Trehalose Content Research Articles

RNAi-Mediated FoxO Silencing Inhibits Reproduction in Locusta migratoria.

FoxO is a downstream target gene of cellular nutrient and growth factors, oxidative stress responses, and insulin signaling pathways. It play a crucial role in insect growth, development, and reproduction. Locusta migratoria is a significant agricultural pest; therefore, the identification of novel control targets for its management is of significant importance. After injecting dsRNA to interfere with FoxO expression, we observed changes in the reproduction-related gene expression and ovary development through RT-qPCR and morphological observation. Simultaneously, the trehalose and glycogen contents were measured following RNAi. The results demonstrate that interference with FoxO significantly downregulates key genes in the Hippo pathway and Notch gene expression. In terms of carbohydrate metabolism, the trehalose content decreases significantly while the glycogen content increases markedly after FoxO silencing. Additionally, FoxO silencing considerably inhibits reproductive-related gene expression, resulting in delayed ovarian development. These findings indicate that FoxO regulates L. migratoria reproduction through the Hippo signaling pathway: when impaired, the reproductive capacity function declines. In addition, FoxO-mediated energy mobilization is involved in the regulation of egg production. These results indicate that the RNAi of FoxO may be a useful control strategy against L. migratoria.

Juvenile hormone controls trehalose metabolism by regulating trehalase 2 activity in ovarian development of Helicoverpa armigera.

Trehalase (Treh) is crucial for ovarian development as it directly regulates the energy supply by hydrolyzing trehalose into glucose. Juvenile hormone (JH) is also essential for ovarian development, but how it affects Treh2 activity remains unclear. This study, which employed Helicoverpa armigera as a model, showed that HaTreh2 transcription and enzymatic activity peaks coincided with the peak of JH titers (the 2 and 3 days after emergence). Compared to the dsGFP control, knockdown of HaTreh2 transcription severely impaired ovarian development. LC-MS/MS and site mutation experiments demonstrated that JH triggered the serine 345 phosphorylation of HaTreh2 via the GPCR-cAMP-PKA pathway, thereby activating its enzymatic activity. Additionally, HaTreh2 is directly bound with trehalose transporter (HaTreT) under JH induction, thus controlling intracellular trehalose and glucose contents as well as the transcription of HaTreT. TreT controls the amount of trehalose, which serves as a substrate for Treh1, entering the cell. Treh2, on the other hand, uses extracellular trehalose as substrate, and the hydrolysis product glucose is further transported into the cell. Here, HaTreh2 regulated the substrate that HaTreh1 can act upon in the cell by directly binding with HaTreT during ovarian development when JH is induced. Therefore, JH systematically regulated trehalose metabolism during ovarian development through regulating the activity of HaTreh2. This study sheds light on the coordinated interplay between JH pathway and sugar metabolism in ovarian development.

Analysis of Frozen Bakery Items Containing Trehalose and its Impact on Quantitative Quality Measures

The frozen dough market has been increasing annually since it reduces cost while ensuring the availability of fresh products. Ice crystal formation inside the dough decreases yeast functionality while thawing releases fluid inside the dough, both leading to an unacceptable product. Trehalose can enhance the viability of yeast cells and reduce ice crystals. Four concentrations of trehalose (0, 10, 20, and 30%) were investigated on the freeze-thaw quality of frozen dough. Samples were prepared, frozen, and thawed before testing for color, moisture, water activity, and texture. Trehalose had no significant impact on color, moisture, or water activity of any sample despite trends in moisture and water activity being present. However, increased trehalose showed decrease in peak load in biscuits and pizza crusts. Therefore, more research is needed but data shows that trehalose could act as an anti-staling agent by reducing crust hardness in frozen biscuits and pizza crusts.

A novel technique for trehalose and sucrose determination in therapeutic monoclonal antibodies using a high-performance liquid chromatography–evaporative light scattering detector

Sucrose and trehalose are commonly used excipients in therapeutic monoclonal antibodies that play a pivotal role in ensuring the safety and stability of drugs. Though it is necessary to control the concentrations of these substances during the quality control of their release, there is currently no comprehensive method for simultaneously determining sucrose and trehalose concentrations. Herein, we established a high-performance liquid chromatography–evaporative light scattering detector (HPLC-ELSD) method and validated it in accordance with the International Council for Harmonization Q2 guidelines. This method utilized the Poroshell 120 HILIC-Z chromatographic column and effectively separated sucrose and trehalose with a detection limit of 0.1 mg/mL. The accuracy recovery rate was within a range of 90%–110%, and the precision relative standard deviations were all less than 5.0% (n = 6). The method thus demonstrated good repeatability and linearity, making it suitable for determining the sucrose and trehalose concentrations in therapeutic monoclonal antibodies.

Lentil protein and trehalose conjugates: Structural interactions and mechanisms for improving multi-level structure and functional characteristics.

This study aimed to improve lentil proteins' (LPs) functionality and nutritional value, specifically addressing their lower water solubility and digestibility. A unique combination of LP-disaccharide interactions was employed. Spectroscopic technologies, which include fluorescence spectra, ultraviolet spectra, and Fourier-transform infrared, investigated the structure of LPs at various concentrations of trehalose. The results indicate that the LP structures and conformation were considerably modified (p<0.05) following trehalose conjugation. The surface charge and hydrophobicity of the trehalose-conjugated LPs (T-LPs) were significantly altered (p<0.05), from -22.7 to -31.4 and 753 and 543 a.u., respectively. Furthermore, the digestibility and solubility of T-LPs increased from 75% to 81.8% and 60% to 66%, respectively. In conclusion, this study showed that combining LPs and trehalose conjugation could improve the quality of conjugates LPs, which could expand their use in manufacturing as the acceptance of plant-based diets increases. PRACTICAL APPLICATION: Currently, lentil proteins (LPs) are used in plant-based protein powders and supplements. Though less popular than soy or pea proteins, LPs are valued for their high protein content and good amino acid profile. LPs are utilized in meat alternatives and high-protein snack products. The application of these products is mainly due to their nutritional benefits rather than functional properties due to their poor water solubility. Increasing the water solubility of LPs could significantly expand their application in various food industries, making LPs a more competitive and functional plant-based protein source. Trehalose-conjugated LPs with better water solubility allow LPs to be used in other food products, such as plant-based protein beverages. Better solubility would enhance the clarity and smoothness of these products, making them more appealing to consumers.

Trehalose metabolism mediates trade-offs between reproduction and survival in beet webworm, Loxostege sticticalis, under heat stress.

Temperature is an important determinant of developmental and reproductive rates in insects. Here, we investigated the physiological responses of adult beet webworm, Loxostege sticticalis L. (Lepidoptera: Crambidae), to three temperatures (16, 23 and 30 °C) focusing on trehalose metabolism. Exposure of moths to 30 °C accelerated eclosion and ovarian development, but shortened the oviposition period and adult longevity, whereas exposure to 16 °C had opposite effects. Transcriptome analysis revealed that vitellogenin (VG) and vitellogenin receptor (VR) genes were up-regulated at 30 °C, as were numerous genes related to energy metabolism, including those involved in the insulin signaling pathway, the tricarboxylic acid (TCA) cycle, and glycolysis. Expression of the trehalose transporter gene TRET1 was also induced at high temperature, primarily in the ovaries, where trehalose content increased, accompanied by lipid degradation in the fat body. Treatment with the trehalase inhibitor validamycin A reduced female fecundity and longevity at 23 °C, but enhanced the expression of genes related to stress resistance and reproduction, mimicking the effect of high temperature. Besides their practical utility for predicting the oviposition behavior and geographic distribution of L. sticticalis in the field, these results elucidate the various physiological roles of trehalose in L. sticticalis during exposure of moths to high temperature and may provide insights into the relationship between stress resistance and reproduction in insects more generally. © 2024 Society of Chemical Industry.

Stress Response of Aphid Population Under Combined Stress of Cadmium and Lead and Its Effects on Development of Harmonia axyridis.

Heavy metal pollution is a serious global environmental issue. It threatens human and ecological health. Heavy metals can accumulate in the soil over extended periods and inevitably transfer through the food chain to herbivorous insects and their natural enemies, leading to various adverse effects. This study aimed to investigate the stress responses and biochemical metabolic changes of aphids and one of aphids' predators, ladybugs, under cadmium (Cd) and lead (Pb) stress by constructing a food chain of Vicia faba L., Megoura crassicauda, and Harmonia axyridis. The results showed that aphids and ladybugs had a notable accumulation of Cd2+ and Pb2+. Insects can adapt to heavy metal stress by regulating their energy metabolism pathways. Glycogen content in the first filial generation (F1) aphids decreased significantly, glucose content in the second filial generation (F2) to the fourth filial generation (F4) adult aphids significantly increased, and trehalose content in the F1 adult aphids increased significantly. Moreover, the relative expression levels of trehalase (TRE) and trehalose-6-phosphate synthase (TPS) in the F1 adult aphids were significantly higher than those in the control group, and the expression levels of TPS genes in the second filial generation to the fifth filial generation (F2 to F5) aphids decreased, suggesting that insects can resist heavy metal stress by regulating trehalose metabolism. The fertility of female aphids in all treatment groups was reduced compared to the control group. Additionally, the relative expression level of vitellogenin (Vg) was down-regulated in all aphid generations except the F1 aphids. There was interaction between heavy metal concentration and aphid generation, and it significantly affected the number of aphids' offspring and the expression of the aphid Vg gene. The developmental duration of the ladybugs from the second to fourth instars was prolonged, and the weight decreased significantly from the prepupa to the adult stages. These results contribute to understanding the effects of Cd2+-Pb2+ accumulation on phytophagous insects and higher trophic levels' natural enemies, laying the foundation for protecting natural enemies and maintaining ecosystem stability.

Combined Analysis of Transcriptome and Metabolome Provides Insights in Response Mechanism under Heat Stress in Avocado (Persea americana Mill.)

Plants generate a range of physiological and molecular responses to sustain their growth and development when suffering heat stress. Avocado is a type of tropical fruit tree with high economic value. Most avocado cultivars delete, wither, or even die when exposed to heat stress for a long time, which seriously restricts the introduction and cultivation of avocados. In this study, samples of a heat-intolerant variety (‘Hass’) were treated under heat stress, and the transcriptomics and metabolomics were analyzed, with the expectation of providing information on the variety improvement and domestication of avocados. The differentially expressed genes identified using transcriptome analysis mainly involved metabolic pathways such as plant hormone signal transduction, plant–pathogen interaction, and protein processing in the endoplasmic reticulum. Combined transcriptome and metabolome analysis indicated that the down-regulation of Hass.g03.10206 and Hass.g03.10205 in heat shock-like proteins may result in the reduced Trehalose and Sinapoyl aldehyde content. Metabolomics analysis results indicated that the decrease in Trehalose and Sinapoyl aldehyde content may be an important factor for heat intolerance. These results provide important clues for understanding the physiological mechanisms of adaptation to heat stress in avocados.

A tomato a day keeps the beetle away - the impact of Solanaceae glycoalkaloids on energy management in the mealworm Tenebrio molitor.

Solanine (SOL), chaconine (CHA), and tomatine (TOM) are plant secondary metabolites produced mainly by the species of Solanaceae family, such as tomato Solanum lycopersicum L. These glycoalkaloids (GAs) have a wide range of biological activity, also in insects. However, their mechanisms of action are not precisely understood. The purpose of the study was to investigate how pure GAs and tomato leaf extract (EXT) affect glycolysis, Krebs cycle and β-oxidation of fatty acid pathways in Tenebrio molitor L. beetle. For this purpose, the larvae were injected with SOL, CHA, TOM, and EXT at two concentrations (10-8 and 10-5M). For experiments, fat body, gut, and heamolymph samples were collected 2 and 24h after injection. Then, the changes in the expression level of phosphofructokinase, citrate synthase, and β-hydroxyacyl-CoA dehydrogenase were measured using the RT-qPCR technique. The catalytic activity of these enzymes and the carbohydrate level in insects after GA treatment were determined by spectrophotometric method. Furthermore, the analysis of the amount of amino acids in tissues was performed with a GC-MS technique. The results obtained show that the GAs changed the activity and expression of the genes encoding key enzymes of crucial metabolic pathways. The effect depends on the type of GA compound, the tissue tested, and the incubation time after treatment. Furthermore, TOM and EXT affected trehalose concentration in the insect hemolymph and led to accumulation of amino acids in the fat body. The observed changes may indicate a protein degradation and/or enhanced catabolism reactions for the production of ATP used in detoxification processes. These results suggest that GAs alter energy metabolism in the mealworm T. molitor. The study contributes to our understanding of the mechanisms of action of secondary metabolites of plants in insects. This knowledge may allow the design of new natural biopesticides against insect pests because proper energy metabolism is necessary for the survival of the organism.

Comparative Study on Growth and Metabolomic Profiles of Six Lactobacilli Strains by Sodium Selenite

Selenium (Se) has garnered increasing attention in the field of nutrition, as it is essential for both humans and animals. Certain microorganisms can enrich inorganic selenium and convert it into organic selenium. The growth and metabolomic profiles of six lactobacilli strains exposed to 50 μg/mL of sodium selenite were performed using gas chromatography tandem time-off light mass spectrometry (GC-TOF-MS) analysis. The addition of selenium significantly increased both the population and weight of the Lacticaseibacillus rhamnosus PS5, Lbs. rhamnosus RT-B, Limosilactobacillus reuteri 3630, and Lmb. reuteri 1663 strains, while those of the other two strains decreased. A total of 271 metabolites were determined, with their concentrations ranked from highest to lowest as follows: organic acids and derivatives, oxygen compounds, lipids and lipid-like molecules, and benzenoids. In certain groups, the concentrations of serine, aspartic acid, trehalose, palmitic acid, methylthreonine, and melibiose increased significantly, whereas glucuronic acid, ribose, ornithine, and methionine were downregulated. The metabolic pathways were significantly associated with ABC transporters, glycine, serine, threonine metabolism, and aminobenzoate degradation and other pathways. Based on these findings, we concluded that the transport, absorption, assimilation, and stress response to selenium by lactobacilli in metabolomic changed. Furthermore, the metabolomic alterations among different types of lactobacilli varied primarily due to their distinct properties.

Improvement of the gel properties of curdlan gel by hydrogen bonding interaction with trehalose

This study investigated the effect of trehalose addition (0.5–2.0%) on the gelation properties of curdlan gels were investigated. The results demonstrated that the optimal gelation performance of curdlan-trehalose composite gel was determined at trehalose concentration of 1.0% with preferable water holding capacity, water absorption rate, and freeze-thaw stability. The gelation behavior of the curdlan-trehalose composite gel showed a more intense network structure with increasing storage modulus (G′), loss modulus (G″), and deceasing tan δ at the trehalose concentration in the range of 0%–1%. And the highest gel strength was also observed in 1% trehalose addition. Moreover, the structures characterized by Fourier transform infrared (FT-IR), X-ray diffraction (XRD), Low field nuclear magnetic resonance (LF-NMR) and scanning electron microscope analysis (SEM) all revealed that the addition of trehalose strengthened the hydrogen bonding interaction with curdlan to create a much denser network of composite gel, which would provide a better mechanistic insight of the curdlan-trehalose gel in practical application potentials in food and medicine.

Assessing the physiological properties of baker's yeast based on single-cell Raman spectrum technology

With rapid progress in the yeast fermentation industry, a comprehensive commercial yeast quality assessment approach integrating efficiency, accuracy, sensitivity, and cost-effectiveness is required. In this study, a new yeast quality assessment method based on single-cell Raman technology was developed and contrasted with traditional methods. The findings demonstrated significant associations (Pearson correlation coefficient of 0.933 on average) between the two methods in measuring physiological indicators, including cell viability and intracellular trehalose content, demonstrating the credibility of the Raman method compared to the traditional method. Furthermore, the sensitivity of the Raman method in viable but non-culturable cells was higher in measuring yeast cell viability (17.9 % more sensitive). According to the accurate quantitative analysis of metabolic activity level (MAL) of yeast cells, the cell vitality was accurately quantified at population and single-cell levels, offering a more comprehensive assessment of yeast fermentation performance. Overall, the single-cell Raman method integrates credibility, feasibility, accuracy, and sensitivity in yeast quality assessment, offering a new technological framework for quality assessments of live-cell yeast products.

PagTPS1 and PagTPS10, the trehalose-6-phosphate synthase genes, increase trehalose content and enhance drought tolerance

Trehalose-6-phosphate synthase (TPS) genes play an active role in the trehalose metabolism pathway that regulates the responses of plants to diverse stresses. However, the functional identification, comparison, and conservatism of TPS genes in the responses of woody plants, especially poplars, to drought stress remain unclear. Here, the trehalose content of 84K (Populus alba × P. glandulosa) poplars was down-regulated and PagTPS and PagTPP genes had diverse response patterns under drought stress. Physicochemical properties, expression patterns, and functions of PagTPS1 and PagTPS10, two class I members of TPS gene family, were identified and compared. Transgenic 84K poplars overexpressing PagTPS1 and PagTPS10 had significantly higher trehalose content with approximately 138% and 123%, respectively, and stronger drought tolerance compared to WT. PagTPS1 and PagTPS10 promoted the expression of TPPA genes and drought-responsive genes. Accordingly, poplars inhibiting PagTPS1 and PagTPS10 expression via RNA interference had lower trehalose content and drought tolerance. Simultaneously, overexpressing PagTPS1 and PagTPS10 improved the trehalose content and drought tolerance of Arabidopsis. Overall, we proposed a model of the effects of PagTPS1 and PagTPS10 as conservative regulators on the responses of plants to drought, which would provide new insights into the functional explorations of TPS genes in plants.

Mesenchymal stem cell cryopreservation with cavitation-mediated trehalose treatment

Dimethylsulfoxide (DMSO) has conventionally been used for cell cryopreservation both in research and in clinical applications, but has long-term cytotoxic effects. Trehalose, a natural disaccharide, has been proposed as a non-toxic cryoprotectant. However, the lack of specific cell membrane transporter receptors inhibits transmembrane transport and severely limits its cryoprotective capability. This research presents a method to successfully deliver trehalose into mesenchymal stem cells (MSCs) using ultrasound in the presence of microbubbles. The optimised trehalose concentration was shown to be able to not only preserve membrane integrity and cell viability but also the multipotency of MSCs, which are essential for stem cell therapy. Confocal imaging revealed that rhodamine-labelled trehalose was transported into cells rather than simply attached to the membrane. Additionally, the membranes were successfully preserved in lyophilised cells. This study demonstrates that ultrasonication with microbubbles facilitated trehalose delivery, offering promising cryoprotective capability without the cytotoxicity associated with DMSO-based methods.

The venom of Habrobracon hebetor induces alterations in host metabolism.

The ability of parasitic wasps to manipulate a host's metabolism is under active investigation. Components of venom play a major role in this process. In the present work, we studied the effect of the venom of the ectoparasitic wasp Habrobracon hebetor on the metabolism of the greater wax moth host (Galleria mellonella). We identified and quantified 45 metabolites in the lymph (cell-free hemolymph) of wax moth larvae on the second day after H. hebetor venom injection, using NMR spectroscopy and liquid chromatography coupled with mass spectrometry. These metabolites included 22 amino acids, nine products of lipid metabolism (sugars, amines and alcohols) and four metabolic intermediates related to nitrogenous bases, nucleotides and nucleosides. An analysis of the larvae metabolome suggested that the venom causes suppression of the tricarboxylic acid cycle, an increase in the number of free amino acids in the lymph, an increase in the concentration of trehalose in the lymph simultaneously with a decrease in the amount of glucose, and destructive processes in the fat body tissue. Thus, this parasitoid venom not only immobilizes the prey but also modulates its metabolism, thereby providing optimal conditions for the development of larvae.

Regulation of trehalose metabolism mediated by validamycin on chitin synthesis in Spodoptera frugiperda

AbstractTrehalase (TRE) is a key enzyme for degrading trehalose, which plays a vital role in the growth and development of insects. Although validamycin, a compound belonging to a class of efficient antibiotics and fungicides, can control pests by suppressing TRE activities, it remains unknown whether it acts on both trehalose and chitin metabolism in Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), a major pest of maize (Zea mays L., Poaceae). This study investigated the changes in trehalose metabolism after validamycin treatment in S. frugiperda and its effects on the downstream chitin synthesis pathway. Compared with the control, S. frugiperda exhibited varying degrees of mortality after treatment with four concentrations of validamycin, showing a dose‐dependent increase in mortality rate. The mortality rates 24 and 48 h after treatment with 0.07 mg μL−1 validamycin were 35.6% and 42.2%, respectively, indicating the effective lethal concentration. Treatment with 0.07 mg μL−1 validamycin led to developmental delay, abnormal molting, and death in S. frugiperda, but it exerted no lasting effects on the survival rate, pupal weight, and phenotype during its subsequent developmental stages. At 24 h after validamycin treatment, TRE1 and TRE2 activities and glucose content decreased significantly, whereas the trehalose content increased significantly. Treatment with validamycin significantly upregulated TRE1 and TRE2 expression after 24 and 48 h and downregulated the mRNA expression of chitin synthase A and B genes. However, after 72 h, chitin content was not significantly affected. Hence, validamycin can destroy the dynamic transformation balance of trehalose and glucose by inhibiting the activities of the two TREs, and further affect the expression of downstream chitin synthase genes. These findings provide a theoretical basis for using TRE inhibitors to control S. frugiperda.

Trehalose mediates salinity-stress tolerance in natural populations of a freshwater crustacean

Salinization poses an increasing problem worldwide, threatening freshwater organisms and raising questions about their ability to adapt. We explored the mechanisms enabling a planktonic crustacean to tolerate elevated salinity. By gradually raising water salinity in clonal cultures from 185 Daphnia magna populations, we showed that salt tolerance strongly correlates with native habitat salinity, indicating local adaptation. A genome-wide association study (GWAS) further revealed a major effect of the Alpha,alpha-trehalose-phosphate synthase (TPS) gene, suggesting that trehalose production facilitates salinity tolerance. Salinity-tolerant animals showed a positive correlation between water salinity and trehalose concentrations, while intolerant animals failed to produce trehalose. Animals with a non-functional TPS gene, generated through CRISPR-Cas9, supported the trehalose role in salinity stress. Our study highlights how a keystone freshwater animal adapts to salinity stress using an evolutionary mechanism known in bacteria, plants, and arthropods.

Regulation of spikelet developmental responses to chilling and freezing stress mediated by differential sugar metabolism in winter wheat

Spring cold stress poses a great threat to wheat reproductive growth, leading to compromised spike development and grain yield. There are two types of cold stress i.e. chilling stress (CS, above zero) and freezing stress (FS, below zero). However, it is unclear whether there is a difference in the mechanism of CS and FS in regulating spikelet development. This study aimed to unravel the underlying regulation in determining the difference for wheat spikelet exposed to CS at 2 °C and FS at −2°C by integrative analyses of transcriptome, metabolome, and physiology. Delayed floret development and shrunken cellular morphology in both CS and FS were observed, even malformation and degradation of anther cells occurred in FS. Kyoto Encyclopedia of gene and genomes (KEGG) analyses revealed that the most abundantly enriched pathways are phytohormone biosynthesis, starch and sucrose metabolism, and phenylpropanoid biosynthesis. Further physiological assays related to the identified pathways were performed. Compared to CS, the signal of abscisic acid (ABA), salicylic acid (SA) and jasmonic acid (JA) was more pronounced, and the signal of auxin (IAA) and gibberellin (GA) was inhibited further in FS. In addition, the contents of glucose, fructose and trehalose were elevated in CS, owing to greater activities of cell wall invertase and sucrose synthase, while the hexose content was decreased owing to lower activities of such enzymes in FS, concomitantly, flavonoid barely changed in CS, but it dramatically amounted in FS. Taken together, the glucose and trehalose pathway, along with induced ABA and SA signal were intensified in CS to maintain growth, while greater flavonoid and promoted JA synthesis were induced in FS for cold survival. Understanding the molecular of growth-defense under cold stress would provide a foundation for the development of breeding strategies.

Ozone induced multigenerational glucose and lipid metabolism disorders in Drosophila

Ozone (O3), a persistent pollutant, poses a significant health threat. However, research on its multigenerational toxicity remains limited. Leveraging the Drosophila model with its short lifespan and advanced genetic tools, we explored the effects of O3 exposure across three generations of fruit flies. The findings revealed that O3 disrupted motility, body weight, stress resistance, and oxidative stress in three generations of flies, with varying effects observed among them. Transcriptome analysis highlighted the disruption of glucose metabolism-related pathways, encompassing gluconeogenesis/glycolysis, galactose metabolism, and carbon metabolism. Hub genes were identified, and RT-qPCR results indicated that O3 decreased their transcription levels. Comparative analysis of their human orthologs was conducted using Comparative Toxicogenomics Database (CTD) and DisGeNET databases. These genes are linked to various metabolic diseases, including diabetes, hypoglycemia, and obesity. The trehalose content was reduced in F0 generation flies but increased in F1-F2 generations after O3 exposure. While the trehalase and glucose levels were decreased across F0-F2 generations. TAG synthesis-related genes were significantly upregulated in F0 generation flies but downregulated in F1-F2 generations. The expression patterns of lipolysis-related genes varied among the three generations of flies. Food intake was increased in F0 generation flies but decreased in F1-F2 generations. Moreover, TAG content was significantly elevated in F0 generation flies by O3 exposure, while it was reduced in F2 generation flies. These differential effects of O3 across three generations of flies suggest a metabolic reprogramming aimed at mitigating the damage caused by O3 to flies. The study affirms the viability of employing the Drosophila model to investigate the mechanisms underlying O3-induced glucose and lipid metabolism disorders while emphasizing the importance of studying the long-term health effects of O3 exposure. Moreover, this research highlights the Drosophila model as a viable tool for investigating the multigenerational effects of pollutants, particularly atmospheric pollutants.

Morphological and Biochemical Changes in the Mediterranean Cereal Cyst Nematode (Heterodera latipons) during Diapause.

The cereal cyst nematode (Heterodera latipons) is becoming an economically important species in global cereal production as it is being identified in many new cereal cultivated areas and causes significant losses. Consequently, understanding its biology becomes crucial for researchers in identifying its vulnerabilities and implementing effective control measures. In the current study, different morphological and biochemical changes of H. latipons cysts containing eggs with infective juveniles from a barley field in Jordan were studied during the summer of 2021, at two sample dates. The first, at the harvest of the cereal crop (June 2021), when the infective second-stage juveniles (J2s) were initiating diapause, and the second, before planting the sequent cereal crop (late October 2021), when the J2s were ending diapause. The studied population was characterized morphologically and molecularly, showing 98.4% molecular similarity to both JOD from Jordan and Syrian "300" isolates of H. latipons. The obtained results and observations revealed that there were dramatic changes in all the investigated features of the cysts and eggs they contained. Morphological changes such as cyst color, sub-crystalline layer, and thickness of the rigid eggshell wall were observed. A slight change in the emergence time of J2s from cysts was observed without any difference in the number of emerged J2s. The results of biochemical changes showed that the total contents of carbohydrates, glycogen, trehalose, glycerol, and protein were higher in cysts collected in October when compared to those cysts collected in June. The SDS-PAGE pattern indicated the presence of a protein with the size of ca. 100 kDa in both sampling dates, whereas another protein (ca. 20 kDa) was present only in the cysts of October. Furthermore, the expression of trehalase (tre) gene was detected only in H. latipons collected in October. The outcomes of this study provide new helpful information that elucidates diapause in H. latipons and may be used for the implementation of new management strategies of cyst nematodes.