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1.
Proc Natl Acad Sci U S A ; 121(26): e2405553121, 2024 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-38889144

RESUMO

The cytoplasm is a complex, crowded environment that influences myriad cellular processes including protein folding and metabolic reactions. Recent studies have suggested that changes in the biophysical properties of the cytoplasm play a key role in cellular homeostasis and adaptation. However, it still remains unclear how cells control their cytoplasmic properties in response to environmental cues. Here, we used fission yeast spores as a model system of dormant cells to elucidate the mechanisms underlying regulation of the cytoplasmic properties. By tracking fluorescent tracer particles, we found that particle mobility decreased in spores compared to vegetative cells and rapidly increased at the onset of dormancy breaking upon glucose addition. This cytoplasmic fluidization depended on glucose-sensing via the cyclic adenosine monophosphate-protein kinase A pathway. PKA activation led to trehalose degradation through trehalase Ntp1, thereby increasing particle mobility as the amount of trehalose decreased. In contrast, the rapid cytoplasmic fluidization did not require de novo protein synthesis, cytoskeletal dynamics, or cell volume increase. Furthermore, the measurement of diffusion coefficients with tracer particles of different sizes suggests that the spore cytoplasm impedes the movement of larger protein complexes (40 to 150 nm) such as ribosomes, while allowing free diffusion of smaller molecules (~3 nm) such as second messengers and signaling proteins. Our experiments have thus uncovered a series of signaling events that enable cells to quickly fluidize the cytoplasm at the onset of dormancy breaking.


Assuntos
Citoplasma , Schizosaccharomyces , Esporos Fúngicos , Trealose , Esporos Fúngicos/metabolismo , Esporos Fúngicos/fisiologia , Schizosaccharomyces/metabolismo , Schizosaccharomyces/fisiologia , Citoplasma/metabolismo , Trealose/metabolismo , Glucose/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Proteínas de Schizosaccharomyces pombe/metabolismo , Proteínas de Schizosaccharomyces pombe/genética , Transdução de Sinais
2.
Sci Rep ; 14(1): 14191, 2024 06 20.
Artigo em Inglês | MEDLINE | ID: mdl-38902334

RESUMO

Feeding behavior, the most fundamental physiological activity, is controlled by two opposing groups of factors, orexigenic and anorexigenic factors. The sulfakinin family, an insect analogue of the mammalian satiety factor cholecystokinin (CCK), has been shown to suppress food intake in various insects. Nevertheless, the mechanisms through which sulfakinin regulates feeding behavior remain a biological question. This study aimed to elucidate the signaling pathway mediated by the anorexigenic peptide sulfakinin in Bombyx mori. We identified the Bombyx mori neuropeptide G protein-coupled receptor A9 (BNGR-A9) as the receptor for sulfakinin through functional assays. Stimulation with sulfakinin triggered a swift increase in intracellular IP3, Ca2+, and a notable enhancement of ERK1/2 phosphorylation, in a manner sensitive to a Gαq-specific inhibitor. Treatment with synthetic sulfakinin resulted in decreased food consumption and average body weight. Additionally, administering synthetic sulfakinin to silkworms significantly elevated hemolymph trehalose levels, an effect markedly reduced by pre-treatment with BNGR-A9 dsRNA. Consequently, our findings establish the sulfakinin/BNGR-A9 signaling pathway as a critical regulator of feeding behavior and hemolymph trehalose homeostasis in Bombyx mori, highlighting its roles in the negative control of food intake and the positive regulation of energy balance.


Assuntos
Bombyx , Comportamento Alimentar , Hemolinfa , Homeostase , Proteínas de Insetos , Trealose , Animais , Bombyx/metabolismo , Bombyx/fisiologia , Trealose/metabolismo , Trealose/análogos & derivados , Trealose/farmacologia , Hemolinfa/metabolismo , Comportamento Alimentar/fisiologia , Proteínas de Insetos/metabolismo , Proteínas de Insetos/genética , Receptores Acoplados a Proteínas G/metabolismo , Neuropeptídeos/metabolismo , Transdução de Sinais
3.
Biophys J ; 123(12): 1690-1704, 2024 Jun 18.
Artigo em Inglês | MEDLINE | ID: mdl-38751113

RESUMO

Alzheimer's disease (AD) is a neurodegenerative disease characterized by dementia and memory loss in the elderly population. The amyloid-ß peptide (Aß) is one of the main pathogenic factors in AD and is known to cause damage to neuronal cellular membranes. There is no cure currently available for AD, and new approaches, including preventive strategies, are highly desirable. In this work, we explore the possibility of protecting neuronal membranes from amyloid-induced damage with naturally existing sugar trehalose. Trehalose has been shown to protect plant cellular membranes in extreme conditions and modify Aß misfolding. We hypothesize that trehalose can protect the neuronal membrane from amyloid toxicity. In this work, we studied the protective effect of trehalose against Aß1-42-induced damage in model lipid membranes (DPPC/POPC/cholesterol) using atomic force microscopy and black lipid membrane electrophysiology. Our results demonstrate that Aß1-42 damaged membranes and led to ionic current leakage across these membranes due to the formation of various defects and pores. The presence of trehalose reduced the ion current across membranes caused by Aß1-42 peptide damage, thus efficiently protecting the membranes. These findings suggest that the trehalose sugar can potentially be useful in protecting neuronal membranes against amyloid toxicity in AD.


Assuntos
Peptídeos beta-Amiloides , Bicamadas Lipídicas , Fragmentos de Peptídeos , Trealose , Trealose/farmacologia , Trealose/metabolismo , Peptídeos beta-Amiloides/metabolismo , Peptídeos beta-Amiloides/química , Fragmentos de Peptídeos/metabolismo , Fragmentos de Peptídeos/química , Bicamadas Lipídicas/química , Bicamadas Lipídicas/metabolismo , Membrana Celular/metabolismo , Membrana Celular/efeitos dos fármacos , Fenômenos Eletrofisiológicos/efeitos dos fármacos
4.
BMC Microbiol ; 24(1): 165, 2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38745279

RESUMO

Globally, drought stress poses a significant threat to crop productivity. Improving the drought tolerance of crops with microbial biostimulants is a sustainable strategy to meet a growing population's demands. This research aimed to elucidate microbial biostimulants' (Plant Growth Promoting Rhizobacteria) role in alleviating drought stress in oil-seed crops. In total, 15 bacterial isolates were selected for drought tolerance and screened for plant growth-promoting (PGP) attributes like phosphate solubilization and production of indole-3-acetic acid, siderophore, hydrogen cyanide, ammonia, and exopolysaccharide. This research describes two PGPR strains: Acinetobacter calcoaceticus AC06 and Bacillus amyloliquefaciens BA01. The present study demonstrated that these strains (AC06 and BA01) produced abundant osmolytes under osmotic stress, including proline (2.21 and 1.75 µg ml- 1), salicylic acid (18.59 and 14.21 µg ml- 1), trehalose (28.35 and 22.74 µg mg- 1 FW) and glycine betaine (11.35 and 7.74 mg g- 1) respectively. AC06 and BA01 strains were further evaluated for their multifunctional performance by inoculating in Arachis hypogaea L. (Groundnut) under mild and severe drought regimes (60 and 40% Field Capacity). Inoculation with microbial biostimulants displayed distinct osmotic-adjustment abilities of the groundnut, such as growth parameters, plant biomass, photosynthetic pigments, relative water content, proline, and soluble sugar in respective to control during drought. On the other hand, plant sensitivity indexes such as electrolyte leakage and malondialdehyde (MDA) contents were decreased as well as cooperatively conferred plant drought tolerance by induced alterations in stress indicators such as catalase (CAT), ascorbate peroxidase (APX), and superoxide dismutase (SOD). Thus, Acinetobacter sp. AC06 and Bacillus sp. BA01 can be considered as osmolyte producing microbial biostimulants to simultaneously induce osmotic tolerance and metabolic changes in groundnuts under drought stress.


Assuntos
Arachis , Secas , Estresse Fisiológico , Arachis/microbiologia , Arachis/crescimento & desenvolvimento , Arachis/metabolismo , Arachis/fisiologia , Prolina/metabolismo , Bacillus amyloliquefaciens/metabolismo , Bacillus amyloliquefaciens/fisiologia , Microbiologia do Solo , Pressão Osmótica , Betaína/metabolismo , Ácidos Indolacéticos/metabolismo , Ácido Salicílico/metabolismo , Acinetobacter/metabolismo , Acinetobacter/crescimento & desenvolvimento , Acinetobacter/fisiologia , Cianeto de Hidrogênio/metabolismo , Trealose/metabolismo
5.
Food Res Int ; 186: 114331, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38729716

RESUMO

Peach fruit is prone to chilling injury (CI) during low-temperature storage, resulting in quality deterioration and economic losses. Our previous studies have found that exogenous trehalose treatment can alleviate the CI symptoms of peach by increasing sucrose accumulation. The purpose of this study was to explore the potential molecular mechanism of trehalose treatment in alleviating CI in postharvest peach fruit. Transcriptome analysis showed that trehalose induced gene expression in pathways of plant MAPK signaling, calcium signaling, and reactive oxygen species (ROS) signaling. Furthermore, molecular docking analysis indicated that PpCDPK24 may activate the ROS signaling pathway by phosphorylating PpRBOHE. Besides, PpWRKY40 mediates the activation of PpMAPKKK2-induced ROS signaling pathway by interacting with the PpRBOHE promoter. Accordingly, trehalose treatment significantly enhanced the activities of antioxidant-related enzymes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and gluathione reductase (GR), as well as the transcription levels AsA-GSH cycle related gene, which led to the reduction of H2O2 and malondialdehyde (MDA) content in peach during cold storage. In summary, our results suggest that the potential molecular mechanism of trehalose treatment is to enhance antioxidant capacity by activating CDPK-mediated Ca2 + -ROS signaling pathway and WRKY-mediated MAPK-WRKY-ROS signaling pathway, thereby reducing the CI in peach fruit.


Assuntos
Antioxidantes , Temperatura Baixa , Frutas , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Prunus persica , Espécies Reativas de Oxigênio , Transdução de Sinais , Trealose , Trealose/farmacologia , Trealose/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Antioxidantes/farmacologia , Antioxidantes/metabolismo , Transdução de Sinais/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Simulação de Acoplamento Molecular , Malondialdeído/metabolismo
6.
Sci Rep ; 14(1): 10243, 2024 05 03.
Artigo em Inglês | MEDLINE | ID: mdl-38702388

RESUMO

The widespread use of multipotent mesenchymal stromal cell-derived secretome (MSC-sec) requires optimal preservation methods. Lyophilization offers benefits like concentrating the secretome, reducing the storage volume, and making storage conditions more flexible. This study evaluated the influence of storage duration and temperature on lyophilized MSC-sec. The conditioned medium from Wharton's jelly MSCs was stored at - 80 °C or lyophilized with or without trehalose. Lyophilized formulations were kept at - 80 °C, - 20 °C, 4 °C, or room temperature (RT) for 3 and 30 months. After storage and reconstitution, the levels of growth factors and cytokines were assessed using multiplex assay. The storage of lyophilized MSC-sec at - 80 °C ensured biomolecule preservation for 3 and 30 months. Following 3 month storage at 4 °C and RT, a notable decrease occurred in BDNF, bNGF, and sVCAM-1 levels. Prolonged 30 month storage at the same temperatures significantly reduced BDNF, bNGF, VEGF-A, IL-6, and sVCAM-1, while storage at - 20 °C decreased BDNF, bNGF, and VEGF- A levels. Trehalose supplementation of MSC-sec improved the outcome during storage at 4 °C and RT. Proper storage conditions were crucial for the preservation of lyophilized MSC-sec composition. Short-term storage at various temperatures maintained over 60% of the studied growth factors and cytokines; long-term preservation was only adequate at -80 °C.


Assuntos
Liofilização , Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/citologia , Humanos , Secretoma/metabolismo , Trealose/metabolismo , Trealose/farmacologia , Citocinas/metabolismo , Células Cultivadas , Meios de Cultivo Condicionados/química , Criopreservação/métodos , Temperatura
7.
BMC Plant Biol ; 24(1): 460, 2024 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-38797833

RESUMO

Trehalose serves as a crucial osmolyte and plays a significant role in stress tolerance. The influence of exogenously added trehalose (1 and 5 mM) in alleviating the chromium (Cr; 0.5 mM) stress-induced decline in growth, photosynthesis, mineral uptake, antioxidant system and nitrate reductase activity in Vigna radiata was studied. Chromium (Cr) significantly declined shoot height (39.33%), shoot fresh weight (35.54%), shoot dry weight (36.79%), total chlorophylls (50.70%), carotenoids (29.96%), photosynthesis (33.97%), net intercellular CO2 (26.86%), transpiration rate (36.77%), the content of N (35.04%), P (35.77%), K (31.33%), S (23.91%), Mg (32.74%), and Ca (29.67%). However, the application of trehalose considerably alleviated the decline. Application of trehalose at both concentrations significantly reduced hydrogen peroxide accumulation, lipid peroxidation and electrolyte leakage, which were increased due to Cr stress. Application of trehalose significantly mitigated the Cr-induced oxidative damage by up-regulating the activity of reactive oxygen species (ROS) scavenging enzymes, including superoxide dismutase (182.03%), catalase (125.40%), ascorbate peroxidase (72.86%), and glutathione reductase (68.39%). Besides this, applied trehalose proved effective in enhancing ascorbate (24.29%) and reducing glutathione content (34.40%). In addition, also alleviated the decline in ascorbate by Cr stress to significant levels. The activity of nitrate reductase enhanced significantly (28.52%) due to trehalose activity and declined due to Cr stress (34.15%). Exogenous application of trehalose significantly improved the content of osmolytes, including proline, glycine betaine, sugars and total phenols under normal and Cr stress conditions. Furthermore, Trehalose significantly increased the content of key mineral elements and alleviated the decline induced by Cr to considerable levels.


Assuntos
Cromo , Estresse Oxidativo , Fotossíntese , Espécies Reativas de Oxigênio , Trealose , Vigna , Trealose/metabolismo , Trealose/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Fotossíntese/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Vigna/efeitos dos fármacos , Vigna/crescimento & desenvolvimento , Vigna/metabolismo , Minerais/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Clorofila/metabolismo , Antioxidantes/metabolismo
8.
PeerJ ; 12: e17332, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38799059

RESUMO

The burgeoning issue of landfill leachate, exacerbated by urbanization, necessitates evaluating its biological impact, traditionally overshadowed by physical and chemical assessments. This study harnesses Caenorhabditis elegans, a model organism, to elucidate the physiological toxicity of landfill leachate subjected to different treatment processes: nanofiltration reverse osmosis tail water (NFRO), membrane bioreactor (MBR), and raw leachate (RAW). Our investigation focuses on the modulation of sugar metabolism, particularly trehalose-a disaccharide serving dual functions as an energy source and an anti-adversity molecule in invertebrates. Upon exposure, C. elegans showcased a 60-70% reduction in glucose and glycogen levels alongside a significant trehalose increase, highlighting an adaptive response to environmental stress by augmenting trehalose synthesis. Notably, trehalose-related genes in the NFRO group were up-regulated, contrasting with the MBR and RAW groups, where trehalose synthesis genes outpaced decomposition genes by 20-30 times. These findings suggest that C. elegans predominantly counters landfill leachate-induced stress through trehalose accumulation. This research not only provides insights into the differential impact of leachate treatment methods on C. elegans but also proposes a molecular framework for assessing the environmental repercussions of landfill leachate, contributing to the development of novel strategies for pollution mitigation and environmental preservation.


Assuntos
Caenorhabditis elegans , Trealose , Poluentes Químicos da Água , Animais , Caenorhabditis elegans/efeitos dos fármacos , Caenorhabditis elegans/genética , Caenorhabditis elegans/metabolismo , Trealose/metabolismo , Estresse Fisiológico/efeitos dos fármacos
9.
Plant Physiol Biochem ; 211: 108620, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38714124

RESUMO

Desiccation is a state of extreme water loss that is lethal to many plant species. Some desert plants have evolved unique strategies to cope with desiccation stress in their natural environment. Here we present the remarkable stress management mechanism of Syntrichia caninervis, a desert moss species which exhibits an 'A' category of desiccation tolerance. Our research demonstrated that desiccation stress triggers autophagy in S. caninervis while inhibiting Programmed Cell Death (PCD). Silencing of two autophagy-related genes, ATG6 and ATG2, in S. caninervis promoted PCD. Desiccation treatment accelerated cell death in ATG6 and ATG2 gene-silenced S. caninervis. Notably, trehalose was not detected during desiccation, and exogenous application of trehalose cannot activate autophagy. These results suggested that S. caninervis is independent of trehalose accumulation to triggered autophagy. Our results showed that autophagy function as prosurvival mechanism to enhance desiccation tolerance of S. caninervis. Our findings enrich the knowledge of the role of autophagy in plant stress response and may provide new insight into understanding of plant desiccation tolerance.


Assuntos
Autofagia , Dessecação , Trealose , Trealose/metabolismo , Apoptose , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Estresse Fisiológico , Regulação da Expressão Gênica de Plantas
10.
Sci Adv ; 10(20): eadn0895, 2024 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-38758793

RESUMO

SUCROSE-NON-FERMENTING1-RELATED PROTEIN KINASE1 (SnRK1), a central plant metabolic sensor kinase, phosphorylates its target proteins, triggering a global shift from anabolism to catabolism. Molecular modeling revealed that upon binding of KIN10 to GEMINIVIRUS REP-INTERACTING KINASE1 (GRIK1), KIN10's activation T-loop reorients into GRIK1's active site, enabling its phosphorylation and activation. Trehalose 6-phosphate (T6P) is a proxy for cellular sugar status and a potent inhibitor of SnRK1. T6P binds to KIN10, a SnRK1 catalytic subunit, weakening its affinity for GRIK1. Here, we investigate the molecular details of T6P inhibition of KIN10. Molecular dynamics simulations and in vitro phosphorylation assays identified and validated the T6P binding site on KIN10. Under high-sugar conditions, T6P binds to KIN10, blocking the reorientation of its activation loop and preventing its phosphorylation and activation by GRIK1. Under these conditions, SnRK1 maintains only basal activity levels, minimizing phosphorylation of its target proteins, thereby facilitating a general shift from catabolism to anabolism.


Assuntos
Proteínas de Arabidopsis , Simulação de Dinâmica Molecular , Proteínas Serina-Treonina Quinases , Fosfatos Açúcares , Trealose , Fosfatos Açúcares/metabolismo , Trealose/análogos & derivados , Trealose/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Fosforilação , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/antagonistas & inibidores , Proteínas de Arabidopsis/química , Ligação Proteica , Arabidopsis/metabolismo , Sítios de Ligação , Fatores de Transcrição
11.
PLoS Biol ; 22(5): e3002299, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38713712

RESUMO

Activation of immune cells requires the remodeling of cell metabolism in order to support immune function. We study these metabolic changes through the infection of Drosophila larvae by parasitoid wasp. The parasitoid egg is neutralized by differentiating lamellocytes, which encapsulate the egg. A melanization cascade is initiated, producing toxic molecules to destroy the egg while the capsule also protects the host from the toxic reaction. We combined transcriptomics and metabolomics, including 13C-labeled glucose and trehalose tracing, as well as genetic manipulation of sugar metabolism to study changes in metabolism, specifically in Drosophila hemocytes. We found that hemocytes increase the expression of several carbohydrate transporters and accordingly uptake more sugar during infection. These carbohydrates are metabolized by increased glycolysis, associated with lactate production, and cyclic pentose phosphate pathway (PPP), in which glucose-6-phosphate is re-oxidized to maximize NADPH yield. Oxidative PPP is required for lamellocyte differentiation and resistance, as is systemic trehalose metabolism. In addition, fully differentiated lamellocytes use a cytoplasmic form of trehalase to cleave trehalose to glucose and fuel cyclic PPP. Intracellular trehalose metabolism is not required for lamellocyte differentiation, but its down-regulation elevates levels of reactive oxygen species, associated with increased resistance and reduced fitness. Our results suggest that sugar metabolism, and specifically cyclic PPP, within immune cells is important not only to fight infection but also to protect the host from its own immune response and for ensuring fitness of the survivor.


Assuntos
Glucose , Hemócitos , Via de Pentose Fosfato , Trealose , Animais , Trealose/metabolismo , Glucose/metabolismo , Hemócitos/metabolismo , Larva/metabolismo , Larva/parasitologia , Drosophila melanogaster/metabolismo , Drosophila melanogaster/parasitologia , Resistência à Doença , Glicólise , Interações Hospedeiro-Parasita , Vespas/metabolismo , Vespas/fisiologia , Diferenciação Celular , Drosophila/metabolismo , Drosophila/parasitologia
12.
Biochem Biophys Res Commun ; 716: 149971, 2024 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-38697009

RESUMO

α,α-trehalose is a well-known sugar that plays a key role in establishing tolerance to environmental stresses in many organisms, except unicellular eukaryotes. However, almost nothing is known about α,ß-trehalose, including their synthesis, function, and even presence in living organisms. In this study, we identified α,ß-trehalose in the resting cyst, a dormancy cell form characterized by extreme tolerance to environmental stresses, of the ciliated protist Colpoda cucullus, using high-performance liquid chromatography (HPLC), and a proton nuclear magnetic resonance (1H NMR). Gene expression analysis revealed that the expression of trehalose-6-phosphate synthase (TPS), glycosyltransferase (GT), alpha-amylase (AMY), and trehalose transporter 1 (TRET1), were up-regulated in encystment, while the expression of α-glucosidase 2 (AG2) and trehalase (TREH) was up-regulated in excystment. These results suggest that α,ß-trehalose is synthesized during encystment process, while and contributes to extreme tolerances to environmental stressors, stored carbohydrates, and energy reserve during resting cyst and/or during excystment.


Assuntos
Cilióforos , Trealose , Cilióforos/metabolismo , Cilióforos/genética , Trealose/metabolismo , Trealose/análogos & derivados , Estresse Fisiológico , Glucosiltransferases/metabolismo , Glucosiltransferases/genética
13.
Molecules ; 29(9)2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38731579

RESUMO

Trehalose is a naturally occurring, non-reducing saccharide widely distributed in nature. Over the years, research on trehalose has revealed that this initially thought simple storage molecule is a multifunctional and multitasking compound protecting cells against various stress factors. This review presents data on the role of trehalose in maintaining cellular homeostasis under stress conditions and in the virulence of bacteria and fungi. Numerous studies have demonstrated that trehalose acts in the cell as an osmoprotectant, chemical chaperone, free radical scavenger, carbon source, virulence factor, and metabolic regulator. The increasingly researched medical and therapeutic applications of trehalose are also discussed.


Assuntos
Trealose , Trealose/farmacologia , Trealose/metabolismo , Humanos , Animais , Fungos/metabolismo , Fungos/efeitos dos fármacos , Bactérias/metabolismo , Bactérias/efeitos dos fármacos , Homeostase/efeitos dos fármacos , Estresse Fisiológico/efeitos dos fármacos
14.
Plant Physiol Biochem ; 211: 108695, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38744088

RESUMO

The presence of sugar in plant tissue can lead to an increase in the osmotic pressure within cells, a decrease in the freezing point of plants, and protection against ice crystal damage to the tissue. Trehalose is closely related to sucrose, which comprises the largest proportion of sugar and has become a hot topic of research in recent years. Our previous studies have confirmed that a key trehalose synthesis gene, TaTPS11, from the cold-resistant winter wheat DM1, could enhance the cold resistance of plants by increasing sugar content. However, the underlying mechanism behind this phenomenon remains unclear. In this study, we cloned TaTPS11-6D, edited TaTPS11-6D using CRISPR/Cas9 technology and transformed 'Fielder' to obtain T2 generation plants. We screened out OE3-3 and OE8-7 lines with significantly higher cold resistance than that of 'Fielder' and Cri 4-3 edited lines with significantly lower cold resistance than that of 'Fielder'. Low temperature storage limiting factors were measured for OE3-3, OE8-7 and Cri 4-3 treated at different temperatures.The results showed that TaTPS11-6D significantly increased the content of sugar in plants and the transfer of sugar from source to storage organs under cold conditions. The TaTPS11-6D significantly increased the levels of salicylic, jasmonic, and abscisic acids while also significantly decreasing the level of gibberellic acid. Our research improves the model of low temperature storage capacity limiting factor.


Assuntos
Temperatura Baixa , Proteínas de Plantas , Triticum , Triticum/genética , Triticum/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Regulação da Expressão Gênica de Plantas , Trealose/metabolismo , Ácido Abscísico/metabolismo , Oxilipinas/metabolismo , Ciclopentanos/metabolismo , Giberelinas/metabolismo , Sacarose/metabolismo
15.
BMC Plant Biol ; 24(1): 472, 2024 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-38811894

RESUMO

Salinity stress, an ever-present challenge in agriculture and environmental sciences, poses a formidable hurdle for plant growth and productivity in saline-prone regions worldwide. Therefore, this study aimed to explore the effectiveness of trehalose and mannitol induce salt resistance in wheat seedlings. Wheat grains of the commercial variety Sakha 94 were divided into three groups : a group that was pre-soaked in 10 mM trehalose, another group was soaked in 10 mM mannitol, and the last was soaked in distilled water for 1 hour, then the pre soaked grains cultivated in sandy soil, each treatment was divided into two groups, one of which was irrigated with 150 mM NaCl and the other was irrigated with tap water. The results showed that phenols content in wheat seedlings increased and flavonoids reduced due to salt stress. Trehalose and mannitol cause slight increase in total phenols content while total flavonoids were elevated highy in salt-stressed seedlings. Furthermore, Trehalose or mannitol reduced salt-induced lipid peroxidation. Salt stress increases antioxidant enzyme activities of guaiacol peroxidase (G-POX), ascorbate peroxidase (APX), and catalase (CAT) in wheat seedlings, while polyphenol oxidase (PPO) unchanged. Trehalose and mannitol treatments caused an increase in APX, and CAT activities, whereas G-POX not altered but PPO activity were decreased under salt stress conditions. Molecular docking confirmed the interaction of Trehalose or mannitol with peroxidase and ascorbic peroxidase enzymes. Phenyl alanine ammonia layase (PAL) activity was increased in salt-stressed seedlings. We can conclude that pre-soaking of wheat grains in 10 mM trehalose or mannitol improves salinity stress tolerance by enhancing antioxidant defense enzyme and/or phenol biosynthesis, with docking identifying interactions with G-POX, CAT, APX, and PPO.


Assuntos
Manitol , Tolerância ao Sal , Plântula , Trealose , Triticum , Triticum/efeitos dos fármacos , Triticum/fisiologia , Triticum/metabolismo , Trealose/metabolismo , Plântula/efeitos dos fármacos , Plântula/fisiologia , Manitol/farmacologia , Tolerância ao Sal/efeitos dos fármacos , Simulação de Acoplamento Molecular , Antioxidantes/metabolismo , Estresse Salino/efeitos dos fármacos , Flavonoides/metabolismo , Fenóis/metabolismo
16.
Int J Biol Macromol ; 271(Pt 1): 132503, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38768913

RESUMO

The cold-adapted bacterium Variovorax sp. PAMC28711 possesses two distinct glycoside hydrolase (GH) families of trehalase, GH15 and GH37. While numerous studies have explored bacterial trehalase, the presence of two different trehalase genes within a single strain has not been reported until now. Interestingly, despite both GH37 and GH15 trehalases serving the same purpose of degrading trehalose, but do not share the sequence similarity. The substrate specificity assay confirmed that Vtre37 and Vtre15 displayed hydrolytic activity on α, α-trehalose. The key catalytic sites were identified as D280 and E469 in Vtre37 and E389 and E554 in Vtre15 through site-directed mutation and confirmed these two enzymes belong to trehalase. In addition, Vtre37 exhibited a relatively high level of enzyme activity of 1306.33 (±53.091) µmolmg-1, whereas Vtre15 showed enzyme activity of 408.39 (±12.503) µmolmg-1. Moreover, Vtre37 performed admirably showing resistance to ethanol (10 %), with high stable at acidic pH range. Furthermore, both prediction and experimental results indicate that validoxylamine A showed a potent inhibitory activity against Vtre37 trehalase with a Ki value of 16.85 nM. Therefore, we postulate that Vtre37 could be utilized as an ethanol enhancer and designed for screening inhibitors related to the trehalose degradation pathway. Additionally, we believe that characterizing these bacterial trehalase contributes to a better understanding of trehalose metabolism and its biological importance in bacteria.


Assuntos
Temperatura Baixa , Comamonadaceae , Trealase , Trealase/metabolismo , Trealase/genética , Trealase/química , Especificidade por Substrato , Comamonadaceae/enzimologia , Comamonadaceae/genética , Domínio Catalítico , Trealose/metabolismo , Trealose/farmacologia , Concentração de Íons de Hidrogênio , Hidrólise , Cinética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/química , Sequência de Aminoácidos , Estabilidade Enzimática , Adaptação Fisiológica
17.
Plant Cell Environ ; 47(7): 2475-2490, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38567814

RESUMO

Phosphorus (P)-hyperaccumulators for phytoextraction from P-polluted areas generally show rapid growth and accumulate large amounts of P without any toxicity symptom, which depends on a range of physiological processes and gene expression patterns that have never been explored. We investigated growth, leaf element concentrations, P fractions, photosynthetic traits, and leaf metabolome and transcriptome response in amphibious P-hyperaccumulators, Polygonum hydropiper and P. lapathifolium, to high-P exposure (5 mmol L-1), with 0.05 mmol L-1 as the control. Under high-P exposure, both species demonstrated good growth, allocating more P to metabolite P and inorganic P (Pi) accompanied by high potassium and calcium. The expression of a cluster of unigenes associated with photosynthesis was maintained or increased in P. lapathifolium, explaining the increase in net photosynthetic rate and the rapid growth under high-P exposure. Metabolites of trehalose metabolism, including trehalose 6-phosphate and trehalose, were sharply increased in both species by the high-P exposure, in line with the enhanced expression of associated unigenes, indicating that trehalose metabolic pathway was closely related to high-P tolerance. These findings elucidated the physiological and molecular responses involved in the photosynthesis and trehalose metabolism in P-hyperaccumulators to high-P exposure, and provides potential regulatory pathways to improve the P-phytoextraction capability.


Assuntos
Regulação da Expressão Gênica de Plantas , Fósforo , Fotossíntese , Folhas de Planta , Polygonum , Fósforo/metabolismo , Polygonum/metabolismo , Polygonum/genética , Polygonum/efeitos dos fármacos , Polygonum/fisiologia , Folhas de Planta/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Trealose/metabolismo , Metaboloma , Transcriptoma
18.
Cryobiology ; 115: 104898, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38663665

RESUMO

Trehalose is widely acknowledged for its ability to stabilize plasma membranes during dehydration. However, the exact mechanism by which trehalose interacts with lipid bilayers remains presently unclear. In this study, we conducted atomistic molecular dynamic simulations on asymmetric model bilayers that mimic the membrane of human red blood cells at various trehalose and water contents. We considered three different hydration levels mimicking the full hydration to desiccation scenarios. Results indicate that the asymmetric distribution of lipids did not significantly influence the computed structural characteristics at full and low hydration. At dehydration, however, the order parameter obtained from the symmetric bilayer is significantly higher compared to those obtained from asymmetric ones. Analysis of hydrogen bonds revealed that the protective ability of trehalose is well described by the water replacement hypothesis at full and low hydration, while at dehydration other interaction mechanisms associated with trehalose exclusion from the bilayer may involve. In addition, we found that trehalose exclusion is not attributed to sugar saturation but rather to the reduction in hydration levels. It can be concluded that the protective effect of trehalose is not only related to the hydration level of the bilayer, but also closely tied to the asymmetric distribution of lipids within each leaflet.


Assuntos
Membrana Eritrocítica , Ligação de Hidrogênio , Bicamadas Lipídicas , Simulação de Dinâmica Molecular , Trealose , Trealose/metabolismo , Trealose/química , Bicamadas Lipídicas/química , Bicamadas Lipídicas/metabolismo , Humanos , Membrana Eritrocítica/metabolismo , Membrana Eritrocítica/química , Água/química , Água/metabolismo , Eritrócitos/metabolismo , Eritrócitos/química , Dessecação
19.
Skin Res Technol ; 30(4): e13666, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38606717

RESUMO

BACKGROUND: It is known that heparinoid, a mucopolysaccharide polysulfate, is effective in improving rough skin and promoting blood circulation as medicines for diseased areas. However, heparinoid has a molecular weight of more than 5000 and cannot penetrate healthy stratum corneum. OBJECTIVE: We tested the efficacy of sulfated oligosaccharides with a molecular weight of less than 2000 on the human skin barrier function and moisturizing function. METHODS: We measured the transepidermal water loss (TEWL) of a three-dimensional human epidermis model cultured for 3 days after topical application of sulfated oligosaccharides, then observed the effects on TEWL suppression. The mRNA levels of proteins involved in intercellular lipid transport and storage in the stratum corneum, and moisture retention were measured using RT-qPCR. RESULTS: An increase in the mRNA levels of the ATP-binding cassette subfamily A member 12 (ABCA12), which transports lipids into stratum granulosum, was confirmed. Increases were also observed in the mRNA levels of filaggrin (FLG), which is involved in the generation of natural moisturizing factors, and of caspase-14, calpain-1 and bleomycin hydrolase, which are involved in the degradation of FLG. Antibody staining confirmed that the application of sodium trehalose sulfate to 3D model skin resulted in more ABCA12, ceramide, transglutaminase1, and FLG than those in controls. In a randomized, placebo-controlled, double-blind study, participants with low stratum corneum water content applied a lotion and emulsion containing sodium trehalose sulfate to their faces for 4 weeks. Sodium trehalose sulfate decreased the TEWL and increased the stratum corneum water content. CONCLUSION: These results suggest that cosmetics containing sodium trehalose sulfate act on the epidermis by increasing barrier factors and moisturizing factors, thereby ameliorating dry skin.


Assuntos
Heparinoides , Trealose , Humanos , Epiderme/metabolismo , Heparinoides/metabolismo , Heparinoides/farmacologia , RNA Mensageiro/metabolismo , Pele/metabolismo , Higiene da Pele , Sódio/metabolismo , Sódio/farmacologia , Trealose/farmacologia , Trealose/metabolismo , Água/metabolismo , População do Leste Asiático
20.
PLoS One ; 19(3): e0299669, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38452127

RESUMO

To investigate the role of sugar metabolism in desiccation-sensitive seeds, we performed a natural desiccation treatment on Phoebe chekiangensis seeds in a room and systematically analyzed the changes in seed germination, sugar compounds, malondialdehyde, and relative electrical conductivity during the seed desiccation. The results revealed that the initial moisture content of P. chekiangensis seed was very high (37.06%) and the seed was sensitive to desiccation, the germination percentage of the seed decreased to 5.33% when the seed was desiccated to 22.04% of moisture content, therefore, the seeds were considered recalcitrant. Based on the logistic model, we know that the moisture content of the seeds is 29.05% when the germination percentage drops to 50% and that it is desirable to keep the seed moisture content above 31.74% during ambient transportation. During seed desiccation, sucrose and trehalose contents exhibited increasing trends, and raffinose also increased during the late stage of desiccation, however, low levels of the non-reducing sugar accumulations may not prevent the loss of seed viability caused by desiccation. Glucose and fructose predominated among sugar compounds, and they showed a slight increase followed by a significant decrease. Their depletion may have contributed to the accumulation of sucrose and raffinose family oligosaccharides. Correlation analysis revealed a significant relationship between the accumulation of sucrose, trehalose, and soluble sugars, and the reduction in seed viability. Sucrose showed a significant negative correlation with glucose and fructose. Trehalose also exhibited the same pattern of correlation. These results provided additional data and theoretical support for understanding the mechanism of sugar metabolism in seed desiccation sensitivity.


Assuntos
Dessecação , Açúcares , Açúcares/metabolismo , Dessecação/métodos , Rafinose/metabolismo , Trealose/metabolismo , Sementes/metabolismo , Germinação , Sacarose/metabolismo , Glucose/metabolismo , Frutose/metabolismo
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