Phosphate Synthase Research Articles

Carbon and nitrogen signaling regulate FLOWERING LOCUS C and impact flowering time in Arabidopsis.

The timing of flowering in plants is modulated by both carbon (C) and nitrogen (N) signaling pathways. In a previous study, we established a pivotal role of the sucrose-signaling trehalose 6-phosphate pathway in regulating flowering under N-limited short-day conditions. In this work, we show that both wild-type Arabidopsis (Arabidopsis thaliana) plants grown under N-limited conditions and knock-down plants of TREHALOSE PHOSPHATE SYNTHASE 1 induce FLOWERING LOCUS C (FLC) expression, a well-known floral repressor associated with vernalization. When exposed to an extended period of cold, a flc mutant fails to respond to N availability and flowers at the same time under N-limited and full-nutrition conditions. Our data suggest that SUCROSE NON-FERMENTING 1 RELATED KINASE 1-dependent trehalose 6-phosphate-mediated C signaling and a mechanism downstream of N signaling (likely involving NIN-LIKE PROTEIN 7) impact the expression of FLC. Collectively, our data underscore the existence of a multi-factor regulatory system in which the C and N signaling pathways jointly govern the regulation of flowering in plants.

Simulation of Defoliation Effects on Relay Strip Intercropping Soybean: Elucidating Foliar Shedding and Leaf-to-Nodule Growth Plasticity.

Extensive foliar shedding in monoculture soybeans post-anthesis negatively impacts yield, whereas relay strip intercropping prolongs leaf area duration, enhancing productivity. However, little is known about the causes of leaf shedding in monoculture and its impact on physiological functions and plasticity of source and sink organs, we conducted a 4-year field experiment and leaf-removal simulations in relay intercropped soybeans. Results revealed that monoculture soybeans experienced severe self-shading and defoliation, while relay intercropping maintained better light conditions, supporting higher leaf area, nodule numbers, and carbon allocation. Increasing leaf removal initially increased leaf area but eventually reduced it. Extensive leaf-removal reduced Rubisco and sucrose phosphate synthase (SPS) activity, as well as sucrose, malate, ATP, and energy charge (EC) in nodules, revealing a trade-off between leaf growth and nodule development. Moderate leaf-removal (L30), however, balanced compensation and consumption, increasing total non-structural carbohydrates (TNC) in roots and N and ureide in leaves and pods. Network analysis showed that L30 improved the synergies of functional traits in leaves and nodules, ultimately benefiting overall plant growth and nutrient accumulation in pods. This study elucidates a mechanism of foliar shedding and highlights how relay strip intercropping optimizes source-sink coordination to enhance photosynthesis and nitrogen fixation.

Aldehyde-based Activation of C2α-lactylthiamin Diphosphate Decarboxylation on Bacterial 1-deoxy-d-xylulose 5-phosphate Synthase.

1-Deoxy-d-xylulose 5-phosphate synthase (DXPS) catalyzes the thiamin diphosphate (ThDP)-dependent formation of DXP from pyruvate (donor substrate) and d-glyceraldehyde 3-phosphate (d-GAP, acceptor substrate) in bacterial central metabolism. DXPS uses a ligand-gated mechanism in which binding of a small molecule "trigger" activates the first enzyme-bound intermediate, C2α-lactylThDP (LThDP), to form the reactive carbanion via LThDP decarboxylation. d-GAP is the natural acceptor substrate for DXPS and also serves a role as a trigger to induce LThDP decarboxylation in the gated step. Additionally, we have shown that O2 and d-glyceraldehyde (d-GA) can induce LThDP decarboxylation. We hypothesize this ligand-gated mechanism poises DXPS to sense and respond to cellular cues in metabolic remodeling during bacterial adaptation. Here we sought to characterize features of small molecule inducers of LThDP decarboxylation. Using a combination of CD, NMR and biochemical methods, we demonstrate that the α-hydroxy aldehyde moiety of d-GAP is sufficient to induce LThDP decarboxylation en route to DXP formation. A variety of aliphatic aldehydes also induce LThDP decarboxylation. The study highlights the capacity of DXPS to respond to different molecular cues, lending support to potential multifunctionality of DXPS and its metabolic regulation by this mechanism.

Exogenous Riboflavin Application at Different Growth Stages Regulates Photosynthetic Accumulation and Grain Yield in Fragrant Rice

Fragrant rice has high market value and is popular among consumers because of its pleasant fragrance. Plant growth regulators and trace elements can influence the yield and physiological indices of fragrant rice. Riboflavin, a vital component of plant vitamins, plays a crucial role in plant growth and development, and it is a water-soluble vitamin that is essential for sustaining normal photosynthesis and metabolic processes. However, the effects of riboflavin on nutrient accumulation and yield formation in fragrant rice have rarely been reported. Therefore, to further increase the yield and quality of fragrant rice, this study investigated the impact of the foliar application of riboflavin at different growth stages on nutrient accumulation and yield formation in two different genotypes of fragrant rice via a pot experiment. The experimental design consisted of three treatment groups, i.e., the T0 treatment group (control), which was sprayed with water; the T1 treatment group, which was sprayed at the booting stage; and the T2 treatment group, which was sprayed at the flowering stage; in all of these groups, 20 mg·L−1 riboflavin solution was used. The results revealed that the yields of the T1 and T2 treatments increased by 6.56–7.25% and 10.52–13.80%, respectively, compared with those of T0, which was attributed mainly to the increase in 1000-grain weight and grain filling. Furthermore, foliar spraying of riboflavin significantly increased the chlorophyll content, which resulted in the increased production of more total starch, soluble sugars, and sucrose and facilitated their transportation to the seeds. Moreover, applying riboflavin directly to the leaves increased the activity levels of the sucrose synthase (SS) and sucrose phosphate synthase (SPS) enzymes. Among the three treatments, the T2 treatment had the most pronounced effect. The results revealed that the foliar application of riboflavin could increase photosynthesis and promote the production of nonstructural carbohydrates, thereby increasing the total aboveground biomass of fragrant rice by 17.00–20.91% and 21.07–72.91% under the T1 and T2 treatments, respectively. Additionally, spraying riboflavin promoted rapid increases in the weight of fragrant rice seeds. In conclusion, foliar spraying of riboflavin during the flowering stage of aromatic rice can effectively enhance photosynthetic accumulation and yield, which is a promising physiological regulation strategy and provides new theoretical guidance for high-yield cultivation practices.

SlMIPS2, a myo-inositol phosphate synthase gene, regulates phosphate homeostasis by influencing SlPHL1 and SlSPX2 levels in tomato seedlings.

Phosphorus (P) is a quintessential macronutrient utilized by plants to support various metabolic processes during growth and development. Recent studies have revealed the pivotal role of inositol hexa-kis/pyrophosphate (InsP6-8), the derivatives of Myo-inositol (MI), in facilitating the interaction between SYG1/PHO81/XPR1 (SPX) and Phosphate starvation response (PHR) proteins. Myo-inositol phosphate synthase (MIPS) catalyzes the first committed step in MI biosynthesis. Although the role of MIPS genes in mediating stress responses in plants is well elucidated, its role in phosphate (Pi) deficiency remains largely unexplored. This study demonstrates that out of the five MIPS genes encoded by the tomato genome, only SlMIPS2 is sharply induced at an early stage of Pi starvation in tomato seedlings. Silencing of SlMIPS2 led to improved seedling growth with enhanced total soluble Pi and total P levels in the silenced plants under high Pi availability. SlMIPS2 silencing also caused a significant reduction in MI and InsP6 content in the tomato seedlings. These seedlings with depleted InsP6 levels accumulated lower levels of SlSPX2 protein. In contrast, stabilized SlPHL1 levels were noticed in these plants, directly implicating this transcription factor in activating phosphate starvation inducible (PSI) genes in the SlMIPS2-silenced seedlings, even under high Pi conditions. The results assign a novel role to SlMIPS2 in regulating cellular InsP6 levels and SPX-PHR interactions to control Pi homeostasis in tomato seedlings.

Baseline survey reveals glyphosate and dicamba resistance in broadleaf weeds before sugar beet trait introduction

Abstract A pre-launch survey of broadleaf weeds was conducted to predict the weed management efficacy of a novel genetically engineered sugar beet with resistance traits for glyphosate, dicamba, and glufosinate. We targeted problematic broadleaf weed species prevalent in the sugar beet system, including kochia, common lambsquarters, Palmer amaranth, and redroot pigweed across sugar beet areas in Colorado, Nebraska, and Wyoming. The results revealed that a significant percentage of kochia populations in Colorado, Nebraska, and Wyoming exhibited resistance to glyphosate (94%, 98%, and 75%, respectively) and dicamba (30%, 42%, and 17%, respectively). Palmer amaranth populations had resistance frequencies for glyphosate and dicamba of 80% and 20% in Colorado and 20% and 3% in Nebraska, respectively. No resistance to the tested herbicides was identified in common lambsquarters or redroot pigweed. Glufosinate resistance was not identified for any species. Kochia and Palmer amaranth populations from Colorado and Nebraska exhibited glyphosate resistance primarily through 5-enolpyruvylshikimate 3-phosphate synthase (EPSPS) gene amplification. However, one glyphosate-resistant kochia population from Wyoming lacked EPSPS gene amplification, indicating the presence of an alternative resistance mechanism. We identified the previously characterized IAA16 G73N substitution in a dicamba-resistant kochia population from Nebraska. However, dicamba-resistant kochia populations from Colorado did not possess this substitution, suggesting an alternative, yet-to-be-determined resistance mechanism. The widespread prevalence of glyphosate and dicamba resistance, coupled with the emergence of novel resistance mechanisms, poses a significant challenge to the long-term efficacy of this novel genetically engineered sugar beet technology. These findings underscore the urgent need for integrated weed management (IWM) strategies that diversify effective herbicide sites-of-action and incorporate alternative weed management practices within cropping systems.

Synergistic Effects of Irrigation and Nitrogen Fertilisation on Maize Photosynthetic Performance and Yield of Rainfed Systems in Drought‐Prone Environments

ABSTRACTMaize, a cereal crop of global significance, encounters cultivation challenges in the subtropical regions of Guangxi, mainly due to variable rainfall and low soil fertility, exacerbating the effects of drought. This study evaluated the effects of irrigation and nitrogen fertilisation on overcoming these challenges and improving maize growth and yield. Between 2020 and 2021, a split‐plot experiment was conducted. The main plots were assigned to two irrigation treatments: irrigated and rainfed. Within each main plot, subplots were treated with different nitrogen levels (0, 150, 200, 250 and 300 kg ha−1). The results showed that nitrogen levels and water regime significantly impacted several key factors, including the net photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), intercellular carbon dioxide concentration (Ci), photosynthetically active radiation (PAR), carbon‐metabolising enzymes and total carbon (TC) content accumulation. Under drought‐like rainfed conditions, the application of nitrogen, RN300 (rainfed application nitrogen 300 kg ha−1), IN250 (irrigated application nitrogen 250 kg ha−1) significantly enhanced the Pn (10.0%), Tr (3.17%), Ci (3.41%) and Gs (2.6%). Additionally, PAR was significantly influenced by the water regime and nitrogen levels. Under IN250, the capture ratio (Ca) increased (2.36%), while the penetration ratio (Pe) and reflectance ratio (Re) decreased by 13.12% and 46.36%, respectively, compared to RN300. The levels of carbon metabolism enzymes (sucrose phosphate synthase and phosphoenolpyruvate carboxylase) and the TC content were higher under RN300 compared to IN250; however, these differences were not statistically significant. Path analysis revealed that thousand kernel weight had the most significant impact on yield under both water regimes. The effect was stronger under irrigated conditions, with a path coefficient of 0.647, compared to 0.459 under rainfed conditions. Correlation analysis indicated that plant height (0.938), stem diameter (0.906), ear diameter (0.928) and ear length (0.803) were positively correlated with nitrogen levels. In conclusion, maize under IN250 exhibited superior photosynthetic performance and carbon accumulation. This suggests that balanced irrigation and nitrogen management can effectively mitigate the adverse impacts of drought on maize, optimising growth and yield sustainably.

Integrated Analysis of Metabolome and Transcriptome Reveals the Effect of Burdock Fructooligosaccharide on the Quality of Chinese Cabbage (Brassica rapa L. ssp. Pekinensis).

Burdock fructooligosaccharide (BFO) is fructose with a low polymerization degree, which could improve the immunity to pathogens, quality, and stress resistance of vegetables. Still, there are no studies on applying BFO in Chinese cabbage. In this study, the effects of exogenous BFO sprayed with different concentrations (0, 5, 10, 20, 30 g·L-1) on the growth and soluble sugar content of Chinese cabbage seedlings were determined. The result showed that 10 g·L-1 was the appropriate spraying concentration. Based on metabolome analysis, a total of 220 differentially accumulated metabolites (DAMs) were found, among which flavonoid metabolites, glucosinolate metabolites, and soluble sugar-related metabolites were the key metabolites involved in improving the quality of Chinese cabbage caused by BFO. Further combination analysis with transcriptome, trans-cinnamate 4-monooxygenase (CYP73A5), and chalcone synthase 1 (CHS1) were more closely associated with the DAMs of flavonoid biosynthesis. Sulfotransferases 18 (SOT18), Branched-chain amino acid amino transferases 6 (BCAT6), and cytochrome P450 monooxygenase (CYP83A1) were the key genes in glucosinolate biosynthesis. Hexokinase (HxK1), beta-glucosidase 8 (BGL08), invertase 3 (INV3), beta-glucosidase 3B (BGL3B), and sucrose phosphate synthase 1 (SPS1) were significantly upregulated, potentially playing crucial roles in the soluble sugar metabolism. In conclusion, these results provided an understanding of the effects of BFO on the expression of genes and the accumulation of metabolites related to quality formation in Chinese cabbage.

An Analysis of the Mechanism About CO2 Enrichment Promoting Carbohydrate Metabolism in Cucumber (Cucumis sativus L.) Leaves.

Elevated CO2 can affect the synthesis and distribution of photosynthetic assimilates. However, the carbohydrate metabolism molecular mechanism of cucumber leaves in response to CO2 enrichment is unclear. Therefore, it is of great significance to investigate the key functional regulatory genes in cucumber. In this study, the growth of cucumber leaves under different CO2 conditions was compared. The results showed that under CO2 enrichment, leaf area increased, the number of mesophyll cells increased, stomata enlarged, and more starch grains accumulated in the chloroplasts. Compared with the control, the starch and soluble sugar content of leaves were maximally increased by 194.1% and 55.94%, respectively; the activities of fructose-1,6-bisphosphatase (FBPase), ADPG pyrophosphorylase (AGPase), starch synthase (SSS), sucrose phosphate synthase (SPS), sucrose synthase (SS) and invertase (Inv) were maximally increased by 36.91%, 66.13%, 33.18%, 21.7%, 54.11%, and 46.01%, respectively. Through transcriptome analysis, a total of 1,582 differential expressed genes (DEGs) were identified, in which the starch and sucrose metabolism pathway was significantly enriched, and 23 genes of carbon metabolism were screened. Through metabolome analysis, a total of 22 differential accumulation metabolites (DAMs) were identified. Moreover, D-glucose and D(+)-glucose were significantly accumulated, showing upregulation 2.4-fold and 2.6-fold, respectively. Through combined analysis of transcriptome and metabolome, it was revealed that seven genes were highly related to D-glucose, and Csa6G153460 (AGPase), Csa5G612840 (β-glucosidase), and Csa4G420150 (4-α-glucanotransferase) were significantly correlated to the carbohydrate regulatory network. Furthermore, the mechanism of CO2 enrichment that promotes carbohydrate metabolism in leaves at the molecular level was revealed. This mechanism advances the development of the cell wall and leaf morphology by activating the expression of key genes and improving enzyme activity.

Comparative transcriptome analysis, identification and expression analysis of sucrose phosphate synthase genes in high and low starch cassava (Manihot esculenta Crantz) genotypes

Comparative transcriptome analysis, identification and expression analysis of sucrose phosphate synthase genes in high and low starch cassava (Manihot esculenta Crantz) genotypes

Improving sugar and respiratory metabolism in pear wounds by postharvest dipping with chitosan and chitooligosaccharide

Chitosan (CTS) and its degradation product, chitooligosaccharide (COS), promote fruit healing by activating phenylpropanoid metabolism. This study investigates their effects on sucrose metabolism in pear wounds. CTS and COS were found to activate neutral invertase, acid invertase, sucrose synthase, and sucrose phosphate synthase, increasing sucrose, glucose, and fructose levels in fruit wounds. They also enhanced sorbitol dehydrogenase activity and promoted sorbitol accumulation. In addition, CTS and COS improved the activities of hexokinase, phosphofructokinase, and pyruvate kinase, increasing phosphoenolpyruvate and ATP production. They activated glucose-6-phosphate dehydrogenase and increased erythrose-4-phosphate, NADPH, and shikimic acid levels. In conclusion, CTS and COS support the formation of the healing closing layer by supplying carbon skeletons, energy, and reducing power through the activation of sugar and respiratory metabolism during the healing process. Compared to CTS, COS was superior in activating the above metabolisms, which is expected to be widely used as a chitin product in postharvest fruit and vegetable preservation and provide new insights into preserving pear freshness.

Loss of Carbamoyl Phosphate Synthetase 1 Potentiates Hepatocellular Carcinoma Metastasis by Reducing Aspartate Level.

Hepatocellular carcinoma (HCC) is one of the most lethal cancers worldwide. Numerous studies have shown that metabolic reprogramming is crucial for the development of HCC. Carbamoyl phosphate synthase 1 (CPS1), a rate-limiting enzyme in urea cycle, is an abundant protein in normal hepatocytes, however, lacking systemic research in HCC. It is found that CPS1 is low-expressed in HCC tissues and circulating tumor cells, negatively correlated with HCC stage and prognosis. Further study reveals that CPS1 is a double-edged sword. On the one hand, it inhibits the activity of phosphatidylcholine-specific phospholipase C to block the biosynthesis of diacylglycerol (DAG), leading to the downregulation of the DAG/protein kinase C pathway to inhibit invasion and metastasis of cancer cells. On the other hand, CPS1 promotes cell proliferation by increasing intracellular S-adenosylmethionin to enhance the m6A modification of solute carrier family 1 member 3 mRNA, a key transporter for aspartate intake. Finally, CPS1 overexpressing adeno-associated virus can dampen HCC progression. Collectively, this results uncovered that CPS1 is a switch between HCC proliferation and metastasis by increasing intracellular aspartate level.

Functional and Proteomic Analyses of a Putative Carbamoyl Phosphate Synthase Large Subunit in Relation to Virulence, Arginine and Pyrimidine Biosynthesis, and Siderophore Production in Erwinia amylovora

The apple is a significant global fruit cultivated extensively worldwide. Fire blight, caused by Erwinia amylovora (Ea), poses a significant threat to global apple production. To control this disease, characterizing the virulence mechanisms/factors is imperative. Carbamoyl phosphate synthase is an essential enzyme in the biosynthesis of arginine and pyrimidine. However, the functions of this protein in Ea remains poorly understood. This study aimed to investigate the functions of the carbamoyl phosphate synthase large subunit in Ea (CarBEa). In a virulence assay using fruitlets, an Ea strain lacking CarBEa exhibited significantly reduced virulence on fruitlets. In the auxotrophy assay, this mutant failed to grow in minimal media lacking both arginine and pyrimidine, but growth was restored when both compounds were supplemented. The comparative proteomic analysis suggests that CarBEa is involved in diverse biological processes, including amino acid and nucleotide metabolism, and inorganic ion transport. Finally, we demonstrated that CarBEa is related to siderophore secretion/production by the chrome azurol S agar plate assay. This report provides valuable insights into the functions of carbamoyl phosphate synthase large subunit, which serves as a potential target for developing efficient anti-virulence substances to control fire blight.

Biocatalytic Ether Lipid Synthesis by an Archaeal Glycerolprenylase

AbstractAlthough ethers are common in secondary natural products, they are an underrepresented functional group in primary metabolism. As such, there are comparably few enzymes capable of constructing ether bonds in a general fashion. However, such enzymes are highly sought after for synthetic applications as they typically operate with higher regioselectivity and under milder conditions than traditional organochemical approaches. To expand the repertoire of well characterized ether synthases, we herein report on a promiscuous archaeal prenyltransferase from the scarcely researched family of geranylgeranylglyceryl phosphate synthases (GGGPSs or G3PSs). We show that the ultrastable Archaeoglobus fulgidus G3PS makes various (E)‐ and (Z)‐configured prenyl glycerol ethers from the corresponding pyrophosphates while exerting perfect control over the configuration at the glycerol unit. Based on experimental and computational data, we propose a mechanism for this enzyme which involves an intermediary prenyl carbocation equivalent. As such, this study provides the fundamental understanding and methods to introduce G3PSs into the biocatalytic alkylation toolbox.

Biocatalytic Ether Lipid Synthesis by an Archaeal Glycerolprenylase.

Although ethers are common in secondary natural products, they are an underrepresented functional group in primary metabolism. As such, there are comparably few enzymes capable of constructing ether bonds in a general fashion. However, such enzymes are highly sought after for synthetic applications as they typically operate with higher regioselectivity and under milder conditions than traditional organochemical approaches. To expand the repertoire of well characterized ether synthases, we herein report on a promiscuous archaeal prenyltransferase from the scarcely researched family of geranylgeranylglyceryl phosphate synthases (GGGPSs or G3PSs). We show that the ultrastable Archaeoglobus fulgidus G3PS makes various (E)- and (Z)-configured prenyl glycerol ethers from the corresponding pyrophosphates while exerting perfect control over the configuration at the glycerol unit. Based on experimental and computational data, we propose a mechanism for this enzyme which involves an intermediary prenyl carbocation equivalent. As such, this study provides the fundamental understanding and methods to introduce G3PSs into the biocatalytic alkylation toolbox.

Postnatal Wing Morph of Pea Aphids Regulates Hemolymph Trehalose Levels.

Trehalose, a nonreducing disaccharide composed of two glucose molecules, functions as a critical energy source in various insect tissues and organs and is the predominant sugar component of the hemolymph. The pea aphid, Acyrthosiphon pisum, exhibits higher hemolymph trehalose levels than other insects. However, the dynamics of hemolymph trehalose levels throughout its life stages remain unclear owing to the challenges associated with obtaining hemolymph from these small insects. Therefore, this study was conducted to quantify hemolymph trehalose levels in A. pisum using a fluorescent trehalose sensor (Tre-C04), which enhances green fluorescent protein fluorescence through the binding of trehalose to a ligand-binding protein fused to the fluorophore. Trehalose levels were successfully quantified in minimal hemolymph samples from individual aphids, with measurements spanning from the first nymphal stage to the adult stage in both the winged and wingless forms of A. pisum. Hemolymph trehalose levels remained relatively stable throughout the life cycle but exhibited a gradual increase with each developmental stage. Notably, adult winged aphids exhibited significantly higher hemolymph trehalose levels than wingless aphids. Given that wing morph determination occurs early in the nymphal stage, these findings suggest that hemolymph trehalose levels are regulated post-wing morph development. Further investigation of the expression of genes associated with trehalose metabolism revealed that trehalose phosphate synthase 2 levels were downregulated in early-stage wingless adults, whereas insulin-related peptide 5 levels were upregulated in wingless aphids. These findings indicate that A. pisum synthesizes trehalose during the winged adult stage to serve as an energy source for flight.

Responses of Growth, Enzyme Activity, and Flower Bud Differentiation of Pepper Seedlings to Nitrogen Concentration at Different Growth Stages

The concentration of nitrogen fertilizer is matched with the nutrient requirements in different growth stages of plants, which coordinates their vegetative and reproductive growth. In this study, the influences of nitrogen concentration before and after initiation of flower bud differentiation (first and second stage, respectively) on pepper seedling quality were studied. The chlorophyll a content, sucrose synthase activity, and sucrose phosphate synthase activity of pepper seedlings grown under moderate nitrogen (15 mmol L−1) in the first stage combined with high nitrogen (25.61 mmol L−1) in the second stage were 15.7%, 39.3%, and 34.6% higher than those of the same nitrogen concentration (15 mmol L−1) in the first and second stages treatment, respectively. The regression model also showed that the values of flower bud diameter, shoot fresh weight, root fresh weight, and glutamine synthetase activity of pepper were high under the condition of moderate nitrogen in the first stage and higher nitrogen in the second stage. In addition, the results of comprehensive evaluation showed that moderate nitrogen (15 mmol L−1) in the first stage and high nitrogen (25.61 mmol L−1) in the second stage treatment ranked first, which improved carbon and nitrogen metabolism, increased biomass accumulation, and promoted the flower bud differentiation and flowering of pepper seedlings.

Impact of nitrogen rates on biosynthesis pathways: A comparative study of diterpene synthases in clerodane diterpenoids and enzymes in benzylisoquinoline alkaloids

Tinospora sagittata is rich in secondary metabolites used in traditional medicine. However, environmental factors impact key enzymes in metabolite synthesis, highlighting the need for improved growth conditions. This study employs transcriptomics and metabolomics to assess nitrogen's impact on enzymes in secondary metabolites biosynthesis pathways. The gene expressions of berberine bridge enzymes (BBEs) like TsBBE2 had peak expression in low nitrogen treatments (A0 and A1) but were absent in higher nitrogen treatments (A2 and A3). Similar trends were observed for other enzymes such as (S)-scoulerine 9-O-methyltransferase (TsCMT3), Tetrahydroberberine oxidase (TsSTOX), and Columbamine O-methyltransferase (TsCoCOMT2-4) in response to nitrogen levels. In examining gene families related to diterpene synthases (diTPS), 1-deoxyxylulose 5-phosphate synthase (TsDXR1) expression increased with higher nitrogen fertilizer, while TsDXR2 peaked at maximal nitrogen levels. Geranylgeranyl diphosphate synthase (TsGGPP3 and TsGGPP5) decreased with nitrogen levels. (−)-kolavenyl diphosphate synthase (KPS) genes had higher expression in treatments, while ent-kaurene synthase (KSL) genes, especially TsKSL1 and TsKSL2, showed higher expression in control conditions with lower nitrogen fertilizer. Metabolite analysis confirmed more upregulated compounds in A3 compared to A0. These findings have practical implications for agriculture and pharmaceuticals, highlighting the link between nitrogen fertilization and specialized metabolism in medicinal plants.

Protein kinase MeSnRK2.3 positively regulates starch biosynthesis by interacting with the transcription factor MebHLH68 in cassava.

Starch biosynthesis involves numerous enzymes and is a crucial metabolic activity in plant storage organs. Sucrose non-fermenting related protein kinase 2 (SnRK2) is an abscisic acid (ABA)-dependent kinase and a significant regulatory enzyme in the ABA signaling pathway. However, whether SnRK2 kinases regulate starch biosynthesis is unclear. In this study, we identified that MeSnRK2.3, an ABA-dependent kinase, was highly expressed in the storage roots of cassava and was induced by ABA. Overexpression of MeSnRK2.3 in cassava significantly increased the starch content in the storage roots and promoted plant growth. MeSnRK2.3 was further found to interact with the cassava basic helix-loop-helix 68 (MebHLH68) transcription factor in vivo and in vitro. MebHLH68 directly bound to the promoters of sucrose synthase 1 (MeSUS1), granule-bound starch synthase I a (MeGBSSIa), and starch-branching enzyme 2.4 (MeSBE2.4), thereby upregulating their transcriptional activities. Additionally, MebHLH68 negatively regulated the transcriptional activity of sucrose phosphate synthase B (MeSPSB). Moreover, phosphorylated MebHLH68 by MeSnRK2.3 up-regulated the transcription activity of MeSBE2.4. These findings demonstrated that the MeSnRK2.3-MebHLH68 module connects the ABA signaling pathway and starch biosynthesis in cassava, thereby providing direct evidence of ABA-mediated participation in the sucrose metabolism and starch biosynthesis pathway.

Molecular descriptors and in silico studies of 4-((5-(decylthio)-4-methyl-4n-1,2,4-triazol-3-yl)methyl)morpholine as a potential drug for the treatment of fungal pathologies

The article explores the polypharmacological profiling of 4-((5-(decylthio)-4-methyl-4H-1,2,4-triazole-3-yl)methyl)morpholine as a potential antimicrobial agent. The study utilized 15148 electronic pharmacophore models of organisms, ranked by the Tversky index. Detailed analysis revealed classical bonding patterns with selected enzymes, identifying key amino acid residues involved in complex formation. Protein target prediction was conducted through various stages using the Galaxy web service, including ligand structure creation, pharmacophore alignment, and target ranking. The activities of the molecules against 1G6C, 2W6O, 3G7F, 3OWU, 4IVR, and 4TZT proteins were compared. Docking studies with PyMOL and Discovery Studio Visualizer revealed binding to thymidine kinase, thiamine phosphate synthase, and biotin carboxylase with promising binding affinities. These interactions suggest potential antibacterial and antiviral effects, warranting further virtual screening and in-depth studies for the development of effective antimicrobial drugs. Calculations of the molecules were made with the gaussian package program. Calculations were made on the 6-31++g** basis set at B3LYP, HF, and M062X levels with Gaussian software. Afterwards, the 0–100 ns interaction of the molecule with the highest activity was examined.