The Vicia micronucleus assay was standardized in an international protocol, ISO 29200, "Assessment of genotoxic effects on higher plants-Vicia faba micronucleus test," for soil or soil materials (e.g., compost, sludge, sediment, waste, and fertilizing materials). The aim of this interlaboratory study on the Vicia micronucleus assay was to investigate the robustness of this in vivo assay in terms of its applicability in different countries where each participant were asked to use their own seeds and reference soil, in agreement with the ISO 29200 standard. The ISO 29200 standard protocol was adopted for this study, and seven laboratories from three countries (France, Italy, and Brazil) participated in the study. Negative and positive controls were correctly evaluated by 100 % of the participants. In the solid-phase test, the micronucleus frequency (number of micronuclei/1,000 cells) varied from 0.0 to 1.8 for the negative control (i.e., Hoagland's solution) and from 5.8 to 85.7 for the positive control (i.e., maleic hydrazide), while these values varied from 0.0 to 1.7 for the negative control and from 14.3 to 97.7 for the positive control in the case of liquid-phase test. The variability in the data obtained does not adversely affect the robustness of the protocol assessed, on the condition that the methodology described in the standard ISO 29200 is strictly respected. Thus, the Vicia micronucleus test (ISO 29200) is appropriate for complementing prokaryotic or in vitro tests cited in legislation related to risk assessment of genotoxicity potential.
Biochar amendments, i.e., the solid product of biomass pyrolysis, reduce soil metal availability, which may lower the toxicity of metal-contaminated soils. A direct link between the decrease in soil metal availability and improved plant development is however often difficult to establish, as biochar may induce undesirable side effects on plant growth, e.g., a modification to plant nutrition. In order to investigate toxicity processes at a cellular level, roots of Vicia faba were exposed for 7 days to three metal-contaminated substrates and one control soil, amended with a 0 or 5% (w/w) addition of a wood-derived biochar. Exposure to pure biochar was also tested. Root tip cells were then observed to count the number of micronuclei as an estimation of DNA damage and the number of cells at mitosis stage. Results showed that biochar amendments led to a significant decrease in soil metal availability (Cd, Cu, Ni, Pb, and Zn) and to enhance root development on acidic substrates. The micronucleus frequency in root tip cells was positively correlated and the number of mitotic cells negatively, to the extractability of Zn in soils and to the concentration of Zn in secondary roots. Exposure to pure biochar caused a lower production of roots than most soil substrates, but led to the lowest number of observed micronuclei. In conclusion, biochar amendments can reduce the genotoxicity associated with the presence of metallic contaminants in soils, thereby potentially improving plant growth.
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