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Plant inoculation with arbuscular mycorrhizal fungi (AMF) is increasingly employed to enhance productivity and sustainability in agricultural ecosystems. In the present study, the potential benefits of AMF inoculation on young grapevines replanted in pots containing vineyard soil with high Cu concentration were evaluated. For this purpose, one-year-old cv. Touriga Nacional grapevines grafted onto 1103P rootstocks were further inoculated with Rhizoglomus irregulare or Funneliformis mosseae, or left non-inoculated, and maintained in a sterilized substrate under greenhouse conditions for three months. After this time, half of the plants were transplanted to containers filled with an Arenosol from a vineyard which had been artificially contaminated or not with 300 mg kg−1 of Cu. At the end of the growing season, soil nutrient concentration, soil dehydrogenase activity and mycorrhizal colonization rate were analyzed. Grapevine performance was assessed by measuring several vegetative growth and physiological parameters as well as nutrient concentrations in leaves and roots. In the non-contaminated soil, R. irregulare- and F. mosseae-inoculated plants had significantly greater root biomass than the non-inoculated ones. However, the opposite effect was observed in the Cu-contaminated soil, where non-inoculated plants performed better regarding shoot and root development. Concerning nutrient levels, an increase in Cu, Mg and Mn concentrations was observed in the roots of plants growing in the contaminated soil, although only Mn was translocated to leaves. This led to a large increase in leaf Mn concentrations, which was significantly higher in non-inoculated and F. mosseae- inoculated plants than in the R. irregulare- inoculated ones. Copper contamination induced a general decrease in leaf N, P and Fe concentrations as well as chlorosis symptoms. The largest decrease in N and P was observed in F. mosseae- inoculated plants, with 73 and 31.2%, respectively. However, these plants were the ones with the least decrease in Fe concentration (10% vs. almost 30% in the other two inoculation treatments). In conclusion, this study indicates that soil Cu levels can modify the outcome of AMF inoculations in young grapevines, disclosing new AMF-plant associations potentially relevant in vineyards with a tradition of Cu-based fungicide application.

& Key message Acacia melanoxylon produces abundant seeds leading to large seed banks in the soil. These seeds display a large viability and their germination is stimulated by heat. To control the populations, it is necessary to remove adults and young individuals, and to prevent seedling establishment after fire occupying the space with rapid growth and high competitive native species. & Context Acacia melanoxylon displays a widespread distribution in South West Europe, and an improved knowledge of its reproductive characteristics is required in order to control its expansion. & Aims This experiment was designed to provide useful indicators for an efficient management of A. melanoxylon populations based on its biological cycle in relation to fire. & Methods We explored the reproductive biology of A. melanoxylon, from seed dissemination-quantifying seed rain over a year, their germination with and without fire-the seedling and sapling banks and the structure of the adult population. We analysed the effects of fire, seed maturation and scarification on the viability of seeds and the stimulation of seed germination in the aerial seed bank and in the different strata of the soil seed bank. & Results Our results indicate that A. melanoxylon produced millions of seeds per ha and per year, half of which germinated and the other half went to the soil seed bank, maintaining the viability many years. The germination was the most critical step in the population dynamics of this species, and fire stimulates germination up to 90%. & Conclusion A. melanoxylon adults and seedlings removal, followed by colonization of rapid growth and high competitive native species that cover the ground very quickly would be a good control action.

Forest fires generate large amounts of ash and biochar, or black carbon (BC), that cover the soil surface, interacting with the soil’s constituents and its seedbank. This study concerns reproductive ecology assessments supported by molecular characterisation to improve our understanding of the effects of fire and fire residues on the germination behaviour of 12 arboreal species with a wide geographic distribution. For this purpose, we analysed the effects of three ash and one BC concentration on the germination of Acacia dealbata Link, A. longifolia (Andrews) Willd., A. mearnsii De Wild., A. melanoxylon R. Br., Pinus nigra Arnold, P. pinaster Aiton, P. radiata D. Don, P. sylvestris L., Quercus ilex L., Q. pyrenaica Willd., Q. robur L., and Q. rubra L. Each tree species was exposed to ash and BC created from its foliage or twigs (except for Q. rubra, which was exposed to ash and BC of Ulex europaeus L.). We monitored germination percentage, the T50 parameter, and tracked the development of germination over time (up to 1 yr). The BC of A. dealbata, P. pinaster, and Q. robur was analysed by pyrolysis-gas chromatography-mass spectrometry (PY-GC-MS) to assess the molecular composition. In six species, ash inhibited the germination, while in another five species, germination was not affected by ash or by BC. In Q. rubra, ash and BC stimulated its germination. This stimulating effect of the BC on Q. rubra is likely to be related to the chemical composition of the ash and BC obtained from Ulex feedstock. The BC of U. europaeus has a very different molecular composition than the other BC samples analysed, which, together with other factors, probably allowed for its germination stimulating effects.