Diversity and botanical composition of native species in the Pampa biome in vineyards cultivated on soils with high levels of copper, zinc and manganese and phytoremediation potential.

Viticulture allows the preservation of native species inside vineyards in the Pampa biome. However, phytosanitary treatments in these areas can increase the levels of Cu, Zn and Mn. The study aimed to (i) verify the influence of Cu, Zn and Mn contents in Pampa biome soils; (ii) identify variables related to Cu, Zn and Mn that most contribute to the variation in richness, diversity, and dry matter production of native vegetation, (iii) investigate the phytoremediation potential of species present in vineyards. Botanical composition, Cu, Zn, Mn available in the soil, and plant nutritional composition in two vineyards (V1 and V2) and native field (NF) were evaluated. Vineyards showed higher Cu, Zn and Mn contents in the soil, resulting in the lowest biomass, richness, and diversity of native species. Mn in tissue was the most important variable in explaining the variation in dry matter. Zn in the soil helped to explain the difference in species richness and diversity. P concentration in tissue was important in elucidating the variation in species diversity. Paspalum plicatulum and Paspalum notatum have potential for phytostabilization of metals in vineyards.

Phytoremediation of Cu-contaminated vineyard soils in Brazil: A compendium of Brazilian pot studies.

Vineyard soils can be contaminated by copper (Cu) due to successive applications of fungicides and organic fertilizers. Soil remediation can be addressed by altering soil properties or selecting efficient Cu-extracting cover crops tolerant to Cu toxicity. Our objectives were to synthesize the Cu-extracting efficiency by plant species tested in Brazil, classify them according to Cu resistance to toxicity, and assess the effect of soil properties on attenuating Cu toxicity. We retrieved results from 41 species and cultivars, totaling 565 observations. Freshly added Cu varied between 50 and 600 mg Cu kg-1 of soil across studies. The partition of Cu removal between the above- and below-ground portions was scaled as a logistic variable to facilitate data synthesis. The data were analyzed using the Adaboost machine learning model. Model accuracy (predicted vs. actual values) reached R2  = 0.862 after relating species, cultivar, Cu addition, clay, SOM, pH, soil test P, and Cu as features to predict the logistic target variable. Tissue Cu concentration varied between 7 and 105 mg Cu kg-1 in the shoot and between 73 and 1340 mg Cu kg-1 in the roots. Among soil properties, organic matter and soil test Cu most influenced the accuracy of the model. Phaseolus vulgaris, Brassica juncea, Ricinus communis, Hordeum vulgare, Sorghum vulgare, Cajanus cajan, Solanum lycopersicum, and Crotolaria spectabilis were the most efficient Cu-extracting cover crops, as shown by positive values of the logistic variable (shoot removal > root removal). Those Cu-tolerant plants showed differential capacity to extract Cu in the long run.

Subcellular distribution and physiological responses of native and exotic grasses from the Pampa biome subjected to excess manganese.

Fungicides containing manganese (Mn) applied to control plant diseases increase the concentration of Mn in soils, which may potentiate Mn toxicity in acid soils. Some species of wild grasses, such as those from the Pampa biome located in South America, or even those introduced into this biome, may possess different mechanisms of tolerance to excess Mn. The present study aimed to evaluate the subcellular distribution and physiological and biochemical responses of exotic and native grasses from the Pampa biome, cultivated in Mn excess. The experiment was conducted in nutrient solution in a greenhouse, in an entirely randomized design, bifactorial 4 × 4, consisting of four Mn concentrations (2 [control], 300, 600 and 900 µM) and four species (two exotic: Avena strigosa and Lolium multiflorum; and two native: Paspalum notatum and Paspalum plicatulum). At 27 days of exposure to the treatments, biomass and growth rates, leaf gas exchange with the environment, photosynthetic pigment concentrations of malondialdehyde and H2O2, antioxidant enzyme activities (SOD and POD), and subcellular distribution of Mn were evaluated. Most of the grasses showed high concentration of Mn in tissues, mainly, in the shoot. In the presence of 900 µM Mn, more than 80% of the absorbed Mn was compartmentalized in the cell walls and vacuoles of the cells. Compartmentalization of Mn excess into metabolically less active organelles is the main tolerance factor in grasses. Physiological and biochemical responses were stimulated in the presence of 300 µM Mn, while 900 µM Mn negatively affected biochemical-physiological responses of grasses. The species L. multiflorum was most sensitive to excess Mn, while P. notatum was the most tolerant.

Physiological and biochemical characterization of copper-toxicity tolerance mechanism in grass species native to Pampa Biome and Atlantic Forest for use in phytoremediation.

Orchards and vineyards account for significant copper (Cu) accumulation in the soil due to frequent Cu fungicide applications to control leaf diseases. Although grass species are distributed in these areas likely because of their physiological mechanisms to combat Cu toxicity-related stress, the aim of the present study is to identify grass species presenting biochemical-physiological responses that feature adaptive Cu toxicity tolerance mechanisms. Three grass species native to the Pampa and Atlantic Forest biomes (Paspalum notatum, P. plicatulum, and P. urvillei) and an exotic species (Cynodon dactylon) were tested. Plants were cultivated in pots filled with 4 kg of typic Hapludalf soil, under two Cu availability, control, and toxicity conditions (80 mg Cu kg soil-1). Photosynthetic parameters, relative growth rate, root dry matter, shoot dry matter, the activity of stress-fighting enzymes (superoxide dismutase and guaiacol peroxidase), root biometry, soluble organic carbon, soil pH, and electrical conductivity were evaluated. P. notatum and P. urvillei have physiological characteristics that allow high translocation factor and Cu accumulation in the root and shoot, and it allows their use in phytoremediation processes due to (1) greater activity of stress-fighting enzymes such as POD in the shoot; (2) to larger diameter roots, which allow greater Cu complexation in them - they are lesser sensitive to stress caused by Cu than the other species; and (3) greater soluble organic carbon exudation in the rhizosphere than species P. plicatulum and C. dactylon, which can complex Cu2+ and reduce the presence of forms toxic to plants.

Potential phytoremediation of Pampa biome native and invasive grass species cohabiting vineyards contaminated with Cu in Southern Brazil.

The objectives were (a) to evaluate whether grasses native to the Pampa biome, Axonopus affinis Chase, Paspalum notatum Flüggé and Paspalum plicatulum Michx, and the invasive grass Cynodon dactylon (L.). Pers have the potential to phytoremediate soil contaminated with Cu (0, 35 and 70 mg Cu kg-1); (b) assess whether the growth of these species is compromised by the excess of Cu available in the soil; and (c) determine the impact of excess Cu on the physiological responses of the studied species. C. dactylon presented the best performance in soil contaminated with 35 mg of Cu kg-1. In C. dactylon, the concentrations of chlorophyll b and carotenoids increased, as did the photosynthetic rate and plant growth. Phytotoxic effects of Cu in soil contaminated with 70 mg of Cu kg-1 were more severe on A. affinis and led to plant death. The other species presented reduced photosynthetic and growth rates, as well as increased activity of antioxidant enzymes such as superoxide dismutase and guaiacol peroxidase. This very same Cu level has decreased photosynthetic pigment concentrations in P. notatum and P. plicatulum. On the other hand, it did not change chlorophyll a and b concentrations in C. dactylon and increased carotenoid concentrations in it. High values recorded for Cu bioaccumulation-in-grass-root factor, mainly in P. plicatulum, have indicated that the investigated plants are potential phytostabilizers. High C. dactylon biomass production-in comparison to other species-compensates for the relatively low metal concentration in its tissues by increasing metal extraction from the soil. This makes C. dactylon more efficient in the phytoremediation process than other species.

The tolerance of grapevine rootstocks to copper excess and to the use of calcium and phosphorus to mitigate its phytotoxicity.

High soil copper (Cu) concentrations in vineyards can cause phytotoxicity to grapevine rootstocks. In order to mitigate toxicity, the use of grapevine rootstock genetic variation and the application of amendments are possible strategies. The aim of this study is to assess the tolerance of grapevine rootstocks to Cu excess and whether phosphorus (P) and calcium (Ca) can reduce phytotoxicity caused by Cu. Grapevine rootstock seedlings were produced from selected stakes: Paulsen 1103 (Vitis berlandieri × Vitis rupestris); SO4 (Vitis berlandieri × Vitis riparia); IAC 572 ((Vitis Riparia × Vitis rupestris) × Vitis caribaea); and Isabel (Vitis labrusca). Seedlings were grown in nutrition solution added with the following treatments: 0.3 µM Cu (control); 60 µM Cu; 60 µM Cu and 62 mg L-1 P; 60 µM Cu and 400 mg L-1 Ca. High Cu concentration caused phytotoxicity in all rootstocks, impairing their growth and decreasing nutrient concentration and photosynthetic activity. P and Ca addition had positive effect on the photosynthetic activity of all rootstocks, although it was not enough to revert growth to levels comparable with controls. Overall, based on the results, the application of P and Ca was not efficient in mitigating Cu phytotoxicity in grapevine plants grown in solution. Isabel was the most sensitive rootstock to Cu phytotoxicity, whereas Paulsen 1103 and SO4 presented more tolerance and can be used, together with other management strategies, in contaminated vineyard areas. Therefore, careful genotype rootstock selection for use in high Cu soils is important, while Ca and P are not efficient mitigators of Cu toxicity.

Root system structure as a criterion for the selection of grapevine genotypes that are tolerant to excess copper and the ability of phosphorus to mitigate toxicity.

Using tolerant genotypes and the correct use of fertilizers can mitigate the negative effect of elevated Cu levels in the growing medium. In this context, the study aimed to evaluate the effects of excess Cu in the root system and the effectiveness of phosphorus (P) in minimizing the phytotoxicity of Cu in three genotypes: IAC 572 [(Vitis riparia x V. rupestris) x V. caribaea], Magnolia (V. rotundifolia) and Paulsen 1103 (V. berlandieri x V. rupestris). The plants were grown in nutrient solutions and were supplemented with the following treatments: 0.3 µM Cu (Control), 60 µM Cu (Cu) and 60 µM Cu and 62 mg L-1 P (Cu + P). Root samples were sectioned for microscopy analyses, and the shoot lengths, shoot and root dry matter, relative growth rates (RGR) and tissue nutrient contents were also evaluated. The roots of the genotypes that were cultivated with high Cu concentrations produced greater numbers of branches and larger diameters, except for Magnolia genotype that was cultivated in a Cu + P solution, which had an organization similar to the control. Excess Cu caused accumulations of phenolic compounds and decreased shoot lengths, dry matter and RGR in the genotypes. In the treatments with excess Cu, there were increases in this element in the tissues, but P decreased the metal concentrations in Magnolia roots. Therefore, Cu accumulations alter the root system development patterns, growth parameters and tissue nutrient contents in the studied genotypes. Magnolia has a higher tolerance and is also the only genotype for which the use of P has been shown to be effective.

Soil tillage affects soybean growth and promotes heavy metal accumulation in seeds.

When soybean is grown in soils with high heavy metal concentrations, it may introduce those contaminants into the human food chain, posing risks to human health. This study evaluated the effect of tilling the soil with high Cu, Zn, and Mn levels on soybean physiology and metal accumulation in seeds. Disturbed and undisturbed soil samples were collected in two different sites: a vineyard with high heavy metal concentration and a grassland area, containing natural vegetation. Two soybean cultivars were sown and grown in the greenhouse. Photosynthetic parameters and biochemical analysis of oxidative stress were performed. Cu, Zn, and Mn in leaves and seeds, dry mass, and weight of seeds were evaluated. Soil structure had a high influence on plant growth and physiology, while soil site had a high impact on heavy metal accumulation in leaves and seeds. Soybean plants that grown in vineyard soils with high heavy metal concentrations, accumulated 50% more Zn in leaves and seeds, 70% more Cu in leaves, and 90% more Cu in seeds, than those plants grown in grassland soils. Besides, Zn concentration in seeds was higher than the permissible limit. Moreover, the disturbance of both vineyard soil and grassland soil was not good for plant growth and physiology, which have increased TBARS and H2O2 concentration in plants, transpiration rate, metal concentration in leaves and seeds. Soil disturbance may have caused organic matter oxidation and changes in the composition and quantity of soil microorganisms and it affects the availability of other nutrients in the soil.

Iron fertilization to enhance tolerance mechanisms to copper toxicity of ryegrass plants used as cover crop in vineyards.

Ryegrass (Lolium perenne L.) is a plant species that can express mechanisms of tolerance to copper (Cu) toxicity. Therefore, the agronomical approach of intercropping system with ryegrass may represent a promising tool to limit the onset of Cu toxicity symptoms in the other intercropped plants species, particularly when an inadequate nutrient availability like iron (Fe) shortage is also concurrently present. This study aimed at assessing the mechanisms involved in the mitigation of Cu phytotoxicity and the stress effects on plant growth, root morphology and nutrition of ryegrass fertilized with two different Fe sources. To this purpose, seedlings of ryegrass were hydroponically grown for 14 days in controlled conditions with 4 different levels of Cu (0.2, 5.0, 25 and 50 µM) and with either 100 µM Fe-EDDHA or Fe-EDTA. Results show that high levels of Cu availability enhanced the root content of organic anions as well as the root exudation. Different Fe fertilizations at the condition of 50 µM Cu induced changes in root phenolic compounds, citrate and fumarate contents and the exudation pattern of phenolic compounds. Differences in plant growth were not observed between the two Fe sources, although Cu concentration in plant tissue fed with Fe-EDTA was lower in the condition of 50 µM Cu. The enhanced root exudation of Cu-complexing organic compounds (including phenolics) in ryegrass plants when exposed to excessive Cu availability could be at the basis of the ameliorated edaphic rhizosphere conditions (lower Cu availability). For this reason, from the agronomical point of view ryegrass plants used in intercropping systems with crops like vine plants could represent a promising strategy to control Cu toxicity in vineyard soils. Further studies under the field conditions must be taken to support present findings.

Potential of vermicompost and limestone in reducing copper toxicity in young grapevines grown in Cu-contaminated vineyard soil.

Foliar fungicide application in grapevines increases the content of heavy metals such as copper (Cu) in vineyard soils, which may reach phytotoxic levels. The application of soil amendments such as limestone and vermicompost may reduce Cu availability and phytotoxicity. The study aimed to assess growth, physiological status and changes in root morphology in young grapevines grown for 12 months in Cu-contaminated soil with and without the application of soil amendments. Samples of a Typic Hapludalf soil were collected in a vineyard with more than 30 years of cultivation. The following treatments were used: 1) control (without amendment), 2) application of organic vermicompost (86.7 g kg-1) and 3) application of limestone (3 Mg ha-1). Grapevines (Paulsen 1103 rootstock) were transplanted and grown for 12 months in PVC soil columns. We assessed parameters of growth, photosynthesis and root morphology. Grapevines grown in soil treated with limestone showed increased growth, dry matter yield and photosynthetic efficiency. The highest Cu concentrations in root tissue were found in grapevines grown in control soil. The application of vermicompost in this study did not alleviate Cu toxicity. Grapevines grown in soil treated with vermicompost showed high manganese (Mn) concentration in shoots before the winter pruning, reflecting the high Mn concentrations in soil solution that caused Mn phytotoxicity, resulting in plant death after the winter pruning. The vermicompost used in this study is not suitable for agronomic use. The use of limestone was an effective strategy to reduce Cu availability and phytotoxicity.

Growth, biochemical response and nutritional status of Angico-Vermelho (Parapiptadenia rigida (Bentham) Brenan) under the application of soil amendment in Cu-contaminated soil.

Forest species Angico-Vermelho (Parapiptadenia rigida (Bentham) Brenan) is an alternative for the revegetation of areas contaminated with high levels of heavy metals such as copper (Cu). However, excess Cu may cause toxicity to plants, which is why the use of soil amendments can facilitate cultivation by reducing the availability of Cu in the soil. The aim of this study was to assess how the use of amendment can contribute to growth and nutritional status as well as reduce oxidative stress in Angico-Vermelho grown in Cu-contaminated soil. Samples of a Typic Hapludalf soil containing high Cu content were used for the application of four amendments (limestone, organic compost, Ca silicate and zeolite), in addition to a control treatment. The treatments were arranged in a completely randomized design, with four replicates. The use of amendments decreased Cu content available in soil and contributed to improve both plant nutritional status and its antioxidant response expressed by enzymatic activity. The application of the amendments, especially zeolite and Ca silicate, increased dry matter yield of Angico-Vermelho. Thus, the results presented here suggest that the use of amendments contributes to improving Cu-contaminated soils and favors revegetation with Angico-Vermelho.