Saline-alkaline stress is a common problem in Akebia trifoliata cultivation. In this study, the enhancing effects of 5-azacytidine (5-AzaC) on the resistance of A. trifoliata to saline-alkaline stress and the underlying mechanisms were investigated. Plant height, stem diameter, biomass, root length, fresh weight of root, and root/shoot ratio of 6-month-old A. trifoliata seedlings were measured after saline-alkaline stress with or without 5-AzaC treatment. Moreover, the contents of photosynthetic pigments, malondialdehyde (MDA), H2O2, sodium, soluble sugar, and proline; activities of superoxide dismutase, peroxidase (POD), and catalase (CAT); and anatomical structures of root, stem, and leaf were assessed. Furthermore, comparative transcriptome sequencing was performed. The results demonstrated that growth and development of A. trifoliata were severely inhibited under saline-alkaline stress, suggesting that the seedlings were exposed to severe oxidative and osmotic stresses. Treatment with exogenous 5-AzaC could significantly relieve the symptoms of saline-alkaline stress in A. trifoliata. Under saline-alkaline stress, 5-AzaC could increase the stem diameter, biomass, root length, fresh weight of root, and root/shoot ratio and minimize damages to the anatomical structure. Moreover, absorption of Na+ was reduced; ionic balance was maintained; POD and CAT activities were significantly improved; proline and soluble sugar contents increased, and H2O2 and MDA contents decreased. Transcriptome analysis revealed that 5-AzaC functioned via regulating KEGG pathways such as plant hormone signal transduction, phenylpropanoid biosynthesis, photosynthesis, amino sugar and nucleotide sugar metabolism, and glutathione metabolism under saline-alkaline stress. Particularly, enhanced expression of genes from the auxin pathway in plant hormone signal transduction; the lignin synthetic pathway in phenylpropanoid biosynthesis; and photosystem II, photosystem I, photosynthetic electron transport, and F-type ATP pathway in photosynthesis may be related to 5-AzaC-induced saline-alkaline resistance. The results provided theoretical references for A. trifoliata cultivation in saline-alkaline soil and application of 5-AzaC to improve saline-alkaline tolerance in plants.

Under saline-alkali stress conditions, inoculation with Rhizophagus irregularis or the application of biochar can both promote plant growth and improve soil physicochemical properties. However, the effects of their combined use on switchgrass growth and soil mechanical properties remain unclear. This study established four treatments: no Ri inoculation and no biochar addition (control, CK), biochar addition alone (BC), Rhizophagus irregularis inoculation alone (Ri), and their combination (RB). The aim was to investigate the effects of these treatments on the biomass, root morphology, and soil mechanical properties of switchgrass under saline-alkali stress. The results showed that compared to the CK treatment, the RB treatment significantly increased the root, stem, leaf, and total biomass of switchgrass by 67.55%, 74.76%, 117.31%, and 82.93%, respectively. Among all treatment groups, RB treatment significantly reduced soil bulk density, soil water-soluble sodium ions (Na+), soil exchangeable sodium percentage (ESP), and sodium adsorption ratio (SAR), while increasing soil porosity. Furthermore, RB treatment significantly improved infiltration rate and shear strength. Compared to the CK treatment, the stable infiltration rate and shear strength under 400 kPa vertical load increased by 70.69% and 22.5 kPa, respectively. In conclusion, the combination of Ri and biochar has the potential to improve soil mechanical properties and increase the biomass of switchgrass under saline-alkali stress.

Cucumbers, cultivated globally on 3.7 million hectares, face yield losses due to salinity, highlighting the need for effective mitigation strategies for degraded soils. Melatonin (MT) has gained significant interest for its ability to relieve plant stress. To explore the regulatory role of exogenous MT in maintaining redox homeostasis in cucumber seedlings under saline-alkali stress (SA), this study employed the cucumber cultivar 'Xinchun No. 4 '. Simulated saline-alkali conditions were applied, and the effects of exogenous MT on seedling growth, reactive oxygen species (ROS) production, the ascorbate-glutathione (AsA-GSH) cycle, and changes in leaf anatomy were systematically assessed. The findings reveal that exposure to 40 mmol center dot L-1 saline-alkali stress significantly impaired cucumber seedling growth, reduced biomass, and led to excessive accumulation of hydrogen peroxide (H2O2) and superoxide anions (O2 center dot ) in the leaves. This, resulted in increased lipid peroxidation (indicated by elevated malondialdehyde (MDA) levels), whichi further compromised the cell membrane. Application of 10 mu mol center dot L-1 MT effectively reduced ROS levels, lowered MDA content, and mitigated electrolyte leakage. MT also enhanced AsA and GSH levels, improved AsA/DHA and GSH/GSSG ratios, and upregulated key AsA-GSH cycle genes (CsAPX, CsAAO, CsMDAR, CsDHAR, CsGR), leading to a significant increase in enzymatic activity. In addition, MT alleviated stress-induced stomatal closure, thereby restoring normal stomatal function. These findings suggest that MT enhances saline-alkali tolerance by mitigating oxidative damage, promoting antioxidant defenses, and effectively preserving stomatal function. Thus, our study points to a sustainable strategy to improve crop resilience in salinized environments via MT application.

The challenge of soil salinization and alkalization, with its significant impact on crop productivity, has raised growing concerns with global population growth and enhanced environmental degradation. Although arbuscular mycorrhizal fungi (AMF) and calcium ions (Ca2+) are known to enhance plant resistance to stress, their combined effects on perennial ryegrass' tolerance to salt and alkali stress and the underlying mechanisms remain poorly understood. This study aimed to elucidate the roles of Arbuscular mycorrhizal (AM) fungus Rhizophagus irregularis and exogenous Ca2+ application in molecular and physiological responses to salt-alkali stress. AM symbiosis and exogenous Ca2+ application enhanced antioxidant enzyme activity and non-enzymatic components, promoting reactive oxygen species (ROS) scavenging and reducing lipid peroxidation while alleviating oxidative damage induced by salt-alkali stress. Furthermore, they enhanced osmotic balance by increasing soluble sugar content (Proportion of contribution of the osmotic adjustment were 34 similar to 38 % in shoots and 30 similar to 37 % in roots) under salt stress and organic acid content (Proportion of contribution of the osmotic adjustment were 32 similar to 36 % in shoots and 37 similar to 42 % in roots) under alkali stress. Changes in organic solute and inorganic cation-anion contents contributed to ion balance, while hormonal regulation played a role in these protective mechanisms. Moreover, the protective mechanisms involved activation of Ca2+-mediated-mediated signaling pathways, regulation of salt-alkali stress-related genes (including LpNHX1 and LpSOS1), increased ATPase activity, elevated ATP levels, enhanced Na+ extrusion, improved K+ absorption capacity, and a reduced Na+/K+ ratio, all contributing to the protection of photosynthetic pigments and the enhancement of photosynthetic efficiency. Ultimately, the combined application of exogenous Ca2+ and AMF synergistically alleviated the inhibitory effects of salt-alkali stress on perennial ryegrass growth. This finding suggested that exogenous Ca2+ may participate in the colonization of perennial ryegrass plants by R. irregularis, while AM symbiosis may activate Ca2+ pathways. Consequently, the combined treatment of AM and Ca2+ is beneficial for enhancing plant regulatory mechanisms and increasing crop yield under salt-alkali stress.

Soil salinization negatively affects plant growth and threatens food security. Halotolerant plant growth-promoting bacteria (PGPB) can alleviate salt stress in plants via diverse mechanisms. In the present study, we isolated salt-tolerant bacteria with phosphate-solubilizing abilities from the rhizosphere of Salix linearistipularis, a halophyte distributed in saline-alkali soils. Strain A103 showed high phosphate solubilization activity and was identified as Enterobacter asburiae based on genome analysis. In addition, it can produce indole-3-acetic acid (IAA), siderophores, and 1-aminocyclopropane-1-carboxylate (ACC) deaminase. Genome mining has also revealed the presence of several functional genes involved in the promotion of plant growth. Inoculation with A103 markedly improved alfalfa growth in the presence of 100 mM NaHCO3. Under alkali stress, the shoot and root dry weights after bacterial inoculation improved by 42.9 % and 21.9 %, respectively. Meanwhile, there was a 35.9-37.1 % increase in the shoot and root lengths after treatment with A103 compared to the NaHCO3-treated group. Soluble sugar content, peroxidase and catalase activities increased in A103-inoculated alfalfa under alkaline stress. A significant decrease in the malondialdehyde content was observed after treatment with strain A103. Metabolomic analysis indicated that strain A103 positively regulated alkali tolerance in alfalfa through the accumulation of metabolites, such as homocarnosine, panthenol, and sorbitol, which could reduce oxidative damage and act as osmolytes. These results suggest that halophytes are valuable resources for bioprospecting halotolerant beneficial bacteria and that the application of halotolerant growth-promoting bacteria is a natural and efficient strategy for developing sustainable agriculture.

Key messageA plant growth regulator, 5-aminolevulinic acid, enhanced the saline-alkali tolerance via photosynthetic, oxidative-reduction, and glutathione metabolism pathways in pepper seedlings.AbstractSaline-alkali stress is a prominent environmental problem, hindering growth and development of pepper. 5-Aminolevulinic acid (ALA) application effectively improves plant growth status under various abiotic stresses. Here, we evaluated morphological, physiological, and transcriptomic differences in saline-alkali-stressed pepper seedlings after ALA application to explore the impact of ALA on saline-alkali stress. The results indicated that saline-alkali stress inhibited plant growth, decreased biomass and photosynthesis, altered the osmolyte content and antioxidant system, and increased reactive oxygen species (ROS) accumulation and proline content in pepper seedlings. Conversely, the application of exogenous ALA alleviated this damage by increasing the photosynthetic rate, osmolyte content, antioxidant enzyme activity, and antioxidants, including superoxide dismutase, catalase, glutathione reductase, and glutathione peroxidase, and reducing glutathione to reduce ROS accumulation and malonaldehyde content. Moreover, the transcriptomic analysis revealed the differentially expressed genes were mainly associated with photosynthesis, oxidation-reduction process, and glutathione metabolism in saline-alkali stress + ALA treatment compared to saline-alkali treatment. Among them, the change in expression level in CaGST, CaGR, and CaGPX was close to the variation of corresponding enzyme activity. Collectively, our findings revealed the alleviating effect of ALA on saline-alkali stress in pepper seedlings, broadening the application of ALA and providing a feasible strategy for utilize saline-alkali soil.

Background and Aims Soil salinization adversely threatens plant survival and food production globally. The mobilization of storage reserves in cotyledons and establishment of the hypocotyl/root axis (HRA) structure and function are crucial to the growth of dicotyledonous plants during the post-germination growth period. Here we report the adaptive mechanisms of wild and cultivated soybeans in response to alkali stress in soil during the post-germination growth period.Methods Differences in physiological parameters, microstructure, and the types, amounts and metabolic pathways of small-molecule metabolites and gene expression were compared and multi-omics integration analysis was performed between wild and cultivated soybean under sufficient and artificially simulated alkali stress during the post-germination growth period in this study.Key Results Structural analysis showed that the cell wall thickness of wild soybean under alkali stress increased, whereas cultivated soybeans were severely damaged. A comprehensive analysis of small-molecule metabolites and gene expression revealed that protein breakdown in wild soybean cotyledons under alkali stress was enhanced, and transport of amino acids and sucrose increased. Additionally, lignin and cellulose syntheses in wild soybean HRA under alkali stress were enhanced.Conclusions Overall, protein decomposition and transport of amino acids and sucrose increased in wild soybean cotyledons under alkali stress, which in turn promoted HRA growth. Similarly, alkali stress enhanced lignin and cellulose synthesis in the wild soybean HRA, which subsequently enhanced cell wall synthesis, thereby maintaining the stability and functionality of the HRA under alkali stress. This study presents important practical implications for the utilization of wild plant resources and sustainable development of agriculture.

Soil salinization is one of the major environmental factors that restrict plant growth and development. Zeaxanthin epoxidase (ZEP) functions in ABA biosynthesis and the xanthophyll cycle and has a vital role in plant responses to various environmental stresses. It was found by quantitative real-time PCR (qRT-PCR) that MhZEP responded to saline-alkali stress and showed the highest expression at 48 h of saline-alkali stress, which was 14.53-fold of 0 h. The MhZEP gene was cloned from the apple rootstock begonia (Malus halliana Koehne) and its protein physicochemical properties were analyzed. Subsequently, the functional characterization of MhZEP (ID: 103403091) was further investigated in Arabidopsis thaliana. The MhZEP contained a complete open reading frame with a length of 1998 bp, and encoded 665 amino acids with an isoelectric point of 7.18. Phylogenetic tree analysis showed that MhZEP was the most homologous and closely related to Glycine max. Compared with wild-type, transgenic plants grew better under saline-alkali stress and the MhZEP-OE line showed higher chlorophyll content, carotenoid content, enzyme activities (POD, SOD, CAT and APX) and K+ content, whereas they had lower chlorosis and Na(+ )content than the wild type (WT), which indicated that they had strong resistance to stress. The expression levels of saline-alkali stress-related genes in A. thaliana MhZEP-OE were examined by qRT-PCR, and it was found that the MhZEP improved the tolerance of A. thaliana to saline-alkali stress tolerance by regulating the expression of carotenoid synthesis genes (MhPSY, MhZDS, MhLYCB and MhVDE) and ABA biosynthesis genes (MhNCED5, MhABI1 and MhCYP707A2). And the potassium-sodium ratio in the cytoplasm was increased to maintain ionic homeostasis by modulating the expression of Na+ transporter genes (MhCHX15 and MhSOS1) and K+ transporter genes (MhHKT1;1, MhNHX1 and MhSKOR1). Moreover, the expression of H+-ATPase genes (MhAHA2 and MhAHA8) was increased to reduce the oxidative damage caused by saline-alkali stress. In summary, MhZEP acted as an essential role in plant resistance to saline-alkali stress, which lays the foundation for further studies on its function in apple.

Key messageSaline-alkali stress induces oxidative damage and photosynthesis inhibition in H. citrina, with a significant downregulation of the expression of photosynthesis- and antioxidant-related genes at high concentration. Soil salinization is a severe abiotic stress that impacts the growth and development of plants. In this study, Hemerocallis citrina Baroni was used to investigate its responsive mechanism to complex saline-alkali stress (NaCl:Na2SO4:NaHCO3:Na2CO3 = 1:9:9:1) for the first time. The growth phenotype, photoprotective mechanism, and antioxidant system of H. citrina were studied combining physiological and transcriptomic techniques. KEGG enrichment and GO analyses revealed significant enrichments of genes related to photosynthesis, chlorophyll degradation and antioxidant enzyme activities, respectively. Moreover, weighted gene co-expression network analysis (WGCNA) found that saline-alkali stress remarkably affected the photosynthetic characteristics and antioxidant system. A total of 29 key genes related to photosynthesis and 29 key genes related to antioxidant enzymes were discovered. High-concentration (250 mmol L-1) stress notably inhibited the expression levels of genes related to light-harvesting complex proteins, photosystem reaction center activity, electron transfer, chlorophyll synthesis, and Calvin cycle in H. citrina leaves. However, most of them were insignificantly changed under low-concentration (100 mmol L-1) stress. In addition, H. citrina leaves under saline-alkali stress exhibited yellow-brown necrotic spots, increased cell membrane permeability and accumulation of reactive oxygen species (ROS) as well as osmolytes. Under 100 mmol L-1 stress, ROS was eliminate by enhancing the activities of antioxidant enzymes. Nevertheless, 250 mmol L-1 stress down-regulated the expression levels of genes encoding antioxidant enzymes, and key enzymes in ascorbate-glutathione (AsA-GSH) cycle as well as thioredoxin-peroxiredoxin (Trx-Prx) pathway, thus inhibiting the activities of these enzymes. In conclusion, 250 mmol L-1 saline-alkali stress caused severe damage to H. citrina mainly by inhibiting photosynthesis and ROS scavenging capacity.

The increasing global phenomenon of soil salinization has prompted heightened interest in the physiological ecology of plant salt and alkali tolerance. Halostachys caspica belonging to Amaranthaceae, an exceptionally salt-tolerant halophyte, is widely distributed in the arid and saline-alkali regions of Xinjiang, in Northwest China. Soil salinization and alkalinization frequently co-occur in nature, but very few studies focus on the interactive effects of various salt and alkali stress on plants. In this study, the impacts on the H. caspica seed germination, germination recovery and seedling growth were investigated under the salt and alkali stress. The results showed that the seed germination percentage was not significantly reduced at low salinity at pH 5.30-9.60, but decreased with elevated salt concentration and pH. Immediately after, salt was removed, ungerminated seeds under high salt concentration treatment exhibited a higher recovery germination percentage, indicating seed germination of H. caspica was inhibited under the condition of high salt-alkali stress. Stepwise regression analysis indicated that, at the same salt concentrations, alkaline salts exerted a more severe inhibition on seed germination, compared to neutral salts. The detrimental effects of salinity or high pH alone were less serious than their combination. Salt concentration, pH value, and their interactions had inhibitory effects on seed germination, with salinity being the decisive factor, while pH played a secondary role in salt-alkali mixed stress.