Regulatory Mechanisms Research Articles

Knockdown of Fzr inhibited the growth of Nilaparvata lugens by blocking endocycle.

The endocycle can generate cells referred to as 'polyploid'. Fizzy-related protein (Fzr) plays an important role in driving the mitosis-to-endocycle transition. The brown planthopper (BPH), Nilaparvata lugens (Stål), a serious insect pest, feeds exclusively on rice. However, polyploidy and its regulatory mechanisms are poorly understood in BPH. Here, we found that the ploidy levels of follicles H (FH) and accessory gland (AG) significantly increased with BPH age when examining the polyploidy of FH and AG of salivary glands. Fzr was identified as an important regulator for polyploidy in BPH salivary gland. Knockdown of Fzr resulted in a decrease in cell size and DNA content in nymph salivary glands. Fzr knockdown transcriptionally upregulated cyclin-dependent kinase 1 (CDK1), CDK2, cyclin A (CycA) and CycB, and downregulated CycD, CycE, Myc and mini-chromosome maintenance protein 2-7 (MCM2-7). Phenotypically, Fzr knockdown significantly suppressed salivary protein production, feeding and survival in BPH nymphs. Our results show that BPH salivary glands exhibit obvious polyploidy, and Fzr positively regulates the endocycle in nymph salivary gland. These findings provide clues for the study of the regulatory mechanisms of insect polyploidy. © 2024 Society of Chemical Industry.

Modulating Polymerase Activity through Light-Oxygen-Voltage Domain Insertion.

Biochemical reaction networks adapt to environmental conditions by sensing chemical or physical stimuli and using tightly controlled mechanisms. While most signals come from molecules, many cells can also sense and respond to light. Among the biomolecular structures that enable light sensing, we selected a light-oxygen-voltage (LOV) domain in a previous study that tested the engineering of novel regulatory mechanisms into a nucleic acid polymerase. In this follow-up study, we studied the activities of previously selected variants in kinetic detail, and we generated additional LOV-polymerase fusion variants based on further insertion criteria. Our results provide mechanistic insights into how LOV domain insertion influences polymerase activity in a light-responsive manner: All active and photoresponsive enzyme variants studied by us to date were partially inhibited (i.e., "turned off") after irradiation with blue light at 470 nm, which can be explained by specific obstructions of the polymerase entry or exit structures (substrate entry channels or product exit channels, or both). Although the effects observed are moderate, we anticipate further engineering strategies that could be used to improve the extent of switchability and possibly to develop a "turn-on mode" insertion.

Cytokinin-responsive RhRR1-RhSCL28 transcription factor module positively regulates petal size by promoting cell division in rose.

Petal size, a crucial trait in the economically important ornamental rose (Rosa hybrida), is synergistically regulated by cell division and cell expansion. Cell division primarily occurs during the early development of petals. However, the molecular mechanism underlying the regulation of petal size is far from clear. In this study, we isolated the transcription factor gene RhSCL28, which is highly expressed at the early stage of rose petal development and is induced by cytokinin. Silencing RhSCL28 resulted in a reduced final petal size and reduced cell number in rose petals. Further analysis showed that RhSCL28 participates in the regulation of cell division by positively regulating the expression of the cyclin genes RhCYCA1;1 and RhCYCB1;2. To explore the potential mechanism for cytokinin-mediated regulation of RhSCL28 expression, we investigated the cytokinin response factor RhRR1 and determined that it positively regulates RhSCL28 expression. Like RhSCL28, silencing RhRR1 also resulted in smaller petals by decreasing cell number. Taken together, these results reveal that the RhRR1-RhSCL28 module positively regulates petal size by promoting cell division in rose.

Inflammatory Signaling via PEIZO1 Engages and Enhances the LPS-Mediated Apoptosis during Clinical Mastitis.

Bovine clinical mastitis is characterized by inflammation and immune responses, with apoptosis of mammary epithelial cells as a cellular reaction to infection. PIEZO1, identified as a mechanotransduction effector channel in nonruminant animals and sensitive to both mechanical stimuli or inflammatory signals like lipopolysaccharide (LPS). However, its role in inflammatory processes in cattle has not been well-documented. The aim of this study was to elucidate the in situ expression of PIEZO1 in bovine mammary gland and its potential involvement in clinical mastitis. We observed widespread distribution and upregulation of PIEZO1 in mammary epithelial cells in clinical mastitis cows and LPS-induced mouse models, indicating a conserved role across species. In vitro studies using mammary epithelial cells (MAC-T) revealed that LPS upregulates PIEZO1. Notably, the effects of PIEZO1 artificial activator Yoda1 increased apoptosis and NLRP3 expression, effects mitigated by PIEZO1 silencing or NLRP3 inhibition. In conclusion, the activation of the PIEZO1-NLRP3 pathway induces abnormal apoptosis in mammary epithelial cells, potentially serving as a regulatory mechanism to combat inflammatory responses to abnormal stimuli.

Super-resolution microscopy to study membrane nanodomains and transport mechanisms in the plasma membrane

Biological membranes are complex, heterogeneous, and dynamic systems that play roles in the compartmentalization and protection of cells from the environment. It is still a challenge to elucidate kinetics and real-time transport routes for molecules through biological membranes in live cells. Currently, by developing and employing super-resolution microscopy; increasing evidence indicates channels and transporter nano-organization and dynamics within membranes play an important role in these regulatory mechanisms. Here we review recent advances and discuss the major advantages and disadvantages of using super-resolution microscopy to investigate protein organization and transport within plasma membranes.

Palmitoyltransferase ZDHHC3 is essential for the oncogenic activity of PML/RARα in acute promyelocytic leukemia.

The oncogenic fusion protein promyelocytic leukemia/retinoic acid receptor alpha (PML/RARα) is critical for acute promyelocytic leukemia (APL). PML/RARα initiates APL by blocking the differentiation and increasing the self-renewal of leukemic cells. The standard clinical therapies all-trans retinoic acid (ATRA) and arsenic trioxide (ATO), which induce PML/RARα proteolysis, have dramatically improved the prognosis of APL patients. However, the emergence of mutations conferring resistance to ATRA and ATO has created challenges in the treatment of APL patients. Exploring pathways that modulate the oncogenic activity of PML/RARα could help develop novel therapeutic strategies for APL, particularly for drug-resistant APL. Herein, we demonstrated for the first time that palmitoylation of PML/RARα was a critical determinant of its oncogenic activity. PML/RARα palmitoylation was found to be catalyzed mainly by the palmitoyltransferase ZDHHC3. Mechanistically, ZDHHC3-mediated palmitoylation regulated the oncogenic transcriptional activity of PML/RARα and APL pathogenesis. The knockdown or overexpression of ZDHHC3 had respective effects on the expression of proliferation- and differentiation-related genes. Consistently, the depletion or inhibition of ZDHHC3 could significantly arrest the malignant progression of APL, particularly drug-resistant APL, whereas ZDHHC3 overexpression appeared to have a promoting effect on the malignant progression of APL. Thus, our study not only reveals palmitoylation as a novel regulatory mechanism that modulates PML/RARα oncogenic activity but also identifies ZDHHC3 as a potential therapeutic target for APL, including drug-resistant APL.

Structural and functional mechanisms of anti-NMDAR autoimmune encephalitis.

Autoantibodies against neuronal membrane proteins can manifest in autoimmune encephalitis, inducing seizures, cognitive dysfunction and psychosis. Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is the most dominant autoimmune encephalitis; however, insights into how autoantibodies recognize and alter receptor functions remain limited. Here we determined structures of human and rat NMDARs bound to three distinct patient-derived antibodies using single-particle electron cryo-microscopy. These antibodies bind different regions within the amino-terminal domain of the GluN1 subunit. Through electrophysiology, we show that all three autoantibodies acutely and directly reduced NMDAR channel functions in primary neurons. Antibodies show different stoichiometry of binding and antibody-receptor complex formation, which in one antibody, 003-102, also results in reduced synaptic localization of NMDARs. These studies demonstrate mechanisms of diverse epitope recognition and direct channel regulation of anti-NMDAR autoantibodies underlying autoimmune encephalitis.

Cannabidiol enhances Atezolizumab efficacy by upregulating PD-L1 expression via the cGAS-STING pathway in triple-negative breast cancer cells.

The treatment of patients with triple negative breast cancer (TNBC) relies on cytotoxic therapy. Currently, atezolizumab and chemotherapy can be combined in patients with TNBC. However, this approach is not effective for all patients with low reactivity to atezolizumab. As there is a lack of alternative treatment options, new anti-cancer drugs are urgently needed to enhance atezolizumab reactivity against TNBC. Recent strategies have focused on regulating the expression of programmed death-ligand 1 (PD-L1) or enhancing immune response activation by combining anti-cancer drugs with immune checkpoint inhibitors (ICIs). Cannabidiol (CBD), a cannabinoid component derived from the cannabis plant, has been reported to have anti-cancer therapeutic potential because of its capacity to induce apoptotic cell death in tumor cells while avoiding cytotoxicity in normal cells. Previous studies have demonstrated the effects of CBD on apoptosis in various cancer cell types. However, the potential role of CBD as an immune modulator in the regulation of PD-L1 expression and anti-cancer immune responses remains to be explored. In this study, we found that CBD stimulated PD-L1 expression in TNBC cells, which significantly induced the CBD-mediated cGAS-STING pathway activation. Taken together, we demonstrated that the combination of CBD and anti-PD-L1 antibody enhances the anti-cancer immune response in vitro and in vivo experiments. Our findings identified the mechanism of PD-L1 regulation by CBD in TNBC cells and suggested that CBD could be a potential candidate for the development of new combinatorial strategies with ICIs in TNBC patients.

‘Now they move you, now they don’t’: transferability, territory and (im)mobilities at the Spanish peripheries

ABSTRACT The analytical emphasis on migration control and external borders has overlooked the mechanisms of state regulation of internal mobility in the peripheral territories of the countries of the European Union. Based on an analysis focused on the Spanish case, and more specifically on the enclaves of Ceuta and Melilla and the Canary Islands, this article introduces the concept of transferability to show the logics and dynamics of intraterritorial migrant (im)mobility between the peripheral enclaves and mainland Spain. The analysis shows the construction – and evolution – of (in)transferable migrant subjects and the complex and changing puzzle of (im)mobility at these peripheral geographies.

Screening of sex-special genes in common long-arm octopus Octopus minor based on gonadal transcriptome sequencing

The common long-arm octopus (Octopus minor) is a commercially important aquaculture species in East Asia, and the male octopus grows faster than the female ones, while the information about sex-regulating mechanisms in octopuses is limited. Therefore, gonadal transcriptome sequencing was performed in O. minor to reveal the molecular mechanisms of sex regulation in cephalopods. Based on the sexuality and gonad development stage, 4 groups of 12 gonad tissues were sampled, and 11 libraries were retained for bioinformatics analysis. A total of 263,749,727 clean reads were obtained. The percentage of clean reads mapped to the O. minor reference genome ranged from 77.20% to 90.59%. A total of 3936 differentially expressed genes (DEGs) were obtained between ovarian and testicular libraries by Venn diagram analysis, including 855 ovarian up-regulated and 3081 testicular up-regulated genes. Four unigenes (transcription factor Sox-8, fermentation-1, forkhead box protein L2, and ribosomal protein large 24) and one pathway (ECM-receptor interaction) were screened to be candidate molecular markers for sex determination. The reliability and accuracy of our analysis were validated via quantitative real-time PCR in 10 randomly selected DEGs. The results of our study enhanced our understanding of the molecular mechanism underlying sex determination and potentially helped to screen the bio-markers for O. minor in different sexes.

Whole-genome analysis of plasma fibrinogen reveals population-differentiated genetic regulators with putative liver roles.

Genetic studies have identified numerous regions associated with plasma fibrinogen levels in Europeans, yet missing heritability and limited inclusion of non-Europeans necessitates further studies with improved power and sensitivity. Compared with array-based genotyping, whole genome sequencing (WGS) data provides better coverage of the genome and better representation of non-European variants. To better understand the genetic landscape regulating plasma fibrinogen levels, we meta-analyzed WGS data from the NHLBI's Trans-Omics for Precision Medicine (TOPMed) program (n=32,572), with array-based genotype data from the Cohorts for Heart and Aging Research in Genomic Epidemiology (CHARGE) Consortium (n=131,340) imputed to the TOPMed or Haplotype Reference Consortium panel. We identified 18 loci that have not been identified in prior genetic studies of fibrinogen. Of these, four are driven by common variants of small effect with reported MAF at least 10 percentage points higher in African populations. Three signals (SERPINA1, ZFP36L2, and TLR10) contain predicted deleterious missense variants. Two loci, SOCS3 and HPN, each harbor two conditionally distinct, non-coding variants. The gene region encoding the fibrinogen protein chain subunits (FGG;FGB;FGA), contains 7 distinct signals, including one novel signal driven by rs28577061, a variant common in African ancestry populations but extremely rare in Europeans (MAFAFR=0.180; MAFEUR=0.008). Through phenome-wide association studies in the VA Million Veteran Program, we found associations between fibrinogen polygenic risk scores and thrombotic and inflammatory disease phenotypes, including an association with gout. Our findings demonstrate the utility of WGS to augment genetic discovery in diverse populations and offer new insights for putative mechanisms of fibrinogen regulation.

Peculiarities of the Ontogenesis of Bacilli During Development from a Vegetative Cell to a Spore

Understanding the development processes of bacteria, in particular spore-forming ones, has both fundamental and applied importance, since at various stages of this process, the cells of microorganisms perform certain functions that can be regulated by influencing certain factors depending on the tasks. Literature data and the results of our research on the influence of the composition of the nutrient medium, pH, temperature, and aeration on sporulation are analyzed in the article. It isshown that the direction of bacterial cells’ development in certain ways is determined by signals that come from the surrounding environment, affect their genome, and determine the ways of cell development – growth or sporulation. Sporogenesis can also be induced by metabolites formed during microorganism development. It is emphasized that the environmental factors that influence the sporulation of bacteria have been studied in sufficient detail. However, the mechanisms of their action remain debatable. Morphological, genetic, and biochemical changes of spore-forming bacteria under the conditions of macrocyclic and microcyclic ways of their development are also highlighted, which makes it possible to correctly understand the functioning of regulatory mechanisms in the ontogenesis of microbial cells. In particular, the data of our research on the dynamics of morphological changes in the ontogenesis of a specific individual bacterial cell are presented. In addition, factors, including specific terminal products of cell metabolism, such as antibiotics, and genetic mechanisms of sporogenesis regulation of various genera and species of bacteria are described in detail. The nature of the vast majority of "sporogenes" has not been clarified, and there are only a few hypotheses regarding the mechanism of their action. However, most of the biological regulators of sporogenesis were found in the culture liquid, which indicates the cellular nature of their action. Therefore, to obtain more convincing data on the regulation of sporogenesis, studies at the cellular level are needed.

Phosphorylation of the DNA damage repair factor 53BP1 by ATM kinase controls neurodevelopmental programs in cortical brain organoids.

53BP1 is a well-established DNA damage repair factor that has recently emerged to critically regulate gene expression for tumor suppression and neural development. However, its precise function and regulatory mechanisms remain unclear. Here, we showed that phosphorylation of 53BP1 at serine 25 by ATM is required for neural progenitor cell proliferation and neuronal differentiation in cortical brain organoids. Dynamic phosphorylation of 53BP1-serine 25 controls 53BP1 target genes governing neuronal differentiation and function, cellular response to stress, and apoptosis. Mechanistically, ATM and RNF168 govern 53BP1's binding to gene loci to directly affect gene regulation, especially at genes for neuronal differentiation and maturation. 53BP1 serine 25 phosphorylation effectively impedes its binding to bivalent or H3K27me3-occupied promoters, especially at genes regulating H3K4 methylation, neuronal functions, and cell proliferation. Beyond 53BP1, ATM-dependent phosphorylation displays wide-ranging effects, regulating factors in neuronal differentiation, cytoskeleton, p53 regulation, as well as key signaling pathways such as ATM, BDNF, and WNT during cortical organoid differentiation. Together, our data suggest that the interplay between 53BP1 and ATM orchestrates essential genetic programs for cell morphogenesis, tissue organization, and developmental pathways crucial for human cortical development.

Integrative analysis of transcriptome and metabolome reveals how ethylene increases natural rubber yield in Hevea brasiliensis

Hevea brasiliensis is an important cash crop with the product named natural rubber (NR) for markets. Ethylene (ET) is the most effective yield stimulant in NR production but the molecular mechanism remains incomplete. Here, latex properties analysis, transcriptome analysis, and metabolic profiling were performed to investigate the mechanism of NR yield increase in four consecutive tappings after ET stimulation. The results revealed that sucrose and inorganic phosphate content correlated positively with dry-rubber yield and were induced upon ET stimulation. Stimulation with ET also led to significant changes in gene expression and metabolite content. Genes involved in phytohormone biosynthesis and general signal transduction as well as 51 transcription factors potentially involved in the ET response were also identified. Additionally, KEGG annotation of differentially accumulated metabolites suggested that metabolites involved in secondary metabolites, amino-acid biosynthesis, ABC transporters, and galactose metabolism were accumulated in response to ET. Integrative analysis of the data collected by transcriptomics and metabolomics identified those differentially expressed genes and differentially accumulated metabolites are mainly involved in amino-acid biosynthesis and carbohydrate metabolism. Correlation analysis of genes and metabolites showed a strong correlation between amino-acid biosynthesis during ET stimulation. These findings provide new insights into the molecular mechanism underlying the ET-induced increase in rubber yield and further our understanding of the regulatory mechanism of ethylene signaling in rubber biosynthesis.

New Insights into the Persistent Shock Resistance of Anaerobic Granular Sludge Based on Quorum Sensing Regulation: A Novel Gene Regulatory Mechanism

New Insights into the Persistent Shock Resistance of Anaerobic Granular Sludge Based on Quorum Sensing Regulation: A Novel Gene Regulatory Mechanism

R2R3 MYB Transcription Factor GhMYB201 Promotes Cotton Fiber Elongation via Cell Wall Loosening and Very-Long-Chain Fatty Acid Synthesis

Cotton fiber is the leading natural textile material, and fiber elongation plays an essential role in the formation of cotton yield and quality. Although a number of components in the molecular network controlling cotton fiber elongation have been reported, a lot of players still need to be functionally dissected to understand the regulatory mechanism of fiber elongation comprehensively. In the present study, an R2R3-MYB transcription factor gene, GhMYB201, was characterized and functionally verified via CRISPR/Cas9-mediated gene editing. GhMYB201 was homologous to Arabidopsis AtMYB60, and both coding genes (GhMYB201At and GhMYB201Dt) were preferentially expressed in elongating cotton fibers. Knocking-out of GhMYB201 significantly reduced the rate and duration of fiber elongation, resulting in shorter and coarser mature fibers. It was found that GhMYB201 could bind and activate the transcription of cell wall loosening genes (GhRDLs) and also β-ketoacyl-CoA synthase genes (GhKCSs) to enhance very-long-chain fatty acid (VLCFA) levels in elongating fibers. Taken together, our data demonstrated that the transcription factor GhMYB201s plays an essential role in promoting fiber elongation via activating genes related to cell wall loosening and VLCFA biosynthesis.

New insights into the dynamics of m6A epitranscriptome: hybrid-seq identifies novel mRNAs of the m6A writers METTL3/14.

Background: N6-methyladenosine (m6A), a prevalent mRNA modification, is dynamically regulated by methyltransferases, including METTL3 and METTL14.Materials & methods: In the current study, we employed a custom hybrid-seq method to identify novel METTL3/14 transcripts, explore their protein-coding capacities and predict the putative role of the METTL isoforms.Results: Demultiplexing of the hybrid-seq barcoded datasets unraveled the expression patterns of the newly identified mRNAs in major malignancies as well as in non-malignant cells, providing a deeper understanding of the methylation pathways. Open reading framequery revealed novel METTL3/14 isoforms, broadening our perspective for the structural diversity within METTL family.Conclusion: Our findings offer significant insights into the intricate transcriptional landscape of METTL3/14, shedding light on the regulatory mechanisms underlying methylation in mRNAs.

Blood pressure variability in elderly and senior hypertensive patients

Background. In recent years the deterioration of the demographic situation has been noted in most countries of the world that is due to the steady increase in population of arterial hypertension and the aging of the population. According to the statistical forecast of the UN by 2025, the number of people over 60 will exceed 1 billion which is 15% of the entire global population. In Ukraine, according to epidemiological studies, the prevalence of hypertension among circulatory system diseases in adults (18 years and older) is 46.8% and almost half of patients with circulatory system diseases have elevated blood pressure. In patients aged 65 years and older, the prevalence of hypertension varies in the range of 53-72%. It has long been proven that the risk of cardiovascular and cerebrovascular complications in hypertensive patients depends not only on the absolute blood pressure level but also on fluctuations in blood pressure over different periods that is blood pressure variability. Blood pressure is not a static parameter but rather undergoes continuous fluctuations over time due to the interaction between environmental factors and behaviour on the one hand and the internal regulatory mechanisms of the cardiovascular system on the other hand. Elevated blood pressure may indicate cardiovascular dysregulation and itself may be a cardiovascular risk factor associated with increased all-cause mortality and cardiovascular mortality, stroke, coronary artery disease, heart failure, end-stage renal disease and incidence of dementia. Aim. The study aimed to improve the system of prevention and diagnosis of elderly and senior hypertensive patients at the ambulatory-polyclinic stage due to the study of the contribution of blood pressure variability. Materials and methods. The group of examinees was formed taking into account 27 elderly and senile hypertensive patients including those combined with coronary artery disease. For this purpose, ambulatory blood pressure monitoring was used. Results. Systolic blood pressure variability (SD) in hypertensive patients was higher than in the control group (p<0.001). So, during the study, SD was 17.9±7.1 mm Hg. while in the control group 12.1±2.6 mm Hg. The frequency of high blood pressure variability detection in elderly and senile hypertensive patients was 51.9% (n=14). High blood pressure variability compared to low blood pressure variability and control was associated with a more pronounced systolic blood pressure variability (during the active monitoring period), and it was equalled 23.7±5.2 mm Hg compared to 14.8±6.5 mm Hg (p<0.001) and 12.1±2.6 mm Hg. (p<0.001). In elderly and senile hypertensive patients with high and low blood pressure variability, the non-dipper group prevailed over the dipper group in the structure of daily blood pressure rhythm (71.4% vs. 28.6% and 84.6% vs. 15.4% respectively). According to the ambulatory blood pressure monitoring data high blood pressure variability group compared to low blood pressure variability and controls was characterised by higher average values of the number of blood pressure indicators. In the group of high blood pressure variability patients, the average daily systolic blood pressure exceeded the control value by 10% (р<0.05). The daily index of systolic blood pressure in the group of high blood pressure variability patients was 111% higher than in patients with low blood pressure variability (p<0.05). The daily index of diastolic blood pressure in the group of high blood pressure variability patients was 140% higher than in patients with low blood pressure variability (p<0.05). The daily index of average blood pressure in the group of high blood pressure variability patients was 191% higher than in patients with low blood pressure variability (p<0.01). Conclusions. 1. High blood pressure variability has occurred in 16 out of 27 (51.9%) hypertensive elderly and senile patients. 2. The non-dipper group has predominated in the structure of blood pressure diurnal rhythm in elderly and senile hypertensive patients with high and low blood pressure variability patients. _________________________________________________________________________________________ Keywords: arterial hypertension; essential hypertension; blood pressure variability; high blood pressure variability; low blood pressure variability; elderly and senile hypertensive patients

Pulse EPR Methods in the Angstrom to Nanometre Scale Shed Light on the Conformational Flexibility of a Fluoride Riboswitch.

Riboswitches control gene regulation upon external stimuli such as environmental factors or ligand binding. The fluoride sensing riboswitch from Thermotoga petrophila is a complex regulatory RNA proposed to be involved in resistance to F- cytotoxicity. The details of structure and dynamics underpinning the regulatory mechanism are currently debated. Here we demonstrate that a combination of pulsed electron paramagnetic resonance (ESR/EPR) spectroscopies, detecting distances in the angstrom to nanometre range, can probe distinct regions of conformational flexibility in this riboswitch. PELDOR (pulsed electron-electron double resonance) revealed a similar preorganisation of the sensing domain in three forms, i.e. the free aptamer, the Mg2+-bound apo, and the F--bound holo form. 19F ENDOR (electron-nuclear double resonance) was used to investigate the active site structure of the F--bound holo form. Distance distributions without a priori structural information were compared with in silico modelling of spin label conformations based on the crystal structure. While PELDOR, probing the periphery of the RNA fold, revealed conformational flexibility of the RNA backbone, ENDOR indicated low structural heterogeneity at the ligand binding site. Overall, the combination of PELDOR and ENDOR with sub-angstrom precision gave insight into structural organisation and flexibility of a riboswitch, not easily attainable by other biophysical techniques.

Exogenous Melatonin Alleviates Osmotic Stress by Enhancing Antioxidant Metabolism, Photosynthetic Maintenance, and Hormone Homeostasis in Forage Oat (Avena sativa) Seedlings

Melatonin (MT) is a multifunctional hormone that enhances crop resilience against various abiotic stresses. However, its regulatory mechanism of osmotic tolerance in forage oats (Avena sativa) plants under water-limited scenarios is still unclear. This study aimed to delineate the impact of MT pretreatment on the morphological, physiological, and biochemical functions of oat seedlings under osmotic stress. Our findings demonstrated that exogenous treatment of MT noticeably elevated leaf area while decreasing the root/shoot ratio of oat seedlings subjected to osmotic stress. Osmotic-induced 38.22% or 48.37% decrease in relative water content could be significantly alleviated by MT pretreatment on day 7 or day 14, respectively. MT treatment also significantly mitigated osmotic-induced decreases in photosynthetic parameters including net photosynthetic rate, stomatic conductance, and intercellular CO2 concentration as well as various chlorophyll fluorescence parameters, which could contribute to enhanced accumulations of free proline and soluble sugars in seedlings after being subjected to a prolonged duration of osmotic stress. Furthermore, MT markedly improved antioxidant enzyme activities including superoxide dismutase, ascorbate peroxidase, catalase, and peroxidase along with the accumulation of ascorbic acid contributing to a significant reduction in reactive oxygen species under osmotic stress. In addition, the MT application induced a 978.12%, 33.54%, or 30.59% increase in endogenous MT, indole acetic acid, or gibberellic acid content under osmotic stress but did not affect the accumulation of abscisic acid. These findings suggest that an optimal concentration of MT (100 μmol·L−1) could relieve osmotic stress via improvement in osmotic adjustment, the enzymatic antioxidant defense system, and endogenous hormonal balance, thereby contributing to enhanced photosynthetic functions and growth of oat seedlings under water-limited conditions.