Abstract
For many woody species, such as Quercus robur, cytokinins in the culture medium are required to maintain in vitro plant material. Among synthetic cytokinins, 6-benzylaminopurine (BAP) and kinetin (KIN) are the most frequently used. In addition to inducing shoots, cytokinins can cause morphophysiological disorders. Therefore, we aimed to investigate the anatomical, biochemical, and physiological alterations and profiles of Q. robur shoots exposed to two cytokinins, applied alone and in combination. Shoots previously established in vitro were transferred to WPM culture media supplemented with BAP at concentrations of 0, 1.25, and 3.50 µM combined with KIN at concentrations of 0, 0.62, and 1.25 µM totaling 9 treatments. Anatomical, physiological, and biochemical analyses were performed after 40 days of culture. BAP induced the formation of new buds with anatomically underdeveloped leaves; induced shoot-tip necrosis, which is considered a response to the inefficient transport of water and nutrients; reduced the thickness of the cell walls of phloem fibers; and decreased the content of phenolic compounds and photosynthetic pigments. These responses were less pronounced with co-exposure to KIN. In contrast, KIN alone stimulated a larger area of secondary xylem and more lignified cell walls. BAP can induce shoots with underdeveloped anatomical and biochemical characteristics. Shoots that grew with KIN alone had stem and leaf anatomical characteristics, indicating greater commitment to cellular differentiation than proliferation. When both cytokinins are combined, KIN can partially mitigate the deleterious effects of BAP on in vitro growth.
Key message
BAP generated shoots with underdeveloped anatomical and biochemical characteristics. KIN can partially mitigate the deleterious effects of BAP on in vitro growth.
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Data availability
All data generated in this study are included as a supplementary file.
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Acknowledgements
The authors acknowledge the Ulam Programme scholarship (PPN/ULM/2019/1/00037) Granted by Polish National Agency for Academic exchange. The authors also acknowledge Andreia Barcelos Passos Lima Gontijo, Lorenzo Toscano Conde, and Ludmila Cristina Oliveira for providing chemical reagents related to plant anatomy, and Tomasz Piotr Wyka for providing the FMS 2+ pulse-modulated chlorophyll fluorimeter. Additional support was provided by the Institute of Dendrology—Polish Academy of Sciences.
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JPRM, MKW, and JMLL performed experiments and analyzes. JPRM wrote the manuscript and performed the statistical analysis. EMK quantified phenolic compounds and their antioxidant activity. MMM performed the PCA analysis. MKW and PC provided the structure and contributed to the design and interpretation of the results.
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Martins, J.P.R., Wawrzyniak, M.K., Ley-López, J.M. et al. 6-Benzylaminopurine and kinetin modulations during in vitro propagation of Quercus robur (L.): an assessment of anatomical, biochemical, and physiological profiling of shoots. Plant Cell Tiss Organ Cult 151, 149–164 (2022). https://doi.org/10.1007/s11240-022-02339-9
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DOI: https://doi.org/10.1007/s11240-022-02339-9