External Environmental Stress Research Articles

Characterization of Optical, Thermal, and Viscoelastic Properties of Pollenkitt in Angiosperm Pollen Using In-Line Digital Holographic Microscopy.

Pollen grains are remarkable material composites, with various organelles in their fragile interior protected by a strong shell made of sporopollenin. The outermost layer of angiosperm pollen grains contains a lipid-rich substance called pollenkitt, which is a natural bioadhesive that helps preserve structural integrity when the pollen grain is exposed to external environmental stresses. In addition, its viscous nature enables it to adhere to various floral and insect surfaces, facilitating the pollination process. To examine the physicochemical properties of aqueous pollenkitt droplets, we used in-line digital holographic microscopy to capture light scattering from individual pollenkitt particles. Comparison of pollenkitt holograms to those modeled using the Lorenz-Mie theory enables investigations into the minute variations in the refractive index and size resulting from changes in local temperature and pollen aging.

Microbial exopolysaccharide composites with inorganic materials and their biomedical applications: A review

Microbial exopolysaccharides (EPS) are high molecular weight carbohydrate-based materials produced and secreted in response to external environmental stress to protect microbial cells. These molecules also contain non-carbohydrate moieties including acetate, pyruvate, phosphate, and succinate along with sugar molecules. EPS has been incorporated into industrial operations as a gelling, emulsifying, and stabilizing agent to increase viscosity. Besides, these molecules have also shown high biocompatibility, immunomodulatory, antitumor, anti-microbial, and anti-viral activity which suggest their possible applications in healthcare. However, these molecules are facing some commercial shortcomings due to low stability, bioactivity, and poor mechanical and tensile strength in comparison to the required standard. Hence use of composites of EPS with other polymers and moieties has been suggested for higher performance. Especially in the context of inorganic moieties like silver, gold nanomaterials, clay, salts, and minerals will add bioactivity and functional sites for the attachment of drug molecules and other compounds. The current review summarizes the higher physical and chemical characteristics of EPS composites with inorganic materials and their immense potential in the biomedical sector.

Rhizobial oxidized 3-hydroxylbutanoyl glycan-based gelatin hydrogels with enhanced physiochemical properties for pH-responsive drug delivery

Rhizobial exopolysaccharide (EPS) is an acidic polysaccharide involved in nitrogen fixation-related signal transduction in the rhizosphere, serving as a structural support for biofilms, and protecting against various external environmental stresses. Rhizobial EPS as a hydrogel biomaterial was used for a pH-responsive drug delivery system combing with gelatins. Pure gelatin (GA) hydrogels have limited practical applications due to their poor mechanical strength and poor thermal stability. We developed new GA hydrogels using oxidized 3-hydroxylbutanoyl glycan (OHbG) as a polymer cross-linking agent to overcome these limitations. OHbG was synthesized from sodium periodate oxidation of 3-hydroxylbutanoyl glycan directly isolated from Rhizobium leguminosarum bv. viciae VF39. The newly fabricated OHbG/GA hydrogels exhibited 21-fold higher compressive stress and 4.7-fold higher storage modulus (G') than GA at the same strain. This result suggested that OHbG provided mechanical improvement. In addition, these OHbG/GA hydrogels showed effective pH-controlled drug release for 5-fluorouracil, self-healable, and self-antioxidant capacity by uronic acids of OHbG. Cell viability tests using HEK-293 cells in vitro also showed that the OHbG/GA hydrogels were non-toxic. This suggests that the new OHbG/GA hydrogels can be used as a potentially novel biomaterial for drug delivery based on its self-healing ability, antioxidant capacity, and pH-responsive drug delivery.

Can "Risk-Sharing" Mechanisms Help Clonal Aquatic Plants Mitigate the Stress of Nanoplastics?

Most aquatic plants applied to ecological restoration have demonstrated a clonal growth pattern. The risk-spreading strategy plays a crucial role in facilitating clonal plant growth under external environmental stresses via clonal integration. However, the effects of different concentrations of nanoplastics (NPs) on the growth traits of clonal aquatic plants are not well understood. Therefore, this study aimed to investigate the impact of NPs exposure on seedlings of parent plants and connected offspring ramets. A dose response experiment (0.1, 1, and 10 mg L-1) showed that the growth of Eichhornia crassipes (water hyacinth) was affected by 100 nm polystyrene nanoplastics after 28 days of exposure. Tracer analysis revealed that NPs are accumulated by parent plants and transferred to offspring ramets through stolon. Quantification analysis showed that when the parent plant was exposed to 10 mg L-1 NPs alone for 28 days, the offspring ramets contained approximately 13 ± 2 μg/g NPs. In the case of connected offspring ramets, leaf and root biomass decreased by 24%-51% and 32%-51%, respectively, when exposed to NP concentrations ranging from 0.1 to 10 mg L-1. Excessive enrichment of NPs had a detrimental effect on the photosynthetic system, decreasing the chlorophyll content and nonphotochemical quenching. An imbalance in the antioxidant defense systems, which were unable to cope with the oxidative stress caused by NP concentrations, further damaged various organs. The root system can take up NPs and then transfer them to the offspring through the stolon. Interference effects of NPs were observed in terms of root activity, metabolism, biofilm composition, and the plant's ability to purify water. However, the risk-spreading strategy employed by parent plants (interconnected offspring ramets) offered some relief from NP-induced stress, as it increased their relative growth rate by 1 to 1.38 times compared to individual plants. These findings provide substantial evidence of the high NP enrichment capacity of E. crassipes for ecological remediation. Nevertheless, we must also remain aware of the environmental risk associated with the spread of NPs within the clonal system of E. crassipes, and contaminated cloned individuals need to be precisely removed in a timely manner to maintain normal functions.

Plant cuticles repress organ initiation and development during skotomorphogenesis in Arabidopsis

After germination in the dark, plants produce a shoot apical hook and closed cotyledons to protect the quiescent shoot apical meristem (SAM), which is critical for seedling survival during skotomorphogenesis. The factors that coordinate these processes, particularly SAM repression, remain enigmatic. Plant cuticles, multilayered structures of lipid components on the outermost surface of the aerial epidermis of all land plants, provide protection against desiccation and external environmental stresses. Whether and how cuticles regulate plant development are still unclear. Here, we demonstrate that mutants of BODYGUARD1 (BDG1) and long-chain acyl-CoA synthetase2 (LACS2), key genes involved in cutin biosynthesis, produce a short hypocotyl with an opened apical hook and cotyledons in which the SAM is activated during skotomorphogenesis. Light signaling represses expression of BDG1 and LACS2, as well as cutin biosynthesis. Transcriptome analysis revealed that cuticles are critical for skotomorphogenesis, particularly for the development and function of chloroplasts. Genetic and molecular analyses showed that decreased HOOKLESS1 expression results in apical hook opening in the mutants. When hypoxia-induced expression of LITTLE ZIPPER2 at the SAM promotes organ initiation in the mutants, the de-repressed expression of cell-cycle genes and the cytokinin response induce the growth of true leaves. Our results reveal previously unrecognized developmental functions of the plant cuticle during skotomorphogenesis and demonstrate a mechanism by which light initiates photomorphogenesis through dynamic regulation of cuticle synthesis to induce coordinated and systemic changes in organ development and growth during the skotomorphogenesis-to-photomorphogenesis transition.

Regulation of chloroplast biogenesis, development, and signaling by endogenous and exogenous cues.

Chloroplasts are one of the defining features in most plants, primarily known for their unique property to carry out photosynthesis. Besides this, chloroplasts are also associated with hormone and metabolite productions. For this, biogenesis and development of chloroplast are required to be synchronized with the seedling growth to corroborate the maximum rate of photosynthesis following the emergence of seedlings. Chloroplast biogenesis and development are dependent on the signaling to and from the chloroplast, which are in turn regulated by several endogenous and exogenous cues. Light and hormones play a crucial role in chloroplast maturation and development. Chloroplast signaling involves a coordinated two-way connection between the chloroplast and nucleus, termed retrograde and anterograde signaling, respectively. Anterograde and retrograde signaling are involved in regulation at the transcriptional level and downstream modifications and are modulated by several metabolic and external cues. The communication between chloroplast and nucleus is essential for plants to develop strategies to cope with various stresses including high light or high heat. In this review, we have summarized several aspects of chloroplast development and its regulation through the interplay of various external and internal factors. We have also discussed the involvement of chloroplasts as sensors of various external environment stress factors including high light and temperature, and communicate via a series of retrograde signals to the nucleus, thus playing an essential role in plants' abiotic stress response.

Postnatal epigenome-mediated aging control and global trends

The epigenome can adequately regulate the on/off states of genes in response to external environmental factors and stress. In recent years, it has been observed that the epigenome, which is modulated through DNA methylation, histone modifications, and chromatin remodeling, changes with age. Alterations in the epigenome lead to the loss of cell-specific epigenome/identity, which in turn triggers a decline in tissue function. In mammals, postnatal epigenomic variations are not only caused by metabolic diseases, such as diabetes or DNA damage, but also by social stress and infectious diseases. Unlike Genome-Wide Association Studies (GWAS), dynamically changing epigenomes, along with their cellular roles, need to be established as objective biomarkers in conjunction with various biological signals, such as walking speed, brain waves, and clinical data. The biological age/aging clock, determined by methylated DNA, has attracted attention, and calorie restriction not only slows the progression of aging, but also seems to suppress it. However, as indicated by gene expression analysis in aging mice, aging is not a linear model, but is represented by nonlinear dynamic changes. Consequently, the development of experimental models and analytical methods that enhance temporal resolution through time-series analysis, tailored to spatial resolution, such as cell distribution and organ specificity, is progressing. Moreover, in recent years, in addition to anti-aging efforts targeting epigenomic variations, global attention has increasingly focused on research and development aimed at rejuvenating treatments, thus leading to the birth of many biotech companies. Aging Hallmarks such as inflammation, stem cells, metabolism, genomic instability, and autophagy, interact closely with the epigenome. Various postnatal and reversible epigenomic controls of aging, including Yamanaka factors (OKSM and OSK), are now entering a new phase. In the future, the development of aging control using diverse modalities, such as mRNA, artificial peptides, and genome editing, is expected, along with an improved molecular understanding of aging and identification of useful biomarkers.

Endolichenic Fungi: A Promising Medicinal Microbial Resource to Discover Bioactive Natural Molecules-An Update.

Lichens are some of the most unique fungi and are naturally encountered as symbiotic biological organisms that usually consist of fungal partners (mycobionts) and photosynthetic organisms (green algae and cyanobacteria). Due to their distinctive growth environments, including hot deserts, rocky coasts, Arctic tundra, toxic slag piles, etc., they produce a variety of biologically meaningful and structurally novel secondary metabolites to resist external environmental stresses. The endofungi that live in and coevolve with lichens can also generate abundant secondary metabolites with novel structures, diverse skeletons, and intriguing bioactivities due to their mutualistic symbiosis with hosts, and they have been considered as strategically significant medicinal microresources for the discovery of pharmaceutical lead compounds in the medicinal industry. They are also of great importance in the fundamental research field of natural product chemistry. In this work, we conducted a comprehensive review and systematic evaluation of the secondary metabolites of endolichenic fungi regarding their origin, distribution, structural characteristics, and biological activity, as well as recent advances in their medicinal applications, by summarizing research achievements since 2015. Moreover, the current research status and future research trends regarding their chemical components are discussed and predicted. A systematic review covering the fundamental chemical research advances and pharmaceutical potential of the secondary metabolites from endolichenic fungi is urgently required to facilitate our better understanding, and this review could also serve as a critical reference to provide valuable insights for the future research and promotion of natural products from endolichenic fungi.

Concrete corrosion in nuclear power plants and other nuclear installations and its mitigation techniques: a review

Abstract Nuclear power plants (NPPs) have been affected by various failures through corrosion which causes economic losses, increased chance of the radiation exposure and environmental risk. One of the major durability issues in reinforced concrete is the corrosion of reinforcement which significantly reduces the life of reinforced concrete. Considering the increasing demand for longer service lives of NPPs, and the high cost involved in building and maintaining it, adequate preventive measures should be followed to minimize the corrosion. This review majorly discusses about the mechanism of corrosion of steel in NPP structures with emphasis on the mechanisms relevant to NPPs, possible reasons for the concrete corrosion as well as potential failure happening in NPPs. The majors reason for the concrete corrosion in nuclear power plants are mainly corrosive external and internal environment, thermal and mechanical stress, moisture content, microorganisms and stray electrical currents. The corrosion of NPPs may result in loss of structural integrity and leakage of radioactive material. The review also discusses about various corrosion prevention and protection techniques against concrete corrosion and concludes with an overview of present methods and possible future perspectives used to enhance the efficiency of concrete corrosion mitigation methods with focus of NPPs.

Characterization and expression analysis of a thaumatin-like protein PpTLP1 from ground cherry Physalis pubescens

Ground cherry, Physalis pubescens, is mainly cultivated as a fruit worldwide and popularly used as a food supplement and traditional Chinese medicine. Plants are challenged by external environmental stress and can initiate resistance to the stress through the regulation of pathogenesis-related (PR) proteins. Among PR proteins, PR-5, a thaumatin-like protein (TLP), was identified in many plants and found to be able to enhance stress resistance. However, PR-5 in ground cherry is not characterized and its expression is yet to be understood. In this study, a PR-5 protein PpTLP1 in P. pubescens was firstly identified. Analysis of the amino acid sequences revealed that PpTLP1 was highly similar to PR-NP24 identified in tomato with a difference in only one amino acid. Expression analysis indicated that the PpTLP1 gene was highly expressed in leaf while the PpTLP1 protein was tissue-specifically accumulated in cherry exocarp. Furthermore, the down-regulation of PpTLP1 in ground cherry was induced by NaCl treatment while the up-regulation was promoted by the infection of Sclerotinia sclerotiorum and Botrytis cinerea. This study will provide a new plant resource containing a TLP in Physalis genus and a novel insight for the improvement of postharvest management of ground cherry and other Solanaceae plants.

Antioxidant peptides, the guardian of life from oxidative stress.

Reactive oxygen species (ROS) are produced during oxidative metabolism in aerobic organisms. Under normal conditions, ROS production and elimination are in a relatively balanced state. However, under internal or external environmental stress, such as high glucose levels or UV radiation, ROS production can increase significantly, leading to oxidative stress. Excess ROS production not only damages biomolecules but is also closely associated with the pathogenesis of many diseases, such as skin photoaging, diabetes, and cancer. Antioxidant peptides (AOPs) are naturally occurring or artificially designed peptides that can reduce the levels of ROS and other pro-oxidants, thus showing great potential in the treatment of oxidative stress-related diseases. In this review, we discussed ROS production and its role in inducing oxidative stress-related diseases in humans. Additionally, we discussed the sources, mechanism of action, and evaluation methods of AOPs and provided directions for future studies on AOPs.

Double Skin Façade Mechanical Systems as Ad-vanced Building Technologies

The double-wall façades (or “double skin façades”) are deter-mined as building mechanical and engineered systems realized by the cavity between the inner curtain and the outer screen, for thermal and acoustic insulation, for ventilation and for the inser-tion of functional devices (such as sunscreens) and, also, plant ducts. The cavity between the two enclosures constitutes a venti-lated cavity that can be used according to certain modes of func-tioning (passive or active type) aimed at controlling external cli-matic and environmental stresses in order to regulate the condi-tions of the interior spaces. The double envelope system is real-ized as an apparatus of mediation and reaction towards envi-ronmental loads, according to the needs of well-being and reduc-tion of energy consumption. In general, the system offers the functioning in the form of a passive solar system, assuming the use and accumulation of solar radiation for the regulation of indoor thermal comfort conditions, and the functioning for the capture and input of air flows. The calibration of solar radiation is integrated with the use of shading devices in order to achieve diffuse lighting conditions in interior spaces. In addition, the application of the double glass surface reduces thermal losses from the interior spaces by reducing the speed of the airflow in contact with the inner curtain, increasing thermal insulation. The system foresees that the ventilated cavity performs various inte-grated functions (for the definition of complex mechanisms of dynamic interaction with the external climatic conditions), both permanent (e.g. for the increase of thermal inertia and acoustic insulation relative to the internal curtain) and temporary (e.g. for the cooling of the same spaces during periods of high tempera-ture).

Biological Sensitization of Blood Immune Cells of Rabbits to External Environmental Stress Factors (Temperature And Seasons)

In this article, the condition and activity of the special immune system index in the blood of Chinchilla and New Zealand rabbits in the extreme climatic conditions of Uzbekistan during the seasons due to temperature fluctuations and nutritional stress are studied. As a result of the study, information on the active physiological state of leukocytes, T-, V-lymphocytes and their subpopulations, helper suppressors and leukocyte T-lymphocyte ratio in blood is presented.

The transcriptional regulator EtrA mediates ompW contributing to the pathogenicity of avian pathogenic Escherichia coli

Avian pathogenic Escherichia coli (APEC) causes avian colibacillosis and leads to high mortality in poultry and huge economic losses. Therefore, it is important to investigate the pathogenic mechanisms of APEC. Outer membrane protein OmpW is involved in the environmental adaptation and pathogenesis of Gram-negative bacteria. OmpW is regulated by many proteins, including FNR, ArcA, and NarL. In previous studies, regulator EtrA is involved in the pathogenicity of APEC and affects the transcript levels of ompW. However, the function of OmpW in APEC and its regulation remain unclear. In this study, we constructed mutant strains with altered etrA and/or ompW genes to evaluate the roles of EtrA and OmpW in the biological characteristics and pathogenicity of APEC. Compared with wild-type strain AE40, mutant strains ∆etrA, ∆ompW, and ∆etrA∆ompW showed significantly lower motility, lower survival under external environmental stress, and lower resistance to serum. Biofilm formation by ∆etrA and ∆etrA∆ompW was significantly enhanced relative to that of AE40. The transcript levels of TNF-α, IL1β, and IL6 were also significantly enhanced in DF-1 cells infected with these mutant strains. Animal infection assays showed that deletion of etrA and ompW genes attenuated the virulence of APEC in chick models, and damage to the trachea, heart, and liver caused by these mutant strains was attenuated relative to that caused by the wild-type strain. RT-qPCR and β-galactosidase assay showed that EtrA positively regulates the expression of the ompW gene. These findings demonstrate that regulator EtrA positively regulates the expression of OmpW, and that they both contribute to APEC motility, biofilm formation, serum resistance, and pathogenicity.

Genome-Wide Analysis of the HD-Zip Gene Family in Chinese Cabbage (Brassica rapa subsp. pekinensis) and the Expression Pattern at High Temperatures and in Carotenoids Regulation

HD-Zip, a special class of transcription factors in high plants, has a role in plant development and responding to external environmental stress. Heat stress has always been an important factor affecting plant growth, quality, and yield. Carotenoid content is also an important factor affecting the color of the inner leaf blades of Chinese cabbage. In this study, the genomes of three Brassicaceae plants were selected: Chinese cabbage (Brassica rapa subsp. pekinensis), Brassica oleracea, and Brassica napus. We identified 93, 96, and 184 HD-Zip genes in the B. rapa, B. oleracea, and B. napus, respectively. The HD-Zip gene family was classified into four subfamilies based on phylogeny: I, II, III, and IV;. The results of cis-acting element analysis suggested that HD-Zip family genes may participate in various biological processes, such as pigment synthesis, cell cycle regulation, defense stress response, etc. Conserved motifs prediction revealed that three motifs exist among the four HD-Zip gene families and that different motifs exhibit significant effects on the structural differences in HD-Zips. Synteny, Ks, and 4DTv results displayed that genome-wide triplication events act in HD-Zip gene family expansion. Transcriptome data showed that 18 genes responded (>1.5-fold change) to heat stress in Chinese cabbage, and 14 of 18 genes were from the HD-Zip I subfamily. Three genes had up-regulation, and eight genes had down-regulation in high-carotenoid-content Chinese cabbage. The BraA09g011460.3C expression level was up-regulated after heat stress treatment and significantly reduced in varieties with high carotenoid content, indicating its potential for heat stress tolerance and carotenoid content regulation. This study provided important gene resources for the subsequent breeding of Chinese cabbage.

Evaluation of the biconditional transformation of land use: a case study of Minhou County in China

ABSTRACT Land use transformation is necessary to achieve the regional long-term socioeconomic development. The recent increase in land demand and external environmental stress in China has led to dramatic land use transformation. In the study, land use transformation in Minhou County in China was investigated from the biconditional perspective of dominant and recessive forms, which included the analysis of the quantity structure, transformation magnitude, transformation speed, spatial structure, efficiency, and function using remote sensing, geographic information system, topography, and socioeconomic data. The results revealed that, in the study of the dominant form, the construction land transformation in the industrialized town developing rapidly with a low-lying slope had the greatest speed, with an average annual growth rate of 8.74%, while the woodland was moving at the lowest rate of −3.62%. In the recessive form, the functional pattern of the construction land, cultivated land production, and the input into the woodland were revealed as the prime factors that impacted land use transformation. The linear growth of the functional index from the construction land, the increasing production index of the cultivated land, and the rebound efficiency of forestry production in Minhou County in China have been continuously influencing the variation in the land use patterns. The findings of the study would promote sustainable development and appropriate utilization of the land resources in the region by providing decision support to the government for scientific and rational planning and strengthening the efficiency of intensive land use.

Current Perspectives on Viable but Non-Culturable Foodborne Pathogenic Bacteria: A Review.

Foodborne diseases caused by foodborne pathogens pose risks to food safety. Effective detection and efficient inactivation of pathogenic bacteria has always been a research hotspot in the field of food safety. Complicating these goals, bacteria can be induced to adopt a viable but non-culturable (VBNC) state under adverse external environmental stresses. When in the VBNC state, pathogens cannot form visible colonies during traditional culture but remain metabolically active and toxic. The resulting false negative results in growth-related assays can jeopardize food safety. This review summarizes the latest research on VBNC foodborne pathogens, including induction conditions, detection methods, mechanism of VBNC formation, and possible control strategies. It is hoped that this review can provide ideas and methods for future research on VBNC foodborne pathogenic bacteria.

Severity of Binge Eating Behavior among Overweight College Students in Taiwan and Associated Factors.

Binge eating (BE) is considered a marker of obesity and overweight and a significant characteristic of feeding and eating disorders. Despite the high prevalence of obesity on college campuses, the issue of BE among college students in Taiwan has received little attention. The aim of this study was to investigate BE behavior among overweight college students in Taiwan and associated factors. This study utilized a cross-sectional survey. A total of 300 overweight college students were recruited through convenience sampling. Data were collected using a self-administered Binge Eating Scale (BES) and a body weight composition monitor (Model No. OMRON, HBF-126) and analyzed using descriptive statistics, correlation analysis, and regression analysis. The average BES score was 10.67 (SD = 6.66, 0-34). With a BES score of 17 as the cut-off point, 17.3% (n = 52) of the participants were found to have moderate or severe BE behavior. Analysis of the demographic and psychosocial data using Spearman's rho rank correlation coefficient revealed that sex, body mass index (BMI), uncontrolled eating, weight loss diets, academic stress, peer competition, interpersonal distress, and unpleasant or major life events were significantly correlated with BE behavior and its probability (rs = -0.14-0.15, p < 0.05). Furthermore, logistic regression analysis indicated that the odds ratio of the BES scores of female participants and those who stated to have experienced uncontrolled eating, weight loss diets, peer competition, and interpersonal distress was 1.05-6.04 times those of male participants and those without such experiences (p < 0.05). The study found that nearly one-fifth of participants presented moderate to severe levels of BE behaviors, and these were significantly correlated with sex and external environmental stress. This study suggests early intervention from campus psychological health personnel to provide proper therapy.

Heat Shock Proteins (Hsps) in Cellular Homeostasis: A Promising Tool for Health Management in Crustacean Aquaculture.

Heat shock proteins (Hsps) are a family of ubiquitously expressed stress proteins and extrinsic chaperones that are required for viability and cell growth in all living organisms. These proteins are highly conserved and produced in all cellular organisms when exposed to stress. Hsps play a significant role in protein synthesis and homeostasis, as well as in the maintenance of overall health in crustaceans against various internal and external environmental stresses. Recent reports have suggested that enhancing in vivo Hsp levels via non-lethal heat shock, exogenous Hsps, or plant-based compounds, could be a promising strategy used to develop protective immunity in crustaceans against both abiotic and biotic stresses. Hence, Hsps as the agent of being an immune booster and increasing disease resistance will present a significant advancement in reducing stressful conditions in the aquaculture system.

Two Arabidopsis MYB-SHAQKYF transcription repressors regulate leaf wax biosynthesis via transcriptional suppression on DEWAX.

Plant cuticular wax accumulation limits nonstomatal transpiration and is regulated by external environmental stresses. DEWAX (DECREASE WAX BIOSYNTHESIS) plays a vital role in diurnal wax biosynthesis. However, how DEWAX expression is controlled and the molecular mechanism of wax biosynthesis regulated by the diurnal cycle remains largely unknown. Here, we identified two Arabidopsis MYB-SHAQKYF transcription factors, MYS1 and MYS2, as new regulators in wax biosynthesis and drought tolerance. Mutations of both MYS1 and MYS2 caused significantly reduced leaf wax, whereas overexpression of MYS1 or MYS2 increased leaf wax biosynthesis and enhanced drought tolerance. Our results demonstrated that MYS1 and MYS2 act as transcription repressors and directly suppress DEWAX expression via ethylene response factor-associated amphiphilic repression motifs. Genetic interaction analysis with DEWAX, SPL9 (SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 9), and CER1 (ECERIFERUM 1) in wax biosynthesis and under drought stresses demonstrated that MYS1 and MYS2 act upstream of the DEWAX-SPL9 module, thus regulating CER1 expression. Expression analysis suggested that the diurnal expression pattern of DEWAX is partly regulated by MYS1 and MYS2. Our findings demonstrate the roles of two unidentified transcription repressors, MYS1 and MYS2, in wax biosynthesis and provide insights into the mechanism of diurnal cycle-regulated wax biosynthesis.