Expression Of Stress-related Genes Research Articles

Molecular mechanism for combined toxicity of micro(nano)plastics and carbon nanofibers to freshwater microalgae Chlorella pyrenoidosa

The understanding of the environmental consequences resulting from the presence of micro(nano)plastics and carbon nanofibers (CNFs) in aquatic ecosystems is currently limited. This research endeavor sought to investigate the underlying molecular mechanisms by which engineered polystyrene-based microplastics (MPs)/nanoplastics (NPs) and CNFs, both individually and in combination, elicit toxic effects on an algal species Chlorella pyrenoidosa. The findings revealed that the combined toxicity of MPs/NPs and CNFs depended on the concentration of the mixture. As the concentration increased, the combined toxicity of MPs/NPs and CNFs was significantly greater than the toxicity of each component on its own. Furthermore, the combined toxicity of NPs and CNFs was higher than that of MPs and CNFs. The study integrated data on cell membrane integrity, oxidative stress, and antioxidant modulation to create an Integrated Biomarker Response index, which demonstrated that the co-exposure of algae to NPs and CNFs resulted in more severe cellular stress compared to exposure to NPs alone. Similarly, the combination of NPs and CNFs caused greater cellular stress than the combination of MPs and CNFs. Additionally, significant changes in the expression of stress-related genes caused by MPs/NPs alone and in combination with CNFs indicated that oxidative stress response, glucose metabolism, and energy metabolism played critical roles in particle-induced toxicity. Overall, this study provides the first insight into the toxicological mechanism of MPs/NPs and CNFs mixtures at the molecular level in freshwater microalgae.

Bacillus cereus enhances salt tolerance of cucumber seedlings by improving antioxidant metabolism and decreasing the ion toxicity

Salt stress poses a significant challenge to plants, exerting a detrimental impact on crop growth and yield. This study investigated the effects of the strain Bacillus cereus on plant growth, ion contents and antioxidant metabolism of cucumber seedlings under salt stress (150 mM NaCl). The results showed that B. cereus could colonize the roots system of cucumber, with peak colonization occurring on the 3rd day. Inoculation with B. cereus effectively alleviated the growth inhibition of cucumber seedlings induced by salt stress, resulting in improvements in plant height, stem diameter, fresh and dry weight. Furthermore, the Na+ content in cucumber seedlings subjected to salt stress was significantly reduced by 38.35 % in leaves and 38.19 % in roots upon B. cereus inoculation, while K+ content increased by 9.88 % and 168.34 % in leaves and roots, respectively. Additionally, B. cereus significantly enhanced antioxidant enzyme activity and antioxidant content, while reduced the levels of H2O2, MDA, and O2−. in cucumber seedlings under salt stress conditions. B. cereus-treated seedlings exhibited an increase in soluble sugar content by 35.27 % and 93.21 % in leaves, and 59.36 % and 128.78 % in roots, respectively. Moreover, B. cereus treatment significantly up-regulated the expression of salt tolerant gene in both cucumber leaves and roots. These results showed that B. cereus enhanced salt tolerance in cucumber seedlings by modulating the antioxidant defense system, maintaining ion homeostasis, and promoting the expression of stress-related genes, ultimately improving the growth of cucumber seedlings.

Effects of Antibiotic Residues on Fish Gut Microbiome Dysbiosis and Mucosal Barrier-Related Pathogen Susceptibility in Zebrafish Experimental Model.

The symbiotic community of microorganisms in the gut plays an important role in the health of the host. While many previous studies have been performed on the interactions between the gut microbiome and the host in mammals, studies in fish are still lacking. In this study, we investigated changes in the intestinal microbiome and pathogen susceptibility of zebrafish (Danio rerio) following chronic antibiotics exposure. The chronic antibiotics exposure assay was performed on zebrafish for 30 days using oxytetracycline (Otc), sulfamethoxazole/trimethoprim (Smx/Tmp), or erythromycin (Ery), which are antibiotics widely used in the aquaculture industry. The microbiome analysis indicated that Fusobacteria, Proteobacteria, Firmicutes, and Bacteroidetes were the dominant phyla in the gut microbiome of the zebrafish used in this study. However, in Smx/Tmp-treated zebrafish, the compositions of Fusobacteria and Proteobacteria were changed significantly, and in Ery-treated zebrafish, the compositions of Proteobacteria and Firmicutes were altered significantly. Although alpha diversity analysis showed that there was no significant difference in the richness, beta diversity analysis revealed a community imbalance in the gut microbiome of all chronically antibiotics-exposed zebrafish. Intriguingly, in zebrafish with dysbiosis in the gut microbiome, the pathogen susceptibility to Edwardsiella piscicida, a representative Gram-negative fish pathogen, was reduced. Gut microbiome imbalance resulted in a higher count of goblet cells in intestinal tissue and an upregulation of genes related to the intestinal mucosal barrier. In addition, as innate immunity was enhanced by the increased mucosal barrier, immune and stress-related gene expression in the intestinal tissue was downregulated. In this study, we provide new insight into the effect of gut microbiome dysbiosis on pathogen susceptibility.

The 14-3-3 Protein BdGF14a Increases the Transcriptional Regulation Activity of BdbZIP62 to Confer Drought and Salt Resistance in Tobacco.

BdGF14a, a 14-3-3 gene from Brachypodium distachyon, induced by salt, H2O2, and abscisic acid (ABA), improved tolerance to drought and salt in tobacco, with a higher survival rate and longer roots under these stresses. Additionally, physiological index analyses showed that the heterologous expression of BdGF14a induced higher expression levels of antioxidant enzymes and their activities, leading to lighter DAB and NBT staining, denoting decreased H2O2 content. Additionally, the lower MDA content and ion leakage indicated enhanced cell membrane stability. Moreover, exogenous ABA resulted in shorter roots and a lower stomatal aperture in BdGF14a transgenic plants. BdGF14a interacted with NtABF2 and regulated the expression of stress-related genes. However, adding an ABA biosynthesis inhibitor suppressed most of these changes. Furthermore, similar salt and drought resistance phenotypes and physiological indicators were characterized in tobacco plants expressing BdbZIP62, an ABRE/ABF that interacts with BdGF14a. And Y1H and LUC assays showed that BdGF14a could enhance the transcription regulation activity of NtABF2 and BdbZIP62, targeting NtNECD1 by binding to the ABRE cis-element. Thus, BdGF14a confers resistance to drought and salinity through interaction with BdbZIP62 and enhances its transcriptional regulation activity via an ABA-mediated signaling pathway. Therefore, this work offers novel target genes for breeding salt- and drought-tolerant plants.

The effect of emulsifier type and oil inclusion on stress-related gene expression of Salmonella typhimurium in oil-in-water emulsion.

Salmonella has been associated with numerous outbreaks from contaminated food products, including emulsions. Emulsions are influenced by emulsifier type and oil presence, which can have varying degrees of stress or protection on bacteria. Although our previous research has shown that emulsifier solutions, rather than emulsions, provide a protective effect on Salmonella typhimurium after thermal treatment, the underlying mechanism remains unclear. This study selected S. typhimurium as the model microorganism and utilized the same emulsifiers (Tween 20, Tween 80, Triton X-100) to create emulsifier solutions and emulsions with the same oil fraction (60% (v/v)) to examine their effect on the expression of nine selected genes (rpoE, rpoH, otsB, proV, fadA, fabA, dnaK, ibpA, ompC) associated with stress response. Specifically, the study observed variations in gene expression under normal and thermal stress at 55°C. After 20-h incubation, Triton X-100 emulsion caused an upregulation of stress-related genes, rpoE, otsB, and fabA, suggesting stressful environment. After thermal treatment, S. typhimurium in Triton X-100 solution showed a longer 5-log reduction time with increased proV and decreased fabA and ompC expression, suggesting enhanced thermal protection compared to its emulsion. Conversely, Tween 80 solution increased fabA and ompC expression, indicating greater membrane fluidity and passive diffusion, potentially reducing thermal resistance. However, according to the upregulation of ibpA, this effect was likely mitigated by the overproduction of heat shock proteins. Notably, Triton X-100 environments exhibited the most significant gene expression changes after heat treatment, whereas Tween 80 without oil was the most inhospitable for bacterial survival. These findings inform bacterial responses under various conditions, aiding food safety strategies.

Stress-Related Gene Expression in Liver Tissues from Laying Hens Housed in Conventional Cage and Cage-Free Systems in the Tropics.

Global egg production is mainly based on cage systems, which have been associated with negative effects on the welfare of birds. Stress factors in restrictive production systems can lead to changes in gene transcription and protein synthesis, ultimately impacting the quality of poultry products. The liver serves various metabolic functions, such as glycogen storage, and plays a crucial role in animals' adaptation to environmental changes. Consequently, both internal and external conditions can influence liver functions. The aim of this study was to evaluate the gene expression of AGP, CRP, NOX4, SOD1, CAT, GPX1, SREBF1, and FXR in the liver of laying hens under two different production systems. Liver tissues from Hy-Line Brown hens housed in conventional cage and cage-free egg production systems at 60 and 80 weeks of production were used. mRNA transcript levels were determined by qPCR using the relative quantification method and ACTB as the reference gene. AGP, SOD1, and SREBF1 gene expressions were significantly higher in the conventional cage group at the 60 weeks of production. Furthermore, the mRNA levels of transcripts related to oxidative stress and lipid metabolism were higher in the group of laying hens housed in conventional cages compared to those in cage-free systems. These results suggest differential gene expression of genes related to oxidative stress in liver tissues from hens housed in conventional cages compared to cage-free systems. The conditions of the egg production system can impact the gene expression of oxidative stress and lipid synthesis genes, potentially leading to changes in the metabolism and performance of hens, including egg quality.

Responses and correlation among ER stress, Ca2+ homeostasis, and fatty acid metabolism in Penaeus vannamei under ammonia stress

The role of endoplasmic reticulum (ER) stress, Ca2+ homeostasis, and fatty acid metabolism in the environmental adaptation of aquatic animals is significant, but further confirmation of the relationship between these factors is needed. This study aimed to investigate the responses and correlations among ER stress, Ca2+ homeostasis, and fatty acid metabolism in Penaeus vannamei under ammonia stress. A total of 640 P. vannamei weighing 3.0 ± 0.4 g were selected and exposed to different total ammonia concentrations (0 mg/L for the control group and 3.80, 7.60, and 11.40 mg/L for the stress groups). The experiment involved a 96 h ammonia stress period to assess indicators related to ER stress, Ca2+ homeostasis, and fatty acid metabolism. The experimental results revealed that after 12 h, exposure to ammonia induced the ER stress response in the hepatopancreas of the shrimp. The groups exposed to concentrations of 3.8 mg/L and 7.6 mg/L exhibited an increase in ER Ca2+ efflux, a decrease in influx, an elevation in mitochondrial Ca2+ influx, an enhanced energy demand within the organism, and substantial consumption of triglycerides. The 11.3 mg/L group exhibited a significant enhancement in fatty acid metabolism. At 24 h, the ER stress response induced by ammonia in the shrimp exhibited a gradual recovery. In the 7.6 mg/L and 11.3 mg/L groups, the ER Ca2+ influx and efflux exhibited significant enhancements, while the mitochondrial Ca2+ influx decreased and the organism's energy demand increased. Moreover, there was a substantial enhancement in fatty acid metabolism. At 48 h, the ER stress response disappeared in each stress group, ER Ca2+ efflux was reduced, triglycerides were consumed, and the body's energy homeostasis was basically restored. At 96 h, a stress response reoccurred in the ER in each stress group, resulting in increased influx of Ca2+ into the ER, augmented energy demand within the organism, and notable enhancement in fatty acid metabolism. Pearson correlation analysis revealed a significant positive correlation between the NH3-N content in the hepatopancreas and the expression of ER stress-related genes, as well as between ER Ca2+ influx/efflux and energy homeostasis/fatty acid metabolism. The findings indicate that the stress induced by ammonia triggers an ER stress response in P. vannamei, resulting in ER Ca2+ efflux and mitochondrial Ca2+ influx, which, in turn, enhances fatty acid metabolism to generate additional energy for adaptation in stressful environments. This study contributes to a deeper understanding of the environmental adaptability of P. vannamei in the context of Ca2+ homeostasis.

Sub-optimal temperatures lead to altered expression of stress-related genes and increased 'Candidatus Liberibacter solanacearum' accumulation in potato psyllid.

The potato psyllid Bactericera cockerelli is the insect vector of the fastidious bacterium 'Candidatus Liberibacter solanacearum'. The bacterium infects both B. cockerelli and plant species, causing zebra chip (ZC) disease of potato and vein-greening disease of tomato. Temperatures are known to influence the initiation and progression of disease symptom in the host plant, and seasonal transitions from moderate to high temperatures trigger psyllid dispersal migration to facilitate survival. 'Ca. L. solanacearum' -infected and uninfected psyllids were reared at previously established 'permissible', optimal, and 'non-permissible' and temperatures of 18°C, 24°C, and 30°C, respectively. Gene expression profiles for 'Ca. L. solanacearum'-infected and -uninfected adult psyllids reared at different temperatures were characterized by Illumina RNA-Seq analysis. Bacterial genome copy number was quantified by real-time quantitative-PCR (qPCR) amplification. Relative gene expression profiles varied in psyllids reared at the three experimental temperatures. Psyllids reared at 18°C and 30°C exhibited greater fold-change increased expression of stress- and 'Ca. L. solanacearum' invasion-related proteins. Quantification by qPCR of bacterial genome copy number revealed that 'Ca. L. solanacearum' accumulation was significantly lower in psyllids reared at 18°C and 30°C, compared to 24°C. Temperature is a key factor in the life history of potato psyllid and multiplication/accumulation of 'Ca. L. solanacearum' in both the plant and psyllid host, influences the expression of genes associated with thermal stress tolerance, among others, and may have been instrumental in driving the co-evolution of the pathosystem.

5-Aminolevulinic acid treatment mitigates pesticide stress in bean seedlings by regulating stress-related gene expression and retrotransposon movements

Overdoses of pesticides lead to a decrease in the yield and quality of plants, such as beans. The unconscious use of deltamethrin, one of the synthetic insecticides, increases the amount of reactive oxygen species (ROS) by causing oxidative stress in plants. In this case, plants tolerate stress by activating the antioxidant defense mechanism and many genes. 5-Aminolevulinic acid (ALA) improves tolerance to stress by acting exogenously in low doses. There are many gene families that are effective in the regulation of this mechanism. In addition, one of the response mechanisms at the molecular level against environmental stressors in plants is retrotransposon movement. In this study, the expression levels of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), and stress-associated protein (SAP) genes were determined by Q-PCR in deltamethrin (0.5 ppm) and various doses (20, 40, and 80 mg/l) of ALA-treated bean seedlings. In addition, one of the response mechanisms at the molecular level against environmental stressors in plants is retrotransposon movement. It was determined that deltamethrin increased the expression of SOD (1.8-fold), GPX (1.4-fold), CAT (2.7-fold), and SAP (2.5-fold) genes, while 20 and 40 mg/l ALA gradually increased the expression of these genes at levels close to control, but 80 mg/l ALA increased the expression of these genes almost to the same level as deltamethrin (2.1-fold, 1.4-fold, 2.6-fold, and 2.6-fold in SOD, GPX, CAT, and SAP genes, respectively). In addition, retrotransposon-microsatellite amplified polymorphism (REMAP) was performed to determine the polymorphism caused by retrotransposon movements. While deltamethrin treatment has caused a decrease in genomic template stability (GTS) (27%), ALA treatments have prevented this decline. At doses of 20, 40, and 80 mg/L of ALA treatments, the GTS ratios were determined to be 96.8%, 74.6%, and 58.7%, respectively. Collectively, these findings demonstrated that ALA has the utility of alleviating pesticide stress effects on beans.

Evaluation of the LDN-0060609 PERK Inhibitor as a Selective Treatment for Primary Open-Angle Glaucoma: An In Vitro Study on Human Retinal Astrocytes.

The term glaucoma encompasses various neurodegenerative eye disorders, among which the most common is primary open-angle glaucoma (POAG). Recently, the essential role of human retinal astrocytes (HRA) in glaucoma progression has been placed in the spotlight. It has been found that placing the endoplasmic reticulum (ER) under stress and activating PERK leads to apoptosis of HRA cells, which inhibits their neuroprotective effect in the course of glaucoma. Therefore, the aim of the present study was to evaluate the effectiveness of the small-molecule PERK inhibitor LDN-0060609 in countering ER stress conditions induced in HRA cells in vitro. The activity of LDN-0060609 was studied in terms of protein and mRNA expression, cytotoxicity, genotoxicity, caspase-3 level and cell cycle progression. LDN-0060609 at 25 μM proved to be a potent inhibitor of the major PERK substrate, p-eIF2α (49% inhibition). The compound markedly decreased the expression of pro-apoptotic ER stress-related genes (ATF4, DDIT3, BAX and Bcl-2). Treatment with LDN-0060609 significantly increased cell viability, decreased genotoxicity and caspase-3 levels, and restored cell cycle distribution in HRA cells with activated ER stress conditions. These findings indicate that the small-molecule PERK inhibitor LDN-0060609 can potentially be developed into a novel anti-glaucoma agent.

Integrative transcriptomic and metabolomic analysis to elucidate the effect of gossypol on Enterobacter sp. GD5.

Gossypol, a yellow polyphenolic compound found in the Gossypium genus, is toxic to animals that ingest cotton-derived feed materials. However, ruminants display a notable tolerance to gossypol, attributed to the pivotal role of ruminal microorganisms in its degradation. The mechanisms of how rumen microorganisms degrade and tolerate gossypol remain unclear. Therefore, in this study, Enterobacter sp. GD5 was isolated from rumen fluid, and the effects of gossypol on its metabolism and gene expression were investigated using liquid chromatography-mass spectrometry (LC-MS) and RNA analyses. The LC-MS results revealed that gossypol significantly altered the metabolic profiles of 15 metabolites (eight upregulated and seven downregulated). The Kyoto Encyclopedia of Genes and Genomes analysis results showed that significantly different metabolites were associated with glutathione metabolism in both positive and negative ion modes, where gossypol significantly affected the biosynthesis of amino acids in the negative ion mode. Transcriptomic analysis indicated that gossypol significantly affected 132 genes (104 upregulated and 28 downregulated), with significant changes observed in the expression of catalase peroxidase, glutaredoxin-1, glutathione reductase, thioredoxin 2, thioredoxin reductase, and alkyl hydroperoxide reductase subunit F, which are related to antioxidative stress. Furthermore, Gene Ontology analysis revealed significant changes in homeostatic processes following gossypol supplementation. Overall, these results indicate that gossypol induces oxidative stress, resulting in the increased expression of antioxidative stress-related genes in Enterobacter sp. GD5, which may partially explain its tolerance to gossypol.

The Beneficial Effect of Brazilian Propolis for Liver Damage through Endoplasmic Reticulum Stress.

There is evidence that propolis exhibits anti-inflammatory, anticancer, and antioxidant properties. We assessed the potential beneficial effects of Brazilian propolis on liver injury in nonalcoholic fatty liver disease (NAFLD). Our findings demonstrate that Brazilian propolis suppresses inflammation and fibrosis in the liver of mice with NAFLD by inhibiting the expression of genes involved in endoplasmic reticulum (ER) stress. Additionally, Brazilian propolis also suppressed the expression of ER stress-related genes in HepG2 cells treated with an excess of free fatty acids, leading to cell apoptosis. A deeper analysis revealed that kaempferol, one of the components present in Brazilian propolis, induces cell proliferation through the mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) pathway and protects against oxidative stress. In conclusion, Brazilian propolis exhibits hepatoprotective properties against oxidative stress by inhibiting ER stress in NAFLD-induced model mice.

Endoplasmic reticulum stress-related gene expression causes the progression of dilated cardiomyopathy by inducing apoptosis.

Background: Previous studies have shown that endoplasmic reticulum stress (ERS) -induced apoptosis is involved in the pathogenesis of dilated cardiomyopathy (DCM). However, the molecular mechanism involved has not been fully characterized. Results: In total, eight genes were obtained at the intersection of 1,068 differentially expressed genes (DEGs) from differential expression analysis between DCM and healthy control (HC) samples, 320 module genes from weighted gene co-expression network analysis (WGCNA), and 2,009 endoplasmic reticulum stress (ERGs). These eight genes were found to be associated with immunity and angiogenesis. Four of these genes were related to apoptosis. The upregulation of MX1 may represent an autocompensatory response to DCM caused by a virus that inhibits viral RNA and DNA synthesis, while acting as an autoimmune antigen and inducing apoptosis. The upregulation of TESPA1 would lead to the dysfunction of calcium release from the endoplasmic reticulum. The upregulation of THBS4 would affect macrophage differentiation and apoptosis, consistent with inflammation and fibrosis of cardiomyocytes in DCM. The downregulation of MYH6 would lead to dysfunction of the sarcomere, further explaining cardiac remodeling in DCM. Moreover, the expression of genes affecting the immune micro-environment was significantly altered, including TGF-β family member. Analysis of the co-expression and competitive endogenous RNA (ceRNA) network identified XIST, which competitively binds seven target microRNAs (miRNAs) and regulates MX1 and THBS4 expression. Finally, bisphenol A and valproic acid were found to target MX1, MYH6, and THBS4. Conclusion: We have identified four ERS-related genes (MX1, MYH6, TESPA1, and THBS4) that are dysregulated in DCM and related to apoptosis. This finding should help deepen understanding of the role of endoplasmic reticulum stress-induced apoptosis in the development of DCM.

Analysis of stress-related gene expression in lobster seeds (Panulirus sp.) post formalin stress test

Indonesia is the potential international-scale lobster economic activity due to the abundant supply of tropical lobster seeds Panulirus sp., however the lobster cultivation sector have still resulted in problems of low survival during the rearing period. This study aimed to analyze the expression levels of the HSP70 gene related to stress response in lobster seeds exposed to formalin dose of LC70 for 1 h. Seeds were obtained from catching lobsters at Ujung Genteng Beach, Sukabumi City, West Java Province, with an average weight of 0.0254±0.004 g and a length of 2.3314±0.1549 cm. The observation parameters tested were the gene expression and survival rates of the post-soaking lobsters. The results showed that formalin immersion at a dose of 220 ppm on juvenile-stage lobster seeds resulted in the death of 70% of the population and the ability to express the stress gene HSP70 at different fitness levels were different with the highest expression level found in fit condition lobster seeds was 2.949±0.420.

Manganese induces oxidative damage in the hippocampus by regulating the expression of oxidative stress-related genes via modulation of H3K18 acetylation.

Prolonged exposure to manganese (Mn) contributes to hippocampal Mn accumulation, which leads to neurodegenerative diseases called manganese poisoning. However, the underlying molecular mechanisms remain unclear and there are no ideal biomarkers. Oxidative stress is the essential mechanisms of Mn-related neurotoxicity. Furthermore, histone acetylation has been identified as being engaged in the onset and development of neurodegenerative diseases. Therefore, the work aims to understand the molecular mechanisms of oxidative damage in the hippocampus due to Mn exposure from the aspect of histone acetylation modification and to assess whether H3K18 acetylation (H3K18ac) modification level in peripheral blood reflect Mn-induced oxidative damage in the hippocampus. Here, we randomly divided 60 male rats into four groups and injected them intraperitoneally with sterile pure water and MnCl2 ⋅4H2 O (5, 10, and 15 mg/kg) for 16 weeks, 5 days a week, once a day. The data confirmed that Mn exposure down-regulated superoxide dismutase activity and glutathione level as well as up-regulated malondialdehyde level in the hippocampus and plasma, and that there was a positive correlation between these indicators in the hippocampus and plasma. Besides, we noted that Mn treatment upregulated H3K18ac modification levels in the hippocampus and peripheral blood and that H3K18ac modification levels correlated with oxidative stress. Further studies demonstrated that Mn treatment decreased the amounts of H3K18ac enrichment in the manganese superoxide dismutase (SOD2) and glutathione transferase omega 1 (GSTO1) gene promoter regions, contributing to oxidative damage in the hippocampus. In short, our results demonstrate that Mn induces oxidative damage in the hippocampus by inhibiting the expression of SOD2 and GSTO1 genes via modulation of H3K18ac. In assessing Mn-induced hippocampal neurotoxicity, oxidative damage in plasma may reflect hippocampal oxidative damage in Mn-exposed groups.

Genome-wide identification of NAC transcription factors and regulation of monoterpenoid indole alkaloid biosynthesis in Catharanthus roseus.

NAC transcription factors (TFs) are crucial to growth and defense responses in plants. Though NACs have been characterized for their role in several plants, comprehensive information regarding their role in Catharanthus roseus, a perennial ornamental plant, is lacking. Homology modelling was employed to identify and characterize NACs in C. roseus. In-vitro propagation of C. roseus plants was carried out using cell suspension and nodal culture and were elicited with two auxin-antagonists, 5-fluoro Indole Acetic Acid (5-F-IAA) and α-(phenyl ethyl-2-oxo)-Indole-Acetic-Acid (PEO-IAA) for the enhanced production of monoterpenoid indole alkaloids (MIAs) namely catharanthine, vindoline, and vinblastine. Analyses revealed the presence of 47 putative CrNAC genes in the C. roseus genome, primarily localized in the nucleus. Phylogenetic analysis categorized these CrNACs into eight clusters, demonstrating the highest synteny with corresponding genes in Camptotheca acuminata. Additionally, at least one defense or hormone-responsive cis-acting element was identified in the promoter region of all the putative CrNACs. Of the two elicitors, 5-F-IAA was effective at 200 µM to elicit a 3.07-fold increase in catharanthine, 2.76-fold in vindoline, and 2.4-fold in vinblastine production in nodal culture. While a relatively lower increase in MIAs was recorded in suspension culture. Validation of RNA-Seq by qRT-PCR showed upregulated expression of stress-related genes (CrNAC-07 and CrNAC-24), and downregulated expression of growth-related gene (CrNAC-25) in elicited nodal culture of C. roseus. Additionally, the expression of genes involved in the biosynthesis of MIAs was significantly upregulated upon elicitation. The current study provides the first report on the role of CrNACs in regulating the biosynthesis of MIAs.

Effects of Clothianidin Pesticide Application on the Strength of Honey Bee Colonies and Stress-Related Genes in the Vicinity of Rice Fields in the Republic of Korea

The honey bee, a crucial organism that provides ecological and economic value to humans, is currently facing threats from various environmental factors including pesticides. Numerous studies have been conducted to demonstrate the risks associated with neonicotinoid pesticides, but research on their occurrence in actual field conditions has not been identified. Therefore, in this study, we observed changes in honey bee (Apis mellifera) colonies located near rice cultivation areas as they progressed beyond the rice pesticide application period. Furthermore, we collected honey bees exposed to the clothianidin and analyzed their stress-related gene expression. The results showed that the foraging behavior of honey bee colonies located near rice cultivation areas did not exhibit significant differences between the treatment sites (Cheongyang and Gimje) and the control site (Wanju) during the experimental period. However, it was observed that the expression levels of stress-related genes in honey bees collected from the treatment group were significantly higher than those in the control. Most of the stress-related genes were associated with detoxification processes in response to pesticides. As a result, pesticide treatment in proximity to rice cultivation areas did not cause direct damage to honey bees but had an indirect impact, suggesting the potential for ongoing chronic damage.

PagMYB73 enhances salt stress tolerance by regulating reactive oxygen species scavenging and osmotic maintenance in poplar

The MYB transcription factor (TF) family, widely recognized for its involvement in multiple biological processes, holds a significant position among plant TF families due to its substantial size. The R2R3-MYB TF has been extensively studied and known for its pivotal role in regulating the synthesis of anthocyanin, lignin, and secondary wall components in plants. However, there is currently limited information available regarding the function of R2R3-MYB genes in response to salt stress specifically in poplar. Here, we focused on a specific R2R3-MYB gene, PagMYB73, which belongs to the 84K poplar (Populus alba × P. glandulosa) species. We aimed to investigate the role of PagMYB73 in the regulatory mechanisms involved in salt stress response. PagMYB73 showed strong induction in leaves under salt stress conditions. Transgenic poplar plants overexpressing PagMYB73 exhibited enhanced salt tolerance, while suppression of the gene resulted in increased sensitivity to high salt levels. PagMYB73 can regulate the expression of several stress-related genes, including PagGSTU19, PagLEA51, PagSODs, PagPODs, and PagP5CS2. It achieves this by binding to the promoter regions of these genes that orchestrate adaptive responses to salt stress by modulating the osmotic balance and reducing the levels of reactive oxygen species (ROS). Indeed, overexpression of PagGSTU19 and PagLEA51 has been shown to enhance the salt tolerance of poplar. In conclusion, PagMYB73 regulates the expression of stress-related genes to improve ROS scavenging ability and osmotic maintenance, thereby improving the salt tolerance of the transgenic poplar.

Ulva prolifera Stress in the Yellow Sea of China: Suppressed Antioxidant Capacity and Induced Inflammatory Response of the Japanese Flounder (Paralichthys olivaceus).

As the largest green macroalgal bloom in the Yellow Sea of China, the overgrowth and degradation of Ulva prolifera (U. prolifera) have a harmful effect on marine organisms and the aquaculture industry. However, the regulation mechanism of U. prolifera stress on the antioxidant capacity and inflammatory response of marine fish is still not completely understood. A 15-day exposure experiment was conducted to evaluate the effects of U. prolifera stress on the antioxidant capacity and inflammatory response of the Japanese flounder (Paralichthys olivaceus) (283.11 ± 6.45 g). The results showed that U. prolifera stress significantly decreased their survival rate. Serum total antioxidant capacity (T-AOC) and non-specific immune-related enzyme activities were significantly impacted under U. prolifera conditions. Moreover, U. prolifera stress significantly decreased T-AOC, superoxide dismutase (SOD) activity, and catalase (CAT) activities in the liver, while malondialdehyde (MDA) contents were significantly increased. Similarly, antioxidant-related gene (cat, nrf2, and keap1) expressions were synchronously downregulated in the liver under U. prolifera stress. Furthermore, U. prolifera stress significantly upregulated pro-inflammatory gene (tnf-α, il-1β, ifn-γ, and p65) expressions and the phosphorylation levels of the p38 and JNK MAPK pathways in the head kidney. In addition, endoplasmic reticulum (ER) stress-related gene and protein expressions were also upregulated in the head kidney. Overall, these results revealed that U. prolifera stress suppressed the antioxidant capacity and induced an inflammatory response in the Japanese flounder. This study could advance the understanding of the adverse effects of U. prolifera stress on marine benthic fish and promote the sustainable development of aquaculture.

Metabolites of Trichoderma harzianum re-balance T-reg/Th-17 cytokine axis in epileptic rats

BackgroundRecurrent seizure is synchronous neuronal network hyperexcitation. Even though many anti-epileptic drugs (AEDs) are available, but have several drawbacks, including multiple drug interactions, suboptimal response rates, significant adverse effects, and a narrow therapeutic index. Trichoderma is a biocontrol agent which maintains stress-related gene expression to adapt to unstable environmental conditions. The current study sought to delineate the ameliorative effects of metabolites of Trichoderma harzianum (T. harzianum) culture filtrate (ThCF) by assessing the functions of T regulatory cells and rebalancing oxidative stress.MethodsExperimental rats were divided into control, epileptic, valproic acid-treated, and T. harzianum cultured filtrate (ThCF). Lipid peroxidation, nitric oxide, and antioxidant defense enzymes were estimated. Moreover, interleukins-6, -10, -17, tumor necrosis factor (TNF-α), and transforming growth factor (TGF-β) were estimated using ELISA kits, in addition to T-reg markers; cytotoxic T-lymphocyte-associated protein 4 (CTLA4), and forkhead box P3 (FOXP3) were estimated by qRT-PCR.ResultsData revealed that the T. harzianum cultured filtrate (ThCF) retarded the lipid oxidation rate and has antioxidant activities, as well as increased levels of GPx, CAT, and SOD. Moreover, ThCF re-balances T-reg/Th-17 cytokines, restoring the pro/anti-inflammatory cytokines and Treg markers, e.g., FOXP3 and CTLA-4, to their normal level.ConclusionTrichoderma harzianum has a potent antioxidant activity with high capacity to scavenge ROS and downregulate all T-reg markers. Therefore, the present data are directed toward the characterization of new active constituents of secondary metabolites of T. harzianum with significant therapeutic functions in several diseases. The promising findings may suggest more clinical and experimental scenarios to reduce AED’s drawbacks and side effects.