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Potato ( Solanum tuberosum L.) is the world's third most popular vegetable in terms of consumption and the fourth most produced. Potatoes can be easily cultivated in different climates and locations around the globe and often in soils contaminated by heavy metals due to industrial activities. This study assessed heavy metal accumulation in different organs of three S. tuberosum L. varieties (Agria, Désirée, and Red Lady) grown in different substrate formulations containing slag and waste from the Caveira polymetallic sulfite mine in Portugal. Results reveal that Cu, Pb, and As accumulation in the different organs of the plant depends on variety and substrate formulation, with tubers exceeding reference values from the literature. Tubers accumulated less Cu (varying between 17.3 and 32 mg/kg), Pb (varying between 5 and 27.6 mg/kg) and As (varying between 4 and 14.8 mg/kg) compared to other plant organs, and the Désirée variety exhibited high Pb (with a maximum of 27.6 mg/kg) accumulation in tubers compared to the remaining varieties. Although the phenological development of plants was not impacted, substrate formulation played a critical role in the plant's metal uptake. The Agria variety presented a lower contamination risk in tubers, but potato cultivation in contaminated soils can present a risk to human health.

Increasing exposure to unfavorable temperatures and water deficit imposes major constraints on most crops worldwide. Despite several studies regarding coffee responses to abiotic stresses, transcriptome modulation due to simultaneous stresses remains poorly understood. This study unravels transcriptomic responses under the combined action of drought and temperature in leaves from the two most traded species: Coffea canephora cv. Conilon Clone 153 (CL153) and C. arabica cv. Icatu. Substantial transcriptomic changes were found, especially in response to the combination of stresses that cannot be explained by an additive effect. A large number of genes were involved in stress responses, with photosynthesis and other physiologically related genes usually being negatively affected. In both genotypes, genes encoding for protective proteins, such as dehydrins and heat shock proteins, were positively regulated. Transcription factors (TFs), including MADS-box genes, were down-regulated, although responses were genotype-dependent. In contrast to Icatu, only a few drought- and heat-responsive DEGs were recorded in CL153, which also reacted more significantly in terms of the number of DEGs and enriched GO terms, suggesting a high ability to cope with stresses. This research provides novel insights into the molecular mechanisms underlying leaf Coffea responses to drought and heat, revealing their influence on gene expression.

Zinc enrichment of edible food products, through the soil and/or foliar application of fertilizers, is a strategy that can increase the contents of some nutrients, namely Zn. In this context, a workflow for agronomic enrichment with zinc was carried out on irrigated Vitis vinifera cv. Syrah, aiming to evaluate the mobilization of photoassimilates to the winegrapes and the consequences of this for winemaking. During three productive cycles, foliar applications were performed with ZnSO 4 or ZnO, at concentrations ranging between 150 and 1350 g.ha -1 . The normal vegetation index as well as some photosynthetic parameters indicated that the threshold of Zn toxicity was not reached; it is even worth noting that with ZnSO 4 , a significant increase in several cases was observed in net photosynthesis (P n ). At harvest, Zn biofortification reached a 1.2 to 2.3-fold increase with ZnSO 4 and ZnO, respectively (being significant relative to the control, in two consecutive years, with ZnO at a concentration of 1350 g.ha -1 ). Total soluble sugars revealed higher values with grapes submitted to ZnSO 4 and ZnO foliar applications, which can be advantageous for winemaking. It was concluded that foliar spraying was efficient with ZnO and ZnSO 4 , showing potential benefits for wine quality without evidencing negative impacts.

Mushroom cultivation presents a viable solution for utilizing agro-industrial byproducts as substrates for growth. This process enables the transformation of low-economic-value waste into nutritional foods. Enhancing the yield and quality of preharvest edible mushrooms, along with effectively preserving postharvest mushrooms, stands as a significant challenge in advancing the industry. Implementing pre- and postharvest strategies for Pleurotus ostreatus (Jacq.) P. Kumm (oyster mushroom) within a circular economy framework involves optimizing resource use, minimizing waste, and creating a sustainable and environmentally friendly production system. This review aimed to analyze the development and innovation of the different themes and trends by bibliometric analysis with a critical literature review. Furthermore, this review outlines the cultivation techniques for Pleurotus ostreatus , encompassing preharvest steps such as spawn production, substrate preparation, and the entire mushroom growth process, which includes substrate colonization, fruiting, harvesting, and, finally, the postharvest. While novel methodologies are being explored for maintaining quality and extending shelf-life, the evaluation of the environmental impact of the entire mushroom production to identify areas for improvement is needed. By integrating this knowledge, strategies can be developed for a more sustainable and circular approach to Pleurotus ostreatus mushroom cultivation, promoting environmental stewardship and long-term viability in this industry.

Climate changes boosted the frequency and severity of drought and heat events, with aggravated when these stresses occur simultaneously, turning crucial to unveil the plant response mechanisms to such harsh conditions. Therefore, plant responses/resilience to single and combined exposure to severe water deficit (SWD) and heat were assessed in two cultivars of the main coffee-producing species: Coffea arabica cv. Icatu and C. canephora cv. Conilon Clone 153 (CL153). Well-watered plants (WW) were exposed to SWD under an adequate temperature of 25/20°C (day/night), and thereafter submitted to a gradual increase up to 42/30°C, and a 14-d recovery period (Rec14). Greater protective response was found to single SWD than to single 37/28°C and/or 42/30°C (except for HSP70) in both cultivars, but CL153-SWD plants showed the larger variations of leaf thermal imaging crop water stress index (CWSI, 85% rise at 37/28°C) and stomatal conductance index (I G , 66% decline at 25/20°C). Both cultivars revealed great resilience to SWD and/or 37/28°C, but a tolerance limit was surpassed at 42/30°C. Under stress combination, Icatu usually displayed lower impacts on membrane permeability, and PSII function, likely associated with various responses, usually mostly driven by drought (but often kept or even strengthened under SWD and 42/30°C). These included the photoprotective zeaxanthin and lutein, antioxidant enzymes (superoxide dismutase, Cu,Zn-SOD; ascorbate peroxidase, APX), HSP70, arabinose and mannitol (involving de novo sugar synthesis), contributing to constrain lipoperoxidation. Also, only Icatu showed a strong reinforcement of glutathione reductase activity under stress combination. In general, the activities of antioxidative enzymes declined at 42/30°C (except Cu,Zn-SOD in Icatu and CAT in CL153), but HSP70 and raffinose were maintained higher in Icatu, whereas mannitol and arabinose markedly increased in CL153. Overall, a great leaf plasticity was found, especially in Icatu that revealed greater responsiveness of coordinated protection under all experimental conditions, justifying low PI Chr and absence of lipoperoxidation increase at 42/30°C. Despite a clear recovery by Rec14, some aftereffects persisted especially in SWD plants ( e.g ., membranes), relevant in terms of repeated stress exposure and full plant recovery to stresses., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Rodrigues, Pais, Leitão, Dubberstein, Lidon, Marques, Semedo, Rakocevic, Scotti-Campos, Campostrini, Rodrigues, Simões-Costa, Reboredo, Partelli, DaMatta, Ribeiro-Barros and Ramalho.)

In recent years, cocoa and dark chocolate have attracted the interest of consumers not only for their sensory characteristics but also for their nutritional properties and positive impact on health. The baobab is a fruit of African origin with a sour and slightly sweet flavour, widely consumed by local communities due to its unique nutritional features. The aim of this work was to evaluate the impact of the concentration of baobab flour in the development of functional dark chocolate, including physical, chemical, nutritional and sensory evaluations. The results presented a positive correlation between the incorporation of baobab flour and the antioxidant activity (up to 2297 mmol TE/100 g), vitamin C content (up to 49.7 mg/100 g), calcium (up to 1052 mg/kg), potassium (up to 10,175 mg/kg), phosphorus (up to 795.9 mg/kg), chlorine (up to 235.4 mg/kg) and sulphur (up to 1158 mg/kg). The sensory evaluation of dark chocolate with 3% baobab presented the highest evaluation on the parameters "texture" and "overall flavour", while the parameter "overall flavour" presented the lowest evaluation on chocolate with 9% baobab. No influence was observed on fatty acid profile, protein, fat and hardness.

Rice ( Oryza sativa L.) is one of the most economically and socially important cereals in the world. Several strategies such as biofortification have been developed in a way eco-friendly and sustainable to enhance crop productivity. This study implemented an agronomic itinerary in Ariete and Ceres rice varieties in experimental fields using the foliar application of selenium (Se) to increase rice nutritional value. At strategic phases of the plant's development (at the end of booting, anthesis, and at the milky grain stage), they were sprayed with sodium selenate (Na 2 SeO 4 ) and sodium selenite (Na 2 SeO 3 ). In the first foliar application plants were sprayed with 500 g Se·ha -1 and in the remaining two foliar applications were sprayed with 300 g Se·ha -1 . The effects of Se in the level of micro and macronutrients in brown grains, the localization of Se in these grains, and the subsequent quality parameters such as colorimetric characteristics and total protein were considered. After grain harvesting, the application of selenite showed the highest enrichment in all grain with levels reaching 17.06 µg g -1 Se and 14.28 µg g -1 Se in Ariete and Ceres varieties, respectively. In the Ceres and Ariete varieties, biofortification significantly affected the K and P contents. Regarding Ca, a clear trend prevailed suggesting that Se antagonizes the uptake of it, while for the remaining elements in general (except Mn) no significant differences were noted. Protein content increased with selenite treatment in the Ariete variety but not in Ceres. Therefore, it was possible to conclude, without compromising quality, that there was an increase in the nutritional content of Se in brown rice grain.

One-third of the world's population is suffering from "hidden hunger" due to micronutrient deficiency. Zinc is acquired through diet, leading its deficiency to the development of disorders such as retarded growth, anorexia, infections, and hypogeusia. Accordingly, this study aimed to develop an agronomic workflow for Zn biofortification on two red winegrapes varieties (cv. Castelão and Syrah) and determine the physicochemical implications for winemaking. Both varieties produced in Setúbal (Portugal) were submitted to four foliar applications of ZnSO 4 or ZnO (900 and 1350 g ha -1 , respectively), during the production cycle. At harvest, Zn biofortification reached a 4.3- and 2.3-fold increase with ZnO 1350 g ha -1 in Castelão and Syrah, respectively (although, with ZnSO 4 1350 g ha -1 both varieties revealed an increase in Zn concentration). On a physiological basis, lower values of NDVI were found in the biofortified grapes, although not reflected in photosynthetic parameters with cv. Syrah shows even a potential benefit with the use of Zn fertilizers. Regarding physical and chemical parameters (density, total soluble solids, dry weight, and color), relative to the control no significant changes in both varieties were observed, being suitable for winemaking. It was concluded that ZnSO 4 and ZnO foliar fertilization efficiently increased Zn concentration on both varieties without a negative impact on quality, but cv. Castelão showed a better index of Zn biofortification and pointed to a potentially higher quality for winemaking.

Drought is a major constraint to plant growth and productivity worldwide and will aggravate as water availability becomes scarcer. Although elevated air [CO 2 ] might mitigate some of these effects in plants, the mechanisms underlying the involved responses are poorly understood in woody economically important crops such as Coffea . This study analyzed transcriptome changes in Coffea canephora cv. CL153 and C. arabica cv. Icatu exposed to moderate (MWD) or severe water deficits (SWD) and grown under ambient (aCO 2 ) or elevated (eCO 2 ) air [CO 2 ]. We found that changes in expression levels and regulatory pathways were barely affected by MWD, while the SWD condition led to a down-regulation of most differentially expressed genes (DEGs). eCO 2 attenuated the impacts of drought in the transcripts of both genotypes but mostly in Icatu, in agreement with physiological and metabolic studies. A predominance of protective and reactive oxygen species (ROS)-scavenging-related genes, directly or indirectly associated with ABA signaling pathways, was found in Coffea responses, including genes involved in water deprivation and desiccation, such as protein phosphatases in Icatu, and aspartic proteases and dehydrins in CL153, whose expression was validated by qRT-PCR. The existence of a complex post-transcriptional regulatory mechanism appears to occur in Coffea explaining some apparent discrepancies between transcriptomic, proteomic, and physiological data in these genotypes.

Inundation, excessive precipitation, or inadequate field drainage can cause waterlogging of cultivated land. It is anticipated that climate change will increase the frequency, intensity, and unpredictability of flooding events. This stress affects 10-15 million hectares of wheat every year, resulting in 20-50% yield losses. Since this crop greatly sustains a population's food demands, providing ca. 20% of the world's energy and protein diets requirements, it is crucial to understand changes in soil and plant physiology under excess water conditions. Variations in redox potential, pH, nutrient availability, and electrical conductivity of waterlogged soil will be addressed, as well as their impacts in major plant responses, such as root system and plant development. Waterlogging effects at the leaf level will also be addressed, with a particular focus on gas exchanges, photosynthetic pigments, soluble sugars, membrane integrity, lipids, and oxidative stress.

Bread wheat ( Triticum aestivum L.) is a major staple crop, and more adapted varieties are needed to ensure productivity under unpredictable stress scenarios resulting from climate changes. In the development of new genotypes, root system traits are essential since roots have a key function in water and nutrient uptake, and root architecture determines the plant's ability to spatially explore the soil resources. Genetic variation in wheat root system may be assessed at the early stages of development. This study evaluates in vitro and at the seedling stage, the genetic diversity of root growth angle (RGA), seminal root number (SRN), and radicle length (RadL) in 30 bread wheat genotypes from different origins and belonging to distinct evolutive or breeding groups. SRN and RadL were analyzed at 1, 2, 3 and 6 days after sowing (DAS) and RGA was measured through the angle between the first pair of seminal roots. A large variability was found in RGA values that ranged from 63° to 122°. Although differences were found between genotypes within the same groups, the narrower angles tended to occur among landraces, while the higher RGA values were observed in advanced lines and Australian varieties. Differences were also observed as regards the SRN (1.0-3.0, 2.7-4.7, 3.2-5.0 and 4.4-6.3 at 1, 2, 3 and 6 DAS, respectively) and RadL (0.1-1.5, 2.1-5.0, 4.0-7.5 and 5.1-13.7 cm at 1, 2, 3 and 6 DAS, respectively). Genetic variability in root traits at seedling stage allows more rapid selection of genotypes better adapted to environmental and soil constraints, necessary to Portuguese Wheat Breeding Program. It will also contribute to the definition of wheat ideotypes with improved performance under Mediterranean climate conditions.

Climate changes with global warming associated with rising atmospheric [CO 2 ] can strongly impact crop performance, including coffee, which is one of the most world's traded agricultural commodities. Therefore, it is of utmost importance to understand the mechanisms of heat tolerance and the potential role of elevated air CO 2 (eCO 2 ) in the coffee plant response, particularly regarding the antioxidant and other protective mechanisms, which are crucial for coffee plant acclimation. For that, plants of Coffea arabica cv. Geisha 3, cv. Marsellesa and their hybrid (Geisha 3 × Marsellesa) were grown for 2 years at 25/20 °C (day/night), under 400 (ambient CO 2 , aCO 2 ) or 700 µL (elevated CO 2 , eCO 2 ) CO 2 L -1 , and then gradually submitted to a temperature increase up to 42/30 °C, followed by recovery periods of 4 (Rec4) and 14 days (Rec14). Heat (37/28 °C and/or 42/30 °C) was the major driver of the response of the studied protective molecules and associated genes in all genotypes. That was the case for carotenoids (mostly neoxanthin and lutein), but the maximal ( α + β ) carotenes pool was found at 37/28 °C only in Marsellesa. All genes (except VDE ) encoding for antioxidative enzymes (catalase, CAT; superoxide dismutases, CuSODs; ascorbate peroxidases, APX) or other protective proteins (HSP70, ELIP, Chape20, Chape60) were strongly up-regulated at 37/28 °C, and, especially, at 42/30 °C, in all genotypes, but with maximal transcription in Hybrid plants. Accordingly, heat greatly stimulated the activity of APX and CAT (all genotypes) and glutathione reductase (Geisha3, Hybrid) but not of SOD. Notably, CAT activity increased even at 42/30 °C, concomitantly with a strongly declined APX activity. Therefore, increased thermotolerance might arise through the reinforcement of some ROS-scavenging enzymes and other protective molecules (HSP70, ELIP, Chape20, Chape60). Plants showed low responsiveness to single eCO 2 under unstressed conditions, while heat promoted changes in aCO 2 plants. Only eCO 2 Marsellesa plants showed greater contents of lutein, the pool of the xanthophyll cycle components (V + A + Z), and β -carotene, compared to aCO 2 plants at 42/30 °C. This, together with a lower CAT activity, suggests a lower presence of H 2 O 2 , likely also associated with the higher photochemical use of energy under eCO 2 . An incomplete heat stress recovery seemed evident, especially in aCO 2 plants, as judged by the maintenance of the greater expression of all genes in all genotypes and increased levels of zeaxanthin (Marsellesa and Hybrid) relative to their initial controls. Altogether, heat was the main response driver of the addressed protective molecules and genes, whereas eCO 2 usually attenuated the heat response and promoted a better recovery. Hybrid plants showed stronger gene expression responses, especially at the highest temperature, when compared to their parental genotypes, but altogether, Marsellesa showed a greater acclimation potential. The reinforcement of antioxidative and other protective molecules are, therefore, useful biomarkers to be included in breeding and selection programs to obtain coffee genotypes to thrive under global warming conditions, thus contributing to improved crop sustainability.

Nowadays, there is a growing concern about micronutrient deficits in food products, with agronomic biofortification being considered a mitigation strategy. In this context, as Zn is essential for growth and maintenance of human health, a workflow for the biofortification of grapes from the Vitis vinifera variety Fernão Pires, which contains this nutrient, was carried out considering the soil properties of the vineyard. Additionally, Zn accumulation in the tissues of the grapes and the implications for some quality parameters and on winemaking were assessed. Vines were sprayed three times with ZnO and ZnSO 4 at concentrations of 150, 450, and 900 g ha -1 during the production cycle. Physiological data were obtained through chlorophyll a fluorescence data, to access the potential symptoms of toxicity. At harvest, treated grapes revealed significant increases of Zn concentration relative to the control, being more pronounced for ZnO and ZnSO 4 in the skin and seeds, respectively. After winemaking, an increase was also found regarding the control (i.e., 1.59-fold with ZnSO 4 -450 g ha -1 ). The contents of the sugars and fatty acids, as well as the colorimetric analyses, were also assessed, but significant variations were not found among treatments. In general, Zn biofortification increased with ZnO and ZnSO 4 , without significantly affecting the physicochemical characteristics of grapes.

As drought threatens crop productivity it is crucial to characterize the defense mechanisms against water deficit and unveil their interaction with the expected rise in the air [CO 2 ]. For that, plants of Coffea canephora cv. Conilon Clone 153 (CL153) and C. arabica cv. Icatu grown under 380 (aCO 2 ) or 700 μL L -1 (eCO 2 ) were exposed to moderate (MWD) and severe (SWD) water deficits. Responses were characterized through the activity and/or abundance of a selected set of proteins associated with antioxidative (e.g., Violaxanthin de-epoxidase, Superoxide dismutase, Ascorbate peroxidases, Monodehydroascorbate reductase), energy/sugar (e.g., Ferredoxin-NADP reductase, NADP-dependent glyceraldehyde-3-phosphate dehydrogenase, sucrose synthase, mannose-6-phosphate isomerase, Enolase), and lipid (Lineolate 13S-lipoxygenase) processes, as well as with other antioxidative (ascorbate) and protective (HSP70) molecules. MWD caused small changes in both genotypes regardless of [CO 2 ] level while under the single imposition to SWD, only Icatu showed a global reinforcement of most studied proteins supporting its tolerance to drought. eCO 2 alone did not promote remarkable changes but strengthened a robust multi-response under SWD, even supporting the reversion of impacts already observed by CL153 at aCO 2 . In the context of climate changes where water constraints and [CO 2 ] levels are expected to increase, these results highlight why eCO 2 might have an important role in improving drought tolerance in Coffea species., (Copyright © 2022 Elsevier GmbH. All rights reserved.)

The baobab tree ( Adansonia digitata L.) is found widely in the forests and savannas of sub-Saharan Africa. The baobab fruit has a sour and slightly sweet taste and is widely consumed by the natives, thus containing a high nutritional value and providing a source of income for rural people. This study aimed to compare the nutritional composition of baobab fruit pulp from different localities in the Namibe province (Angola). Twenty samples of baobab pulp were collected in markets of the four municipalities of Namibe. The results obtained showed that there is some geographic location dependence on nutritional and functional composition. The municipality of Camucuio showed samples with higher fibre content (56.62 g/100 g) and vitamin C (288.9 mg/100 g). Samples from the Virei municipality stood out for their antioxidant activity (1936 mmol TE/100 g), high K content (42.4 mg/g) and higher values of protein (2.42 g/100 g). The samples collected in the municipality of Bibala stood out for their high contents of carbohydrates (28.1 g/100 g), total phenolic compounds (972 mg GAE/100 g) and Ca (3.80 mg/g). Despite the differences in origin, the high nutritional value of baobab fruit has the potential to improve the diet of thousands of people in Africa qualitatively.

As the productivity and quality of tomato fruits are responsive to Mg applications, without surpassing the threshold of toxicity, the assessment of potential levels of Mg accumulation in tissues, as well as the interactions with Ca and physicochemical properties, prompt this study. An agronomic workflow for Mg enrichment, consisting of six foliar applications of MgSO 4 with four concentrations (0%, 0.25%, 1% and 4%), equivalent to 0, 43.9, 175.5 and 702 g ha -1 , was applied on two tomato ( Lycopersicum esculentum L.) genotypes (Heinz1534 and Heinz9205). During fruit development, leaf gas exchange was screened, with only minor physiological deviations being found. At harvest, Mg contents among tissues and the interactions with Ca were analyzed, and it was found that in both varieties a higher Mg/Ca ratio prevailed in the most external part of the fruit sprayed with 4% MgSO 4 . However, Mg distribution prevailed relatively near the epidermis in H1534, while in H9205 the higher contents of this nutrient occurred in the core of the fruit, which indicated a decrease of the relative proportion of Ca. The morphologic (height and diameter), physical (dry weight and density) and colorimetric parameters, and the total soluble solids of fruits, did not reveal significant changes in both tomato varieties. It was further concluded that foliar application until 4% MgSO 4 does not have physiological impacts in the fruit's quality of both varieties, but in spite of the different patterns of Mg accumulation in tissues, if the mean value in the whole fruit is considered, this nutrient prevails in H1534. This study thus suggests that variety H1534 can be used to attain tomato fruits with added value, providing an option of further processing to achieve food products with functional properties, ultimately proving a beneficial option to producers, the food processing industry and consumers. Moreover, the study reinforces the importance of variety choice when designing enrichment workflows.

Calcium is essential for plants, yet as its mobility is limited, the understanding of the rate of Ca 2+ accumulation and deposition in tissues of tubers, as well as the interactions with other critical nutrients prompted this study. To assess the interactions and differential accumulation of micro and macronutrients in the tissues of tubers, Solanum tuberosum L. varieties Agria and Rossi were cultivated and, after the beginning of tuberization, four foliar sprayings (at 8-10 day intervals) with CaCl 2 (3 and 6 kg ha -1 ) or Ca(NO 3 ) 2 (2 and 4 kg ha -1 ) solutions were performed. It was found that both fertilizers increased Ca accumulation in tubers (mostly in the parenchyma tissues located in the center of the equatorial region). The functioning of the photosynthetic apparatus was not affected until the 3rd application but was somewhat affected when approaching the end of the crop cycle (after the 4th application), although the lower dose of CaCl 2 seemed to improve the photochemical use of energy, particularly when compared with the greater dose of Ca(NO 3 ) 2 . Still, none of these impacts modified tuber height and diameter. Following the increased accumulation of Ca, in the tubers of both varieties, the mean contents of P, K, Na, Fe, and Zn revealed different accumulation patterns. Moreover, accumulation of K, Fe, Mn, and Zn prevailed in the epidermis, displaying a contrasting pattern relative to Ca. Therefore, Ca accumulation revealed a heterogeneous trend in the different regions analyzed, and Ca enrichment of tubers altered the accumulation of other nutrients.

This study evaluated the elemental composition of 25 herbal tea plants commonly used in infusions by Portuguese consumers and the contribution to the elemental daily intake of some essential elements. Hydrocotyle asiatica (L.), Matricaria chamomilla (L.), and Melissa officinalis (L.) samples are a rich source of K with around 6.0 mg g -1 while the Asteraceae Silybum marianum (L.) and Echinacea angustifolia (DC.) exhibited 4.9 and 5.6 mg g -1 Ca, respectively. The highest concentrations of S and Zn were noted in Hydrocotyle asiatica (L.), while the highest concentration of Sr was found in Cassia angustifolia (Vahl.). In general, a large variability in the concentrations among different families and plant organs had been observed, except Cu with levels around 30 μg g -1 . The principal component analysis (PCA) showed positive correlations between Zn and S and Sr and Ca, also revealing that Hydrocotyle asiatica (L.), Echinacea angustifolia (DC.), Silybum marianum (L.), and Cassia angustifolia (Vahl.) samples, stands out about all other samples regarding the enrichment of macro and micronutrients. The elemental solubility of macronutrients in the infusion is greater than the micronutrient solubility, despite the contribution to the recommended daily intake was weak. As a whole, Cynara scolymus (L.) and Hibiscus sabdariffa (L.) are the species with the best elemental solubilities, followed by Hydrocotyle asiatica (L.). No harmful elements, such as As and Pb, were observed in both the raw material and the infusions.

Grapes and wine are widely consumed in the world, yet their mineral content can be influenced by many factors such as the mineral composition of soils, viticulture practices and environmental conditions. In this context, considering the importance of Zn in the human physiology, the enrichment of Moscatel and Castelão grapes (white and red variety, respectively) with this nutrient prompted this study; further assessment of tissue deposition and some implications for wine production. Using two foliar fertilizers (ZnO or ZnSO 4 , at 150, 450 and 900 g ha -1 ), decreases in net photosynthesis and stomatal conductance occurred in both varieties, suggesting that the physiological threshold of Zn toxicity was reached without visible symptoms. Following foliar spraying with both fertilizers, the content of Zn in leaves of the Castelão and Moscatel varieties showed higher values in all treatments relative to the control. Moreover, in grapes this tendency occurred only in Castelão. Concerning Cu, Fe, Ca, K, S and P, some significant differences also happened in leaves and grapes among treatments. At harvest, the indexes of Zn enrichment in grapes increased between 2.14- and 8.38-fold and between 1.02- and 1.44-fold in Castelão and Moscatel varieties, respectively. Zinc in the dried skin of Castelão only increased with ZnO and ZnSO 4 sprayed at 900 g ha -1 (ca. 2.71- and 1.5-fold relative to the control, respectively), but in Moscatel a clear accumulation trend could not be found. The dry weight of grapes ranged (in %) between 16 and 23 (but did not vary significantly among treatments of each variety or in each treatment between varieties), and total soluble solids (e.g., mainly soluble sugars and proteins) and color parameters showed some significant variations. Through winemaking, the contents of Zn increased in both varieties (1.34- and 3.57-fold, in Castelão and Moscatel, respectively) and in all treatments, although non-significantly in Castelão. It is concluded that, to increase the contents of Zn in grapes without reaching the threshold of toxicity, ZnO or ZnSO 4 can be used for foliar spraying of Castelão and Moscatel varieties until 900 g ha -1 and that winemaking augments the level of this nutrient.

Bread wheat ( Triticum aestivum ) is a major crop worldwide, and it is highly susceptible to heat. In this work, grain production and composition were evaluated in Portuguese T. aestivum germplasm (landraces and commercial varieties), which was subjected to heat after anthesis (grain filling stage). Heat increased the test weight (TW) in Nabão, Grécia and Restauração, indicating an improved flour-yield potential. Mocho de Espiga Branca (MEB) and Transmontano (T94) showed higher thousand-kernel weight (TKW). Gentil Rosso presented increased soluble sugars, which are yeast substrates in the bread-making process. Ardila stood out for its protein increase under heat. Overall SDS was unaffected by higher temperature, but increased in T94, indicating a better dough elasticity for bread-making purposes. Under heat, lipid content was maintained in most genotypes, being endogenous fatty acids (FAs) key players in fresh bread quality. Lipid unsaturation, evaluated through the double bond index (DBI), also remained unaffected in most genotypes, suggesting a lower flour susceptibility to lipoperoxidation. In Grécia, heat promoted a higher abundance of monounsaturated oleic (C18:1) and polyunsaturated linoleic (C18:2) acids, which are essential fatty acids in the human diet. This work highlighted a great variability in most parameters both under control conditions or in response to heat during grain filling. Cluster analysis of traits revealed a lower susceptibility to heat during grain filling in Ardila, Restauração, and Ruivo, in contrast to MEQ, which seems to be more differentially affected at this stage. Characterization and identification of more favorable features under adverse environments may be relevant for agronomic, industrial, or breeding purposes, in view of a better crop adaptation to changing climate and an improved crop sustainability in agricultural systems more prone to heat stress.

Colour is one of the most relevant organoleptic attributes that directly affects consumers' acceptance and food selection. However, as food colouring pigments are generally unstable and become modified during processing, in order to maintain or restore product colour uniformity, colourants are added to food products around the world. In this context, although they are still widely used, synthetic food colorants, due to their potential hazards, are being replaced by those obtained from natural origins. Indeed, numerous side effects and toxicities, at both the medium and long-terms-namely allergic reactions, and behavioral and neurocognitive effects-have been related to the use of synthetic colourants, whereas their naturally-derived counterparts seem to provide a somewhat high-quality and effective contribution as a health promoter. In order to further understand the implications of the use of synthetic and naturally derived food colourants, this review aims to provide a synoptical approach to the chemical characteristics, properties, uses and side effects on health of those which are currently allowed and applied during food processing.

The aim of this study is to evaluate the elemental composition of fifteen algae-based supplements commonly sold in the Portuguese market, by energy dispersive X-ray fluorescence. Despite the fact that the majority of Kelp samples were a good source of iodine, the levels observed might well contribute to an excess in the human body, which can cause dysfunction of the thyroid gland. Furthermore, the presence of lead in Sea spaghetti, Arame, Hijiki and Wakame caused a considerable risk to public health vis a vis possible ingestion of a high daily dose. Regarding arsenic, great variability was observed in all the samples with concentrations equal to or above 60 μg/g in the case of Arame, KelpJ and Hijiki. Although algae mainly accumulate organic arsenic, some also contain high levels of its inorganic form, as is commonly pointed out for Hijiki. Thus, regular ingestion of these supplements must also take into account the mentioned facts. There is no doubt that these supplements are also good sources of other nutrients, but the lack of accurate regulations and control should alert consumers to avoid indiscriminate use of these types of products.

Climate change scenarios pose major threats to many crops worldwide, including coffee. We explored the primary metabolite responses in two Coffea genotypes, C. canephora cv. Conilon Clone 153 and C. arabica cv. Icatu, grown at normal (aCO 2 ) or elevated (eCO 2 ) CO 2 concentrations of 380 or 700 ppm, respectively, under well-watered (WW), moderate (MWD), or severe (SWD) water deficit conditions, in order to assess coffee responses to drought and how eCO 2 can influence such responses. Primary metabolites were analyzed with a gas chromatography time-of-flight mass spectrometry metabolomics platform (GC-TOF-MS). A total of 48 primary metabolites were identified in both genotypes (23 amino acids and derivatives, 10 organic acids, 11 sugars, and 4 other metabolites), with differences recorded in both genotypes. Increased metabolite levels were observed in CL153 plants under single and combined conditions of aCO 2 and drought (MWD and SWD), as opposed to the observed decreased levels under eCO 2 in both drought conditions. In contrast, Icatu showed minor differences under MWD, and increased levels (especially amino acids) only under SWD at both CO 2 concentration conditions, although with a tendency towards greater increases under eCO 2 . Altogether, CL153 demonstrated large impact under MWD, and seemed not to benefit from eCO 2 in either MWD and SWD, in contrast with Icatu.

Metamitron (MET) is a fruitlet thinning compound for apple trees, needing better understanding of its action on leaf energy metabolism, depending on nighttime temperature. A trial under environmental controlled conditions was set with 'Golden Reinders' potted trees, under 25/7.5 and 25/15 °C (diurnal/nighttime temperature), with (MET, 247.5 ppm) or without (CTR) application, and considering the monitoring of photosynthetic and respiration components from day 1 (D1) to 14 (D14). Net photosynthesis (P n ) decline promoted by MET after D1 was not stomatal related. Instead, non-stomatal constraints, reflected on the photosynthetic capacity (A max ), included a clear photosystem (PS) II inhibition (but barely of PSI), as shown by severe reductions in thylakoid electron transport at PSII level, maximal (F v /F m ) and actual (F v '/F m ') PSII photochemical efficiencies, estimate of quantum yield of linear electron transport (Y (II) ), and the rise in PSII photoinhibition status (F s /F m ' and PI Chr ) and uncontrolled energy dissipation (Y (NO) ). To P n inhibition also contributed the impact in RuBisCO along the entire experiment, regardless of night temperature, here reported for the first time. Globally, MET impact on the photosynthetic parameters was usually greater under 7.5 °C, with maximal impacts between D4 and D7, probably associated to a less active metabolism at lower temperature. Cellular energy metabolism was further impaired under 7.5 °C, through moderate inhibition of NADH-dependent malate dehydrogenase (MDH) and pyruvate kinase (PK) enzymes involved in respiration, in contrast with the increase of dark respiration in MET 7.5 until D7. The lower impact on PK and MDH under 15 °C and a likely global higher active metabolism at that temperature would agree with the lowest sucrose levels in MET 15 at D4 and D7. Our findings showed that MET alters the cell energy machinery in a temperature dependent manner, affecting the sucrose balance mainly at 15 °C, justifying the observed greater thinning potential., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

Edible wild plants are part of the ethnobotanical and gastronomic heritage of different geographical areas. Corema album (L.) D. Don is an endemic species of the dune systems of the Atlantic coast of the Iberian Peninsula. The aerial parts of Corema album are a source of nutrients and antioxidants. The Corema album white berry (Portuguese crowberry) is rich in calcium, iron, and zinc. The plant also shows high phenolic content and antioxidant capacity associated with the leaves, fruit, and flowers. The presence of organic acids, namely phenolic acids, such as hydroxycinnamic acids, and long chain polyunsaturated fatty acids (PUFAs) omega-3 and omega-6 has also been confirmed. Toxicity studies evaluated by cell viability tests with human intestinal epithelium model cells (Caco-2) have shown that, at low concentrations, plant extracts may present beneficial effects., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Growing water restrictions associated with climate changes constitute daunting challenges to crop performance. This study unveils the impacts of moderate (MWD) or severe (SWD) water deficit, and their interaction with air [CO2], on the photosynthetic apparatus of Coffea canephora Pierre ex A. Froehner cv. Conilon Clone 153 (CL153) and Coffea arabica L. cv. Icatu. Seven year-old potted plants grown under 380 (aCO2) or 700 μl l -1 (eCO2) [CO2] gradually reached predawn water potentials between -1.6 and -2.1 MPa (MWD), and below -3.5 MPa (SWD). Under drought, stomata closure was chiefly related to abscisic acid (ABA) rise. Increasing drought severity progressively affected gas exchange and fluorescence parameters in both genotypes, with non-stomatal limitations becoming gradually dominating, especially regarding the photochemical and biochemical components of CL153 SWD plants. In contrast, Icatu plants were highly tolerant to SWD, with minor, if any, negative impacts on the potential photosynthetic functioning and components (e.g., Amax, Fv/Fm, electron carriers, photosystems (PSs) and ribulose-1,5-bisphosphate carboxylase oxygenase (RuBisCO) activities). Besides, drought-stressed Icatu plants displayed increased abundance of a large set of proteins associated with the photosynthetic apparatus (PSs, light-harvesting complexes, cyclic electron flow, RuBisCO activase) regardless of [CO2]. Single eCO2 did not promote stomatal and photosynthetic down-regulation in both genotypes. Instead, eCO2 increased photosynthetic performance, moderately reinforced photochemical (PSs activity, electron carriers) and biochemical (RuBisCO, ribulose-5-phosphate kinase) components, whereas photoprotective mechanisms and protein abundance remained mostly unaffected. In both genotypes, under MWD, eCO2 superimposition delayed stress severity and promoted photosynthetic functioning with lower energy dissipation and PSII impacts, whereas stomatal closure was decoupled from increases in ABA. In SWD plants, most impacts on the photosynthetic performance were reduced by eCO2, especially in the moderately drought affected CL153 genotype, although maintaining RuBisCO as the most sensitive component, deserving special breeder's attention to improve coffee sustainability under future climate scenarios., (© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permission@oup.com.)

The contamination of abandoned mining areas is a problem worldwide that needs urgent attention. Phytoremediation emerges as a successful method to extract different contaminants from the soil. In this context, Eucalyptus globulus plants growing in soils artificial contaminated with arsenic (As) were used to access its phytoremediation capabilities. The effects of As on photosynthetic performance were monitored through different physiological parameters, whereas the uptake and translocation of As and the putative effects on calcium, iron, potassium, and zinc levels on plants were evaluated by X-ray fluorescence analysis. Root system is the major accumulator organ, while the translocation to the above-ground organs is poor. In the end of the experiment, the root biomass of plants treated with 200 μg As mL -1 is 27% and 49.7% lower than equivalent biomass from plants treated with 100 μg As mL -1 and control plants, respectively. Each plant can accumulate 8.19 and 8.91 mg As after a 6-month period, when submitted to 100 As and 200 As, respectively. It seems to exist an antagonistic effect of As on Zn root uptake by E. globulus . In general, the tested concentrations do not influence negatively plant metabolism, indicating that this species is suitable for plantation in contaminated areas.

Understanding the effect of extreme temperatures and elevated air (CO 2 ) is crucial for mitigating the impacts of the coffee industry. In this work, leaf transcriptomic changes were evaluated in the diploid C. canephora and its polyploid C. arabica , grown at 25 °C and at two supra-optimal temperatures (37 °C, 42 °C), under ambient (aCO 2 ) or elevated air CO 2 (eCO 2 ). Both species expressed fewer genes as temperature rose, although a high number of differentially expressed genes (DEGs) were observed, especially at 42 °C. An enrichment analysis revealed that the two species reacted differently to the high temperatures but with an overall up-regulation of the photosynthetic machinery until 37 °C. Although eCO 2 helped to release stress, 42 °C had a severe impact on both species. A total of 667 photosynthetic and biochemical related-DEGs were altered with high temperatures and eCO 2 , which may be used as key probe genes in future studies. This was mostly felt in C. arabica , where genes related to ribulose-bisphosphate carboxylase (RuBisCO) activity, chlorophyll a-b binding, and the reaction centres of photosystems I and II were down-regulated, especially under 42°C, regardless of CO 2 . Transcriptomic changes showed that both species were strongly affected by the highest temperature, although they can endure higher temperatures (37 °C) than previously assumed.

An agronomic itinerary for Se biofortification of two rice cultivars (Ariete and Ceres) through foliar fertilization with sodium selenate and sodium selenite with different concentrations (25, 50, 75 and 100 g Se.ha -1 ), was implemented in experimental fields. The selenium toxicity threshold was not exceeded, as shown by the eco-physiological data obtained through leaf gas exchanges. The highest Se enrichment in paddy grains was obtained with selenite for both cultivars, especially at the highest doses, i.e., 75 and 100 g Se.ha -1 , with approximately a 5.0-fold increase compared with control values. In paddy grains, Zn was the most affected element by the treatments with Se with decreases up to 54%. When comparing the losses between rough and polished grains regardless of the cultivars, Se species and concentrations, it was observed that only Cu, Mg and Zn exhibited losses <50%. The remaining elements generally had losses >70%. The loss of Se is more pronounced in Ceres cultivar than in Ariete but rarely exceeds 50%. The analysis by µ-EDXRF showed that, in Ariete cultivar, Se is mostly homogeneously distributed in the grain regardless of any treatments, while in Ceres cultivar, the Se distribution seems to favor accumulation in the periphery, perhaps in the bran.

This study aimed to assess the efficiency of Ca enrichment in tubers of three genotypes of Solanum tuberosum L., through foliar spraying with CaCl 2 and Ca(NO 3 ) 2 solutions. In this context, soil heterogeneity of three potato-growing fields, as well as the implications of Ca accumulation among tissues and some quality parameters were assessed. Three potato varieties (Agria, Picasso and Rossi) were grown in three production fields and during the life cycle, four pulverizations with calcium chloride (3 and 6 kg ha -1 ) or calcium nitrate (0.5, 2 and 4 kg ha -1 ) were applied. For screening the potential phytotoxicity, using Agria as a test system, the potential synthesis of photoassimilates was determined, and it was found that after the 3rd Ca application, leaf gas exchanges were moderately (net photosynthesis), to strongly (stomatal conductance) affected, although without impact on Ca accumulation in tubers. At harvest, the average Ca biofortification index varied between 5-40%, 40-35% and 4.3-13% in Agria, Picasso and Rossi, respectively. Moreover, the equatorial region of the tubers in general showed that Ca accumulation prevailed in the epidermis and, in some cases, in inner areas of the potato tubers. Biofortified tubers with Ca also showed some significant changes in total soluble solids and colorimetric parameters. It is concluded that Ca enrichment of potato tubers through foliar spraying complemented the xylem mass flow of Ca from roots, through phloem redistribution. Both fertilizers showed similar efficiency, but Rossi revealed a lower index of Ca accumulation, eventually due to different metabolic characteristics. Although affected by Ca enrichment, potato tubers maintained a high quality for industrial processing.

This study aimed to assess the implications of Zn enrichment in wheat grains as a function of contrasting genotypes, edaphic conditions and foliar fertilizers. Triticum aestivum L. varieties Roxo and Paiva were grown in four production fields, and sprayed with ZnSO 4 (0, 16.20 and 36.40 kg/ha) Zn-EDTA (0, 6.30 and 12.60 kg/ha) and Tecnifol Zinc (0, 3.90 and 7.80 kg/ha). The heterogeneous edaphic conditions of the wheat fields were chemically characterized, it being found that soil properties determine different Zn accumulation in the grains of both genotypes. Foliar spraying enhanced to different extents Zn content in the grains of both genotypes, but the average of enrichment indexes varied among the wheat fields. Zinc mostly accumulated in the embryo and vascular bundle and to a lesser extent in the endosperm. Grain yield and test weight sprayed by ZnSO 4 gave the highest values in both genotypes, but the opposite was found for Zn-EDTA. Considering the color parameters, lightness and red-green transitions were found to be a conjunction of genotype characteristics, fertilization types and edaphic conditions prevailing in each field. It is concluded that the index of Zn enrichment in wheat grains is a docket of edaphic conditions, genotype and type of fertilization.

As atmospheric [CO 2 ] continues to rise to unprecedented levels, understanding its impact on plants is imperative to improve crop performance and sustainability under future climate conditions. In this context, transcriptional changes promoted by elevated CO 2 (eCO 2 ) were studied in genotypes from the two major traded coffee species: the allopolyploid Coffea arabica (Icatu) and its diploid parent, C. canephora (CL153). While Icatu expressed more genes than CL153, a higher number of differentially expressed genes were found in CL153 as a response to eCO 2 . Although many genes were found to be commonly expressed by the two genotypes under eCO 2 , unique genes and pathways differed between them, with CL153 showing more enriched GO terms and metabolic pathways than Icatu. Divergent functional categories and significantly enriched pathways were found in these genotypes, which altogether supports contrasting responses to eCO 2 . A considerable number of genes linked to coffee physiological and biochemical responses were found to be affected by eCO 2 with the significant upregulation of photosynthetic, antioxidant, and lipidic genes. This supports the absence of photosynthesis down-regulation and, therefore, the maintenance of increased photosynthetic potential promoted by eCO 2 in these coffee genotypes.

In worldwide production, rice is the second-most-grown crop. It is considered a staple food for many populations and, if naturally enriched in Se, has a huge potential to reduce nutrient deficiencies in foodstuff for human consumption. This study aimed to develop an agronomic itinerary for Se biofortification of Oryza sativa L. (Poaceae) and assess potential physicochemical deviations. Trials were implemented in rice paddy field with known soil and water characteristics and two genotypes resulting from genetic breeding (OP1505 and OP1509) were selected for evaluation. Plants were sprayed at booting, anthesis and milky grain phases with two different foliar fertilizers (sodium selenate and sodium selenite) at different concentrations (25, 50, 75 and 100 g Se·ha -1 ). After grain harvesting, the application of selenate showed 4.9-7.1 fold increases, whereas selenite increased 5.9-8.4-fold in OP1509 and OP1505, respectively. In brown grain, it was found that in the highest treatment selenate or selenite triggered much higher Se accumulation in OP1505 relatively to OP1509, and that no relevant variation was found with selenate or selenite spraying in each genotype. Total protein increased exponentially in OP1505 genotype when selenite was applied, and higher dosage of Se also increased grain weight and total protein content. It was concluded that, through agronomic biofortification, rice grain can be enriched with Se without impairing its quality, thus highlighting its value in general for the industry and consumers with special needs.

This study unveils the single and combined drought and heat impacts on the photosynthetic performance of Coffea arabica cv. Icatu and C. canephora cv. Conilon Clone 153 (CL153). Well-watered (WW) potted plants were gradually submitted to severe water deficit (SWD) along 20 days under adequate temperature (25/20°C, day/night), and thereafter exposed to a gradual temperature rise up to 42/30°C, followed by a 14-day water and temperature recovery. Single drought affected all gas exchanges (including A max ) and most fluorescence parameters in both genotypes. However, Icatu maintained F v /F m and RuBisCO activity, and reinforced electron transport rates, carrier contents, and proton gradient regulation (PGR5) and chloroplast NADH dehydrogenase-like (NDH) complex proteins abundance. This suggested negligible non-stomatal limitations of photosynthesis that were accompanied by a triggering of protective cyclic electron transport (CEF) involving both photosystems (PSs). These findings contrasted with declines in RuBisCO and PSs activities, and cytochromes ( b 559 , f , b 563 ) contents in CL153. Remarkable heat tolerance in potential photosynthetic functioning was detected in WW plants of both genotypes (up to 37/28°C or 39/30°C), likely associated with CEF in Icatu. Yet, at 42/30°C the tolerance limit was exceeded. Reduced A max and increased C i values reflected non-stomatal limitations of photosynthesis, agreeing with impairments in energy capture ( F 0 rise), PSII photochemical efficiency, and RuBisCO and Ru5PK activities. In contrast to PSs activities and electron carrier contents, enzyme activities were highly heat sensitive. Until 37/28°C, stresses interaction was largely absent, and drought played the major role in constraining photosynthesis functioning. Harsher conditions (SWD, 42/30°C) exacerbated impairments to PSs, enzymes, and electron carriers, but uncontrolled energy dissipation was mitigated by photoprotective mechanisms. Most parameters recovered fully between 4 and 14 days after stress relief in both genotypes, although some aftereffects persisted in SWD plants. Icatu was more drought tolerant, with WW and SWD plants usually showing a faster and/or greater recovery than CL153. Heat affected both genotypes mostly at 42/30°C, especially in SWD and Icatu plants. Overall, photochemical components were highly tolerant to heat and to stress interaction in contrast to enzymes that deserve special attention by breeding programs to increase coffee sustainability in climate change scenarios., (Copyright © 2020 Dubberstein, Lidon, Rodrigues, Semedo, Marques, Rodrigues, Gouveia, Armengaud, Semedo, Martins, Simões-Costa, Moura, Pais, Scotti-Campos, Partelli, Campostrini, Ribeiro-Barros, DaMatta and Ramalho.)

Human malnutrition due to micronutrient deficiencies, particularly with regards to Zinc (Zn) and Selenium (Se), affects millions of people around the world, and the enrichment of staple foods through biofortification has been successfully used to fight hidden hunger. Rice ( Oryza sativa L.) is one of the staple foods most consumed in countries with high levels of malnutrition. However, it is poor in micronutrients, which are often removed during grain processing. In this study, we have analyzed the transcriptome of rice flag leaves biofortified with Zn (900 g ha -1 ), Se (500 g ha -1 ), and Zn-Se. Flag leaves play an important role in plant photosynthesis and provide sources of metal remobilization for developing grains. A total of 3170 differentially expressed genes (DEGs) were identified. The expression patterns and gene ontology of DEGs varied among the three sets of biofortified plants and were limited to specific metabolic pathways related to micronutrient mobilization and to the specific functions of Zn (i.e., its enzymatic co-factor/coenzyme function in the biosynthesis of nitrogenous compounds, carboxylic acids, organic acids, and amino acids) and Se (vitamin biosynthesis and ion homeostasis). The success of this approach should be followed in future studies to understand how landraces and other cultivars respond to biofortification., (Copyright © 2020 Roda, Marques, Batista-Santos, Esquível, Ndayiragije, Lidon, Swamy, Ramalho and Ribeiro-Barros.)

Soils and different vegetable species in Costa de Caparica (Portugal) are subject to the intensive use of inorganic fertilizers and pesticides. Thus, the concentrations of As, Cu, Fe, Mn, Pb, and Zn were evaluated. Lettuce, spinach, and potatoes collected in station 9 cause reason for concern due to their high Pb concentrations close to 20 μg g -1 which is probably related to an intensive use of copper and iron sulphate fertilizers. Additionally, the consumption of Portulaca oleracea collected in stations 3 and 4 must be avoided due to the high concentrations of Zn, and even Cu. The derived estimated daily intake (EDI) dose of Zn will be a risk to human consumption if P. oleracea was the single basis of a soup, although the addition of other ingredients might lower the tolerable upper intake (TUI) value of 39 mg/day of P. oleracea, to admissible levels, i.e., not exceeding 25 mg/day. Pumpkin collected in station 1 contained 44.1 μg g -1 Cu and a TUI value of 9.8 mg/day, when the recommendation must not exceed 5.0 mg/day. In this context, it is strongly advised to not include this vegetable in household menus.

The Neves-Corvo mining complex (MC) situated in southern Portugal exploits one of the most world's important copper deposits. Agricultural soils surrounding the MC, used by the inhabitants for crop production, contain excessive amounts of As, Cu, Pb, and Zn. Thus, a potential risk to human consumption exists if edible plants grow on these substrata. Arsenic and Pb were not detected in edible samples collected near the MC and 5 km away, but in the leaves-structural or adsorbed onto the surface. In general, Zn was the most mobile element in both contaminated and reference areas as seen by the bioaccumulation factors (BAF). The tolerable upper intake (TUI) values for Cu are a reason of concern, since in 57.1% of the cases, the TUI values are above the recommended upper limit of 5 mg/day, in the case of Ficus carica, Cucurbita pepo, and Phaseolus vulgaris, whereas in 28.6% of the cases, the TUI values are near this limit (C. pepo and Citrus x sinensis). The consumption of such vegetables from these areas must be banned or strongly reduced, since long-term accumulation of Cu can cause a chronic toxicity in humans.

The understanding of acclimation strategies to low temperature and water availability is decisive to ensure coffee crop sustainability, since these environmental conditions determine the suitability of cultivation areas. In this context, the impacts of single and combined exposure to drought and cold were evaluated in three genotypes of the two major cropped species, Coffea arabica cv. Icatu, Coffea canephora cv. Apoatã, and the hybrid Obatã. Crucial traits of plant resilience to environmental stresses have been examined: photosynthesis, lipoperoxidation and the antioxidant response. Drought and/or cold promoted leaf dehydration, which was accompanied by stomatal and mesophyll limitations that impaired leaf C-assimilation in all genotypes. However, Icatu showed a lower impact upon stress exposure and a faster and complete photosynthetic recovery. Although lipoperoxidation was increased by drought (Icatu) and cold (all genotypes), it was greatly reduced by stress interaction, especially in Icatu. In fact, although the antioxidative system was reinforced under single drought and cold exposure (e.g., activity of enzymes as Cu,Zn-superoxide dismutase, ascorbate peroxidase, APX, glutathione reductase and catalase, CAT), the stronger increases were observed upon the simultaneous exposure to both stresses, which was accompanied with a transcriptional response of some genes, namely related to APX. Complementary, non-enzyme antioxidant molecules were promoted mostly by cold and the stress interaction, including α-tocopherol (in C. arabica plants), ascorbate (ASC), zeaxanthin, and phenolic compounds (all genotypes). In general, drought promoted antioxidant enzymes activity, whereas cold enhanced the synthesis of both enzyme and non-enzyme antioxidants, the latter likely related to a higher need of antioxidative capability when enzyme reactions were probably quite repressed by low temperature. Icatu showed the wider antioxidative capability, with the triggering of all studied antioxidative molecules by drought (except CAT), cold, and, particularly, stress interaction (except ASC), revealing a clear stress cross-tolerance. This justified the lower impacts on membrane lipoperoxidation and photosynthetic capacity under stress interaction conditions, related to a better ROS control. These findings are also relevant to coffee water management, showing that watering in the cold season should be largely avoided., Competing Interests: The authors have declared that no competing interests exist.

Climate changes, mostly related to high temperature, are predicted to have major negative impacts on coffee crop yield and bean quality. Recent studies revealed that elevated air [CO 2 ] mitigates the impact of heat on leaf physiology. However, the extent of the interaction between elevated air [CO 2 ] and heat on coffee bean quality was never addressed. In this study, the single and combined impacts of enhanced [CO 2 ] and temperature in beans of Coffea arabica cv. Icatu were evaluated. Plants were grown at 380 or 700 μL CO 2 L -1 air, and then submitted to a gradual temperature rise from 25°C up to 40°C during ca. 4 months. Fruits were harvested at 25°C, and in the ranges of 30-35 or 36-40°C, and bean physical and chemical attributes with potential implications on quality were then examined. These included: color, phenolic content, soluble solids, chlorogenic, caffeic and p -coumaric acids, caffeine, trigonelline, lipids, and minerals. Most of these parameters were mainly affected by temperature (although without a strong negative impact on bean quality), and only marginally, if at all, by elevated [CO 2 ]. However, the [CO 2 ] vs. temperature interaction strongly attenuated some of the negative impacts promoted by heat (e.g., total chlorogenic acids), thus maintaining the bean characteristics closer to those obtained under adequate temperature conditions (e.g., soluble solids, caffeic and p -coumaric acids, trigonelline, chroma, Hue angle, and color index), and increasing desirable features (acidity). Fatty acid and mineral pools remained quite stable, with only few modifications due to elevated air [CO 2 ] (e.g., phosphorous) and/or heat. In conclusion, exposure to high temperature in the last stages of fruit maturation did not strongly depreciate bean quality, under the conditions of unrestricted water supply and moderate irradiance. Furthermore, the superimposition of elevated air [CO 2 ] contributed to preserve bean quality by modifying and mitigating the heat impact on physical and chemical traits of coffee beans, which is clearly relevant in a context of predicted climate change and global warming scenarios.

World coffee production has faced increasing challenges associated with ongoing climatic changes. Several studies, which have been almost exclusively based on temperature increase, have predicted extensive reductions (higher than half by 2,050) of actual coffee cropped areas. However, recent studies showed that elevated [CO 2 ] can strongly mitigate the negative impacts of heat stress at the physiological and biochemical levels in coffee leaves. In addition, it has also been shown that coffee genotypes can successfully cope with temperatures above what has been traditionally accepted. Altogether, this information suggests that the real impact of climate changes on coffee growth and production could be significantly lower than previously estimated. Gene expression studies are an important tool to unravel crop acclimation ability, demanding the use of adequate reference genes. We have examined the transcript stability of 10 candidate reference genes to normalize RT-qPCR expression studies using a set of 24 cDNAs from leaves of three coffee genotypes (CL153, Icatu, and IPR108), grown under 380 or 700 μL CO 2 L -1 , and submitted to increasing temperatures from 25/20°C (day/night) to 42/34°C. Samples were analyzed according to genotype, [CO 2 ], temperature, multiple stress interaction ([CO 2 ], temperature) and total stress interaction (genotype, [CO 2 ], and temperature). The transcript stability of each gene was assessed through a multiple analytical approach combining the Coeficient of Variation method and three algorithms (geNorm, BestKeeper, NormFinder). The transcript stability varied according to the type of stress for most genes, but the consensus ranking obtained with RefFinder, classified MDH as the gene with the highest mRNA stability to a global use, followed by ACT and S15 , whereas α -TUB and CYCL showed the least stable mRNA contents. Using the coffee expression profiles of the gene encoding the large-subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase ( RLS ), results from the in silico aggregation and experimental validation of the best number of reference genes showed that two reference genes are adequate to normalize RT-qPCR data. Altogether, this work highlights the importance of an adequate selection of reference genes for each single or combined experimental condition and constitutes the basis to accurately study molecular responses of Coffea spp. in a context of climate changes and global warming.

Modeling studies have predicted that coffee crop will be endangered by future global warming, but recent reports highlighted that high [CO2] can mitigate heat impacts on coffee. This work aimed at identifying heat protective mechanisms promoted by CO2 in Coffea arabica (cv. Icatu and IPR108) and Coffea canephora cv. Conilon CL153. Plants were grown at 25/20°C (day/night), under 380 or 700 μL CO2 L(-1), and then gradually submitted to 31/25, 37/30, and 42/34°C. Relevant heat tolerance up to 37/30°C for both [CO2] and all coffee genotypes was observed, likely supported by the maintenance or increase of the pools of several protective molecules (neoxanthin, lutein, carotenes, α-tocopherol, HSP70, raffinose), activities of antioxidant enzymes, such as superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), catalase (CAT), and the upregulated expression of some genes (ELIP, Chaperonin 20). However, at 42/34°C a tolerance threshold was reached, mostly in the 380-plants and Icatu. Adjustments in raffinose, lutein, β-carotene, α-tocopherol and HSP70 pools, and the upregulated expression of genes related to protective (ELIPS, HSP70, Chape 20, and 60) and antioxidant (CAT, CuSOD2, APX Cyt, APX Chl) proteins were largely driven by temperature. However, enhanced [CO2] maintained higher activities of GR (Icatu) and CAT (Icatu and IPR108), kept (or even increased) the Cu,Zn-SOD, APX, and CAT activities, and promoted a greater upregulation of those enzyme genes, as well as those related to HSP70, ELIPs, Chaperonins in CL153, and Icatu. These changes likely favored the maintenance of reactive oxygen species (ROS) at controlled levels and contributed to mitigate of photosystem II photoinhibition at the highest temperature. Overall, our results highlighted the important role of enhanced [CO2] on the coffee crop acclimation and sustainability under predicted future global warming scenarios.

The actinorhizal tree Casuarina glauca tolerates extreme environmental conditions, such as high salinity. This species is also able to establish a root-nodule symbiosis with N2-fixing bacteria of the genus Frankia. Recent studies have shown that C. glauca tolerance to high salt concentrations is innate and linked to photosynthetic adjustments. In this study we have examined the impact of increasing NaCl concentrations (200, 400 and 600mM) on membrane integrity as well as on the control of oxidative stress in branchlets of symbiotic (NOD+) and non-symbiotic (KNO3+) C. glauca. Membrane selectivity was maintained in both plant groups at 200mM NaCl, accompanied by an increase in the activity of antioxidative enzymes (superoxide dismutase, ascorbate peroxidase, glutathione reductase and catalase). Regarding cellular membrane lipid composition, linolenic acid (C18:3) showed a significant decline at 200mM NaCl in both NOD+ and KNO3+ plants. In addition, total fatty acids (TFA) and C18:2 also decreased in NOD+ plants at this salt concentration, resulting in malondialdehyde (MDA) production. Such initial impact at 200mM NaCl is probably due to the fact that NOD+ plants are subjected to a double stress, i.e., salinity and low nitrogen availability. At 400mM NaCl a strong reduction of TFA and C18:3 levels was observed in both plant groups. This was accompanied by a decrease in the unsaturation degree of membrane lipids in NOD+. However, in both NOD+ and KNO3+ lipid modifications were not reflected by membrane leakage at 200 or 400mM, suggesting acclimation mechanisms at the membrane level. The fact that membrane selectivity was impaired only at 600mM NaCl in both groups of plants points to a high tolerance of C. glauca to salt stress independently of the symbiotic relation with Frankia., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

This analysis is focused on the effect of the abrupt decline of oil prices on biofuels, particularly second-generation ethanol. The efforts to decrease the production costs of biofuels, especially cellulosic ethanol (CE), will be greatly threatened if current oil prices remain low, especially since production is not slowing. Only huge state subsidies could alleviate this threat, but the challenge is to persuade citizens that this sacrifice is worthwhile., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The tropical coffee crop has been predicted to be threatened by future climate changes and global warming. However, the real biological effects of such changes remain unknown. Therefore, this work aims to link the physiological and biochemical responses of photosynthesis to elevated air [CO2 ] and temperature in cultivated genotypes of Coffea arabica L. (cv. Icatu and IPR108) and Coffea canephora cv. Conilon CL153. Plants were grown for ca. 10 months at 25/20°C (day/night) and 380 or 700 μl CO2 l(-1) and then subjected to temperature increase (0.5°C day(-1) ) to 42/34°C. Leaf impacts related to stomatal traits, gas exchanges, C isotope composition, fluorescence parameters, thylakoid electron transport and enzyme activities were assessed at 25/20, 31/25, 37/30 and 42/34°C. The results showed that (1) both species were remarkably heat tolerant up to 37/30°C, but at 42/34°C a threshold for irreversible nonstomatal deleterious effects was reached. Impairments were greater in C. arabica (especially in Icatu) and under normal [CO2 ]. Photosystems and thylakoid electron transport were shown to be quite heat tolerant, contrasting to the enzymes related to energy metabolism, including RuBisCO, which were the most sensitive components. (2) Significant stomatal trait modifications were promoted almost exclusively by temperature and were species dependent. Elevated [CO2 ], (3) strongly mitigated the impact of temperature on both species, particularly at 42/34°C, modifying the response to supra-optimal temperatures, (4) promoted higher water-use efficiency under moderately higher temperature (31/25°C) and (5) did not provoke photosynthetic downregulation. Instead, enhancements in [CO2 ] strengthened photosynthetic photochemical efficiency, energy use and biochemical functioning at all temperatures. Our novel findings demonstrate a relevant heat resilience of coffee species and that elevated [CO2 ] remarkably mitigated the impact of heat on coffee physiology, therefore playing a key role in this crop sustainability under future climate change scenarios., (© 2015 John Wiley & Sons Ltd.)

Casuarina glauca is an actinorhizal tree which establishes root-nodule symbiosis with N2-fixing Frankia bacteria. This plant is commonly found in saline zones and is widely used to remediate marginal soils and prevent desertification. The nature of its ability to survive in extreme environments and the extent of Frankia contribution to stress tolerance remain unknown. Thus, we evaluated the ability of C. glauca to cope with salt stress and the influence of the symbiosis on this trait. To this end, we analysed the impact of salt on plant growth, mineral contents, water relations, photosynthetic-related parameters and non-structural sugars in nodulated vs. non-nodulated plants. Although the effects on photosynthesis and stomatal conductance started to become measurable in the presence of 200 mM NaCl, photochemical (e.g., photosynthetic electron flow) and biochemical (e.g., activity of photosynthetic enzymes) parameters were only strongly impaired when NaCl levels reached 600 mM. These results indicate the maintenance of high tissue hydration under salt stress, probably associated with enhanced osmotic potential. Furthermore, the maintenance of photosynthetic assimilation potential (A(max)), together with the increase in the quantum yield of down-regulated energy dissipation of PSII (Y(NPQ)), suggested a down-regulation of photosynthesis instead of photo-damaging effects. A comparison of the impact of increasing NaCl levels on the activities of photosynthetic (RubisCO and ribulose-5 phosphate kinase) and respiratory (pyruvate kinase and NADH-dependent malate dehydrogenase) enzymes vs. photosynthetic electron flow and fluorescence parameters, revealed that biochemical impairments are more limiting than photochemical damage. Altogether, these results indicate that, under controlled conditions, C. glauca tolerates high NaCl levels and that this capacity is linked to photosynthetic adjustments., (Copyright © 2015 Elsevier Masson SAS. All rights reserved.)

Coffee is one of the world's most traded agricultural products. Modeling studies have predicted that climate change will have a strong impact on the suitability of current cultivation areas, but these studies have not anticipated possible mitigating effects of the elevated atmospheric [CO2] because no information exists for the coffee plant. Potted plants from two genotypes of Coffea arabica and one of C. canephora were grown under controlled conditions of irradiance (800 μmol m(-2) s(-1)), RH (75%) and 380 or 700 μL CO2 L(-1) for 1 year, without water, nutrient or root development restrictions. In all genotypes, the high [CO2] treatment promoted opposite trends for stomatal density and size, which decreased and increased, respectively. Regardless of the genotype or the growth [CO2], the net rate of CO2 assimilation increased (34-49%) when measured at 700 than at 380 μL CO2 L(-1). This result, together with the almost unchanged stomatal conductance, led to an instantaneous water use efficiency increase. The results also showed a reinforcement of photosynthetic (and respiratory) components, namely thylakoid electron transport and the activities of RuBisCo, ribulose 5-phosphate kinase, malate dehydrogenase and pyruvate kinase, what may have contributed to the enhancements in the maximum rates of electron transport, carboxylation and photosynthetic capacity under elevated [CO2], although these responses were genotype dependent. The photosystem II efficiency, energy driven to photochemical events, non-structural carbohydrates, photosynthetic pigment and membrane permeability did not respond to [CO2] supply. Some alterations in total fatty acid content and the unsaturation level of the chloroplast membranes were noted but, apparently, did not affect photosynthetic functioning. Despite some differences among the genotypes, no clear species-dependent responses to elevated [CO2] were observed. Overall, as no apparent sign of photosynthetic down-regulation was found, our data suggest that Coffea spp. plants may successfully cope with high [CO2] under the present experimental conditions.

Roasted coffee samples of the two major trade species (Coffea arabica and C. canephora) were studied to identify sensory descriptors that might be used to determine blends production and evaluation, following the expectations of consumers. Coffee beans were roasted at 220 + 10 °C, for 7, 9, and 11 min, and the sensory profiles of the beverages were assessed. From descriptive analysis the eigenvalues allowed the identification of two principal components (PCs), being the variance between samples 68.9% and 21.1%. In the first PC the characteristic odor, astringency, body, bitter flavor, burned aroma, and residual, typical, and burned tastes prevailed. The correlation coefficient between the second PC and citric acid flavor and aroma reached 0.96 and 0.78, respectively. It was concluded that in beverages of these species, the descriptors of both components can be separated according to bean roasting time. Considering roasting time, the overall quality was also rated.

To attain chemical descriptors responsible for sensory characteristics linked to the botanical origin of five Brazilian coffee genotypes, a chemical survey was carried out. Highest and lowest amounts of caffeine were found in Apoatã and Obatã. Coffea dewevrei showed the lowest contents of 5-CQA and 3-CQA. 3,5-diCQA was higher for Apoatã and minimum values were detected in Icatu. Apoatã showed the highest contents of 3,4-diCQA and 4,5-diCQA, Catuaí, Icatu and Obatã revealed lower values of 4,5-diCQA. Among hydroxycinnamic acids ferulic acid prevailed in all genotypes, with lower values in Icatu and Obatã. 3,4-DCA remained significantly higher in Apoatã, C. dewevrei and Catuaí. Caffeic acid and p-coumaric acid did not vary significantly. It was concluded that Apoatã was most adequate for the decaffeinated industry; Catuaí, Icatu and Obatã were identified as producers of good beverages. Apoatã and C. dewevrei seemed to share closer parental origins to Coffea canephora cv. Robusta, whereas Icatu, Obatã and Catuaí showed higher similarities to Coffea arabica.

Arabica and Robusta coffee beans were roasted at 220 ± 10°C for 7, 9 and 11 min to identify chemical descriptors in the beverages. The pH of the beverages showed the lowest value in the medium roasting level. In each degree of browning, the soluble solids content remained slightly higher in Arabica drinks. The contents of caffeine did not vary, but trigonelline decreased with burning up intensity. Chlorogenic acids also decreased with increasing roasting time. The 5-O-caffeoylquinic acid prevailed in Arabica and Robusta beverages, but the isomers of dicaffeoylquinic and feruolilquínic acids remained higher in Robusta. It was concluded that trigonelline and total caffeoylquinic, fatty dicaffeoylquinic and fatty feruolilquínic acids detached the beverages according to roasting intensity. Caffeine and pH allowed drinks separation between both species. Soluble solids take apart Arabica and Robusta drinks in each degree of roasting. All the individual groups of chlorogenic acids also explained 90% of the variance among samples.

Low positive temperature (chilling) is frequently linked to the promotion of oxidative stress conditions, and is of particular importance in the coffee plant due to its severe impact on growth, development, photosynthesis and production. Nevertheless, some acclimation ability has been reported within the Coffea genus, and is possibly related to oxidative stress control. Using an integrated biochemical and molecular approach, the characterization of the antioxidative system of genotypes with different cold acclimation abilities was performed. Experiments were carried out using 1.5-year-old coffee seedlings of Coffea canephora cv. Apoatã, C. arabica cv. Catuaí, C. dewevrei and 2 hybrids, Icatu (C. arabicaxC. canephora) and Piatã (C. dewevreixC. arabica) subjected to a gradual cold treatment and a recovery period. Icatu showed the greatest ability to control oxidative stress, as reflected by the enhancement of several antioxidative components (Cu,Zn-SOD and APX activities; ascorbate, alpha-tocopherol and chlorogenic acids (CGAs) contents) and lower reactive oxygen species contents (H(2)O(2) and OH). Gene expression studies show that GRed, DHAR and class III and IV chitinases might also be involved in the cold acclimation ability of Icatu. Catuaí showed intermediate acclimation ability through the reinforcement of some antioxidative molecules, usually to a lesser extent than that observed in Icatu. On the other hand, C. dewevrei showed the poorest response in terms of antioxidant accumulation, and also showed the greatest increase in OH values. The difference in the triggering of antioxidative traits supports the hypothesis of its importance to cold (and photoinhibition) tolerance in Coffea sp. and could provide a useful probe to identify tolerant genotypes., (Copyright 2009 Elsevier GmbH. All rights reserved.)