Alkaline Stress Research Articles

Co-application of organic amendments and natural biostimulants on plants enhances wheat production and defense system under salt-alkali stress

Soil alkalinity and salinity are major challenges to wheat production in arid regions. Eco-friendly amendments (organic matter and bio-stimulants) offer promising solutions, but their combined effects are underexplored. This study assessed the effects of organic amendments (vermicompost, compost, and chicken manure) combined with foliar bio-stimulants (licorice root, ginger rhizome, moringa leaf extract (MLE), and potassium humate) on wheat under salt and alkalinity stress. Organic amendments combined with bio-stimulants significantly improved wheat yields by enhancing chlorophyll content, proline levels, photosynthetic pigments, water uptake, and enzyme activities. Vermicompost outperformed compost and chicken manure in improving plant physico-biochemical properties. The combination of vermicompost and MLE was most effective in increasing plant height, leaf area, and photosynthetic rate by 97, 126, and 136%, respectively, while also enhancing catalase, peroxidase, and superoxide dismutase by 65, 97, and 185%, respectively. Consequently, this resulted in 64% increase in straw yield and 27% increase in grain yield compared to controls. Additionally, nutrient uptake (N, P, and K) significantly increased, while sodium uptake decreased. Integrating vermicompost with MLE can significantly enhance wheat productivity under abiotic stress, offering a sustainable solution to improve crop resilience in arid environments. Further research is required to understand the mechanisms and optimize bio-stimulant use in agriculture.

Exogenous Calcium Enhances Castor Tolerance to Saline–Alkaline Stress by Regulating Antioxidant Enzyme Activity and Activating Ca2+ and ROS Signaling Crosstalk

Saline–alkaline stress is a major factor limiting agricultural development, with calcium (Ca2+) playing a role in regulating plant tolerance through multiple signaling pathways. However, the specific mechanisms by which Ca2+ mediates saline–alkaline stress tolerance at the molecular level remain incompletely understood. This study investigates the effects of exogenous Ca2+ application on enhancing plant tolerance to saline–alkaline stress, focusing on its impact on the antioxidant system and Ca2+ and reactive oxygen species (ROS) signaling pathways. Through physiological assays and transcriptomic analyses, we evaluated oxidative damage markers, antioxidant enzyme activities, and the expression of key Ca2+ and ROS signaling genes. The results showed that saline–alkaline stress significantly elevated ROS levels, which led to increased membrane lipid peroxidation and induced upregulation of antioxidant response elements in castor roots. Exogenous calcium treatment reduced ROS accumulation by increasing superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) activities and decreasing malondialdehyde (MDA) levels, demonstrating a marked improvement in the antioxidant system. Transcriptomic analysis identified CAT2 (LOC107261240) as the primary target gene associated with increased CAT activity in response to exogenous calcium. Additionally, the upregulation of specific Ca2+ channels, Ca2+ sensors, ROS receptors, and antioxidant-related genes with calcium treatment highlights the critical role of Ca2+–ROS signaling crosstalk in enhancing stress tolerance. Protein–protein interaction analysis identified APX3 and other hub genes involved in Ca2+–ROS signaling transduction and the regulation of antioxidant activity. These findings enhance our understanding of calcium’s complex regulatory roles in plant abiotic stress responses, offering new theoretical insights for improving crop resilience in agriculture.

Ulva clathrata Enzymatic Hydrolysate Protects Wheat from Salt–Alkaline Stress by Regulating ABA Biosynthesis

Ulva clathrata Enzymatic Hydrolysate Protects Wheat from Salt–Alkaline Stress by Regulating ABA Biosynthesis

Integration of transcriptome, histopathology, and physiological indicators reveals regulatory mechanisms of largemouth bass (Micropterus salmoides) in response to carbonate alkalinity stress

The use of saline-alkali water resources is an important initiative for freshwater aquaculture. Largemouth bass (Micropterus salmoides) shows some salinity tolerance, but little is known about its alkali adaptation mechanism. First, we determined the 96-h 50 % lethal dose of alkaline carbonate (96-h LC50) for largemouth bass (41.41 mmol/L). Then, largemouth bass were subjected to 96-h carbonate alkalinity treatments and their responses were determined on the basis of histopathological, biochemical, and transcriptome analyses. The experiment included a control group (CA0, 0.49 mmol/L) and two treatment groups (CA20, 20 mmol/L; CA45, 45 mmol/L). The gill and liver tissues of the CA45 group exhibited severe deformation, including a thickened matrix, gill cell detachment, hepatocyte swelling, increased vacuolization, and localized necrotic lesions. In the CA20 group, some gill filaments were curled and deformed, hepatocytes had migrated, and tissue damage was minor. Both groups showed a lower percentage of apoptotic cells in gill and liver tissues. Serum antioxidant capacity first increased and then decreased, whereas energy metabolism and hormone indexes significantly increased under alkaline carbonate stress. Liver injury in the CA45 group resulted in significantly lower levels of cortisol, lactate dehydrogenase, and total protein compared with those in the CA20 group (P < 0.05). Transcriptome analyses of gill tissues from the CA45 group revealed significant alterations in pathways related to steroid biosynthesis (sqlea, lss, msmo1, tm7sf2), terpenoid backbone biosynthesis (hmgcs1, fdps, hmgcra, mvda), and ECM-receptor interaction and focal adhesion (col1a, col2a1b, col9a, col6a) due to alkalinity stress. These results indicate that liver damage under high alkalinity stress inhibits energy supply, while gill cell dysregulation triggers gill lesions.

Changes in Physiological Homeostasis in the Gills of Litopenaeus vannamei Under Carbonate Alkalinity Stress and Recovery Conditions

Carbonate alkalinity (CA) is the major toxic factor that interferes with the survival and growth of shrimp in saline–alkaline water. Gills are the main entry organ for CA toxicity in shrimp. In this study, low-salinity cultured Litopenaeus vannamei were exposed to 5 mmol/L CA stress for 7 days and then recovered for 7 days to explore the physiological changes in the gills under CA stress and recovery conditions at multiple biological levels. The results showed that CA stress increased the activities of antioxidative biochemical indexes (T-AOC, T-SOD, and POD) and the relative expression levels of romo1, nrf2, and gpx genes, while it decreased the relative expression levels of the sod and hsp70 genes. In addition, CA stress also increased the relative expression levels of genes involved in endoplasmic reticulum (ER) stress (bip, ire1, and xbp1), immunity (alf, crus, pen-3, lys, and propo), apoptosis (casp-3), detoxification metabolism (cyp450 and gst), and osmotic adjustment (ca, nka-α, nka-β, vatp, nhe, clc, aqp, tip4, and ccp). Although changes in some of the physiological indexes were reversed after the CA stress was relieved, they still could not effectively recover to the control level. These results reveal that CA stress has a negative impact on physiological homeostasis in the shrimp gills by inducing oxidation and ER stress and by interfering with immunity, apoptosis, detoxification, and osmotic adjustment.

Enhancing strawberry resilience to saline, alkaline, and combined stresses with light spectra: impacts on growth, enzymatic activity, nutrient uptake, and osmotic regulation

BackgroundThis study examines the effects of various complementary light spectra on the growth, development, antioxidant activity, and nutrient absorption in strawberry plants under stress conditions. Light-emitting diodes (LEDs) were used to provide specific wavelengths, including monochromatic blue (460 nm), monochromatic red (660 nm), a dichromatic mix of blue and red (1:3 ratio), full-spectrum white light (400–700 nm), and ambient light as a control (no LED treatment). The stress treatments applied were: control (no stress), salinity (80 mM NaCl), alkalinity (40 mM NaHCO₃), and a combined salinity/alkalinity condition.ResultsOur results indicated that complementary light spectra, especially red and blue/red, helped mitigate the adverse effects of stress on plant growth and development. These spectra improved plant tolerance by enhancing the activity of polyphenol oxidase and peroxidase enzymes and increasing starch accumulation in the leaves. Furthermore, under stress conditions, red and blue-red light significantly boosted fruit anthocyanin levels. Although stress elevated antioxidant activity, supplementary light reduced this activity by alleviating stress compared to ambient light. While stress led to increased Na and Cl ion concentrations in leaves, treatments with blue, red, and blue-red light minimized these harmful effects and promoted the absorption of beneficial ions such as K, Mg, Fe, and Cu.ConclusionsAdjusting light quality significantly influences the morphology and physiology of strawberry plants, underscoring the role of specific light spectra in promoting optimal growth under stress conditions.Clinical trial numberNot applicable

Identifying Calmodulin and Calmodulin-like Protein Members in Canavalia rosea and Exploring Their Potential Roles in Abiotic Stress Tolerance.

Calmodulins (CaMs) and calmodulin-like proteins (CMLs) belong to families of calcium-sensors that act as calcium ion (Ca2+) signal-decoding proteins and regulate downstream target proteins. As a tropical halophyte, Canavalia rosea shows great resistance to multiple abiotic stresses, including high salinity/alkalinity, extreme drought, heat, and intense sunlight. However, investigations of calcium ion signal transduction involved in the stress responses of C. rosea are limited. The CaM and CML gene families have been identified and characterized in many other plant species. Nevertheless, there is limited available information about these genes in C. rosea. In this study, a bioinformatic analysis, including the gene structures, conserved protein domains, phylogenetic relationships, chromosome distribution, and gene synteny, was comprehensively performed to identify and characterize CrCaMs and CrCMLs. A spatio-temporal expression assay in different organs and environmental conditions was then conducted using the RNA sequencing technique. Additionally, several CrCaM and CrCML members were then cloned and functionally characterized using the yeast heterogeneous expression system, and some of them were found to change the tolerance of yeast to heat, salt, alkalinity, and high osmotic stresses. The results of this study provide a foundation for understanding the possible roles of the CrCaM and CrCML genes, especially for halophyte C. rosea's natural ecological adaptability for its native habitats. This study also provides a theoretical basis for further study of the physiological and biochemical functions of plant CaMs and CMLs that are involved in tolerance to multiple abiotic stresses.

The Key Enzymes of Carbon Metabolism and the Glutathione Antioxidant System Protect Yarrowia lipolytica Yeast Against pH-Induced Stress.

In this study, we first thoroughly assayed the response of the key enzymes of energy metabolism and the antioxidant system in Yarrowia lipolytica yeast at extreme pH. The activity of the tricarboxylic acid cycle enzymes, namely NAD-dependent isocitrate dehydrogenase, aconitate hydratase, NAD-dependent malate dehydrogenase, and fumarate hydratase, NADPH-producing enzymes of glucose-6-P dehydrogenase and NADP-dependent isocitrate dehydrogenase, and the enzymes of the glutathione system was assessed. All the enzymes that were tested showed a significant induction contrary to some decrease in the aconitate hydratase activity with acidic and alkaline stress. It is probable that a change in the enzyme activity in the mitochondria matrix is involved in the regulation of the cellular metabolism of Y. lipolytica, which allows the species to prosper at an extreme ambient pH. It distinguishes it from any other type of ascomycete. A close relationship between the induction of the Krebs cycle enzymes and the key enzymes of the glutathione system accompanied by an increased level of reduced glutathione was shown. The assumption that the increased activity of the Krebs cycle dehydrogenases and promotion of the pentose phosphate pathway at pH stress launches a set of events determining the adaptive response of Y. lipolytica yeast.

Selection and immobilization of the optimal phytase-producing probiotic: a comparative in silico analyses of five autochthonous lactic acid bacteria

Phytic acid and its salts are the major storage form of both phosphate and myo-inositol, especially in cereals. Phytic acid chelates multivalent cations, which decreases the solubilized form of these nutrients and their dietary bioavailability for absorption and assimilation. Increasing micronutrient bioavailability from the economic phytate-rich food can be achieved by subjecting this food to fermentation by phytase-producing bacteria. A total of 8 lactic acid bacteria were isolated from different fermented food samples. Among the bacteria tested for their ability to produce phytase, only five isolates were selected as promising producers. The results revealed that isolate No. 4 produced the highest phytase levels of 411 U/mL. The five isolates were differentially able to tolerate acidic, alkaline, heat, surfactant, osmotic, bile, and pancreatic enzyme stresses, with the superiority of isolate No. 4, which demonstrated the most desirable probiotic potentials as confirmed by principal component analysis (PCA), heat map, and network analysis. Isolate No. 4 was identified by 16S rRNA sequencing and later confirmed as Pediococcus pentosaceus strain NMP4762Ch under the accession number MZ413646. Moreover, the active cells of P. pentosaceus were immobilized in alginate; the reusability of the immobilized active cells resulted in continuous production with an optimum phytase activity of 432.0 U/mL after 14 days, which decreased with time to reach 142.5 U/mL after 56 days. The maximum shelf stability was also observed, 557.5 U/mL, after 14 days and declined with time, reaching 133.5 U/mL by the end of the 56th day. Since humans don’t naturally produce phytase, utilizing probiotics for phytase production is important to combat mineral deficiencies.

The Identification of Proteomic Signatures Associated with Alkaline Tolerance in the Skin Mucus of Crucian Carp (Carassius auratus).

The skin is covered by a protective mucus layer, which is essential to the innate defense mechanism of fish. Investigating the response of skin mucus to various toxic stresses is crucial for enhancing its ability to tackle environmental challenges and developing strategies to mitigate toxic effects. Alkalinity stress assays (50 mmol/L NaHCO3) were conducted on crucian carp (Carassius auratus) from Lake Dali Nur (pH = 9.6) and Ping Xiang red crucian carp from freshwater (pH = 7) over 7 days. The expression of skin mucous proteins was analyzed using the liquid chromatography (LC)-spectrometry (MS)/MS Analysis-Data-independent acquisition (DIA) mode. A total of 12,537 proteins were identified across 20 samples from four groups, with 12,025 quantified. In the alkaline water population, high alkali stress resulted in the up-regulation of 139 proteins and the down-regulation of 500 proteins. In contrast, the freshwater population showed an increase in 112 proteins and a decrease in 120; both populations had a total of 23 genes up-regulated and 21 down-regulated. The protein regulatory network for the alkaline water group included 3146 pairwise interactions among 464 nodes, with only 20 being differentially expressed proteins. Conversely, the freshwater group's network comprised just 1027 specific interactions across 337 nodes, with 6 corresponding to differentially expressed proteins. A common protein regulatory network responding to high alkali stress was extracted and visualized for both populations. Based on their regulatory relationships and expression levels, these proteins are hypothesized to play similar roles under high alkali stress. Notably, the alpha-globin fragment and keratin type I cytoskeletal 13-like proteins showed markedly up-regulated expression, with the alpha-globin fragment increasing nearly a thousandfold from an extremely low level. This suggests it could serve as a potential biomarker for alkali tolerance, warranting further investigation.

Removal of nitrate nitrogen by aerobic denitrification granular sludge under strongly alkaline conditions: Performance and mechanism

Aerobic granular sludge (AGS) technology faces challenges in treating strongly alkaline wastewater because high pH affects sludge settleability and microbial metabolic activity. In this study, aerobic denitrification granular sludge (ADGS) was cultivated with strongly autoaggregating aerobic denitrifying bacteria as the sludge source at an influent pH of 11.0 under aerobic conditions. The formation characteristics of ADGS and its pollutant removal efficiency under strongly alkaline conditions were explored. Compared with ADGS cultivated under neutral conditions, ADGS cultivated under strongly alkaline conditions resulted in superior particle formation rates, settleability, particle strengths and particle stabilities. Nitrate nitrogen was almost completely removed at a pH of 11.0 under aerobic conditions. Protein, polysaccharides and humic acid in extracellular polymeric substances (EPS) under strongly alkaline conditions were involved in the ADGS formation process. In addition, the presence of metallic elements in the EPS and inorganic nuclei in the sludge facilitated the formation of ADGS. The strongly alkaline environment enriched the alkali-resistant aerobic denitrifying bacteria, such as unclassified_f__Rhodobacteraceae, Tessaracoccus, Halomonas, and Pseudofulvimonas, as well as specialized alkali-resistant genera. Furthermore, the strongly alkaline environment increased the abundance of key enzymes involved in carbon metabolism and upregulated the expression of nitrogen metabolism genes in the ADGS to maintain their metabolic activity. In addition, the microorganisms upregulated genes that encoded reverse transporter protein genes (mnhACEFG, nhaACP, and phaACDEFG) and K+ uptake protein genes (trkAHG) and secreted greater amounts of acidic functional groups and free amino groups to help maintain intracellular and extracellular osmolality and acid–base homeostasis under strongly alkaline stress. This research provides a possible option and innovative strategy for the management of highly alkaline nitrate-containing wastewater.

TaWRKY55-TaPLATZ2 module negatively regulate saline-alkali stress tolerance in wheat.

Saline-alkaline soils are a major environmental problem that limit plant growth and crop productivity. Plasma membrane H+-ATPases and the salt overly sensitive (SOS) signaling pathway play important roles in plant responses to saline-alkali stress. However, little is known about the functional genes and mechanisms regulating the transcription of H+-ATPases and SOS pathway genes under saline-alkali stress. In the present study, we identified that the plant AT-rich sequence and zinc-binding (TaPLATZ2) transcription factor are involved in wheat response to saline-alkali stress by directly suppressing the expression of TaHA2/TaSOS3. The knockdown of TaPLATZ2 enhances salt and alkali stress tolerance, while overexpression of TaPLATZ2 leads to salt and alkali stress sensitivity in wheat. In addition, TaWRKY55 directly upregulated the expression of TaPLATZ2 during saline-alkali stress. Through knockdown and overexpression of TaWRKY55 in wheat, TaWRKY55 was shown to negatively modulate salt and alkali stress tolerance. Genetic analyses confirmed that TaPLATZ2 functions downstream of TaWRKY55 in response to salt and alkaline stresses. These findings provide a TaWRKY55-TaPLATZ2-TaHA2/TaSOS3 regulatory module that regulates wheat responses to saline-alkali stress.

The response mechanism of alkalophilic Nitzschia sp. NW129 to low alkalinity-A study combining physiological and transcriptional analysis

Global aquatic acidification significantly threatens alkaline lake ecosystems. The mechanisms by which alkaliphilic microalgae, the key producers in these ecosystems, respond to reduced environmental alkalinity remain poorly understood. Here, we investigated the responses of alkalophilic Nitzschia sp. NW129 to low alkalinity (pH 9.2) through integrated physiological-biochemical and transcriptomic analyses. Relative to the control (pH 11.5), we observed a 60.1 % decrease in polysaccharide content, while total lipids and proteins increased by 1.74-fold and 2-fold, respectively. Transcriptome analysis revealed up-regulation of genes encoding carbonic anhydrase (CA) and malic enzyme (ME), along with those involved in glycolysis and fatty acid (FA) synthesis, compensating for carbon supply and shifting carbon flux from carbohydrate synthesis to lipid accumulation. Enhanced expression of TCA cycle genes and those encoding F-ATP synthase and inorganic pyrophosphatase (PPase) provided sufficient energy for cellular homeostasis, further facilitated by the up-regulated expression of ATP-dependent V-ATPase and ABC transporter genes. Temporal analysis revealed that the expression of genes involved in protein synthesis pathways was up-regulated on days 1 and 4 but notably down-regulated on day 2, suggesting protein degradation at this time to balance energy supply for adaptation. Despite these coping shifts, impairments in photosynthetic energy dissipation and electron transport, along with transcriptional changes including down-regulating cell cycle and inducing apoptotic pathways, ultimately caused a substantial reduction in biomass. These findings provide a basic understanding of the response mechanisms of alkalophilic microalgae to low alkalinity stress, which should aid to develop strategies to improve microalgal tolerance against acidification.

Proline metabolism is essential for alkaline adaptation of Nile tilapia (Oreochromis niloticus)

BackgroundSaline-alkaline water aquaculture has become a key way to mitigate the reduction of freshwater aquaculture space and meet the increasing global demand for aquatic products. To enhance the comprehensive utilization capability of saline-alkaline water, it is necessary to understand the regulatory mechanisms of aquatic animals coping with saline-alkaline water. In this study, our objective was to elucidate the function of proline metabolism in the alkaline adaptation of Nile tilapia (Oreochromis niloticus).ResultsExpose Nile tilapia to alkaline water of different alkalinity for 2 weeks to observe changes in its growth performance and proline metabolism. Meanwhile, to further clarify the role of proline metabolism, RNA interference experiments were conducted to disrupt the normal operation of proline metabolic axis by knocking down pycr (pyrroline-5-carboxylate reductases), the final rate-limiting enzyme in proline synthesis. The results showed that both the synthesis and degradation of proline were enhanced under carbonate alkalinity stress, and the environmental alkalinity impaired the growth performance of tilapia, and the higher the alkalinity, the greater the impairment. Moreover, environmental alkalinity caused oxidative stress in tilapia, enhanced ion transport, ammonia metabolism, and altered the intensity and form of energy metabolism in tilapia. When the expression level of the pycr gene decreased, the proline metabolism could not operate normally, and the ion transport, antioxidant defense system, and energy metabolism were severely damaged, ultimately leading to liver damage and a decreased survival rate of tilapia under alkalinity stress.ConclusionsThe results indicated that proline metabolism plays an important role in the alkaline adaptation of Nile tilapia and is a key regulatory process in various biochemical and physiological processes.

Biochemical and Transcriptomic Analyses Reveal Key Salinity and Alkalinity Stress Response and Tolerance Pathways in Salix linearistipularis Inoculated with Trichoderma

Soil salinization and alkalinization are pervasive environmental issues that severely restrict plant growth and crop yield. Utilizing plant growth-promoting rhizobacteria (PGPR) is an effective strategy to enhance plant tolerance to saline–alkaline stress, though the regulatory mechanisms remain unclear. This study employed biochemical and RNA-Seq methods to uncover the critical growth-promoting effects of Trichoderma spp. on Salix linearistipularis under saline–alkaline stress. The results showed that, during saline–alkaline stress, inoculation with Trichoderma sp. M4 and M5 significantly increased the proline and soluble sugar contents in Salix linearistipularis, enhanced the activities of SOD, POD, CAT, and APX, and reduced lipid peroxidation levels, with M4 exhibiting more pronounced effects than M5. RNA-Seq analysis of revealed that 11,051 genes were upregulated after Trichoderma sp. M4 inoculation under stress conditions, with 3532 genes primarily involved in carbon metabolism, amino acid biosynthesis, and oxidative phosphorylation—processes that alleviate saline–alkaline stress. Additionally, 7519 genes were uniquely upregulated by M4 under stress, mainly enriched in secondary metabolite biosynthesis, amino acid metabolism, cyanamide metabolism, and phenylpropanoid biosynthesis. M4 mitigates saline–alkaline stress-induced damage in Salix linearistipularis seedlings by reducing oxidative damage, enhancing organic acid and amino acid metabolism, and activating phenylpropanoid biosynthesis pathways to eliminate harmful ROS. This enhances the seedlings’ tolerance to saline–alkaline stress, providing a basis for studying fungi–plant interactions under such conditions.

Maize Endophytic Plant Growth-Promoting Bacteria Peribacillus simplex Can Alleviate Plant Saline and Alkaline Stress.

Soil salinization is currently one of the main abiotic stresses that restrict plant growth. Plant endophytic bacteria can alleviate abiotic stress. The aim of the current study was to isolate, characterize, and assess the plant growth-promoting and saline and alkaline stress-alleviating traits of Peribacillus simplex M1 (P. simplex M1) isolates from maize. One endophytic bacterial isolate, named P. simplex M1, was selected from the roots of maize grown in saline-alkali soil. The P. simplex M1 genome sequence analysis of the bacteria with a length of 5.8 Mbp includes about 700 genes that promote growth and 16 antioxidant activity genes that alleviate saline and alkaline stress. P. simplex M1 can grow below 400 mM NaHCO3 on the LB culture medium; The isolate displayed multiple plant growth-stimulating features, such as nitrogen fixation, produced indole-3-acetic acid (IAA), and siderophore production. This isolate had a positive effect on the resistance to salt of maize in addition to the growth. P. simplex M1 significantly promoted seed germination by enhancing seed vigor in maize whether under normal growth or NaHCO3 stress conditions. The seeds with NaHCO3 treatment exhibited higher reactive oxygen species (ROS) levels than the maize in P. simplex M1 inoculant on maize. P. simplex M1 can colonize the roots of maize. The P. simplex M1 inoculant plant increased chlorophyll in leaves, stimulated root and leaf growth, increased the number of lateral roots and root dry weight, increased the length and width of the blades, and dry weight of the blades. The application of inoculants can significantly reduce the content of malondialdehyde (MDA) and increase the activity of plant antioxidant enzymes (Catalase (CAT), Superoxide Dismutase (SOD), and Peroxidase (POD)), which may thereby improve maize resistance to saline and alkaline stress. Conclusion: P. simplex M1 isolate belongs to plant growth-promoting bacteria by having high nitrogen concentration, indoleacetic acid (IAA), and siderophore, and reducing the content of ROS through the antioxidant system to alleviate salt alkali stress. This study presents the potential application of P. simplex M1 as a biological inoculant to promote plant growth and mitigate the saline and alkaline effects of maize and other crops.

Effects of saline-alkali stress on cotton growth and physiochemical expression with cascading effects on aphid abundance.

Environmental stresses, such as soil salinity or alkalinity, usually affect crop growth and secondary plant metabolism, with follow on effects on foliar-feeding insects. Nevertheless, the underlying mechanism of how saline-alkali stress affects the key cotton pest Aphis gossypii Glover is poorly understood. In this study, we first considered effects of three types of saline-alkali stress (i.e., salinity alone, alkalinity alone - both at different concentration - and their mixed effects) on cotton plants. We then measured impacts of stress on (1) above and below plant growth traits (e.g., plant height, leaf area, root volume), (2) levels of nutrients and secondary metabolites in cotton leaves, and (3) feeding behavior, life-table parameters, and population growth of A. gossypii. We then used a path analysis to evaluate cascading effects of changes in plant growth (due to stress) and changes in levels of nutrients or secondary metabolites on growth of individual cotton aphids and aphid populations. We found either salinity or alkalinity stresses significantly reduced cotton growth, increased the content of tannin, soluble sugars, and proline in the leaves, and suppressed aphid growth and development, (including longevity, fecundity, and intrinsic rate of increase) and aphid population growth. Alkalinity had stronger effects on these traits than did salinity. This work provides insights into the bottom-up interaction mechanism by which these environmental stresses mediate aphid infestation levels in the cotton agricultural ecosystem.

Physiological and molecular responses of juvenile silver crucian carp (Carassius gibelio) to long-term high alkaline stress: Growth performance, histopathology, and transcriptomic analysis

Salty-alkaline waters are widely distributed globally, and how to effectively develop saline-alkaline water to improve the utilization rate of water resources has become a global challenge. This study examined the survival performance of juvenile silver crucian carp (Carassius gibelio) under alkaline gradient acclimation conditions and explored the regulatory mechanisms for their adaptation to high alkalinity. Compared to directly immersing in high concentrations, alkaline gradient acclimation significantly improved the survival rate of juvenile silver crucian carp. Long-term high alkaline stress caused significant reduction in the growth performance of juvenile silver crucian carp. After alkaline stress, the enzyme activities of Na+-K+-ATP (NKA) and Ca2+-Mg2+-ATP (CMA) in the gills were significantly decreased. Meanwhile, histopathology of the gills showed structural changes, indicating that the physiological functions of juvenile silver crucian carp were impaired under alkaline conditions. Transcriptomic analysis found that the expression of genes related to metabolism and immune response pathways in the gill and kidney underwent significant changes. Genes related to carbohydrate metabolism were upregulated while those related to protein metabolism were downregulated, indicating that alkaline stress caused the organism to utilize carbohydrates rather than proteins as much as possible to meet enhanced metabolism. The HIF pathway that maintains the body's survival was significantly up-regulated, at the expense of high energy consumption. Dysregulation of genes related to the formation of neutrophil extracellular traps (NETosis) pathway indicated that the immune function of the fish was suppressed. Overall, this study revealed the physiological and molecular responses of juvenile silver crucian carp to alkaline stress, deepening our understanding of their adaptation strategies and the potential impact of alkaline environments on aquatic life.

Development and validation of a stability-indicating HPLC method for assay of tonabersat in pharmaceutical formulations

A stability-indicating reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed to assay tonabersat and assess its stability in pharmaceutical formulations. Chromatographic separation was achieved using a Kinetex® C18 column (2.6 µm, 150 x 3 mm, 100 Å) at 50 °C, with a 20 µL injection volume. A linear gradient of acetonitrile in water (5 – 33.5 %) was applied for 1 min, followed by a gradual increase to 100 % over 26 min at a flow rate of 0.5 mL/min. Tonabersat and its degradation products were detected at 275 nm and 210 nm, respectively. The optimized method was used to evaluate the stability of tonabersat in lipid-based pharmaceutical formulations at 5 ± 3 °C, 25 ± 2°C/60 ± 5 % RH, and 40 ± 2 °C/75 ± 5 % RH over 3 months. The method was validated as per ICH guidelines and demonstrated linearity in the range of 5 – 200 µg/mL (R2 = 0.99994) with good accuracy (98.25 – 101.58 % recovery) and precision (% RSD < 2.5 %). The limits of detection and quantitation were 0.8 µg/mL and 5 µg/mL, respectively. Forced degradation studies showed significant degradation on exposure to alkaline (90.33 ± 0.80 %), acidic (70.60 ± 1.57 %), and oxidative stress (33.95 ± 0.69 %) at 70 °C, but no degradation was observed on exposure to thermal or photolytic stress. No chemical degradation was observed in either formulation on storage. Thus, the method was sensitive, specific, and suitable for stability testing of tonabersat in pharmaceutical formulations.

Promoting effects of bioplastics and sludge anaerobic co-fermentation for carboxylates production with pH regulation: Insights into the plastic structure, microbial metabolic traits, and adaptive mechanism

Biodegradable plastics (BPs) are presenting new challenges for their reutilization. This work found that volatile fatty acids (VFAs) production by co-fermentation of BPs with waste activated sludge (WAS) reached 4–37 times of the WAS fermentation alone, which was further amplified by pH regulation (especially alkaline regulation). Moreover, the VFAs composition is highly associated with BPs category. By contrast, the traditional plastic showed a limited effect on the VFAs yield and composition. Alkaline regulation enhanced the breakdown of BPs’ ester bonds and boosted WAS disintegration, increasing bioavailable substrates. The hydrolytic-acidogenic anaerobes (i.e., Serpentinicella and Proteiniclasticum) and the major metabolic processes participated in the transformation of BPs and WAS to VFAs were upregulated under alkaline conditions. Further exploration unveiled that quorum sensing and peptidoglycan synthesis played important roles in counteracting alkaline stress and maintaining microbial activity for effective VFAs generation. The works demonstrated the effectiveness of pH-regulated anaerobic co-fermentation for BPs valorization.