Comparative analysis of Cd and Zn impacts on root distribution and morphology of Lolium perenne and Trifolium repens: implications for phytostabilization

Backgrounds and aims The phytostabilization potential of plants is a direct function of their root systems. An experimental design was developed to investigate the impact of Cd and Zn on the root distribution and morphology of Lolium perenne and Trifolium repens. Methods Seedlings were transplanted into columns filled with washed quartz and irrigated daily with Cdor Zn-containing nutrient solutions during 1 month. Root biomass, root length density (RLD) and diameter were subsequently quantified as a function of depth. Pot experiments were also performed to quantify metal, lignin and structural polysaccharides concentrations as well as cell viability. Results Lolium perenne accumulated Cd and Zn in the roots whereas T. repens was unable to restrict heavy metal translocation. Cadmium and Zn reduced rooting depth and RLDbut induced thick shoot-borne roots in L. perenne. Cd-induced root swelling was related to lignification occurring in the exodermis and parenchyma of central cylinder. Hemicelluloses and lignin did not play a key role in root metal retention. Cadmium slightly reduced mean root cell viability whereas Zn increased this parameter in comparison to Cd. Conclusions Even though plant species like Lolium perenne and Trifolium repens may appear suitable for a phytostabilization scheme based on their shoot metal tolerance, exposure to toxic heavy metals drastically impairs their root distribution. This could jeopardize the setting up of phytostabilization trials. The metal-induced alterations of root system properties are clearly metal- and speciesspecific. At sites polluted with multiple metals, it is therefore recommended to first test their impact on the root system of multiple plant species so as to select the most appropriate species for each site

Seed Priming of Trifolium repens L. Improved Germination and Early Seedling Growth on Heavy Metal-Contaminated Soil

Abstract Seed priming effects on Trifolium repens were analysed both in Petri dishes and in two soils (one unpolluted soil and a soil polluted with Cd and Zn). Priming treatments were performed with gibberellic acid 0.1 mM at 22 °C during 12 h or with polyethylene glycol (−6.7 MPa) at 10 °C during 72 h. Both priming treatments increased the germination speed and the final germination percentages in the presence of 100 μM CdCl2 or 1 mM ZnSO4. Flow cytometry analysis demonstrated that the positive effect of priming was not related with any advancement of the cell cycle in embryos. Seed imbibition occurred faster for primed seeds than for control seeds. X-ray and electronic microscopy analysis suggested that circular depressions on the seed coat, in addition to tissue detachments inside the seed, could be linked to the higher rate of imbibition. Priming treatments had no significant impact on the behaviour of seedlings cultivated on nonpolluted soil while they improved seedling emergence and growth on polluted soil. The two priming treatments reduced Zn accumulation. Priming with gibberellic acid increased Cd accumulation by young seedlings while priming with polyethylene glycol reduced it. Priming improved the light phase of photosynthesis and strengthened the antioxidant system of stressed seedlings. Optimal priming treatment may thus be recommended as efficient tools to facilitate revegetation of former mining area

Morphological changes induced by heavy metals in dandelion (Taraxacum officinale Web.) growing on mine soils.

Purpose Heavy metal accumulation produces significant physiological and biochemical responses in vascular plants. Plants growing on abandoned mine sites are of particular interest, since they are genetically tolerant to high metal concentrations. In this work, we examined the effect of heavy metals (HMs) on the morphology of T. officinale growing in pots with mine soils, with the following objectives: (1) to determine the evolution of HM concentration in leaves and roots over 3 years of cultivation; (2) to highlight possible damage at anatomical and cytological level. Materials and methods Wild specimens of Taraxacum officinale Web., with their soil clod, were gathered from three sites with different contamination levels by heavy metals (Cd, Cr, Cu, Fe, Pb, Zn) in the abandoned Imperina Valley mine (Northeast Italy). A control plant was also gathered from a non-contaminated site nearby. Plants were cultivated in pots at the botanical garden of the University of Florence (HBF), and appeared macroscopically not affected by toxic signals (reduced growth, leaf necrosis) possibly induced by soil HM concentration. Leaves and roots taken at the same growing season were observed by light microscopy and transmission electron microscopy. Results and discussion Light microscopy observations show a clear difference in the cellular organisation of noncontaminated and contaminated samples. The unpolluted samples present a well-organised palisade tissue and spongy photosynthetic parenchyma. Samples from contaminated sites, instead, present a palisade parenchyma less organised, and a reduction of leaf thickness proportional to HM concentration. The poor structural organisations, and the reduced foliar thickness of the contaminated plants, are related to soil contamination. Differences in root micromorphology concern the cortical parenchyma. Moreover, all the samples examined present mycorrhiza. Ultrastructure observations of the parenchyma cells show mitochondrial structure alteration, with lacking or reduced cristae of the internal membrane at increasing metal content. Instead, chloroplast organisation does not present significant differences, particularly in number and compartmentalization of thylakoids. Conclusions Although macromorphology does not present evidence of phytotoxicity, the recorded observations of the micromorphological characteristics of leaves and roots, show a suffering state of the plants, strictly related to HM content. Leaching reduced partly the HM content of the soil, therefore decreasing their phytotoxic effect. A gradual restoration of leaf organisation suggests that somewhat resilience occurred in plants. Moreover, the presence of stress-tolerant mycorrhizal fungi could contribute to reduce metal toxicity

Mechanisms of metal-phosphates formation in the rhizosphere soils of pea and tomato: environmental and sanitary consequences

International audiencePurpose At the global scale, soil contamination with persistent metals such as lead (Pb), zinc (Zn), and copper (Cu) induces a serious threat of entering the human food chain. In the recent past, different natural and synthetic compounds have been used to immobilizemetals in soil environments. However, the mechanisms involved in amendment-induced immobilization of metals in soil remained unclear. The objective of the present work was therefore to determine the mechanisms involved in metal-phosphates formation in the rhizospheric soils of pea and tomato currently cultivated in kitchen gardens. Materials and methods Pea and tomato were cultivated on a soil polluted by past industrial activities with Pb and Zn under two kinds of phosphate (P) amendments: (1) solid hydroxyapatite and (2) KH2PO4. The nature and quantities of metal-P formed in the rhizospheric soils were studied by using the selective chemical extractions and employing the combination of X-ray fluorescence micro-spectroscopy, scanning electron microscopy, and electron microprobe methods. Moreover, the influence of soil pH and organic acids excreted by plant roots on metal-P complexes formation was studied. Results and discussion Our results demonstrated that P amendments have no effect on metal-P complex formation in the absence of plants. But, in the presence of plants, P amendments cause Pb and Zn immobilization by forming metal-P complexes. Higher amounts of metal-P were formed in the pea rhizosphere compared to the tomato rhizosphere and in the case of soluble P compared to the solid amendment. The increase in soil-metal contact time enhanced metal-P formation. Conclusions The different forms of metal-P formed for the different plants under two kinds of P amendments indicate that several mechanisms are involved in metal immobilization. Metal-P complex formation in the contaminated soil depends on the type of P amendment added, duration of soil-plant contact, type of plant species, and excretion of organic acids by the plant roots in the rhizosphere