Search

Twin-scale explants of Iris hollandica cv. 'Bronze Queen' produced callus when cultured on a modified Murashige and Skoog medium supplemented with 9.0, 18.1, 27.2 or 36.2 μM 24-dichlorophenoxyacetic acid (2,4-D). Induction of embryogenic callus was obtained at 9.0 or 18.1 μM 2,4-D, while higher concentrations of the auxin (27.2 or 36.2 μM) were not effective. Embryo-like structures developed on the surface of embryogenic calli. Somatic embryo maturation was achieved when the bipolar structures produced were cultured on medium without auxin. To improve embryo germination, the developing embryos were transferred to medium supplemented with 0.95 pM abscisic acid (ABA). in this medium, mature embryos developed into normal plantlets. Histological studies confirmed that the structures exhibiting bipolar differentiation were somatic embryos. Regeneration of I. hollandica cv. 'Bronze Queen' plants through somatic embryogenesis was thus established. [ABSTRACT FROM AUTHOR]

The aim of this systematic review was to determine whether there is a root canal filling for deciduous teeth equally or more effective than zinc oxide-eugenol cement (ZOE). Six clinical trials selected for inclusion were independently reviewed by two researchers. Only two showed statistically significant different success rates between the test and the control groups. One found that an iodoform paste with calcium hydroxide (IP + Ca) performed better than ZOE, and the other found that ZOE performed similarly to IP + Ca. The other four studies compared ZOE with an iodoform paste (IP), a calcium hydroxide cement (Ca(OH)2), or IP + Ca. In these trials, the success rates in the ZOE groups were slightly lower than in the other groups. There seems to be no convincing evidence to support the superiority of any material over ZOE, and both ZOE and IP + Ca appear to be suitable as root canal fillings for deciduous teeth. [ABSTRACT FROM AUTHOR]

Objective: To compare the efficacy of silver diamine fluoride (SDF) and atraumatic restorative treatment (ART) in arresting caries lesions. Variables such as treatment time, adverse effects/parental aesthetic perception, anxiety and patients' behavior were also evaluated., Materials and Methods: Children (3.53 ± 1.03 years) with dentin caries lesions on the occlusal surface of primary molars were randomized into test (SDF) and control (ART) groups. To determine the presence of caries, dmf-t and ICDAS indexes were used. Caries lesions were also classified according to activity (active or inactive). The time required to perform treatments was recorded and the children's anxiety was assessed by a Facial Image Scale. The adverse events/aesthetic perception were registered by the operator and caregivers. The Frankl Behavioral Scale assessed patients' behavior. The chi-square/Fisher's exact and Mann-Whitney tests were used for comparisons between the groups., Results: Of the 118 participants who received treatment (SDF, n = 59; ART, n = 59), after 12-month follow-up, 91.5% (43/47) of caries lesions in the SDF and 90.2% (46/51) in the ART group were arrested (p = 1.000). After 24 months, 72% (18/25) were arrested in the SDF group and 95.2% (20/21) in the ART. The treatment times were 6.08 ± 1.72 and 13.58 ± 4.83, for SDF and ART, respectively (p < 0.001). No statistically significant difference of adverse effects, aesthetic perception, anxiety, and patients' behavior were found between the groups., Conclusions: SDF was similar to ART in arresting caries lesion but required less time for treatment. The anxiety, adverse effects/aesthetic perception and patients' behavior were also similar between the groups.   CLINICAL RELEVANCE: The use of SDF may be a faster and less invasive alternative for the treatment of caries lesions. Clinical trial registration number and date of registration: NCT03063307. September 2016., Competing Interests: Declarations. Ethics approval and consent to participate: All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. The study was approved by the Research Ethics Committee of the Clementino Fraga Filho Hospital of the Federal University of Rio de Janeiro (UFRJ) (protocol 4.169.258/CAAE:56374216.2.0000.5257). Written informed consent was taken from the guardians of all participants. Competing interests: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The Mediterranean basin is highly susceptible to climate change, with soil salinization and the increase in average temperatures being two of the main factors affecting crop productivity in this region. Following our previous studies on describing the detrimental effects of heat and salt stress co-exposure on tomato plants, this study aimed to understand if substrate supplementation with a combination of arbuscular mycorrhizal fungi (AMF) and biochar could mitigate the negative consequences of these stresses. Upon 21 days of exposure, stressed tomato plants grown under supplemented substrates showed increased tolerance to heat (42 °C for 4 h/day), salt (100 mM NaCl), and their combination, presenting increased biomass and flowering rate. The beneficial effects of AMF and biochar were associated with a better ionic balance (i.e. lower sodium accumulation and higher uptake of calcium and magnesium) and increased photosynthetic efficiency. Indeed, these plants presented higher chlorophyll content and improved CO 2 assimilation rates. Biochemical data further supported that tomato plants grown with AMF and biochar were capable of efficiently modulating their defence pathways, evidenced by the accumulation of proline, ascorbate, and glutathione, coupled with a lower dependency on energy-costly enzymatic antioxidant players. In summary, the obtained data strongly point towards a beneficial role of combined AMF and biochar as sustainable tools to improve plant growth and development under a climate change scenario, where soil salinization and heat peaks often occur together., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Castanea sativa Miller, a high-valuable crop for Mediterranean countries, is facing frequent and prolonged periods of heat and drought, severely affecting chestnut production. Aiming to tackle this problem, this study unraveled the influence of mycorrhizal association with the fungi Paxillus involutus (Batsch) on young chestnut plants' responses to combined heat (42 °C; 4 h/day) and drought (no irrigation until soil moisture reached 25%) over 21 days of stress exposure. Heat stress had no harmful effects on growth, photosynthesis, nor induced oxidative stress in either mycorrhizal (MR) or non-mycorrhizal (NMR) chestnut plants. However, drought (alone or combined) reduced the growth of NMR plants, affecting water content, leaf production, and foliar area, while also hampering net CO 2 assimilation and carbon relations. The mycorrhizal association, however, mitigated the detrimental effects of both stresses, resulting in less susceptibility and fewer growth limitations in MR chestnut plants, which were capable of ensuring a proper carbon flow. Evaluation of the oxidative metabolism revealed increased lipid peroxidation and hydrogen peroxide levels in NMR plants under water scarcity, supporting their higher susceptibility to stress. Conversely, MR plants activated defense mechanisms by accumulating antioxidant metabolites (ascorbate, proline and glutathione), preventing oxidative damage, especially under the combined stress. Overall, drought was the most detrimental condition for chestnut growth, with heat exacerbating stress susceptibility. Moreover, mycorrhizal association with P. involutus substantially alleviated these effects by improving growth, water relations, photosynthesis, and activating defense mechanisms. Thus, this research highlights mycorrhization's potential to enhance C. sativa resilience against climate change, especially at early developmental stages., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Glyphosate (GLY), the most widely used herbicide, has been regarded as an emergent environmental contaminant due to its constant and cumulative use, with potential harm to non-target organisms, such as crops, disrupting cells' redox balance. Therefore, plants need to fine-tune their antioxidant (AOX) mechanisms to thrive under GLY-contaminated environments. Proline overaccumulation is a common response in plants exposed to GLY, yet its role in GLY-induced toxicity remains unclear. Thus, this study explores whether Pro overaccumulation in response to GLY is perceived as a downstream tolerance mechanism or an early-warning stress signal. To investigate this, Arabidopsis thaliana T-DNA mutant lines for Pro biosynthetic (P5CS1) and catabolic genes (ProDH) were used and screened for their GLY susceptibility. Upon seedlings' exposure to GLY (0.75 mg L -1 ) for 14 days, the herbicide led to reduced biomass in all genotypes, accompanied by Pro overaccumulation. Mutants with heightened Pro levels (prodh) exhibited the greatest biomass reduction, increased lipid peroxidation (LP), and hydrogen peroxide (H 2 O 2 ) levels, accompanied by a compromised performance of the AOX system. Conversely, p5cs1-4, mutants with lower Pro levels, demonstrated an enhanced AOX system activation, not only with increased levels of glutathione (GSH) and ascorbate (AsA), but also with increased activity of both ascorbate peroxidase (APX) and catalase (CAT). These findings suggest that Pro overaccumulation under GLY exposure is associated with stress sensitivity rather than tolerance, highlighting its potential as an early-warning signal for GLY toxicity in non-target plants and for detecting weed resistance., (© 2024 Scandinavian Plant Physiology Society.)

Progesterone (PROG) has been detected at various concentrations in the environment and has adverse effects on humans and wildlife. This work evaluated the impact of PROG in Solanum nigrum L. plants, its removal capacity, and how 2,4-epibrassinolide (24-EBL) affects this process. Three treatments were used: (1) control, (2) irrigation with 0.8 µM PROG, and (3) treatment with 0.8 µM PROG after a pre-treatment with a foliar application of 1 µM 2,4-EBL (PROG/24EBL). After 20 days of treatment, no PROG was detected in the nutrient solution or plant tissues, indicating that the PROG was removed and metabolized. Lipid peroxidation significantly decreased in response to PROG in shoots and roots, and this effect was even more significant for both organs of the PROG/24EBL plants. Additionally, both treatments in both organs showed a decrease in H 2 O 2 levels, and both steroid hormones increased the plants' antioxidant system at both the biochemical and gene expression levels. In conclusion, S. nigrum can swiftly remove PROG without affecting its growth, and the use of 24-EBL synergistically decreases oxidative damage by increasing the activity of the antioxidant system and enhancing plant PROG removal ability., Competing Interests: Competing interestThe authors have no relevant financial or nonfinancial interests to disclose., (© The Author(s) 2024.)

The equilibrium of the marine ecosystem is currently threatened by several constraints, among which climate change and anthropogenic activities stand out. Indeed, these factors favour the growth of macroalgae, which sometimes end up stranded on the beaches at the end of their life cycle, forming what is known as beach wrack. Despite its undeniable important ecological role on beaches, as it is an important source of organic matter (OM), and provides food and habitat for several invertebrates, reptiles, small mammals, and shorebirds, the overaccumulation of beach wrack is often associated with the release of greenhouse gases, negatively impacting tourist activities, and generating economic expenses for its removal. Although currently beach wrack is mainly treated as a waste, it can be used for numerous potential applications in distinct areas. This review aimed at providing a solid point of view regarding the process of wrack formation, its spatiotemporal location, as well as its importance and risks. It also contains the current advances of the research regarding sustainable alternatives to valorise this organic biomass, that range from bioenergy production to the incorporation of wrack in agricultural soils, considering a circular economy concept. Although there are some concerns regarding wrack utilisation, from its variable availability to a possible soil contamination with salts and other contaminants, this review comprises the overall beneficial effects of the incorporation of this residue particularly in the organic agricultural model, strengthening the conversion of this wasted biomass into a valuable resource., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

The occurrence of drought in soils, particularly in those contaminated by metals, poses a current threat to crops, as these factors can interact and induce unique stress responses. Therefore, this study mainly focused on understanding the crosstalk between drought and copper (Cu) stress in the physiology of the barley (Hordeum vulgare L.) plant. Using a bifactorial experimental design, seedlings were grown in a natural soil under the following treatments: plants continuously irrigated in uncontaminated soil for 14 days (control); plants continuously irrigated in Cu-contaminated soil (115 mg Cu kg -1 ) for 14 days (Cu); plants only irrigated during the initials 7 days of growth in uncontaminated soil (drought); plants co-exposed to Cu and drought (combined). After 14 days of growth, the results revealed that drought prevented Cu bioaccumulation in barley roots, which were still severely affected by the metal, both individually and in combination with the water deficit. Furthermore, individual and combined exposure to these stressors resulted in impaired photosynthetic performance in barley plants. Despite the increased activation of enzymatic and non-enzymatic antioxidant defence mechanisms, particularly in the green organs, the plants co-exposed to both stress factors still showed higher oxidative damage, severely impacting biomass production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Masson SAS.)

Abiotic stress combinations, such as high temperatures and soil/water salinization, severely threaten crop productivity worldwide. In this work, an integrative insight into the photosynthetic metabolism of tomato plants subjected to salt (100 mM NaCl) and/or heat (42 °C; 4 h/day) was performed. After three weeks, the stress combination led to more severe consequences on growth and photosynthetic pigments than the individual stresses. Regarding the photochemical efficiency, transcript accumulation and protein content of major actors (CP47 and D1) were depleted in all stressed plants, although the overall photochemical yield was not negatively affected under the co-exposure. Gas-exchange studies revealed to be mostly affected by salt (single or combined), which harshly compromised carbon assimilation. Additionally, transcript levels of stress-responsive genes (e.g., HsfA1 and NHX2) were differentially modulated by the single and combined treatments, suggesting the activation of stress-signature responses. Overall, by gathering an insightful overview of the main regulatory hub of photosynthesis, we show that the impacts on the carbon metabolism coming from the combination of heat and salinity, two major conditioners of crop yields, were not harsher than those of single stresses, indicating that the growth impairment might be attributed to a proficient distribution of resources towards defense mechanisms., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Background: Fluoride varnish (FV) is widely recommended for caries prevention in preschool children, despite its anticaries benefits being uncertain and modest. Dentists often report using clinical practice guidelines (CPGs) as a source of scientific information., Aim: To identify and analyze recommendations for clinical practice on the use of FV for caries prevention in preschool children and to assess the methodological quality of the CPG on this topic., Design: Two researchers independently used 12 search strategies and searched the first five pages of Google Search™ and three guideline databases for recommendations freely available to health professionals on the use of FV for caries prevention in preschoolers. Then, they retrieved and recorded recommendations that met the eligibility criteria and extracted the data. A third researcher resolved disagreements. Each included CPG was appraised using the AGREE II instrument., Results: Twenty-nine documents were included. Recommendations varied according to age, patients' caries risk, and application frequency. Of the six CPGs, only one scored above 70% in the AGREE II overall assessment., Conclusion: Recommendations on the use of FV lacked scientific evidence, and CPGs were of poor quality. Application of FV is widely recommended despite recent evidence showing an uncertain, modest, and possibly not clinically relevant anticaries benefit. Dentists should be aware that it is necessary to critically appraise CPGs since they may be of poor quality., (© 2023 BSPD, IAPD and John Wiley & Sons Ltd.)

Increased genetic risk for melanoma can occur in the context of germline pathogenic variants in high-penetrance genes, such as CDKN2A and CDK4 , risk variants in low- to moderate-penetrance genes ( MC1R and MITF ), and possibly due to variants in emerging genes, such as ACD , TERF2IP, and TERT . We aimed to identify germline variants in high- and low- to moderate-penetrance melanoma risk genes in Brazilian patients with clinical criteria for familial melanoma syndrome. We selected patients with three or more melanomas or melanoma patients from families with three tumors (melanoma and pancreatic cancer) in first- or second-degree relatives. Genetic testing was performed with a nine-gene panel ( ACD , BAP1 , CDK4 , CDKN2A , POT1 , TERT , TERF2IP , MC1R, and MITF ). In 36 patients, we identified 2 (5.6%) with germline pathogenic variants in CDKN2A and BAP1 and 4 (11.1%) with variants of uncertain significance in the high-penetrance genes. MC1R variants were found in 86.5%, and both red hair color variants and unknown risk variants were enriched in patients compared to a control group. The low frequency of germline pathogenic variants in the high-penetrance genes and the high prevalence of MC1R variants found in our cohort show the importance of the MC1R genotype in determining the risk of melanoma in the Brazilian melanoma-prone families.

Allelopathy, the inhibition of neighbouring plant growth by certain plants, can be particularly useful if applied in a targeted way for weed management. So, this study aimed to assess and characterize the herbicidal activity of fresh and dried leaves from young Eucalyptus globulus Labill. trees applied as a soil amendment. For this, fresh and dried leaves (FL and DL, respectively) were incorporated into the soil at different concentrations (0, 1, 5, and 10% w/w), where Portulaca oleracea L. seeds were sown. After 5 weeks of exposure, results revealed that the soil incorporation of DL at 10% (w/w) presented the strongest herbicidal properties, inhibiting seed germination by 63% and inducing the loss of cell viability. To unravel the possible mode of action and the main targets at both cellular and subcellular levels, an in vitro experiment was performed. Purslane seeds were sown in a nutritive medium containing different dilutions of an aqueous extract prepared with dried eucalyptus leaves. After 5 days of exposure, germinated seedlings were processed for transmission electron microscopy and histological analyses as well as for reactive oxygen species (ROS) in vivo detection by confocal laser scanning microscopy. Results revealed that the allelochemical release from DL induced ROS overproduction, resulting in the loss of cell integrity and organization, which was characterized by damage to several cellular sub-structures, along with enhanced accumulation of lipid droplets. Overall, the incorporation of DL into the soil can represent a sustainable alternative to reduce synthetic herbicide application and subsequent environmental contamination., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Mafalda Pinto and Fernanda Fidalgo reports financial support was provided by Fundação para a Ciência e Tecnologia. Mafalda Pinto reports financial support was provided by European Union's Horizon 2020 Research and Innovation program., (Copyright © 2023 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

As available tools for crop disease management are scarce, new, effective, and eco-friendly solutions are needed. So, this study aimed at assessing the antibacterial activity of a dried leaf Eucalyptus globulus Labill. aqueous extract (DLE) against Pseudomonas syringae pv. tomato ( Pst ), Xanthomonas euvesicatoria ( Xeu ), and Clavibacter michiganensis michiganensis ( Cmm ). For this, the inhibitory activity of different concentrations of DLE (0, 15, 30, 45, 60, 75, 90, 105, 120, 135, and 250 g L -1 ) was monitored against the type strains of Pst , Xeu , and Cmm through the obtention of their growth curves. After 48 h, results showed that the pathogen growth was strongly inhibited by DLE, with Xeu the most susceptible species (15 g L -1 MIC and IC 50 ), followed by Pst (30 g L -1 MIC and IC 50 ), and Cmm (45 and 35 g L -1 MIC and IC 50 , respectively). Additionally, using the resazurin assay, it was possible to verify that DLE considerably impaired cell viability by more than 86%, 85%, and 69% after Pst , Xeu , and Cmm were incubated with DLE concentrations equal to or higher than their MIC, respectively. However, only the treatment with DLE at 120 g L -1 did not induce any hypersensitive response in all pathogens when treated bacterial suspensions were infiltrated onto tobacco leaves. Overall, DLE can represent a great strategy for the prophylactic treatment of tomato-associated bacterial diseases or reduce the application of environmentally toxic approaches.

Agricultural soil degradation is occurring at unprecedented rates, not only as an indirect effect of climate change (CC) but also due to intensified agricultural practices which affect soil properties and biodiversity. Therefore, understanding the impacts of CC and soil degradation on plant physiology is crucial for the sustainable development of mitigation strategies to prevent crop productivity losses. The amino acid proline has long been recognized for playing distinct roles in plant cells undergoing osmotic stress. Due to its osmoprotectant and redox-buffering ability, a positive correlation between proline accumulation and plants' tolerance to abiotic stress has been pointed out in numerous reviews. Indeed, proline quantification is used systematically by plant physiologists as an indicator of the degree of tolerance and a measurement of the antioxidant potential in plants under stressful conditions. Moreover, the exogenous application of proline has been shown to increase resilience to several stress factors, including those related to soil degradation such as salinity and exposure to metals and xenobiotics. However, recent data from several studies often refer to proline accumulation as a signal of stress sensitivity with no clear correlation with improved antioxidant activity or higher stress tolerance, including when proline is used exogenously as a stress reliever. Nevertheless, endogenous proline levels are strongly modified by these stresses, proving its involvement in plant responses. Hence, one main question arises-is proline augmentation always a sign of improved stress resilience? From this perspective, the present review aims to provide a more comprehensive understanding of the implications of proline accumulation in plants under abiotic stress induced by soil degradation factors, reinforcing the idea that proline quantification should not be employed as a sole indicator of stress sensitivity or resilience but rather complemented with further biochemical and physiological endpoints.

This study aimed to understand the morphophysiological responses and primary metabolism of tomato seedlings subjected to mild levels of nitrogen and/or water deficit (50% N and/or 50% W). After 16 days of exposure, plants grown under the combined deficit showed similar behavior to the one found upon exposure to single N deficit. Both N deficit treatments resulted in a significantly lower dry weight, leaf area, chlorophyll content, and N accumulation but in a higher N use efficiency when compared to control (CTR) plants. Moreover, concerning plant metabolism, at the shoot level, these two treatments also responded in a similar way, inducing higher C/N ratio, nitrate reductase (NR) and glutamine synthetase (GS) activity, expression of RuBisCO encoding genes as well as a downregulation of GS2.1 and GS2.2 transcripts. Interestingly, plant metabolic responses at the root level did not follow the same pattern, with plants under combined deficit behaving similarly to W deficit plants, resulting in enhanced nitrate and proline concentrations, NR activity, and an upregulation of GS1 and NR genes than in CTR plants. Overall, our data suggest that the N remobilization and osmoregulation strategies play a relevant role in plant acclimation to these abiotic stresses and highlight the complexity of plant responses under a combined N+W deficit., Competing Interests: The authors declare no conflict of interest.

This study aims to perform a broad analysis of the antioxidant (AOX) responses of young tomato plants exposed to single and combined mild nitrogen (N) and water deficits through the evaluation of oxidative biomarkers, non-enzymatic and enzymatic AOX components. 'Micro-Tom' seedlings were subjected to four treatments: control (CTR; 100%N + 100%W), N deficit (N; 50%N), water deficit (W; 50%W), and combined deficits (N + W; 50%N + 50%W). An enhancement of several non-enzymatic and enzymatic components was found in plants subjected to N + W deficit, which presented higher anthocyanins accumulation (up to 103%) as well as higher levels of superoxide dismutase (SOD) transcripts at root level and of ascorbate peroxidase (APX) and catalase (CAT) transcripts at shoot level. This increase in the gene expression was also translated in augmented SOD (up to 202%), APX (up to 155%) and CAT (up to 108%) activity compared to CTR plants and the single deficits. Overall, tomato plants were able to employ defense strategies to cope with this combined deficit, as demonstrated by the higher total AOX capacity (up to 87%) compared to the single deficits, which contributed to the maintenance of their redox homeostasis, with unchanged values of lipid peroxidation and hydrogen peroxide compared with CTR plants.

Glyphosate (GLY) and flazasulfuron (FLA) are two non-selective herbicides commonly applied together. However, research focused on their single and combined ecotoxicological impacts towards non-target organisms is still inconclusive. Therefore, this study aimed to test their single effects on soil's habitat and retention functions, and to unravel their combined impacts to earthworms and terrestrial plants. For this, ecotoxicological assays were performed with plants (Medicago sativa), oligochaetes (Eisenia fetida) and collembola (Folsomia candida). Soil elutriates were also prepared and tested in macrophytes (Lemna minor) and microalgae (Raphidocelis subcapitata). FLA (82-413 µg kg -1 ) reduced earthworms' and collembola's reproduction and severely impaired M. sativa growth, being much more toxic than GLY (up to 30 mg kg -1 ). In fact, the latter only affected plant growth (≥ 9 mg kg -1 ) and earthworms (≥ 13 mg kg -1 ), especially at high concentrations, with no effects on collembola. Moreover, only elutriates from FLA-contaminated soils significantly impacted L. minor and R. sucapitata. The experiments revealed that the co-exposure to GLY and FLA enhanced the toxic effects of contaminated soils not only on plants but also on earthworms'. However, such increase in toxicity was dependent on GLY residual concentrations in soils. Overall, this work underpins that herbicides risk assessment should consider herbicides co-exposures, since the evaluation of single exposures is not representative of current phytosanitary practices and of the potential effects under field conditions, where residues of different compounds may persist in soils., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Plants must adapt to cope with adverse environmental conditions that affect their growth and development. To overcome these constraints, they can alter their developmental patterns by modulating cellular processes and activating stress-responsive signals. Alongside the activation of the antioxidant (AOX) system, a high number of genes are expressed, and proteins must be distributed to the correct locations within the cell. The endomembrane system and associated vesicles thus play an important role. Several pathways have been associated with adverse environmental conditions, which is the case for the exocyst-positive organelle-EXPO. The present work, using Arabidopsis mutants with T-DNA insertions in the gene EXO70 , essential for EXPO vesicles formation, was designed to characterise the anatomical (morphology and root length), biochemical (quantification of stress markers and antioxidant system components), and molecular responses (gene expression) to abiotic stresses (saline, drought, oxidative, and metal-induced toxicity). The results obtained showed that mutant plants behave differently from the wild type (WT) plants. Therefore, in the exo70 mutant, morphological changes were more noticeable in plants under stress, and the non-enzymatic component of the antioxidant system was activated, with no alterations to the enzymatic component. Furthermore, other defence strategies, such as autophagy, did not show important changes. These results confirmed the EXPO as an important structure for tolerance/adaptation to stress.

Aluminum (Al) toxicity limits crops growth and production in acidic soils. Compared to roots, less is known about the toxic effects of Al in leaves. Al subcellular compartmentalization is also largely unknown. Using rye (Secale cereale L.) Beira (more tolerant) and RioDeva (more sensitive to Al) genotypes, we evaluated the patterns of Al accumulation in leaf cell organelles and the photosynthetic and metabolic changes to cope with Al toxicity. The tolerant genotype accumulated less Al in all organelles, except the vacuoles. This suggests that Al compartmentalization plays a role in Al tolerance of Beira genotype. PSII efficiency, stomatal conductance, pigment biosynthesis, and photosynthesis metabolism were less affected in the tolerant genotype. In the Calvin cycle, carboxylation was compromised by Al exposure in the tolerant genotype. Other Calvin cycle-related enzymes, phoshoglycerate kinase (PGK), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), triose-phosphate isomerase (TPI), and fructose 1,6-bisphosphatase (FBPase) activities decreased in the sensitive line after 48 h of Al exposure. Consequentially, carbohydrate and organic acid metabolism were affected in a genotype-specific manner, where sugar levels increased only in the tolerant genotype. In conclusion, Al transport to the leaf and compartmentalization in the vacuoles tolerant genotype's leaf cells provide complementary mechanisms of Al tolerance, protecting the photosynthetic apparatus and thereby sustaining growth., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Alexandra de Sousa reports financial support was provided by Fundação para a Ciência eTecnologia, Shereen M. Korny., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Aqueous extracts from Eucalyptus globulus leaves contain a wide variety of specialized metabolites, mainly polyphenols and appreciable amounts of volatile compounds, which are responsible for their diverse biological activities, such as antioxidant, antimicrobial, and allelopathic features. For this reason, several studies have been conducted to explore the composition of E. globulus leaf extracts for multiple therapeutic and commercial applications. However, so far, the available bibliographic reports only refer to the chemical composition of extracts prepared with leaves from mature trees, leaving much to clarify about the composition of juvenile eucalyptus leaf extracts. Furthermore, there is no consensus regarding the type of leaves, fresh or dried ones, to be used in the extraction procedure, considering the highest recovery of biologically active compounds. In this sense, this study aimed to characterize the chemical composition of aqueous extracts prepared with fresh and dried leaves from young and mature E. globulus trees. For this, leaf biomass from young and mature E. globulus trees was collected in three distinct places from a forest area, and after oven-drying a portion of the leaves, an extraction in hot water was carried out, followed by GC-MS and HPLC-MS/MS analyses. The results revealed that the maturity of eucalyptus trees and biomass drying significantly influenced the volatile and non-volatile composition of the aqueous extracts. Accordingly, while fresh leaf extracts of young trees had great levels of hydrolysable tannins, extracts prepared with fresh leaves from mature trees presented a wide range of terpenes. When dried leaf material was used, extracts had notorious contents of amino acids derivatives, C 13 norisoprenoids, fatty and other organic acids. Overall, this study showed, for the first time, that plant maturity (young vs mature) and pre-processing (fresh vs dried) of foliar biomass of E. globulus trees need to be considered in the preparation of leaf aqueous extracts depending on the desired purposes, since major changes in what regards biologically active compounds were found., 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., (Copyright © 2022 Pinto, Soares, Pereira, Rodrigues, Fidalgo and Valente.)

This work aimed to evaluate the potential of Fe 2 O 3 nanoparticles (nano-Fe 2 O 3 ) to alleviate potentially toxic elements (PTEs) - induced stress in barley plants (Hordeum vulgare L.), focusing on bioaccumulation patterns and on plant growth and redox homeostasis. To achieve this goal, plants grew in two agricultural soils, contaminated by different levels of PTEs, collected from an industrial area, previously amended, or not, with 1% (w/w) nano-Fe 2 O 3 . After 14 d of growth, biometric parameters were evaluated, along with the analysis of PTEs bioaccumulation and biochemical endpoints. After exposure to contaminated soils, plant development was greatly affected, as evidenced by significant decreases in root length and biomass production. However, upon co-treatment with nano-Fe 2 O 3 , lower inhibitory effects on biometric parameters were observed. Regarding the oxidative damage, both soils led to increases in lipid peroxidation and superoxide anion concentration, though hydrogen peroxide levels were only increased in the most contaminated soil. In general, these changes in the oxidative stress markers were accompanied by an upregulation of different antioxidant mechanisms, whose efficiency was even more powerful upon soil amendment with nano-Fe 2 O 3 , thus lowering PTEs-induced oxidative damage. Altogether, the present study revealed that nano-Fe 2 O 3 can protect the growth of barley plants under contaminated soils., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Currently, salinity and heat are two critical threats to crop production and food security which are being aggravated by the global climatic instability. In this scenario, it is imperative to understand plant responses to simultaneous exposure to different stressors and the cross-talk between underlying functional mechanisms. Thus, in this study, the physiological and biochemical responses of tomato plants ( Solanum lycopersicum L.) to the combination of salinity (100 mM NaCl) and heat (42 °C; 4 h/day) stress were evaluated. After 21 days of co-exposure, the accumulation of Na + in plant tissues was superior when salt-treated plants were also exposed to high temperatures compared to the individual saline treatment, leading to the depletion of other nutrients and a harsher negative effect on plant growth. Despite that, neither oxidative damage nor a major accumulation of reactive oxygen species took place under stress conditions, mostly due to the accumulation of antioxidant (AOX) metabolites alongside the activation of several AOX enzymes. Nonetheless, the plausible allocation of resources towards the defense pathways related to oxidative and osmotic stress, along with severe Na toxicity, heavily compromised the ability of plants to grow properly when the combination of salinity and heat was imposed.

Genetic predisposition accounts for nearly 10% of all melanoma cases and has been associated with a dozen moderate- to high-penetrance genes, including CDKN2A, CDK4, POT1 and BAP1. However, in most melanoma-prone families, the genetic etiology of cancer predisposition remains undetermined. The goal of this study was to identify rare genomic variants associated with cutaneous melanoma susceptibility in melanoma-prone families. Whole-exome sequencing was performed in 2 affected individuals of 5 melanoma-prone families negative for mutations in CDKN2A and CDK4, the major cutaneous melanoma risk genes. A total of 288 rare coding variants shared by the affected relatives of each family were identified, including 7 loss-of-function variants. By performing in silico analyses of gene function, biological pathways, and variant pathogenicity prediction, we underscored the putative role of several genes for melanoma risk, including previously described genes such as MYO7A and WRN, as well as new putative candidates, such as SERPINB4, HRNR, and NOP10. In conclusion, our data revealed rare germline variants in melanoma-prone families contributing with a novel set of potential candidate genes to be further investigated in future studies., Competing Interests: The authors have declared that no competing interests exist.

Diclofenac (DCF) is a very common pharmaceutical that, due to its high use and low removal rate, is considered a prominent contaminant in surface and groundwater worldwide. In this study, Solanum lycopersicum L. cv. Micro-Tom (tomato) was used to disclose the role of glutathione (GSH)-related enzymes, as GSH conjugation with DCF is a well reported detoxification mechanism in mammals and some plant species. To achieve this, S. lycopersicum plants were exposed to 0.5 and 5 mg L -1 of DCF for 5 weeks under a semi-hydroponic experiment. The results here obtained point towards an efficient DCF detoxification mechanism that prevents DCF bioaccumulation in fruits, minimizing any concerns for human health. Although a systemic response seems to be present in response to DCF, the current data also shows that its detoxification is mostly a root-specific process. Furthermore, it appears that GSH-mediated DCF detoxification is the main mechanism activated, as glutathione-S-transferase (GST) activity was greatly enhanced in roots of tomato plants treated with 5 mg L -1 DCF, accompanied by increased glutathione reductase activity, responsible for GSH regeneration. By applying a targeted gene expression analysis, we provide evidence, for the first time, that SlGSTF4 and SlGSTF5 genes, coding for GSTs from phi class, were the main players driving the conjugation of this contaminant. In this sense, and even though tomato plants appear to be somewhat tolerant to DCF exposure, research on GST activity can prove to be instrumental in remediating DCF-contaminated environments and improving plant growth under such conditions., (Copyright © 2021 Elsevier Masson SAS. All rights reserved.)