, Volume 236, Issue 6, pp 1775–1790 | Cite as

Physiological responses and endogenous cytokinin profiles of tissue-cultured ‘Williams’ bananas in relation to roscovitine and an inhibitor of cytokinin oxidase/dehydrogenase (INCYDE) treatments

  • Adeyemi O. Aremu
  • Michael W. Bairu
  • Ondřej Novák
  • Lenka Plačková
  • Marek Zatloukal
  • Karel Doležal
  • Jeffrey F. Finnie
  • Miroslav Strnad
  • Johannes Van StadenEmail author
Original Article


The effect of supplementing either meta-topolin (mT) or N 6-benzyladenine (BA) requiring cultures with roscovitine (6-benzylamino-2-[1(R)-(hydroxymethyl)propyl]amino-9-isopropylpurine), a cyclin-dependent kinase (CDK) and N-glucosylation inhibitor, and INCYDE (2-chloro-6-(3-methoxyphenyl)aminopurine), an inhibitor of cytokinin (CK) degradation, on the endogenous CK profiles and physiology of banana in vitro was investigated. Growth parameters including multiplication rate and biomass were recorded after 42 days. Endogenous CK levels were quantified using UPLC–MS/MS while the photosynthetic pigment and phenolic contents were evaluated spectrophotometrically. The highest regeneration rate (93 %) was observed in BA + roscovitine while mT + INCYDE plantlets produced most shoots. Treatment with BA + roscovitine had the highest shoot length and biomass. Although not significant, there was a higher proanthocyanidin level in BA + roscovitine treatments compared to the control (BA). The levels of total phenolics and flavonoids were significantly higher in mT + roscovitine treatment than in the mT-treated regenerants. The presence of roscovitine and/or INCYDE had no significant effect on the photosynthetic pigments of the banana plantlets. Forty-seven aromatic and isoprenoid CKs categorized into nine CK-types were detected at varying concentrations. The presence of mT + roscovitine and/or INCYDE increased the levels of O-glucosides while 9-glucosides were higher in the presence of BA. Generally, the underground parts had higher CK levels than the aerial parts; however, the presence of INCYDE increased the level of CK quantified in the aerial parts. From a practical perspective, the use of roscovitine and INCYDE in micropropagation could be crucial in the alleviation of commonly observed in vitro-induced physiological abnormalities.


Cyclin-dependent kinase Cytokinin metabolism Micropropagation Musa spp. Photosynthesis Plant secondary metabolites 



Analysis of variance


N 6-Benzyladenine


N 6-Benzyladenine-9-glucoside


N 6-Benzyladenosine


N 6-Benzyladenosine-5′-monophosphate


Cyanidin chloride equivalents


Cyclin-dependent kinase


Catechin equivalents




Cytokinin oxidase/dehydrogenase








cis-Zeatin riboside


cis-Zeatin riboside-5′-monophosphate


cis-Zeatin-O-glucoside riboside








Dihydrozeatin riboside


Dihydrozeatin riboside-5′-monophosphate


Dihydrozeatin-O-glucoside riboside


Duncan’s multiple range test


Gallic acid equivalents


Immunoaffinity chromatography




N 6-Isopentenyladenine


N 6-Isopentenyladenine-9-glucoside


N 6-Isopentenyladenosine


N 6-Isopentenyladenosine-5′-monophosphate








Kinetin riboside


Kinetin riboside-5′-monophosphate


Multiple reaction monitoring


Murashige and Skoog medium








meta-Topolin riboside




meta-Topolin-O-glucoside riboside








ortho-Topolin riboside




ortho-Topolin-O-glucoside riboside


Plant growth regulator


Photosynthetic photon flux density




Plant tissue culture




para-Topolin riboside




para-Topolin-O-glucoside riboside








trans-Zeatin riboside


trans-Zeatin riboside-5′-monophosphate


trans-Zeatin-O-glucoside riboside


Ultra performance liquid chromatography



The University of KwaZulu-Natal (Pietermaritzburg), South Africa and Centre of the Region Haná for Biotechnological and Agricultural Research, Palacký University (Olomouc), Czech Republic for providing financial support (Grant No. ED0007/01/01).


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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Adeyemi O. Aremu
    • 1
  • Michael W. Bairu
    • 1
  • Ondřej Novák
    • 2
  • Lenka Plačková
    • 2
    • 3
  • Marek Zatloukal
    • 2
    • 3
  • Karel Doležal
    • 2
    • 3
  • Jeffrey F. Finnie
    • 1
  • Miroslav Strnad
    • 2
    • 3
  • Johannes Van Staden
    • 1
    Email author
  1. 1.Research Centre for Plant Growth and Development, School of Life SciencesUniversity of KwaZulu-NatalPietermaritzburgSouth Africa
  2. 2.Laboratory of Growth Regulators, Institute of Experimental Botany AS CRPalacký UniversityOlomoucCzech Republic
  3. 3.Faculty of Science, Centre of the Region Haná for Biotechnological and Agricultural ResearchPalacký UniversityOlomoucCzech Republic

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