Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 121, Issue 3, pp 579–590 | Cite as

Physiological and biochemical effects of a tetrahydropyranyl-substituted meta-topolin in micropropagated Merwilla plumbea

  • Stephen O. Amoo
  • Adeyemi O. Aremu
  • Mack Moyo
  • Taofik O. Sunmonu
  • Lucie Plíhalová
  • Karel Doležal
  • Johannes Van Staden
Original Paper


In comparison to the most active cytokinins (CKs) previously reported for the micropropagation of Merwilla plumbea, we examined the effect of meta-topolin tetrahydropyran-2-yl (mTTHP—a novel aromatic CK derivative) on in vitro adventitious shoot production, rooting and photosynthetic pigment content of regenerated plants. Its carry-over effect on ex vitro growth, photosynthetic performance and antioxidant enzyme system of this bulbous medicinal plant was also investigated. The treatments with mTTHP and meta-topolin riboside (mTR) gave the highest number of adventitious shoots when compared to thidiazuron (TDZ) application and the control. The highest rooting frequency was observed in mTTHP treatments. Unlike in mTTHP treatments, an increase in mTR or TDZ concentration beyond 0.5 µM resulted in a significant decrease in the concentrations of all the photosynthetic pigments quantified. After 6 months of ex vitro growth, regenerated plants from 0.5 µM mTTHP treatment had the highest significant total leaf area, total leaf fresh weight and bulb size compared to all mTR and TDZ-treated plants. Plants regenerated from mTTHP or mTR treatments demonstrated a high capacity for energy dissipation in comparison to TDZ-regenerated plants with low photochemical quenching, PSII quantum efficiency and non-photochemical quenching. Despite a significant increase in the antioxidant enzyme activities, malondialdehyde concentration was significantly high in the leaves of TDZ-regenerated plants compared to other CK treatments. This finding indicated a high production of reactive oxygen species beyond the scavenging efficiency of the antioxidant enzymes leading to oxidative stress and subsequent low biomass accumulation in TDZ-derived plants.


Antioxidant enzymes Chlorophyll fluorescence Medicinal plants Thidiazuron Topolins 







Relative electron transport rate


Maximum photochemical efficiency of PSII




meta-Methoxytopolin 9-tetrahydropyran-2-yl or 6-(3-methoxybenzylamino-9-tetrahydropyran-2ylpurine


Murashige and Skoog


meta-Topolin riboside


meta-Topolin tetrahydropyran-2-yl or 6-(3-hydroxylbenzylamino)-9-tetrahydropyran-2-ylpurine


Non-photochemical quenching


Plant growth regulator




Photosynthetic photon flux


Photosystem II


Plant tissue culture


Photochemical quenching


Superoxide dismutase






Actual quantum yield of PSII



The Claude Leon Foundation and University of KwaZulu-Natal provided financial support. This work was also supported by the Centre of the Region Hana for Biotechnological and Agricultural Research, Palacký University (Olomouc), Czech Republic (Grant No. ED0007/01/01), the Ministry of Education, Youth and Sports, Czech Republic (Grant L01204 from the National Program of Sustainability) as well as by IGA of Palacký University (Grant IGA_PrF_2014006). We thank Prof. R.P. Beckett (School of Life Sciences, University of KwaZulu-Natal) for his support relating to the use of chlorophyll fluorometer. Mrs Alison Young and her staff (Botanical Garden, University of KwaZulu-Natal, Pietermaritzburg) are gratefully acknowledged for the maintenance of the greenhouse facilities.


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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Stephen O. Amoo
    • 1
    • 3
  • Adeyemi O. Aremu
    • 1
  • Mack Moyo
    • 1
  • Taofik O. Sunmonu
    • 1
  • Lucie Plíhalová
    • 2
  • Karel Doležal
    • 2
  • Johannes Van Staden
    • 1
  1. 1.Research Centre for Plant Growth and Development, School of Life SciencesUniversity of KwaZulu-Natal PietermaritzburgScottsvilleSouth Africa
  2. 2.Department of Chemical Biology and Genetics, Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Institute of Experimental BotanyAcademy of Sciences of Czech RepublicOlomouc-HoliceCzech Republic
  3. 3.Agricultural Research CouncilRoodeplaat Vegetable and Ornamental Plant InstitutePretoriaSouth Africa

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