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CaMYB121-CaABF2 negative feedback loop modulates CaNHX2 expression to confer salt tolerance in pepper.

Salt stress is a major abiotic factor that severely restricts pepper (Capsicum annuum) production. Although abscisic acid (ABA) is vital for salt tolerance, the transcriptional regulatory networks governing ABA-mediated salt defense remain largely unknown. Here, we uncovered a negative feedback loop between CaMYB121 and CaABF2.1/2 that modulates the expression of CaNHX2.1/2/3, thereby enhancing salt tolerance in pepper plants. RNA-seq analysis revealed that CaMYB121 displayed an expression pattern consistent with that of CaNHX2 after salt treatment. Silencing CaMYB121 markedly reduced salt tolerance and inhibited root growth. Mechanistically, CaMYB121 directly binds to the CaNHX2 promoter to activate transcription, thereby promoting salt resilience. Salt stress also robustly triggered ABA signaling genes, with CaABF2.1/2 displaying expression patterns closely mirroring those of CaMYB121. Transient silencing of CaABF2.1/2 results in phenotypes similar to those observed with CaMYB121 suppression. Notably, CaMYB121 activates CaABF2.1/2 transcription by binding to its promoters, whereas CaABF2.1/2 represses CaMYB121 expression by directly targeting its promoter, forming a self-regulating feedback loop that prevents excessive defense activation. Collectively, our findings reveal a CaMYB121-CaABF2 feedback circuit that dynamically balances growth and defense to optimize salt tolerance in pepper plants.

Salt Tolerance

Genome-wide identification of CXE gene family in soybean and functional characterization of GmCXE31 in lipid biosynthesis and salt tolerance.

GmCXE31 negatively regulates salt tolerance and lipid synthesis in soybean, and the cxe31-edited lines improve soybean yield and seed quality. Carboxylesterases (CXEs), as essential lipid hydrolases of the α/β-hydrolase fold superfamily, are critical for plant stress responses, hormone signaling and secondary metabolism. The key candidate gene GmCXE31 was previously identified in our laboratory through a genome‑wide association study (GWAS) of soybean lipid‑related traits. In the present study, we further identified 60 GmCXE family genes in soybean. Phylogenetic analysis clustered them into 11 conserved subfamilies. Cis-acting element analysis showed their promoters are enriched with elements related to abiotic stress, growth and hormone signaling, suggesting potential roles in soybean development and stress adaptation. GmCXE31 is highly expressed in seedling roots and responsive to strigolactones (SLs) and salt stress. Functional assays revealed that GmCXE31 negatively regulates soybean salt tolerance: its overexpression reduced salt tolerance in Arabidopsis and soybean under 150 mM NaCl stress, while its knockout enhanced this trait. Lipid profiling revealed GmCXE31-edited lines had higher seed oil content, elevated oleic/linoleic acid ratio and lower saturated fatty acid proportion, which was achieved by regulating lipid synthesis-related genes like GmNFYA. Agronomic trait analysis showed GmCXE31-edited lines had increased nodule number, plant height and single-plant yield at maturity, with opposite phenotypes in overexpression lines. In conclusion, this study elucidates the multifaceted roles of GmCXE31 in coordinating soybean salt tolerance, lipid metabolism and agronomic traits, providing theoretical and genetic resources for salt-tolerant and high-quality soybean molecular breeding.

Glycine max

miR9772, a Triticum-specific miRNA involved in regulating wheat salt tolerance and grain size.

Salt stress severely impairs crop productivity worldwide. MicroRNAs (miRNAs) are a class of endogenous small noncoding RNAs, which played the crucial role in regulating plant growth, development as well as stress responses at the posttranscriptional level. However, the significance of miRNA on salt response in wheat is not well understood at present. In this study, we identified a salt-responsive miRNA from wild emmer wheat, miR9772, which appears to be specific to Triticum species. Under salt stress, the expression of miR9772 was significantly induced and upregulated. Functional analyses revealed that overexpression of miR9772 increased salt sensitivity in wheat, whereas silencing of miR9772 using Short Tandem Target Mimic (STTM) technology markedly enhanced salt tolerance, demonstrated its crucial role in regulating wheat's salt response. Furthermore, we revealed that miR9772 could target on CYP76C4 to decline its expression abundance to affect wheat's salt resistance. Additionally, agronomic and yield-related traits of transgenic wheat lines based on field experiments showed that miR9772-silenced lines exhibited larger grain size and higher grain yield per plant, indicating that miR9772 simultaneously regulated the salt tolerance and grain development. Collectively, this study provided a new target for improving wheat salt tolerance without yield penalty through genome editing breeding.

Triticum

Effect of transgene on salt tolerance of tobacco.

To explore the effects of salt-tolerance gene accumulation on salt tolerance in transgenic plant, we used four types of plant expression vector (N27, N28, N29, and N30) carrying mtlD, mtlD + gutD, mtlD + gutD + BADH, mtlD + gutD + BADH + sacB genes respectively, to transform tobacco through Agrobacterium-mediated method. Transgenic lines were identified through polymerase chain reaction (PCR) detection. Transgenic lines and non-transgenic plant (CK) were subjected to 6‰ sodium chloride solution stress; then, fluorescence quantitative PCR (FQ-PCR) and salt tolerance indexes were used to assess characteristics. PCR showed the exogenous genes had been integrated into the tobacco genome. FQ-PCR showed under clean water treatment the target genes were expressed in all transgenic plants at the transcriptional level. The transcript abundances of target genes changed with the number of genes increased, and improved following salt stress. Comparative analyses of salt tolerance indexes showed height growth, biomass (except for N29), chlorophyll content, net photosynthetic rate, Fv/Fm, and PI of all transgenic plants and CK were lower under salt stress than under clean water treatment, to varying degrees. However, the descent ratio was smaller in transgenic plants. A comprehensive evaluation of multiple salt-tolerance indicators performed using the membership function method showed the average salt tolerance of each vector transgenic line was higher than that of CK, and salt tolerance was greater in transgenic polyvalent gene lines than in transgenic monovalent gene lines. The average salt tolerance was N29 > N28 > N30 > N27 > CK. This study provides a theoretical and practical reference for salt tolerance breeding in other plants.

Plants, Genetically Modified

Transcription factor LbUBC positively regulates salt gland development and salt tolerance by directly binding to the LbTTG1 promoter and repressing its transcription.

KEY MESSAGE:: LbUBC enhances salt tolerance by promoting salt gland development via repressing LbTTG1, revealing a synergisticregulatory mechanism in Limonium bicolor. In the context of increasingly severe soil salinization, salt-tolerant genetic resources from halophytes show great application potential. In particular, the recretohalophyte Limonium bicolor, which possesses specialized salt gland structures, has become a key model for deciphering the molecular mechanisms underlying salt tolerance and salt gland development. In this study, using LbTTG1-overexpressing and -silenced lines, we demonstrate that LbTTG1 negatively regulates salt-gland development and salt tolerance. Through yeast one-hybrid, EMSA, and dual-luciferase assays, Lb7G33228 (LbUBC) was screened and verified as an upstream transcriptional regulator of LbTTG1. LbUBC enhances salt tolerance in L. bicolor by positively regulating salt-gland development, verified using LbUBC silence and overexpression strains. Interestingly, LbUBC represses the expression of its downstream target LbTTG1, thereby releasing the inhibitory effect of LbTTG1 on salt-gland development. In this manner, LbUBC positively regulates salt-gland development, achieving a dynamic balance in the regulation of salt-gland development and salt tolerance in L. bicolor. This study reveals a synergistic regulatory mechanism involving multiple genes, offering new insights for comprehensively dissecting the molecular regulatory network of salt-gland development.

Salt Tolerance

Identification and analysis of HD-ZIP transcription factors that regulate salt gland development and salt tolerance in Limonium bicolor.

Soil salinity severely constrains agricultural production. Elucidating the salt-tolerance mechanisms of halophytes can provide innovative approaches for improving the salt tolerance of crop plants. In this study, we performed genome-wide identification and analysis of 36 LbHDZ genes encoding homeodomain-leucine zipper (HD-ZIP) transcription factors in Limonium bicolor, a typical recretohalophyte that excretes excess salt ions through specialized salt glands. Expression profiling across different stages of salt gland development, as well as in various tissues under salt stress, indicated that multiple LbHDZ genes are involved in regulating salt gland development and salt tolerance. Among these genes, LbHDZ14 (a member of the HD-ZIP II subfamily) exhibited sustained high expression during the critical period of salt gland formation, while its transcript levels were significantly downregulated in leaves and roots under salt stress. Subsequent experiments demonstrated that LbHDZ14 is localized in the nucleus and negatively regulates salt gland density and salt tolerance by directly binding to the promoter of LbGDSL, a positive regulator of salt gland development. In conclusion, this study reveals the expression patterns of LbHDZ genes in L. bicolor, characterizes the functional mechanism of LbHDZ14, further elucidates the regulatory network underlying salt gland development, and provides candidate genes for enhancing crop salt tolerance.

Plumbaginaceae

[Clinical application of the radiolabeled bile salt tolerance test].

The labeled bile salt tolerance test is the measure of the decrease in plasmatic radioactivity after intravenous injection of carboxyl-14C-labeled chenic acid. The label is distributed in the blood, taken up by hepatocytes and then secreted in the bile. The decrease in plasmatic radioactivity during the 4 h following the injection follows a bi-exponential curve. It has been studied in 6 normal subjects, 4 patients equipped with "T tube", 3 cases of acute viral hepatitis, 4 cases of hepatic steatosis, and 6 cases of hepatic cirrhosis. The first slope (b1) represents the hepatic uptake of the label. It is lowered in cases of viral hepatitis and in cirrhosis. The second slope (b2) represents hepato-biliary secretion of the label. It is lowered in patients equipped with a "T tube". From 100 min after the injection, the plasma concentration of radioactivity remains constant. This is the residual value (R), and it is very low in normal subjects. It is increased in cases of acute viral hepatitis and cirrhosis, indicating displacement of a fraction of the bile salt pool into peripheral blood. After a standard meal, the R value is not modified in the normal subject. In cases of steatosis and cirrhosis, a temporary peak may be seen, indicating recirculation of the label towards the periphery due to a porto-systemic shunt or a hepatocyte lesion.

Chenodeoxycholic Acid

Desert-derived Ensifer sp. SA403 enhances potato salt tolerance by reshaping rhizosphere microbiome functions and host responses.

Soil salinization increasingly threatens global food security, and potato (Solanum tuberosum L.), a moderately salt-sensitive crop, is particularly vulnerable to saline soils. Plant growth-promoting rhizobacteria (PGPR) offer a promising strategy to improve crop performance, yet how PGPR interact with native microorganisms to enhance potato salt tolerance remains poorly understood. In this study, we identified a desert-derived PGPR strain, Ensifer sp. SA403, which substantially enhanced potato performance under high salinity across sterile, non-sterile and field conditions. Physiologically, inoculation with SA403 reduced shoot Na⁺ accumulation and increased the K⁺/Na⁺ ratio; notably, these effects were markedly stronger in non-sterile substrates than under sterile conditions, indicating that SA403-mediated ion homeostasis relies on cooperation with the resident microbiota rather than on the strain acting alone. Metagenomic profiling indicated that SA403 strain reshaped rhizosphere communities, significantly enriching beneficial taxa such as Priestia and Bradyrhizobium, and upregulated functional pathways involved in glutathione and sulfur metabolism. Furthermore, host transcriptomic analyses showed that SA403 modulated plant responses to salt stress, with differentially expressed genes enriched in jasmonic acid signaling, ethanolamine metabolism and amino-acid biosynthesis pathways. Field trials on saline soils confirmed that SA403 significantly increased seedling emergence and tuber weight. Together, our results demonstrate that SA403 functions as a biological mediator that optimizes rhizosphere microecology and coordinates ion balance and host signaling to enhance potato salt tolerance. These findings support the potential of SA403 as a robust PGPR-based tool for sustainable potato production on saline soils.

Rhizosphere

Salt tolerance of Rhizobium species in broth cultures.

Salt tolerance of five rhizobia strains was examined in broth cultures. Five levels of NaCl concentration were used and the optical density was taken as a measure for the vigour of bacterial growth. Rhizobium leguminosarum and R. meliloti were tolerant to high levels of salinity and growth curves in saline broth showed a similar pattern to the control level. Rhizobium japonicum, cowpea Rhizobium, and R. trifolii were intolerant to salt and showed a strong growth retardation with increasing salt concentration. Growth was inhibited at high levels of salinity. It is suggested that rhizobia sensitivity to salts may be partly responsible to the inhibition of nitrogen fixation by legumes growing under salt stress.

Culture Media

Genome-wide identification of the Glutathione Peroxidase (GPX) gene family in Taxodium distichum and functional characterization of TdGPX9 in enhancing salt tolerance.

This study systematically identified 10 TdGPX genes in Taxodium distichum, demonstrating that the nucleocytoplasmic-localized TdGPX9 plays a pivotal role in salt stress response. Overexpression of TdGPX9 significantly enhances salt tolerance by strengthening the antioxidant defense system and improving root system plasticity under stress. Taxodium distichum is a premier coniferous species renowned for its exceptional waterlogging and salinity tolerance, serving as a vital forest resource for coastal afforestation and wetland ecological restoration. Within the physiological framework of plant stress resistance, the glutathione peroxidase (GPX) family represents a cornerstone of the antioxidant enzymatic system, playing a critical role in scavenging reactive oxygen species and maintaining cellular redox homeostasis. In this study, 10 TdGPX genes were identified via a comprehensive genome-wide analysis and mapped across eight chromosomes. These genes possess a highly conserved Thioredoxin_like domain, with structural and motif analyses revealing a well-maintained arrangement of conserved motifs within each subgroup. The promoter analysis identified a sophisticated regulatory network enriched with cis-acting elements responsive to light, phytohormones, and abiotic stresses, suggesting their integration into diverse signaling pathways. Expression profiling across various tissues and embryonic developmental stages further highlighted the versatile roles of TdGPX members in plant growth and organogenesis. Notably, qRT-PCR analysis identified the nucleocytoplasmic-localized TdGPX9 as a primary respondent to salinity. Functional validation demonstrated that TdGPX9 overexpression significantly enhances salt tolerance in transgenic Arabidopsis and T. distichum callus by strengthening the antioxidant defense system. Furthermore, TdGPX9 promoted root system plasticity under stress, as evidenced by increased lateral root density. These findings provide a systematic basis for understanding the redox-regulatory mechanisms in baldcypress and offer vital genetic resources for improving forest resilience in coastal wetland ecosystems.

Salt Tolerance

Identification of BoRR gene family in cauliflower: roles in curd development and salt tolerance.

BACKGROUND: Cauliflower, as an important vegetable crop, the research on its curd formation mechanism and stress-responsive gene networks is of great significance for improving its quality, yield and abiotic stress tolerance. The response regulator (RR) gene family plays a crucial role in the regulation of various life processes of many organisms. In this research, a comprehensive analysis of the BoRR gene family in cauliflower was carried out. RESULTS: A Total of 57 BoRR genes were identified in cauliflower and classified into seven subtypes (type A/B-I/B-II/B-IV/C/B-PRR/Clock PRR) based on sequence homology. Chromosomal mapping showed even distribution across genomes, while physicochemical analysis revealed diverse protein properties (134-915 amino acids, pI 4.51-9.19) with predominant nuclear localization. Structural analyses found all BoRR proteins contain REC-type domains, with subtype-specific features: type A has REC_typeA_ARR, type B harbors REC_typeB_ARR domains, and Clock PRR shows circadian-related psREC_RR domains. Exon numbers range from 2 to 10, with type A BoRR genes having shorter CDS lengths. Collinearity analysis identified 28 pairs of gene duplicates (26 inter-chromosomal). Comparative analysis showed 133 collinear pairs with Brassica napus, 96 with Brassica. rapa, and only 1 with monocots specie (rice and maize). Promoter analysis identified hormone-responsive motifs (ABRE, TGACG), development-related elements (ARE), and stress-responsive sequences (e.g., MBS for drought tolerance) in the promoters of BoRR genes. GO enrichment linked BoRR genes to phosphorelay signaling, cytokinin/ethylene response, and developmental processes like meristem maintenance. Expression profiling during curd development showed type A genes (BoRR23/27/34/38/45) up-regulated in vegetative-reproductive transition, BoRR3/6/12/32/54 in curd enlargement, and several genes like BoRR49 in flower bud differentiation. Salt stress (1.5% NaCl) induced transient expression in 8 of 9 selected BoRR genes at day 1 after treatment. qRT-PCR validated their roles in developmental regulation and salt tolerance. CONCLUSION: This study provides valuable insights into the BoRR gene family in cauliflower, laying a foundation for further understanding its genetic mechanisms and potentially guiding efforts to enhance curd quality and salt tolerance in cauliflower.

Salt Tolerance

GhDMT7-mediated DNA methylation dynamics enhance starch and sucrose metabolism pathways to confer salt tolerance in cotton.

This study provides a comprehensive analysis of the impact of DNA methylation in cotton under salt stress conditions, elucidating its effects on gene expression and biological processes. Here, we determined the structures of the DNA methylation landscape across the cotton genome subjected to salt stress using whole-genome bisulfite sequencing (WGBS) and RNA-seq methodologies. We identified 4938 differentially methylated regions (DMRs) correlated with alterations in gene expression. Salt stress induced significant shifts in DNA methylation patterns, particularly in CHH contexts, suggesting context-dependent epigenetic regulation. DMRs were found to be implicated in diverse biological processes and pathways, encompassing protein metabolism, cellular homeostasis, starch and sucrose metabolism, and plant hormone signaling, all pivotal for cotton's adaptation to salt stress. Furthermore, RNA-seq analysis confirmed the impact of DNA methylation on gene expression, uncovering 9642 salt stress-responsive differentially expressed genes (DEGs). These DEGs exhibited enrichment in pathways such as carbohydrate metabolism, cell wall synthesis, and defense response, underscoring the intricate interplay between methylation and gene regulation in stress response. Moreover, the study investigated the role of the key DNA methyltransferase gene GhDMT7 in modulating cotton's response to salt stress, revealing that its downregulation enhanced cotton's salt tolerance, potentially attributed to decreased DNA methylation levels, reduced membrane damage, and enhanced antioxidant capacity. These findings elucidate the role of DNA methylation in abiotic stress resilience and provide insights for crop improvement.

Gossypium

A retrotransposon insertion upstream of Arabidopsis thaliana CRK8 receptor-like kinase modulates a trade-off between pathogen defense and salt tolerance.

In response to necrotrophic fungal pathogens, plants often display quantitative disease resistance (QDR), an immune response with complex genetic determinants. Due to their diversity and small phenotypic effect, the genetic bases of QDR are challenging to characterize. Here, we used genome-wide association mapping in Arabidopsis thaliana natural populations to identify novel determinants of QDR against the fungal pathogen Sclerotinia sclerotiorum. We found that presence-absence polymorphism of the AT4TE56270 Copia transposable element (TE) upstream of the cysteine-rich receptor-like kinase 8 (CRK8) gene is associated with QDR. The presence of the TE associates with higher CRK8 expression in healthy and inoculated plants and increased QDR. The constitutive knockdown of CRK8 reduced QDR, hydrogen peroxide production, and the expression of defense genes upon inoculation. Transcriptome analysis revealed altered defense pathways and salt responses in CRK8 mutants, including impaired glutathione and camalexin biosynthesis, likely contributing to disease susceptibility. Mutants in CRK8 showed altered seed germination on salt, and the absence of AT4TE56270 is associated with enhanced seed germination under salt stress in A. thaliana natural populations. These results reveal a trade-off between salt tolerance and defense against S. sclerotiorum associated with presence-absence polymorphism of a TE.

Arabidopsis

PtoeIF5A1: A Pleiotropic Regulator of Development, PCD, and Salt Tolerance in Populus tomentosa.

Eukaryotic translation initiation factor 5A (eIF5A) is a highly conserved protein family unique to eukaryotes, yet its functional characterization in woody plants remains limited. In this study, we identified four eIF5A genes (PtoeIF5A1-PtoeIF5A4) from the genome of Populus tomentosa, a fast-growing tree species indigenous to China, and characterized their expression patterns and functional roles through bioinformatics analysis, quantitative real-time PCR, stable overexpression in Arabidopsis thaliana, and transient expression in Nicotiana benthamiana leaves. Our results demonstrated that all PtoeIF5A proteins contain a conserved OB-fold domain and multiple phosphorylation sites, with PtoeIF5A1 showing predominant expression in roots and secondary xylem. Functional assays revealed that PtoeIF5A1 overexpression accelerated inflorescence stem elongation and early flowering in Arabidopsis, induced visible chlorosis and programmed cell death (PCD) in tobacco leaves, and significantly enhanced salt tolerance under NaCl treatment. Collectively, these findings establish PtoeIF5A1 in poplar as a pleiotropic regulator integrating developmental cues, programmed cell death, and stress responses; and as a valuable genetic resource for breeding stress-resilient woody plants.

Populus tomentosa

On the mechanism of salt tolerance. Production of glycerol and heat during growth of Debaryomyces hansenii.

As glycerol was suggested as an osmotic agent in the salt tolerant Debaryomyces hansenii the concentrations of total, intracellular, and extracellular glycerol produced by this yeast was followed during growth in 4 mM, 0.68 M, and 2.7 M NaCl media. The total amount of glycerol was not directly proportional to biomass production but to the cultural salinity with maximum concentrations just prior to or at the beginning of the stationary phase. In all cultures the cells lost some glycerol to the media, at 2.7 M NaCl the extracellular glycerol even amounted maximally to 80% of the total. A distinct maximum of intracellular glycerol, related to dry weight or cell number, appeared during the log phase at all NaCl concentrations. As the intracellular calculated glycerol concentrations amounted to 0.2 M, 0.8 M, and 2.6 M in late log phase cells at 4mM, 0.68 M, and 2.7 M NaCl, respectively, whereas the corresponding analysed values for the glycerol concentrations of the media were 0.7 mM, 2.5 mM, and 3.0 mM, glycerol contributes to the osmotic balance of the cells. During the course of growth all cultures showed a decreasing heat production related to cell substance produced, most pronounced at 2.7 M NaCl. At 2.7 M NaCl the total heat production amounted to--1690 kJ per mole glucose consumed in contrast to--1200 and--1130 kJ at 4 mM and 0.68 M NaCl, respectively. The Ym-values were of an inverse order, being 129, 120, and 93 at 4 mM, 0.68 M, and 2.7 M NaCl respectively.

Ascomycota

Multi-omics analyses provide insights into the molecular basis for salt tolerance of Phyla nodiflora.

The perennial herbaceous plant, Phyla nodiflora (Verbenaceae), which possesses natural resistance to multiple abiotic stresses, is widely used as a pioneer species in island ecological restoration. Due to the lack of information about its genome, the mechanism underlying its tolerance to environmental stresses, such as salinity, is almost entirely unknown. Here, we report on the high-quality genome of P. nodiflora that is 403.07 Mb in size, and which was assembled and anchored onto 18 pseudo-chromosomes. Genomic synteny revealed that P. nodiflora underwent two whole genome duplication events, which promoted the expansion of genes related to environmental adaptation and the biosynthesis of secondary metabolites. An integrated genomic and transcriptomic analysis suggested that salt stress tolerance in P. nodiflora is associated with the expansion and activated expression of genes related to abscisic acid (ABA) homeostasis and signaling. The expansion of ZEP family genes may contribute to the consistent increase in ABA levels under salt stress. Lysine acetylomic analysis revealed that exposure to salt led to widespread protein deacetylation, with these proteins primarily involved in signal transduction, carbohydrate transport and metabolism, and transcription regulation. Deacetylation of glutathione S-transferase increased enzymatic activities in response to salt-induced oxidative stress. Collectively, the genomic, transcriptomic, and lysine acetylomic analyses provide profound insight into the molecular basis of the adaptation of P. nodiflora to salt stress, and will be helpful to engineer salt-tolerant plants for ecological restoration.

Salt Tolerance

Hydroxylamine sensitivity and salt tolerance in screening and identifying mycobacteria.

Hydroxylamine sensitivity offered no advantage over p-nitrobenzoic acid in distinguishing between tubercle bacilli and other (opportunist and non-significant) mycobacteria. It was of limited use in identification procedures. Salt tolerance assisted in the identification of strains which may be Mycobacterium triviale and M. chelonei var borstelense.

Bacteriological Techniques