Abstract
The pseudometallophyte Rumex acetosella L. occupies habitats with normal and high soil concentrations of zinc (Zn), lead (Pb), and copper (Cu). It remains unclear if the plants respond to the toxic metals by altering their morphology and increasing the resilience of their cells. We compared plants growing on soils contaminated with Zn/Pb (populations Terézia, Lintich), or Cu (populations Špania Dolina, Staré Hory), with those from non-contaminated soil (Dúbravka) in Slovakia, and analysed leaf structure, physiology, and metal contents by light and electron microscopy, element localization by energy-dispersive X-ray analysis (EDX) in scanning electron microscope, and by specific fluorescence dyes. In control population, the epidermis of the amphistomatic leaves of R. acetosella contained capitate glandular trichomes, consisting of four head (secretory), two stalk, and two basal cells. The ultrastructure of secretory cells revealed fine wall ingrowths bordered by plasma membrane protruding into the cytoplasm. The metallicolous populations had higher contents of Zn and Cu in the epidermal and glandular cells, and a higher density of both stomata and trichomes. Extensive cell wall labyrinth was present in the trichome secretory cells. Their abnormal number and elevated metal contents might indicate effects of heavy metals, especially of Cu, on mitosis and cell plate formation. Differences in leaf physiology were indicated by significantly higher cytoplasmic tolerance to Zn and Cu in metallicolous populations and by structural properties of glandular heads suggesting secretion of toxic metals. Our findings are suggestive of plant reactions to metal stress, which facilitate the populations to occupy the metal-contaminated sites.
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Acknowledgements
We thank Ivo Vávra, Institute of Electrical Engineering, Slovak Academy of Sciences, for providing the opportunity to use the JEOL 1200 electron microscope in his laboratory. CIUS is a member of EuroBioImaging and of VLSI.
Funding
This work was supported by the Grant Agency VEGA (grant no. 25/5086/05), Grant Agency APVV (grant no. APVV-0432-06), Aktion Österreich–Slowakei (grant no. 46s5], and the Austrian OEAD (grant PL 07_2018).
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Fig. S1
Cross sections of R. acetosella leaves from Zn/Pb site Terézia (a) and Cu site Staré Hory (b) with stoma (s) and glandular trichome (g) in the epidermal layer, palisade (pp) and spongy (sp) parenchyma and vascular bundles (vb) in the mesophyll. Scale bars = 100 μm (PNG 1204 kb)
Fig. S2
Ultrastructure of leaf mesophyll cells of Rumex acetosella plants from the control site Dúbravka (a,b), Zn/Pb site Terézia (c) and Cu site Špania Dolina (d). a-d large vacuoles (v) containing electron-dense inclusions (i), chloroplasts (ch) adjacent to the cell walls may contain starch grains (sg). b Sections of mesophyll cells showing irregular distribution of the vacuolar inclusions (i): numerous in the cell (1) and absent from the neighbouring cell (2). Scale bars = 1 μm (JPG 1454 kb)
Fig. S3
Higher magnification of the detail from the Fig. 4e showing parts of the stalk and basal cells of the trichome from Špania Dolina (Cu site), with Golgi body (G) and rough endoplasmic reticulum (RER) close to the cell wall crossed by plasmodesmata (pd). m mitochondria, v vacuole. Scale bar = 0.5 μm (JPG 1135 kb)
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Čiamporová, M., Nadubinská, M., Banásová, V. et al. Structural traits of leaf epidermis correspond to metal tolerance in Rumex acetosella populations growing on metal-contaminated soils. Protoplasma 258, 1277–1290 (2021). https://doi.org/10.1007/s00709-021-01661-x
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DOI: https://doi.org/10.1007/s00709-021-01661-x