Salicylic Acid Priming Improves Cotton Seedling Heat Tolerance through Photosynthetic Pigment Preservation, Enhanced Antioxidant Activity, and Osmoprotectant Levels.

The escalating global temperatures associated with climate change are detrimental to plant growth and development, leading to significant reductions in crop yields worldwide. Our research demonstrates that salicylic acid (SA), a phytohormone known for its growth-promoting properties, is crucial in enhancing heat tolerance in cotton (Gossypium hirsutum). This enhancement is achieved through modifications in various biochemical, physiological, and growth parameters. Under heat stress, cotton plants typically show significant growth disturbances, including leaf wilting, stunted growth, and reduced biomass. However, priming cotton plants with 1 mM SA significantly mitigated these adverse effects, evidenced by increases in shoot dry mass, leaf-water content, and chlorophyll concentrations in the heat-stressed plants. Heat stress also prompted an increase in hydrogen peroxide levels-a key reactive oxygen species-resulting in heightened electrolyte leakage and elevated malondialdehyde concentrations, which indicate severe impacts on cellular membrane integrity and oxidative stress. Remarkably, SA treatment significantly reduced these oxidative stresses by enhancing the activities of critical antioxidant enzymes, such as catalase, glutathione S-transferase, and ascorbate peroxidase. Additionally, the elevated levels of total soluble sugars in SA-treated plants enhanced osmotic regulation under heat stress. Overall, our findings reveal that SA-triggered protective mechanisms not only preserve photosynthetic pigments but also ameliorate oxidative stress and boost plant resilience in the face of elevated temperatures. In conclusion, the application of 1 mM SA is highly effective in enhancing heat tolerance in cotton and is recommended for field trials before being commercially used to improve crop resilience under increasing global temperatures.

Ascorbic Acid Improves Tomato Salt Tolerance by Regulating Ion Homeostasis and Proline Synthesis.

In this study, processing tomato (Solanum lycopersicum L.) 'Ligeer 87-5' was hydroponically cultivated under 100 mM NaCl to simulate salt stress. To investigate the impacts on ion homeostasis, osmotic regulation, and redox status in tomato seedlings, different endogenous levels of ascorbic acid (AsA) were established through the foliar application of 0.5 mM AsA (NA treatment), 0.25 mM lycorine (LYC, an inhibitor of AsA synthesis; NL treatment), and a combination of LYC and AsA (NLA treatment). The results demonstrated that exogenous AsA significantly increased the activities and gene expressions of key enzymes (L-galactono-1,4-lactone dehydrogenase (GalLDH) and L-galactose dehydrogenase (GalDH)) involved in AsA synthesis in tomato seedling leaves under NaCl stress and NL treatment, thereby increasing cellular AsA content to maintain its redox status in a reduced state. Additionally, exogenous AsA regulated multiple ion transporters via the SOS pathway and increased the selective absorption of K+, Ca2+, and Mg2+ in the aerial parts, reconstructing ion homeostasis in cells, thereby alleviating ion imbalance caused by salt stress. Exogenous AsA also increased proline dehydrogenase (ProDH) activity and gene expression, while inhibiting the activity and transcription levels of Δ1-pyrroline-5-carboxylate synthetase (P5CS) and ornithine-δ-aminotransferase (OAT), thereby reducing excessive proline content in the leaves and alleviating osmotic stress. LYC exacerbated ion imbalance and osmotic stress caused by salt stress, which could be significantly reversed by AsA application. Therefore, exogenous AsA application increased endogenous AsA levels, reestablished ion homeostasis, maintained osmotic balance, effectively alleviated the inhibitory effect of salt stress on tomato seedling growth, and enhanced their salt tolerance.

Decoding Microbial Responses to Ammonia Shock Loads in Biogas Reactors through Metagenomics and Metatranscriptomics.

The presence of elevated ammonia levels is widely recognized as a significant contributor to process inhibition in biogas production, posing a common challenge for biogas plant operators. The present study employed a combination of biochemical, genome-centric metagenomic and metatranscriptomic data to investigate the response of the biogas microbiome to two shock loads induced by single pulses of elevated ammonia concentrations (i.e., 1.5 g NH4+/LR and 5 g NH4+/LR). The analysis revealed a microbial community of high complexity consisting of 364 Metagenome Assembled Genomes (MAGs). The hydrogenotrophic pathway was the primary route for methane production during the entire experiment, confirming its efficiency even at high ammonia concentrations. Additionally, metatranscriptomic analysis uncovered a metabolic shift in the methanogens Methanothrix sp. MA6 and Methanosarcina flavescens MX5, which switched their metabolism from the acetoclastic to the CO2 reduction route during the second shock. Furthermore, multiple genes associated with mechanisms for maintaining osmotic balance in the cell were upregulated, emphasizing the critical role of osmoprotection in the rapid response to the presence of ammonia. Finally, this study offers insights into the transcriptional response of an anaerobic digestion community, specifically focusing on the mechanisms involved in recovering from ammonia-induced stress.

The sweetpotato ß-amylase gene IbBAM1.1 enhances drought and salt stress resistance by regulating ROS homeostasis and osmotic balance.

Abiotic stressors, such as drought and high salinity, seriously affect plant growth, productivity, and quality. Maintaining reactive oxygen species (ROS) homeostasis and osmotic balance plays a crucial role in abiotic stress tolerance. ß-amylase (BAM) hydrolyzes α-1,4-glycosidic bonds by releasing maltose from starch in the regulation of soluble sugars. However, the function and mechanism of BAMs related to abiotic stress resistance remain unclear in sweetpotato (Ipomoea batatas (L.) Lam.). In this study, we isolated a novel ß-amylase gene IbBAM1.1, which was strongly induced by PEG6000, NaCl, and maltose treatments in sweetpotato variety Yanshu25. Overexpression of IbBAM1.1 conferred enhanced tolerance to the drought and high salinity stressors in Arabidopsis thaliana. The activity of ß-amylase and the degradation of starch were promoted under drought or salt stress. Accordingly, the contents of osmoprotectants, including maltose and proline were significantly higher in the transgenic lines than those in wild type (WT) plants. Less ROS, such as H2O2 and O2-, accumulated in the overexpressing lines than in WT plants. Superoxide dismutase activity was strongly enhanced and the level of malondialdehyde was lower under the drought or salt treatment in transgenic plants. Taken together, these results demonstrate that IbBAM1.1 acted as a positive regulator, at least in part, by regulating the level of osmoprotectants to balance the osmotic pressure and activate the scavenging system to maintain ROS homeostasis in the plants.

Differences in Ionic, Enzymatic, and Photosynthetic Features Characterize Distinct Salt Tolerance in Eucalyptus Species.

In the face of rising salinity along coastal regions and in irrigated areas, molecular breeding of tolerant crops and reforestation of exposed areas using tolerant woody species is a two-way strategy. Thus, identification of tolerant plants and of existing tolerance mechanisms are of immense value. In the present study, three Eucalyptus ecotypes with potentially differential salt sensitivity were compared. Soil-grown Eucalyptus plants were exposed to 80 and 170 mM NaCl for 30 days. Besides analysing salt effects on ionic/osmotic balance, and hydrolytic enzymes, plants were compared for dynamics of light-induced redox changes in photosynthetic electron transport chain (pETC) components, namely plastocyanin (PC), photosystem I (PSI) and ferredoxin (Fd), parallel to traditional chlorophyll a fluorescence-based PSII-related parameters. Deconvoluted signals for PC and Fd from PSI allowed identification of PC and PSI as the prime salinity-sensitive components of pETC in tested Eucalyptus species. Eucalyptus loxophleba portrayed efficient K+-Na+ balance (60-90% increased K+) along with a more dynamic range of redox changes for pETC components in old leaves. Young leaves in Eucalyptus loxophleba showed robust endomembrane homeostasis, as underlined by an increased response of hydrolytic enzymes at lower salt concentration (~1.7-2.6-fold increase). Findings are discussed in context of salinity dose dependence among different Eucalyptus species.

A decade of research on the second messenger c-di-AMP.

Cyclic dimeric adenosine 3',5'-monophosphate (c-di-AMP) is an emerging second messenger in bacteria and archaea that is synthesized from two molecules of ATP by diadenylate cyclases and degraded to pApA or two AMP molecules by c-di-AMP-specific phosphodiesterases. Through binding to specific protein- and riboswitch-type receptors, c-di-AMP regulates a wide variety of prokaryotic physiological functions, including maintaining the osmotic pressure, balancing central metabolism, monitoring DNA damage and controlling biofilm formation and sporulation. It mediates bacterial adaptation to a variety of environmental parameters and can also induce an immune response in host animal cells. In this review, we discuss the phylogenetic distribution of c-di-AMP-related enzymes and receptors and provide some insights into the various aspects of c-di-AMP signaling pathways based on more than a decade of research. We emphasize the key role of c-di-AMP in maintaining bacterial osmotic balance, especially in Gram-positive bacteria. In addition, we discuss the future direction and trends of c-di-AMP regulatory network, such as the likely existence of potential c-di-AMP transporter(s), the possibility of crosstalk between c-di-AMP signaling with other regulatory systems, and the effects of c-di-AMP compartmentalization. This review aims to cover the broad spectrum of research on the regulatory functions of c-di-AMP and c-di-AMP signaling pathways.

Osmotic adjustments support growth of poplar cultured cells under high concentrations of carbohydrates.

KEY MESSAGE: Poplar callus maintained a specific difference in osmotic potential with respect to media when supplemented with different carbohydrate concentrations. This balance in osmotic potential guaranteed the growth capacity. Osmotic stress is caused by several abiotic factors such as drought, salinity, or freezing. However, the threshold of osmotic potential that allows the growth under stress conditions has not been thoroughly studied. In this study, different levels of osmotic stress in Populus alba (L.) callus have been induced with the addition of mannitol or sorbitol in the medium (from 0 to 500 mM). The key factor for preserving the growth was observed to be the restoration of a constant difference in osmotic potential between callus and medium for all the tested conditions. The osmotic adjustments were primarily achieved with the uptake of mannitol or sorbitol from the media considering their chemical properties instead of their biological functions. The decrease in water content (from - 1 to - 10% after 21 days) and mineral elements, such as potassium, calcium, and magnesium, together with the alterations in cell morphology, did not show negative effects on growth. The activity of sorbitol dehydrogenase was detected for the first time in poplar (+ 4.7 U l-1 in callus treated with sorbitol compared to control callus). This finding suggested the importance of choosing carefully the molecules used to exert osmotic stress for separating the dual function of carbohydrates in osmotic adjustments and cell metabolism.

Effects of exercise on cations/anions in blood serum of English Thoroughbred horses / Efeito do exercício sobre cátions/ânions em soro sanguíneo de equinos da raça Puro-Sangue-Inglês

English Thoroughbred horses, are widespread in Mexico and due to the lack of data on their exercise physiology, it is important to conduct exercise tests in order to obtain information the effects of exercise on more essential cations/anions in blood serum, as these horses are submitted to constant efforts. The study was carried out with 150 blood samples of English Thoroughbred horses clinically healthy. The blood sample collection was performed during three periods: 1) rest, 2) 30min after exercise (speed race of 12km/h for 30min with no rest) and 3) 60min after exercise. Mean values were calculated for cations (sodium and potassium) and anions (chloride and bicarbonate). The resulting data set was analyzed using Gaussian distribution and descriptive statistics. Confidence intervals of 95% were established. The linear relationships between ions were quantified, and an analysis of variance was performed to compare the mean values between groups. The concentrations of the described analytes are consistent with values reported by international literature. The comparison between groups, revealed that during exercise, sodium ion did not show changes 30min after exercise and increase 60min after. Potassium ion showed increase 30min after exercise and decrease 60min after. Chloride ion showed a decrease 30min after exercise, to recover gradually 60min after. Meanwhile, bicarbonate ion showed increase 30min after exercise, decreasing slightly in the final stage. Negative correlation between bicarbonate ion and chloride ion were determined. It was concluded that exercise tests are useful for the determination of acid-base balance and osmotic balance, and their main role is to evaluate the athletic ability of horses.Considering that chloride ion excretion and metabolic adjustments of potassium ion and bicarbonate ion are superior to water loss, compared to the normal osmolarity of blood serum. The results found can be used to structure an adequate replacement program of electrolytes lost in sweat.(AU)

Equinos da raça Puro-Sangue-Inglês são difundidos no México e, devido à falta de dados sobre sua fisiologia do exercício, é importante fazer testes de exercício para obter informações sobre os efeitos do exercício em cátions/ânions mais essenciais no soro do sangue, pois esses equinos são submetidos a esforços constantes. O estudo foi realizado com 150 amostras de sangue de equinos Puro-Sangue-Inglês, clinicamente saudáveis. A coleta de sangue foi realizada em três períodos: 1) descanso, 2) 30min após o exercício (corrida de velocidade de 12km/h por 30min, sem descanso) e 3) 60min após o exercício. Os valores médios foram calculados para cátions (sódio e potássio) e ânions (cloreto e bicarbonato). O conjunto de dados resultante foi analisado utilizando-se distribuição gaussiana e estatística descritiva. Intervalos de confiança de 95% foram estabelecidos. As relações lineares entre os íons foram quantificadas, e uma análise de variância foi realizada para se compararem os valores médios entre grupos. As concentrações dos analitos descritos são consistentes com os valores relatados na literatura internacional. A comparação entre os grupos revelou que, durante o exercício, o sódio íon não mostrou alterações 30min após o exercício e aumentou 60min após. O potássio íon mostrou aumento 30min após o exercício e diminuiu 60min após. O cloreto íon mostrou uma diminuição 30min após o exercício, para recuperar gradualmente 60min depois. O bicarbonato íon mostrou aumento 30min após o exercício, diminuindo ligeiramente no estágio final. Correlação negativa entre bicarbonato íon e cloreto íon foi encontrada. Concluiu-se que os testes de exercício são úteis para a determinação do equilíbrio ácido-base e do equilíbrio osmótico, e seu principal papel é avaliar a capacidade atlética dos equinos. Considerando-se que a excreção de cloro íon e ajustes metabólicos de potássio íon e bicarbonato íon são superiores à perda de água, comparada à osmolaridade normal do soro sanguíneo,os resultados encontrados podem ser usados para estruturar um programa adequado de reposição de eletrólitos perdidos no suor.(AU)

Mitigation of Salinity Stress in Plants by Arbuscular Mycorrhizal Symbiosis: Current Understanding and New Challenges.

Modern agriculture is facing twin challenge of ensuring global food security and executing it in a sustainable manner. However, the rapidly expanding salinity stress in cultivable areas poses a major peril to crop yield. Among various biotechnological techniques being used to reduce the negative effects of salinity, the use of arbuscular mycorrhizal fungi (AMF) is considered to be an efficient approach for bio-amelioration of salinity stress. AMF deploy an array of biochemical and physiological mechanisms that act in a concerted manner to provide more salinity tolerance to the host plant. Some of the well-known mechanisms include improved nutrient uptake and maintenance of ionic homeostasis, superior water use efficiency and osmoprotection, enhanced photosynthetic efficiency, preservation of cell ultrastructure, and reinforced antioxidant metabolism. Molecular studies in past one decade have further elucidated the processes involved in amelioration of salt stress in mycorrhizal plants. The participating AMF induce expression of genes involved in Na+ extrusion to the soil solution, K+ acquisition (by phloem loading and unloading) and release into the xylem, therefore maintaining favorable Na+:K+ ratio. Colonization by AMF differentially affects expression of plasma membrane and tonoplast aquaporins (PIPs and TIPs), which consequently improves water status of the plant. Formation of AM (arbuscular mycorrhiza) surges the capacity of plant to mend photosystem-II (PSII) and boosts quantum efficiency of PSII under salt stress conditions by mounting the transcript levels of chloroplast genes encoding antenna proteins involved in transfer of excitation energy. Furthermore, AM-induced interplay of phytohormones, including strigolactones, abscisic acid, gibberellic acid, salicylic acid, and jasmonic acid have also been associated with the salt tolerance mechanism. This review comprehensively covers major research advances on physiological, biochemical, and molecular mechanisms implicated in AM-induced salt stress tolerance in plants. The review identifies the challenges involved in the application of AM in alleviation of salt stress in plants in order to improve crop productivity.

Responses of marine mussel Mytilus galloprovincialis (Bivalvia: Mytilidae) after infection with the pathogen Vibrio splendidus.

Bivalve molluscs possess effective cellular and humoral defence mechanisms against bacterial infection. Although the immune responses of mussels to challenge with pathogenic vibrios have been largely investigated, the effects at the site of injection at the tissue level have not been so far evaluated. To this aim, mussels Mytilus galloprovincialis were herein in vivo challenged with Vibrio splendidus to assess the responses induced in hemolymph and posterior adductor muscle (PAM), being the site of bacterial infection. The number of living intra-hemocyte bacteria increased after the first hour post-injection (p.i.), suggesting the occurrence of an intense phagocytosis, while clearance was observed within 24 h p.i. A recruitment of hemocytes at the injection site was found in mussel PAM, together with marked morphological changes in the volume of muscular fibers, with a recovery of muscle tissue organization after 48 h p.i. A concomitant impairment in the osmoregulatory processes were observed in PAM by an initial inhibition of aquaporins and increased immunopositivity of Na+/K+ ATPase ionic pump, strictly related to the histological alterations and hemocyte infiltration detected in PAM. Accordingly, an intense cell turnover activity was also recorded following the infection event. Overall, results indicated the hemolymph as the system responsible for the physiological adaptations in mussels to stressful factors, such as pathogenicity, for the maintenance of homeostasis and immune defence. Also, the osmotic balance and cell turnover can be used as objective diagnostic criteria to evaluate the physiological state of mussels following bacterial infection, which may be relevant in aquaculture and biomonitoring studies.

Trichoderma harzianum mitigates salt stress in cucumber via multiple responses.

Trichoderma harzianum T-soybean plays an important role in controlling soybean root rot disease. However, the mechanism by which it improves plant tolerance to salt stress is not clear. In this study, we investigated the possible mechanism of T-soybean in mitigating the damage caused by salt stress in Cucumis sativus L plants. Our results suggest that T-soybean improved salt tolerance of cucumber seedlings by affecting the antioxidant enzymes including peroxidase (POD) (EC 1.11.1.6), polyphenol oxidase (PPO) (EC 1.14.18.1), phenylalanine ammonia-lyase (PAL) (EC 4.3.1.5), catalase (CAT) (EC 1.11.1.6), superoxide dismutase (SOD) (EC 1.15.1.1), ascorbate peroxidase (APX) (EC 1.11.1.11), and glutathione reductase (GR) (EC 1.6.4.2), by increasing the levels of proline, soluble sugars, soluble protein, ascorbic acid (AsA) and chlorophyll as well as improving root activity. Treatment with T-soybean improved the ratio of glutathione (GSH)/oxidized glutathione (GSSG) and AsA/dehydroascorbate (DHA), and up-regulated the expression of CsAPX and CsGR genes involved in the AsA-GSH cycle. In addition, treatment with T-soybean increased the K+ content and K+/Na+ ratio while decreased the Na+ concentration and ethylene level. In summary, the improved salt tolerance of cucumber plants may be due to multiple mechanisms of T-soybean, such as the increase in reactive oxygen species (ROS) scavenging, as well as maintaining osmotic balance and metabolic homeostasis under salt stress.

Exogenous myo-inositol alleviates salinity-induced stress in Malus hupehensis Rehd.

Myo-inositol mediates various physiological processes and stress responses. Here, we investigated its role in Malus hupehensis Rehd. plants when grown hydroponically under saline conditions. Salt-stressed plants showed reduced growth and marked declines in photosynthetic activity and chlorophyll concentrations. However, pretreatment with 50 µM myo-inositol significantly alleviated those inhibitions and enabled plants to maintain their photosynthetic capacity. In addition to changing stomatal behavior, exogenous myo-inositol inhibited ROS accumulation and Na+ uptake. In contrast, activities of antioxidant systems were enhanced, and expression was elevated for genes involved in Na+ uptake (e.g., HKT1, NHX1, SOS1, and SOS2). This exogenous application also provoked the accumulation of sugars or sugar alcohols, which partially contributed to the maintenance of osmotic balance, and the scavenging of ROS, either directly or indirectly. In summary, myo-inositol appears to alleviate the salt-induced inhibition of physiological processes for M. hupehensis, not only by supporting the plant's antioxidant defense system but also by mediating Na+ and K+ homeostasis and the osmotic balance.

MicroRNA-132 controls water homeostasis through regulating MECP2-mediated vasopressin synthesis.

Fine-tuning of the body's water balance is regulated by vasopressin (AVP), which induces the expression and apical membrane insertion of aquaporin-2 water channels and subsequent water reabsorption in the kidney. Here we demonstrate that silencing of microRNA-132 (miR-132) in mice causes severe weight loss due to acute diuresis coinciding with increased plasma osmolality, reduced renal total and plasma membrane expression of aquaporin-2, and abrogated increase in AVP levels. Infusion with synthetic AVP fully reversed the antagomir-132-induced diuresis, and low-dose intracerebroventricular administration of antagomir-132 similarly caused acute diuresis. Central and intracerebroventricular antagomir-132 injection both decreased hypothalamic AVP mRNA levels. At the molecular level, antagomir-132 increased the in vivo and in vitro mRNA expression of methyl-CpG-binding protein-2 (MECP2), which is a miR-132 target and which blocks AVP gene expression by binding its enhancer region. In line with this, treatment of hypothalamic N6 cells with a high-salt solution increased its miR-132 levels, whereas it attenuated endogenous Mecp2 mRNA levels. In conclusion, we identified miR-132 as a first miRNA regulating the osmotic balance by regulating the hypothalamic AVP gene mRNA expression.

Hemolymph metabolites and osmolality are tightly linked to cold tolerance of Drosophila species: a comparative study.

Drosophila, like most insects, are susceptible to low temperatures, and will succumb to temperatures above the freezing point of their hemolymph. For these insects, cold exposure causes a loss of extracellular ion and water homeostasis, leading to chill injury and eventually death. Chill-tolerant species are characterized by lower hemolymph [Na(+)] than chill-susceptible species and this lowered hemolymph [Na(+)] is suggested to improve ion and water homeostasis during cold exposure. It has therefore also been hypothesized that hemolymph Na(+) is replaced by other 'cryoprotective' osmolytes in cold-tolerant species. Here, we compared the hemolymph metabolite profiles of five drosophilid species with marked differences in chill tolerance. All species were examined under 'normal' thermal conditions (i.e. 20°C) and following cold exposure (4 h at 0°C). Under benign conditions, total hemolymph osmolality was similar among all species despite chill-tolerant species having lower hemolymph [Na(+)]. Using NMR spectroscopy, we found that chill-tolerant species instead have higher levels of sugars and free amino acids in their hemolymph, including classical 'cryoprotectants' such as trehalose and proline. In addition, we found that chill-tolerant species maintain a relatively stable hemolymph osmolality and metabolite profile when exposed to cold stress while sensitive species suffer from large increases in osmolality and massive changes in their metabolic profiles during a cold stress. We suggest that the larger contribution of classical cryoprotectants in chill-tolerant Drosophila plays a non-colligative role for cold tolerance that contributes to osmotic and ion homeostasis during cold exposure and, in addition, we discuss how these comparative differences may represent an evolutionary pathway toward more extreme cold tolerance of insects.

Variations of natremia in sea kraits (Laticauda spp.) kept in seawater and fresh water.

Marine tetrapods evolved specific excretory structures (e.g. salt glands) that maintain salt concentrations within a narrow range of variation. However, recent investigations showed that in some lineages (sea snakes), individuals dehydrate in seawater and cannot equilibrate their hydromineral balance without access to fresh water. How these marine species cope with salt gain is therefore puzzling. We sampled two species of amphibious sea kraits (Laticauda saintgironsi and L. laticaudata) in the field. We also experimentally investigated patterns of salt regulation, specifically variations in natremia (plasma sodium) and body mass (net water flow), in individuals transferred first to fresh water and then to seawater. Our results show that free-ranging sea kraits display hypernatremia (up to 205mmol·l(-1)). Experimental data showed that natremia markedly decreased in snakes exposed to fresh water and increased when they were transferred to saltwater, thereby demonstrating a marked flexibility in their relation to environmental conditions. A literature survey indicated that all free-ranging marine snake species usually display hypernatremia despite having functional salt glands. Overall, sea snakes exhibit a marked tolerance to salt load compared to other marine tetrapods and apparently trigger substantial salt excretion only once natremia exceeds a high threshold. We hypothesise that this high tolerance significantly decreases energetic costs linked to salt gland functioning.