Alterations of root architecture and cell wall modifications in Tilia cordata Miller (Linden) growing on mining sludge.

Trees are considered good candidates for phytoremediation of soils contaminated with trace elements (TE), e.g. mine tailings. Using two year-old Tilia cordata plants, we demonstrated the nature and the scale of root architecture, especially root apices, as an indicator of mining sludge toxicity and plant capability to cope with these stress conditions. The novelty of our research is the analysis of the root response to substrate with extremely high concentrations of numerous toxic TE, and the 3D illustration of the disorders in root apex architecture using a clarity technique for confocal microscopy. The analysis demonstrates (1) a marked reduction in the size of the root apex zones (2) the occurrence of vascular tissues abnormally close to the root apex (3) collapse of the internal tissues in many root apices. Simultaneously, at the cellular level we observed some signs of a defensive response - such as a common increase of cell wall (CW) thickness and the formation of local CW thickenings - that enlarge the CW capacity for TE sequestration. However, we also detected harmful effects. Among others, a massive deposition of TE in the middle lamella which caused major damage - probably one of the reasons why the inner tissues of the root apex often collapsed – and the formation of incomplete CWs resulting in the occurrence of extremely large cells. Moreover, many cells of the root apex exhibited degenerated protoplasts. All these alterations indicate the harsh conditions for lime growth and survival and simultaneously, the manifestation of a defensive response. The obtained results allowed us to conclude that analysis of the nature and scale of structural alterations in roots can be useful indicators of plant ability to cope with stress conditions, e.g. in prospect of using the examined plants for reclamation of soils contaminated with TE. Image 1 • Clearing technique excellently documented the nature of root architecture alterations. • Root architecture analysis clearly shows the extremely high toxicity of the substrate. • Mining sludge exposure caused an increase of CW capacity for TE sequestration. • CW thickening occurred not only in the laboratory but also in similar to field conditions. • Presence of incomplete CW indicates a strong cell dysfunction. The nature and scale of alterations in root apex architecture can be considered as an indicator of mining sludge toxicity and the degree of plant resistance to contaminated substrate. [ABSTRACT FROM AUTHOR]