Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 136, Issue 1, pp 15–27 | Cite as

Structural and ultrastructural differences between field, micropropagated and acclimated leaves and stems of two Leucospermum cultivars (Proteaceae)

  • Emma SuárezEmail author
  • Carmen Alfayate
  • Juan Felipe Pérez-Francés
  • Juan Alberto Rodríguez-Pérez
Original Article


The anatomy of field, in vitro and acclimatized shoots (leaves and stems) of two cultivars of Leucospermum (L. cordifolium ‘Flame Spike’ and L. ‘Tango’) was compared using light, scanning and transmission electron microscopy. Field plants showed several scleromorphic anatomical structures related to excess solar radiation such as: cuticle thickness, subepidermal collenchyma and sclerenchyma. Furthermore, a large quantity of phenolic deposits present in the cell lumen of various tissues is also a scleromorphic feature. The special conditions during in vitro culture result in plantlets with abnormal morphology and anatomy. These disorders are associated with the gaseous environment in the culture vessels, low irradiance in the incubation chamber and the addition of sucrose, nutrients and growth regulators to the culture medium. After transfer from in vitro to ex vitro conditions, substantial changes in leaf and stem anatomy were observed, above all in cuticle thickness, epidermal characteristics (stomatal and trichome index, and stomatal and pore size), differentiation of leaf mesophyll, chloroplast structure, and amount and localization of phenolic deposits. These changes allowed the plants to adapt to the new environmental conditions. The study of anatomical features of in vitro shoots facilitated adapting the acclimation protocol to predict which plantlet would survive the critical acclimation stage.


Adaptation In vitro tissue culture Leucospermum Plant anatomy Proteaceae Tissue features 


Author contributions

ES and CA planned and designed the research, JAR-P contributed plant material, ES performed the experiments and collected data. ES, CA, JFP-F and JAR-P analysed the data, ES wrote the manuscript and CA supervised the writing. All the co-authors reviewed the manuscript before submission.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Emma Suárez
    • 1
    Email author
  • Carmen Alfayate
    • 2
  • Juan Felipe Pérez-Francés
    • 1
  • Juan Alberto Rodríguez-Pérez
    • 3
  1. 1.Pharmacy Section, Department of Botany, Ecology and Plant Physiology, Faculty of Health SciencesUniversity of La LagunaTenerifeSpain
  2. 2.Biology Section, Department of Biochemistry, Microbiology, Cell Biology and Genetics, Faculty of SciencesUniversity of La LagunaTenerifeSpain
  3. 3.Agricultural Engineering Section, Department of Agricultural, Nautical, Civil and Marine Engineering, Higher Polytechnic School of EngineeringUniversity of La LagunaTenerifeSpain

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