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In vitro culture of Hibiscus rosa-sinensis L.: Influence of iron, calcium and BAP on establishment and multiplication

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Some factors influencing in vitro cultures of potted Hibiscus rosa-sinensis L. using nodal cuttings were investigated. A protocol using a modified MS medium helped to overcome chlorosis, shoot tip necrosis (STN) and leaf drop. These disorders have been caused by mineral imbalance associated with calcium and iron deficiency. STN and leaf drop were overcome by increasing calcium level from 3 mM (MS standard concentration) to 9 mM, and this increase, in addition, enhanced shoot dry weight and shoot extension. The chlorophyll content and leaf area increased by increasing the iron concentration 3-fold from 98 μM to 295 μM. Furthermore, substituting Fe-EDTA with Fe-EDDHA resulted in an increase in chlorophyll content, leaf area and shoot extension. The most suitable multiplication medium for H. rosa-sinensis L. was demonstrated to be a modified MS medium containing 2.2 μM BAP and increased concentrations of calcium at 9 mM and iron at 295 μM provided as Fe-EDDHA. The shoots were rooted in half-strength modified MS medium containing 2.7 μM NAA. Acclimatization was successful with all shoots with or without roots.

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Chlorophyll content index


Ethylenediamine di-2-hydroxy-phenyl acetate ferric


Ethylenediamine tetraacetate ferric sodium iron


Indole-3-butyric acid


α-Naphtaleneacetic acid


Photosynthetically active radiation


Shoot tip necrosis


Phenyl-N1-1,2,3,-thiadiazol-5-ylurea (Thidiazuron)


McCown Woody Plant Medium


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The study was funded by a PhD grant from the University of Copenhagen, Faculty of Life Sciences, and the authors would like to thank Graff Kristensen A/S, Sabro, Denmark for providing the plant material for the study.

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Correspondence to Brian Christensen.

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Christensen, B., Sriskandarajah, S., Serek, M. et al. In vitro culture of Hibiscus rosa-sinensis L.: Influence of iron, calcium and BAP on establishment and multiplication. Plant Cell Tiss Organ Cult 93, 151–161 (2008). https://doi.org/10.1007/s11240-008-9354-4

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  • Calcium chloride
  • Fe-EDDHA
  • Fe-EDTA
  • Iron chlorosis
  • Micropropagation