NO is an important regulatory molecule in eukaryotes. Much of its effect is ascribed to the action of NO as a signalling molecule. However, NO can also directly modify proteins thus affecting their activities. Although the signalling functions of NO are relatively well recognized in plants, very little is known about its potential influence on the structural integrity of plant cells. In this study, the reorganization of the actin cytoskeleton, and the recycling of wall polysaccharides in plants via the endocytic pathway in the presence of NO or NO-modulating substances were analysed. The actin cytoskeleton and endocytosis in maize (Zea mays) root apices were visualized with fluorescence immunocytochemistry. The organization of the actin cytoskeleton is modulated via NO levels and the extent of such modulation is cell-type specific. In endodermis cells, actin cables change their orientation from longitudinal to oblique and cellular cross-wall domains become actin-depleted/depolymerized. The reaction is reversible and depends on the type of NO donor. Actin-dependent vesicle trafficking is also affected. This was demonstrated through the analysis of recycled wall material transported to newly-formed cell plates and BFA compartments. Therefore, it is concluded that, in plant cells, NO affects the functioning of the actin cytoskeleton and actin-dependent processes. Mechanisms for the reorganization of the actin cytoskeleton are cell-type specific, and such rearrangements might selectively impinge on the functioning of various cellular domains. Thus, the dynamic actin cytoskeleton could be considered as a downstream effector of NO signalling in cells of root apices.

Aims Ectomycorrhizal fungi can improve poplar growth and tolerance to heavy metal stress, and may be useful during the afforestation and phytoremediation of polluted regions with poplar trees. In this study, we determined the effects of the symbiotic interaction between Populus × canescens trees and Paxillus involutus strains different in their tolerance to lead. Methods In vitro inoculated and non-inoculated plants were treated with 0.75 mM Pb(NO₃)₂. The root colonization rate of the two fungal strains, as well as their impacts on poplar health and lead accumulation were examined. Results Based on the colonization level, the roots were classified into one of three categories: non-mycorrhized, changed (ie, fungal cells were present on the root surface, but the Hartig net did not fully develop), and fully mycorrhized. The lead-tolerant P. involutus strain colonized roots better than the non-tolerant strain (ie, changed and fully mycorrhized roots). Moreover, plants inoculated with the tolerant fungal strain grew better than the control plants (217 % increase in dry weight over the controls), and accumulated lead in the roots and stems. Conclusions Inoculation of P. × canescens trees with a Pb-tolerant strain of P. involutus improves host plant growth and may increase Pb phytostabilization potential.

Lead is a dangerous pollutant that accumulates in plant tissues and causes serious damage to plant cell macromolecules. However, plants have evolved numerous tolerance mechanisms, including ectomycorrhizae, to maintain cellular Pb2+ at the lowest possible level. When those mechanisms are successful, Pb-exposed plants should exhibit no negative phenotypic changes. However, actual molecular-level plant adjustments at Pb concentrations below the toxicity threshold are largely unknown, similar to the molecular effects of protective ectomycorrhizal root colonization. In this study, we (1) determined the molecular adjustments in plants exposed to Pb but without visible Pb stress symptoms and (2) examined ectomycorrhizal root colonization (the role of fungal biofilters) with respect to molecular-level Pb perception by plant root cells. Biochemical, microscopic, proteomic and metabolomic studies were performed to determine the molecular status of Populus × canescens microcuttings grown in agar medium enriched with 0.75 mM Pb(NO3)2. Noninoculated and inoculated with Paxillus involutus poplars were analyzed in two independent comparisons of the corresponding control and Pb treatments. After six weeks of growth, Pb caused no negative phenotypic effects. No Pb-exposed poplar showed impaired growth or decreased leaf pigmentation. Proteomic signals of intensified Pb sequestration in the plant cell wall and vacuoles, cytoskeleton modifications, H+-ATPase-14-3-3 interactions, and stabilization of protein turnover in chronically Pb-exposed plants co-occurred with high metabolomic stability. There were no differentially abundant root primary metabolites; only a few differentially abundant root secondary metabolites and no Pb-triggered ROS burst were observed. Our results strongly suggest that proteome adjustments targeting Pb sequestration and ROS scavenging, which are considerably similar but less intensive in ectomycorrhizal poplars than in control poplars due to the P. involutus biofilter (as confirmed in a mineral study), were responsible for the metabolomic and phenotypic stability of poplars exposed to chronic mild Pb stress.