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The anthropogenic input of potentially toxic elements (PTEs) from industry, agrochemicals, etc., into the environment are of great concern. Models derived from pedotransfer functions can provide estimates of the levels of PTEs based on soil attributes. Based on the importance of these models in studies in contaminated areas, we assessed the concentrations of the reactive contents of Ba, Cu, Cr, Ni, Pb, and Zn in soils cultivated with vegetable crops in the state of São Paulo, Brazil. We also evaluated the influence of chemical and physical soil attributes on their reactivity and availability. The reactive contents of PTEs represent the fraction of PTEs easily sorbed at the adsorptions sites of organic matter, iron hydroxides, or clay. This fraction can supply information about the PTE content that is more or less readily released into the soil solution. The reactive and available fraction was extracted with 0.43 M HNO 3 and 0.01 M CaCl 2 , respectively. The proportion of reactivity of metal pools decreased in the order of Ba>Zn > Cu > Pb > Ni > Cr. The empirical models were able to predict the relationship between the reactive fractions, the pseudototal content, and the soil attributes. The available concentrations of Cr, Cu, Ni, and Pb in the soils were lower than the limit of quantification, while 3% of the Ba content and 1% of the Zn content were available in the soil solution in relation to their pseudototal content, suggesting low mobility of these elements in the soil.

Although bottom ash (BA) [or mixtures of bottom and fly ash (FA)] from clean biomass fuels is currently used as liming agent, additive for compost, and fertilizer on agricultural and forest soils in certain European countries, in several other countries most of the ashes are currently disposed in landfills. This is due to both a lack of a proper classification of the materials and of regulatory barriers.Chemical characterization including analysis of an array of potentially toxic elements (PTEs) proved that over 100,000 tons of BA currently landfilled every year in Portugal actually complied with legal limits for PTEs for soil fertilizers applied in other countries. Pot experiments were conducted, testing three dosages of BA and FA (1, 2.5, and 5%, in weight) in three mining soils with different properties. Additions of ash materials to soils led to an increase in the pore water pH relative to control pots (0% of ash added) and had a clear impact on DOC and on the solubilization of both macro- and micronutrients (notably Cu).The results from the case study using BA and FA from a Portuguese biomass thermal power plant demonstrate that it is imperative to further develop a regulatory framework to alleviate technological and environmental barriers for biomass ash utilization as raw material for fertilizers and/or soil liming agent, in accordance with the goals of the circular economy. A more harmonized view on how to assess the merits and risks of the re-use of these materials is also needed.

In this study, we compared the size of the mobile Hg pool in soil to those obtained by extractions using 2 M HNO3, 5 M HNO3, and 2 M HCl. This was done to evaluate their suitability to be used as proxies in view of Hg uptake by ryegrass. Total levels of Hg in soil ranged from 0.66 to 70 mg kg(-1) (median 17 mg kg(-1)), and concentrations of Hg extracted increased in the order: mobile Hg < 2 M HNO3 < 5 M HNO3 < 2 M HCl. The percentage of Hg extracted relative to total Hg in soil varied from 0.13 to 0.79 % (for the mobile pool) to 4.8-82 % (for 2 M HCl). Levels of Hg in ryegrass ranged from 0.060 to 36 mg kg(-1) (median 0.65 mg kg(-1), in roots) and from 0.040 to 5.4 mg kg(-1) (median 0.34 mg kg(-1), in shoots). Although results from the 2 M HNO3 extraction appeared to the most comparable to the actual total Hg levels measured in plants, the 2 M HCl extraction better expressed the variation in plant pools. In general, soil tests explained between 66 and 86 % of the variability of Hg contents in ryegrass shoots. Results indicated that all methods tested here can be used to estimate the plant total Hg pool at contaminated areas and can be used in first tier soil risk evaluations. This study also indicates that a relevant part of Hg in plants is from deposition of soil particles and that splashing of soil can be more significant for plant contamination than actual uptake processes. Graphical Abstract Illustration of potential mercury soil-plant transfer routes.

As plants constitute the foundation of the food chain, concerns have been raised about the possibility of toxic concentrations of metals and metalloids being transported from plants to the higher food chain strata. In this perspective, the use of important phytotoxicity endpoints may be of utmost significance in assessing the hazardous nature of metals and metalloids and also in developing ecological soil screening levels. The current study aimed to investigate the role of glutathione (GSH) and its associated enzymes in the metabolic adaptation of two grass species namely Eriophorum angustifolium Honck. and Lolium perenne L. to metals and metalloids stress in the vicinity of a chemical industrial complex (Estarreja, Portugal). Soil and plant samples were collected from contaminated (C) and non-contaminated (reference, R) sites, respectively, near and away from the Estarreja Chemical Complex, Portugal. Soils (from 0 to 10 and 10 to 20 cm depths) were analyzed for pH, organic carbon, and metals and metalloids concentrations. Plant samples were processed fresh for physiological and biochemical estimations, while oven-dried plant samples were used for metals and metalloids determinations following standard methodologies. Both soils and plants from the industrial area exhibited differential concentrations of major metals and metalloids including As, Cu, Hg, Pb, and Zn. In particular, L. perenne shoot displayed significantly higher and lower concentrations of Pb and As, respectively at contaminated site (vs. E. angustifolium). Irrespective of sites, L. perenne shoot exhibited significantly higher total GSH pool, oxidized glutathione (GSSG) and oxidized protein (vs. E. angustifolium). Additionally, severe damages to photosynthetic pigments, proteins, cellular membrane integrity (in terms of electrolyte leakage), and lipid peroxidation were also perceptible in L. perenne shoot. Contrarily, irrespective of the sites, activities of catalase and GSH-regenerating enzyme, GSH reductase, and GSH-metabolizing enzymes such as GSH peroxidase and GSH sulfotransferase were significantly higher in shoot of E. angustifolium. Despite the higher total GSH content, L. perenne is vulnerable to multi-metals-induced stress in comparison to E. angustifolium as depicted by increased GSH- and protein oxidation, low reactive oxygen radical-processing potential (exhibited in terms of low catalase activity) and poor GSH pool utilization efficiency (in terms of lower GSH-associated enzymes activities). The outcome of the present study may be significant for understanding vital GSH-mediated metals and metalloids tolerance mechanisms in plants as well as their unsuitability for animal consumption due to higher metals and metalloids burdens.

The tropical plant Bixa orellana L. (annatto) produces an array of natural products, including the pigment bixin used in the food and cosmetics industries. In order to understand the biochemical and molecular basis of the biosynthesis of these natural products, a reliable method for isolating high yields of high-quality RNA is required. Here we described a successful and reproducible method for isolation and purification of high-quantity and high-quality RNA from different tissues of annatto. This protocol overcomes the usual problems associated with large amounts of polyphenols, polysaccharides, pigments, and other secondary metabolites that are not easily removed by conventional extraction procedures. Furthermore, the proposed protocol can be easily carried out in any laboratory and it could also be extended to isolate RNA from other plant species showing similar abundance of compounds that interfere with RNA extractions. The yield and quality of the RNA were monitored by spectrophotometric analysis, separation on agarose gel, Reverse Transcription-Polymerase Chain Reaction (RT-PCR), and construction of a cDNA library.

Aphidius colemani Viereck is among the main natural enemies used for biological control of Aphis gossypii Glover. The objective of the present study was to evaluate the development of A. colemani and the alterations caused by the parasitism in the host A. gossypii in different temperatures and to estimate the thermal requirements of the parasitoid. The experiments were carried out in controlled environmental chambers at 16, 19, 22, 25, 28 and 31 +/- 1 degrees C, 70 +/- 10% RH, and 12h photophase. Second-instar nymphs of A. gossypii were parasitized once and kept individually in glass tubes (2.5 cm x 8.5 cm), containing leaf disc of cucumber (2 cm) and 1% water/agar solution. The development time of A. colemani, from oviposition to mummies (11.9, 9.8, 7.7, 6.4 and 6.4 days) and from oviposition to adult (19.4, 16.2, 12.6, 10.5 and 10.7 days) decreased with the increase of the temperature from 16 degrees C to 25 degrees C. The rates of mummies and the emergence of the parasitoid, and its longevity also decreased with the increase of the temperature. Mummies were not produced at 31 degrees C. The lower temperature threshold of A. colemani was 5.94 degrees C and its thermal constant was 200 degrees-day. The alterations caused by the parasitoid in the A. gossypii host were minimized at 31 degrees C, where 98% of the host did not show symptoms of parasitism and produced nymphs. The temperature of 22 degrees C was optimal for the development time of A. colemani.

The parasitoids Lysiphlebus testaceipes (Cresson) and Aphidius colemani Viereck play an important role on the regulation of aphid populations on chrysanthemum grown in greenhouses. This study evaluated the interaction of biological control and plant resistance by the development and the parasitism of L. testaceipes and A. colemani on Aphis gossypii Glover. The aphid host was kept on commercial cultivars of chrysanthemum susceptible Yellow Snowdon (YS) and resistant White Reagan (WR). The experiment was carried out in climatic chamber at 22 +/- 1 masculineC, 70 +/- 10% RH, and photophase 12h. Nymphs of 2nd and 3rd instars of A. gossypii attacked only once by the parasitoids were used in the experiment. The development time was 15.0 and 12.9 days for L. testaceipes and 17.0 and 16.3 days for A. colemani kept on A gossypii on cvs. YS and WR, respectively. The parasitoid L. testaceipes showed greater parasitism rate (68.4%) on A. gossypii on cv. YS than on WR (50%), and for A. colemani was not found significative difference on parasitism (46.8 and 35% to YS and WR, respectively). The emergence rates of L. testaceipes and A. colemani were 91.6% and 85.3% on A gossypii reared on YS, and 78.1% and 100% on WR, respectively. The longevity of A colemani and the parasitism rate of L. testaceipes were affected in a negative way by the cv. WR. The results indicated a positive interaction of biological control with parasitoids and the resistant cv. WR on A. gossypii control in protected cultivation.