Expression Of Stress-responsive Genes Research Articles

C-repeat binding factor (CBF) identification and expression pattern to abiotic stress response in Casuarina equisetifolia and overexpression of CeCBF4 increase stress tolerance in Arabidopsis

Casuarina equisetifolia stands out as a highly valuable tree species crucial for industries and ecological restoration projects in tropical and subtropical regions, owing to its exceptional salt resistance. However, its growth is hampered in cold and drought-prone areas. Understanding the genetic basis for abiotic stress tolerance is imperative for enhancing stress-resistant breeding efforts of C. equisetifolia. However, the CBF genes are renowned for their pivotal roles in cold response and broad resilience to other adverse environments, such as drought, and high salinity. In this study, five CeCBFs were identified from the genome of C. equisetifolia. CeCBF1 and CeCBF3 belong to Clade I group, which diverged early from DREBIII members, while CeCBF2, CeCBF4, and CeCBF5 are classified into Clade II, a group innovated from Clade I. These genes lack introns in their sequences. The proteins encoded by CeCBFs feature classic AP2 domains and CBF signature sequences, localize in the nucleus and exhibit transactivation activity. Additionally, stress-related, abscisic acid, and jasmonic acid responsive elements are present on the CeCBFs promoters. Expression analysis revealed that CeCBFs were prodominantly induced by cold, drought, and salinity treatments, except for CeCBF1, which showed repression specifically under low temperature. Moreover, CeCBF1, CeCBF4, and CeCBF5 were inhibited by abscisic acid, while CeCBF1 showed reduced transcript levels in response to methyl jasmonate treatment. Furthermore, overexpression of CeCBF4 enhanced tolerance to low temperature, depriving of water, and salt by up-regulating stress-responsive genes expression and promoting proline accumulation, albeit with a trade-off in growth and productivity. Hence, CeCBFs emerge as potential regulators contributing significantly to stress tolerance mechanisms in C. equisetifolia.

Exploring Photoreceptor Gene Expression and Seasonal Physiology in Mediterranean Swordfish (Xiphias gladius).

Mature and immature female swordfish show different gonadal expression patterns of melatonin synthesis-related and opsin genes, indicating that seasonality affects gonadal circadian genes, especially through opsin modulation. The high energy demands of reproduction may cause mature female swordfish to become more susceptible to environmental stressors/changes, potentially affecting their health post-reproduction. The modulation of melatonin and opsin expression may also influence swordfish biology, impacting basal metabolism and stress responses. This study aimed to investigate the hepatic expression patterns of genes involved in melatonin and opsins activity after the reproductive period, comparing mature and immature females. Liver samples of swordfish females, collected from the Central Adriatic Sea, were analyzed to evaluate the expression of a melatonin receptor (mel1b), acetylserotonin O-methyltransferase (asmt), opsins (sws, VA opsin and opsin4), and stress response (sod1, sod2 and hsp4b) genes, while histological analysis focused on hepatic melanomacrophages, melanomacrophage centers, and lipid content. The expression of all genes analyzed did not differ between mature and immature females except for sws, which was significantly downregulated in mature females. The overall health conditions of mature and immature females appeared comparable, as suggested by the histological analysis and the absence of significant differences in the expression of stress response genes. Although the study used a limited number of samples and lacked a comparison between the reproductive and non-reproductive periods, the preliminary results suggest that opsins may play a role in modulating physiological processes beyond reproduction.

'Re-Wilding' an Animal Model With Microbiota Shifts Immunity and Stress Gene Expression During Infection.

The frequency of emerging disease is growing with ongoing human activity facilitating new host-pathogen interactions. Novel infection outcomes can also be shaped by the host microbiota. Caenorhabditis elegans nematodes experimentally colonised by a wild microbiota community and infected by the widespread animal pathogen, Staphylococcus aureus, have been shown to suffer higher mortality than those infected by the pathogen alone. Understanding the host responses to such microbiota-pathogen ecological interactions is key to pinpointing the mechanism underlying severe infection outcomes. We conducted transcriptomic analyses of C. elegans colonised by its native microbiota, S. aureus and both in combination. Correlations between altered collagen gene expression and heightened mortality in co-colonised hosts suggest the microbiota modified host resistance to infection. Furthermore, microbiota colonised hosts showed increased expression of immunity genes and variable expression of stress response genes during infection. Changes in host immunity and stress response could encompass both causes and effects of severe infection outcomes. 'Re-wilding' this model nematode host with its native microbiota indicated that typically commensal microbes can mediate molecular changes in the host that are costly when challenged by a novel emerging pathogen.

The role of gibberellin synthase gene VvGA2ox7 acts as a positive regulator to salt stress in Arabidopsis thaliana.

Soil salinity is an important environmental component affecting plant growth and yield, but high-salinity soils are a major constraint to the development of the grape industry. Previous studies have provided lines of evidence that gibberellins (GAs) play a significant regulatory role in plant responses to salt stress. However, it remains unclear whether GA2ox, a key enzyme that maintains the balance of bioactive gibberellins and intermediates in plants, is involved in the mechanism of salt stress tolerance in grapes. In this study, we found that GA2ox7 positively modulates salt stress via its ectopic expression in Arabidopsis thaliana. The GA2ox7 gene cloned from grape was a hydrophilic protein, its CDS length was 1002bp. Besides, VvGA2ox7 protein contained DIOX_N and 2OG-FeII_Oxy domains and was localized at the nucleus and cytoplasm. Yeast two-hybrid (Y2H) showed VvARCN1, VvB5R, VvRUB2, and VvCAR11 might be potential interaction proteins of VvGA2ox7. Compared with the wild type, overexpression of VvGA2ox7 in Arabidopsis thaliana enhanced antioxidant enzymatic activities and proline, chlorophyll, and ABA contents, and decreased relative electrical conductivity, malondialdehyde, and GA3 contents. Moreover, overexpression of VvGA2ox7 positively regulated the expression of salt stress response genes (KAT1, APX1, LEA, P5CS1, AVP1, CBF1), indicating that the VvGA2ox7 overexpression improved the salt stress tolerance of plants. Taken together, this investigation indicates that VvGA2ox7 may act as a positive regulator in response to salt stress and provide novel insights for a deeper understanding of the role of VvGA2ox7 in grapes.

The Adhesion GPCR ADGRL2/LPHN2 Can Protect Against Cellular and Organismal Dysfunction.

The most common trigger of sepsis and septic shock is bacterial lipopolysaccharide (LPS). Endothelial cells are among the first to encounter LPS directly. Generally, their function is closely linked to active endothelial NO Synthase (eNOS), which is significantly reduced under septic conditions. LPS treatment of endothelial cells leads to their activation and apoptosis, resulting in loss of integrity and vascular leakage, a hallmark of septic shock. Hence, therapies that prevent endothelial leakage or restore the endothelial barrier would be invaluable for patients. Adhesion GPCRs (aGPCRs) have been largely overlooked in this context, although particularly one of them, ADGRL2/LPHN2, has been implicated in endothelial barrier function. Our study shows that overexpression of ADGRL2 protects endothelial cells from LPS-induced activation, apoptosis, and impaired migration. Mechanistically, ADGRL2 preserves eNOS activity by shifting its binding from Caveolin-1 to Heat Shock Protein 90. Furthermore, ADGRL2 enhances antioxidative responses by increasing NRF2 activity. Notably, we found that this function may be evolutionarily conserved. In the absence of lat-2, a homolog of ADGRL2 in Caenorhabditis elegans, worms show higher ROS levels and altered stress response gene expression. Additionally, lat-2 mutants have a significantly reduced lifespan, altogether indicating a protective role of ADGRL2 against oxidative stress across species.

Exploring genotypic variation and gene expression associated to cadmium accumulation in bread wheat

Cadmium (Cd) contamination poses significant risks to agricultural productivity and human health, particularly through its accumulation in staple crops such as bread wheat (Triticum aestivum L.). This study evaluated Cd accumulation and tolerance among six bread wheat cultivars exposed to six Cd concentrations (0, 2.5, 5, 10, 15, 20, and 25 mg kg−1 soil). Phenotypic assessments and quantitative real-time PCR (qRT-PCR) were conducted to analyze the expression patterns of TaNRAMP and TaZIP genes in various tissues and developmental stages of wheat, which play crucial roles in Cd uptake and transport. Results demonstrated significant variability in Cd accumulation. The Barat cultivar exhibited the lowest accumulation in grain (ranging from 0.21 to 8.8 mg kg−1) and the highest tolerance. In contrast, Kavir and Pishtaz displayed elevated Cd levels in both grain and straw, while Parsi accumulated more Cd in straw at lower concentrations (56.9 mg kg−1 in Cd concentration of 10 mg kg−1 soil). The gene expression analysis revealed that most cultivars showed increased expression of TaNRAMP genes, particularly TaNRAMP2 in Cd concentration of 10 mg kg−1 soil, which facilitates Cd uptake from the soil, and TaZIP genes, such as TaZIP4 and TaZIP7, involved in transporting Cd within the plant. Notably, the expression of TaZIP1 was significantly lower in cultivars with high Cd accumulation, suggesting a potential regulatory mechanism for Cd tolerance. Furthermore, cultivars exhibiting higher Cd levels correlated with increased expression of stress-responsive genes, indicating a broader response to Cd stress. These findings highlight Barat’s potential for bread-making applications due to its low Cd accumulation, while Morvarid and Pishtaz which show reduced Cd content in the straw even under high Cd exposure are better suited for animal feed. This research underscores the genetic variability of wheat cultivars in response to Cd stress and provides essential insights into the molecular mechanisms underlying Cd accumulation, offering valuable information for breeding programs aimed at developing Cd-tolerant varieties to ensure food security in contaminated regions.

Homocysteine S-Methyltransferase 3 Positively Regulates Cadmium Tolerance in Maize.

The increasing contamination of agricultural soils with cadmium (Cd) poses a significant threat to human health and global food security. Plants initiate a series of mechanisms to reduce Cd toxicity. However, the response of maize to Cd toxicity remains poorly understood. In this study, we identified that ZmHMT3, which encodes a homocysteine S-methyltransferases family protein, acted as a regulator of Cd tolerance in maize. Subcellular localization and in situ PCR exhibited that ZmHMT3 was localized in the cytoplasm and predominantly expressed in the phloem. Overexpression of ZmHMT3 enhanced Cd tolerance and reduced Cd concentration in both shoots and roots. In contrast, ZmHMT3 mutants attenuated Cd tolerance but did not change shoot Cd concentration. Heterologous overexpression of ZmHMT3 in rice enhanced Cd tolerance and reduced grain Cd concentration. Transcriptome analysis revealed that ZmHMT3 upregulated the expression of stress-responsive genes, especially glutathione S-transferases (GSTs) and transcription factors, including MYBs, NACs and WRKYs, and modulates the expression of different ATP-binding cassette (ABC) transporters, thereby enhancing Cd tolerance. Collectively, these findings highlight the pivotal role of ZmHMT3 in Cd tolerance and as a candidate gene for improving Cd tolerance in elite maize varieties.

Melatonin Ameliorates Cadmium Toxicity in Tobacco Seedlings by Depriving Its Bioaccumulation, Enhancing Photosynthetic Activity and Antioxidant Gene Expression.

Soil remediation for cadmium (Cd) toxicity is essential for successful tobacco cultivation and production. Melatonin application can relieve heavy metal stress and promote plant growth; however, it remains somewhat unclear whether melatonin supplementation can remediate the effects of Cd toxicity on the growth and development of tobacco seedlings. Herein, we evaluated the effect of soil-applied melatonin on Cd accumulation in tobacco seedlings, as well as the responses in growth, physiological and biochemical parameters, and the expression of stress-responsive genes. Our results demonstrate that melatonin application mitigated Cd stress in tobacco, and thus promoted plant growth. It increased root fresh weight, dry weight, shoot fresh weight and dry weight by 58.40%, 163.80%, 34.70% and 84.09%, respectively, compared to the control. Physiological analyses also showed significant differences in photosynthetic rate and pigment formation among the treatments, with the highest improvements recorded for melatonin application. In addition, melatonin application alleviated Cd-induced oxidative damage by reducing MDA content and enhancing the activities of enzymatic antioxidants (CAT, SOD, POD and APX) as well as non-enzymatic antioxidants (GSH and AsA). Moreover, confocal microscopic imaging confirmed the effectiveness of melatonin application in sustaining cell integrity under Cd stress. Scanning Electron Microscopy (SEM) observations illustrated the alleviative role of melatonin on stomata and ultrastructural features under Cd toxicity. The qRT-PCR analysis revealed that melatonin application upregulated the expression of photosynthetic and antioxidant-related genes, including SNtChl, q-NtCSD1, NtPsy2 and QntFSD1, in tobacco leaves. Together, our results suggest that soil-applied melatonin can promote tobacco tolerance to Cd stress by modulating morpho-physiological and biochemical changes, as well as the expression of relevant genes.

A C2H2-Type Zinc Finger Protein from Mentha canadensis, McZFP1, Negatively Regulates Epidermal Cell Patterning and Salt Tolerance

C2H2-type zinc finger protein (C2H2-ZFP) transcription factors play evident roles in regulating plant growth and development and abiotic stress responses. However, the role of C2H2-ZFP from Mentha canadensis remains uncertain. We identified the multifunctional C2H2-ZFP gene McZFP1 from M. canadensis based on phylogenetic analysis. The McZFP1 gene was highly expressed in stems, responding to abiotic stress and phytohormone treatments. McZFP1 localized in the nucleus and showed no transcriptional autoactivation activity in yeast. McZFP1 overexpression in Arabidopsis thaliana significantly reduced the number of trichomes and root hairs, root hair length, and salt stress tolerance. Further study revealed that McZFP1 overexpression increased the expression of negative regulator genes and decreased that of positive regulator genes to inhibit plant trichome and root hair development. Malondialdehyde accumulation was promoted, but the proline content and catalase, superoxide dismutase, and peroxidase activities were reduced and the expression of stress response genes was inhibited in McZFP1 overexpression lines under salt treatment, thereby compromising plant salt tolerance. Overall, these results indicate that McZFP1 is a novel C2H2-ZFP transcription factor that plays negative roles in trichome and root hair development and salt stress tolerance.

MYB transcription factors, their regulation and interactions with non-coding RNAs during drought stress in Brassica juncea

BackgroundBrassica juncea(L.) Czern is an important oilseed crop affected by various abiotic stresses like drought, heat, and salt. These stresses have detrimental effects on the crop’s overall growth, development and yield. Various Transcription factors (TFs) are involved in regulation of plant stress response by modulating expression of stress-responsive genes. The myeloblastosis (MYB) TFs is one of the largest families of TFs associated with various developmental and biological processes such as plant growth, secondary metabolism, stress response etc. However, MYB TFs and their regulation by non-coding RNAs (ncRNAs) in response to stress have not been studied in B. juncea. Thus, we performed a detailed study on the MYB TF family and their interactions with miRNAs and Long non coding RNAs.ResultsComputational investigation of genome and proteome data presented a comprehensive picture of the MYB genes and their protein architecture, including intron-exon organisation, conserved motif analysis, R2R3 MYB DNA-binding domains analysis, sub-cellular localization, protein-protein interaction and chromosomal locations. Phylogenetically, BjuMYBs were further classified into different subclades on the basis of topology and classification in Arabidopsis. A total of 751 MYBs were identified in B. juncea corresponding to 297 1R-BjuMYBs, 440 R2R3-BjuMYBs, 12 3R-BjuMYBs, and 2 4R-BjuMYBs types. We validated the transcriptional profiles of nine selected BjuMYBs under drought stress through RT-qPCR. Promoter analysis indicated the presence of drought-responsive cis-regulatory components. Additionally, the miRNA-MYB TF interactions was also studied, and most of the microRNAs (miRNAs) that target BjuMYBs were involved in abiotic stress response and developmental processes. Regulatory network analysis and expression patterns of lncRNA-miRNA-MYB indicated that selected long non-coding RNAs (lncRNAs) acted as strong endogenous target mimics (eTMs) of the miRNAs regulated expression of BjuMYBs under drought stress.ConclusionsThe present study has established preliminary groundwork of MYB TFs and their interaction with ncRNAs in B. juncea and it will help in developing drought- tolerant Brassica crops.

Use of transcriptomics and genomics to assess the effect of disinfectant exposure on the survival and resistance of Escherichia coli O157:H7, a human pathogen.

Disinfectants are essential for biosecurity, preventing the persistence and spread of zoonotic pathogens on farms and subsequent human infections. In this study, transcriptomics and genomics were utilised to assess the effect of disinfectant exposure on pathogenic Escherichia coli. The exposure of E. coli O157:H7 to sub-optimal concentrations of commonly used farm disinfectants elicited changes in both the transcriptome and genome. The transcriptomics identified upregulation of >300 genes and downregulation of >100 genes with functions, which included stress response, metabolism, transcription, transportation, membrane-associated and virulence genes. The phage shock protein (psp) operon was highly upregulated in response to a quaternary ammonium compound (QAC)-containing disinfectant, which has not previously been associated with a response to chemical stress. Disinfectant-adapted isolates generated by exposure to sub-lethal disinfectants levels demonstrated resistance to several common antibiotics and decreased sensitivity to biocides. Whole genome sequencing of the mutant strains indicated that they had acquired mutations in the genes associated with the upregulation of the multiple antibiotic resistance (MAR) efflux system (lon protease and marR) and topoisomerase genes (gyrA and gyrB). The disinfectant-adapted isolates also exhibited increased expression of transcription, respiration and several pH stress response genes localised in the "acid fitness island." This study demonstrated that sub-optimal disinfectant concentrations allow E. coli O157:H7 to adapt and survive disinfection and develop antibiotic resistance. These changes could have implications for disease treatment and elimination on farms. Although E. coli O157:H7 and farm disinfectants were the focus of this study, we believe these findings are also applicable to other settings, including hospitals.

Osmotic signaling releases PP2C-mediated inhibition of Arabidopsis SnRK2s via the receptor-like cytoplasmic kinase BIK1

Osmotic stress and abscisic acid (ABA) signaling are important for plant growth and abiotic stress resistance. Activation of osmotic and ABA signaling downstream of the PYL-type ABA receptors requires the release of SnRK2 protein kinases from the inhibition imposed by PP2Cs. PP2Cs are core negative regulators that constantly interact with and inhibit SnRK2s, but how osmotic signaling breaks the PP2C inhibition of SnRK2s remains unclear. Here, we report that an Arabidopsis receptor-like cytoplasmic kinase, BIK1, releases PP2C-mediated inhibition of SnRK2.6 via phosphorylation regulation. The dominant abi1-1 ABA-signaling mutation (G180D) disrupts PYL-PP2C interactions and disables PYL-initiated release of SnRK2s; in contrast, BIK1 releases abi1-1-mediated inhibition of SnRK2.6. BIK1 interacts with and phosphorylates SnRK2.6 at two tyrosine residues, which are critical for SnRK2.6 activation and function. Phosphorylation of the two tyrosine residues may affect the docking of the tryptophan “lock” of PP2C into SnRK2.6. Moreover, the bik1 mutant is defective in SnRK2 activation, stress-responsive gene expression, ABA accumulation, growth maintenance, and water loss under osmotic stress. Our findings uncover the critical role of BIK1 in releasing PP2C-mediated inhibition of SnRK2s under osmotic stress.

Multi-functional PGPR Serratia liquefaciens confers enhanced resistance to lead stress and bacterial blight in soybean (Glycine max L.)

Lead toxicity and bacterial blight disease caused by Pseudomonas savastanoi pv. glycinea have destructive impacts on soybean growth and productivity. Plant growth-promoting rhizobacteria have been used as an eco-friendly approach for augmenting crop growth and stress resistance. The current study investigated the efficacy of Serratia liquefaciens ZM6 strain in enhancing soybean resistance to lead (Pb) stress and bacterial blight. Two pot experiments were performed. In the first pot experiment, soybean plants were inoculated with S. liquefaciens ZM6 and grown under variable Pb stress levels (0, 200 and 400 µM of Pb(NO3)2). In the second experiment, S. liquefaciens-inoculated soybean plants were infected with P. savastanoi pv. glycinea, and disease severity was assessed two weeks post infection. The results revealed that S. liquefaciens strain resisted Pb stress up to 400 µM Pb(NO3)2 and exhibited the highest levels of solubilized phosphate, solubilized zinc, siderophore, indole acetic acid, exopolysaccharide, trehalose and antioxidant enzymes at 400 µM Pb compared to the other treatments. Moreover, Pb stress (200 and 400 µM) significantly decreased the growth, yield, nutrient uptake, gas exchange, and contents of chlorophyll, soluble proteins, sugars, and phenolics of soybean plants. Pb stress also induced the levels of proline, glycine betaine, Pb, oxidative stress markers, antioxidant enzymes, ascorbate, glutathione and expression of stress-responsive genes (CAT, APX, POD, Fe-SOD, CHS7, CHI1A, PAL, IFS2, P5CS and WRKY54) in soybean plants. On the other hand, S. liquefaciens application markedly boosted the growth, yield and levels of nutrients, gas-exchange, chlorophyll, osmolytes, antioxidant enzymes and expression of stress-tolerant genes of Pb-stressed soybean plants. The bacterial inoculation significantly diminished oxidative stress indicators and Pb content in stressed plants. Inoculation of soybean plants with S. liquefaciens also caused significant reductions in blight disease symptoms in P. savastanoi pv. glycinea-infected plants, indicating the efficiency of this strain in controlling harmful blight disease. Overall, this study demonstrated S. liquefaciens ZM6 effectiveness in enhancing soybean resistance to Pb stress and bacterial blight.

Stress-Responsive Gene Expression, Metabolic, Physiological, and Agronomic Responses by Consortium Nano-Silica with Trichoderma against Drought Stress in Bread Wheat

The exploitation of drought is a critical worldwide challenge that influences wheat growth and productivity. This study aimed to investigate a synergistic amendment strategy for drought using the single and combined application of plant growth-promoting microorganisms (PGPM) (Trichoderma harzianum) and biogenic silica nanoparticles (SiO2NPs) from rice husk ash (RHA) on Saudi Arabia’s Spring wheat Summit cultivar (Triticum aestivum L.) for 102 DAS (days after sowing). The significant improvement was due to the application of 600 ppm SiO2NPs and T. harzianum + 600 ppm SiO2NPs, which enhanced the physiological properties of chlorophyll a, carotenoids, total pigments, osmolytes, and antioxidant contents of drought-stressed wheat plants as adaptive strategies. The results suggest that the expression of the studied genes (TaP5CS1, TaZFP34, TaWRKY1, TaMPK3, TaLEA, and the wheat housekeeping gene TaActin) in wheat remarkably enhanced wheat tolerance to drought stress. We discovered that the genes and metabolites involved significantly contributed to defense responses, making them potential targets for assessing drought tolerance levels. The drought tolerance indices of wheat were revealed by the mean productivity (MP), stress sensitivity index (SSI), yield stability index (YSI), and stress tolerance index (STI). We employed four databases, such as BAR, InterPro, phytozome, and the KEGG pathway, to predict and decipher the putative domains in prior gene sequencing. As a result, we discovered that these genes may be involved in a range of important biological functions in specific tissues at different developmental stages, including response to drought stress, proline accumulation, plant growth and development, and defense response. In conclusion, the sole and/or dual T. harzianum application to the wheat cultivar improved drought tolerance strength. These findings could be insightful data for wheat production in Saudi Arabia under various water regimes.

AtC3H3, an Arabidopsis Non-TZF Gene, Enhances Salt Tolerance by Increasing the Expression of Both ABA-Dependent and -Independent Stress-Responsive Genes.

Salinity causes widespread crop loss and prompts plants to adapt through changes in gene expression. In this study, we aimed to investigate the function of the non-tandem CCCH zinc-finger (non-TZF) protein gene AtC3H3 in response to salt stress in Arabidopsis. AtC3H3, a gene from the non-TZF gene family known for its RNA-binding and RNase activities, was up-regulated under osmotic stress, such as high salt and drought. When overexpressed in Arabidopsis, AtC3H3 improved tolerance to salt stress, but not drought stress. The expression of well-known abscisic acid (ABA)-dependent salt stress-responsive genes, namely Responsive to Desiccation 29B (RD29B), RD22, and Responsive to ABA 18 (RAB18), and representative ABA-independent salt stress-responsive genes, namely Dehydration-Responsive Element Binding protein 2A (DREB2A) and DREB2B, was significantly higher in AtC3H3-overexpressing transgenic plants (AtC3H3 OXs) than in wild-type plants (WT) under NaCl treatment, indicating its significance in both ABA-dependent and -independent signal transduction pathways. mRNA-sequencing (mRNA-Seq) analysis using NaCl-treated WT and AtC3H3 OXs revealed no potential target mRNAs for the RNase function of AtC3H3, suggesting that the potential targets of AtC3H3 might be noncoding RNAs and not mRNAs. Through this study, we conclusively demonstrated that AtC3H3 plays a crucial role in salt stress tolerance by influencing the expression of salt stress-responsive genes. These findings offer new insights into plant stress response mechanisms and suggest potential strategies for improving crop resilience to salinity stress.

Polyamine impact on physiology of early stages of reef-building corals-insights from rearing experiments and RNA-Seq analysis.

Polyamines are involved in various functions related to the cellular-level responses. To assess effects of polyamines on marine organisms, rearing experiments and comprehensive gene expression analyses were conducted on Acropora digitifera and Acropora sp.1, representative reef-building corals along the west-central coast of Okinawa, Japan, to evaluate effects of putrescine. Concentrations of putrescine ≥ 1mM dissolved tissues of juvenile polyps and increased mortality of planula larvae. RNA-Seq analysis of juvenile polyps exposed to putrescine at the stage before effects became visible revealed dynamic fluctuations in gene expression in the putrescine-treated samples, with increased expression of stress-responsive genes (e.g. NAD-dependent protein deacylase sirtuin-6) and the polyamine transporter Slc18b1-like protein. These results also suggest that putrescine affects expression of genes related to ribosomes and translation. This study provides important insights into roles of polyamines and future directions regarding physiological responses of corals.

Sex differences in the development of experimental diabetic retinopathy

This study aimed to characterize the role of female sex in the pathogenesis of diabetic retinopathy. In the retinae of female Ins2Akita-diabetic mice (F-IA), ovariectomized female Ins2Akita-diabetic mice (F-IA/OVX), male Ins2Akita-diabetic mice (M-IA), and female STZ-diabetic mice (F-STZ), the formation of reactive metabolites and post-translational modifications, damage to the neurovascular unit, and expression of cellular stress response genes were analyzed. Compared to the male diabetic retina, the concentrations of the glycation adduct fructosyl-lysine, the Maillard product 3-deoxyglucosone, and the reactive metabolite methylglyoxal were significantly reduced in females. In females, there was also less evidence of diabetic damage to the neurovascular unit, as shown by decreased pericyte loss and reduced microglial activation. In the male diabetic retina, the expression of several members of the crystallin gene family (Cryab, Cryaa, Crybb2, Crybb1, and Cryba4) was increased. Clinical data from type 1 diabetic females showed that premenopausal women had a significantly lower prevalence of diabetic retinopathy compared to postmenopausal women stratified for disease duration and glycemic control. These data emphasize the importance of estradiol in protecting the diabetic retina and highlight the pathogenic relevance of sex in diabetic retinopathy.

Expression of stress responsive genes enables Limosilactobacillus reuteri to cross-protection against acid, bile salt, and freeze-drying.

Limosilactobacillus reuteri effectively colonizing the gut, secretes antimicrobial compounds and strengthens immune system function. Considering these health benefits, increasing its stress assessments efficiency could improve its commercial viability. In this work, the resistance of L. reuteri FP41 to acid, bile salts, and freeze-drying was examined. The findings showed that strain FP41 demonstrated a strong resistance to acid/bile salt stresses. The transcriptome revealed a significant up-regulation of various stress response genes, including those related to membrane integrity, glutamine metabolism, OsmC family protein, ABC transporters, and chaperonin. Subsequent research demonstrated that overexpression of three stress response-specific proteins, including glutamate decarboxylase GatD, osmotically induced bacterial protein OsmC, and membrane protein component CsbD, significantly increased the survival rate of L. reuteri Z204 under acid/bile salts stress. Notably, overexpression of the OsmC, CsbD, and GatD proteins also enhanced the survival of L. reuteri after freeze-drying. The development of a unique cross-protection method is highlighted in this study, that might significantly increase cellular resistance to acid, bile salts, and cold stresses. This finding could significantly impact the way that L. reuteri is employed in industrial manufacturing processes.

Selection of Reference Genes and HSP17.9A Expression Profiling in Heat-Stressed Grapevine Varieties

Background: “Touriga Franca” (TF) and “Touriga Nacional” (TN) are grapevine varieties cultivated in the ‘Douro Superior’ subregion (Northern Portugal) that experience stressful environmental conditions during the summer. Objectives: Aiming to profile the expression of stress-responsive genes by quantitative real-time PCR (qPCR) in TF and TN plants growing naturally, three candidate reference genes were first tested under controlled conditions. Methods: To simulate a summer’s day, TF and TN in vitro plants were exposed to 32 °C–3 h (heat acclimation) and 42 °C–1 h (severe heat stress, HS) followed by two recovery periods (32 °C–3 h and 24 °C–24 h). Leaf samples were collected at the end of each phase. Control plants were kept at 24 °C. Results: Among the candidate reference genes, the UBC and VAG pair showed the highest stability. The suitability of these genes for qPCR was validated by heat shock protein 17.9A (HSP17.9A) gene profiling. The HSP17.9A expression was up-regulated in both varieties and all experimental phases except in TF control plants. TN showed the highest HSP17.9A relative expression ratio after severe HS. Conclusions: TN responded faster than TF to the induced heat shocks. The UBC, VAG, and HSP17.9A genes revealed to be suitable for further qPCR assays in TF and TN grapevine varieties.

Links between mutations in functionally separate arms of mitochondrial complex I and responses to volatile anesthetics.

Individuals with mitochondrial defects, especially those in Complex I of the electron transport chain, exhibit behavioral hypersensitivity and toxicity to volatile anesthetics. In Drosophila melanogaster, mutation of ND23 (NDUFS8 in mammals), which encodes a subunit of the matrix arm of Complex I, sensitizes flies to toxicity from isoflurane but not an equipotent dose of sevoflurane. Also, in ND23 flies, both anesthetics activate expression of stress response genes, but to different extents. Here, we investigated the generality of these findings by examining flies mutant for ND2 (ND2 in mammals), which encodes a subunit of the membrane arm of Complex I. The serial anesthesia array was used to expose ND2del1 and ND2360114 flies to precise doses of isoflurane, sevoflurane, and oxygen. Behavioral sensitivity was assessed by a climbing assay and toxicity by percent mortality within 24 h of exposure. Changes in expression were determined by qRT-PCR of RNA isolated from heads at 0.5 h after anesthetic exposure. Unlike ND2360114, ND2del1 did not affect behavioral sensitivity to isoflurane or sevoflurane. Furthermore, sevoflurane in hyperoxia as well as anoxia caused mortality of ND2del1 but not ND2360114 flies. Finally, the mutations had different effects on induction of stress response gene expression by the anesthetics. Mutations in different arms of Complex I resulted in different behavioral sensitivities and toxicities to isoflurane and sevoflurane, indicating that (i) the anesthetics have mechanisms of action that involve arms of Complex I to different extents and (ii) the lack of behavioral hypersensitivity does not preclude susceptibility to anesthetic toxicity.