Petit Havana SR1) during in vitro induction of shoot organogenesis. T explants harbored the maize gene Zm-p60.1β-glucosidase. Higher levels of shoot regeneration were observed on T explants in the early stages of cultivation. In WT explants, the content of cytokinin (CK)-O- and N-glucosides increased. In T explants, a higher content of Z-9-riboside and Z-9-riboside-5′-monophosphate and higher CKX activity during the early stage of cultures were found. A positive correlation was obtained for bioactive CK content and the organogenic response in T explants. Our results indicate a connection between the organogenic capacity of tobacco explants, metabolism of endogenous CKs and uptake of exogenous CKs from the cultivation medium.


β-glucosidase Cytokinin oxidase/dehydrogenase Dihydrozeatin Organogenesis in vitro trans-zeatin 



N 6-benzyladenine




Cytokinin oxidase/dehydrogenase


2,4-dichlorophenoxyacetic acid
















Induction medium containing trans-zeatin


Induction medium containing dihydrozeatin


Growth regulator-free medium


Subcultivation from zeatin induction to growth regulator-free medium


Subcultivation from dihydrozeatin induction to growth regulator-free medium


Indole-3-butyric acid


N 6-(Δ2-isopentenyl)adenine


Gene coding isopentenyltransferase


N 6-(Δ2-isopentenyl)adenine-7-glucoside


Transgenic tobacco


Wild-type tobacco










trans-zeatin-7-glucoside (abbreviations for cytokinins according to Kamínek et al. 2000)



This study was funded by the Ministry of Education for Youth and Sports of the Czech Republic (1M0603 and LC06034), the Czech Science Foundation (P506/11/0774), the Grant Agency of the Academy of Sciences of the Czech Republic (IAA 600380701) and the Grant Agency of MUAF (IGA 4/4).


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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Marek Klemš
    • 1
  • Zdeňka Slámová
    • 1
  • Václav Motyka
    • 2
  • Jiří Malbeck
    • 2
  • Alena Trávníčková
    • 2
  • Ivana Macháčková
    • 2
  • Josef Holík
    • 3
  • Stanislav Procházka
    • 1
  1. 1.Department of Plant BiologyMendel UniversityBrnoCzech Republic
  2. 2.Institute of Experimental BotanyAcademy of Sciences of the Czech RepublicLysolaje, Prague 6Czech Republic
  3. 3.Isotope LaboratoryInstitute of Experimental Botany, Academy of Sciences of the Czech RepublicPrague 4Czech Republic

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