Inhibition Of Biosynthesis Research Articles

The Role of Polyamines in pH Regulation in the Extracellular Calcifying Medium of Scleractinian Coral Spats.

This study aims to elucidate a novel mechanism for elevating the pH within the calicoblastic extracellular calcifying medium (pHECM) of corals and demonstrate the potential contribution of calcifying organisms to CO2 sequestration. Departing from traditional models that attribute the increase in pHECM primarily to H+ expulsion via Ca2+-ATPase, we emphasize the significant role of polyamines. These ubiquitous biogenic amines conveyed by calicoblastic cells through polyamine transporters demonstrate a remarkable affinity for CO2. Their ability to form stable carbamate complexes is pivotal in facilitating carbonate ion transport, which is crucial for pH regulation and skeletal structure formation. In this study, a polyamine transporter inhibitor and a polyamine biosynthesis inhibitor in conjunction with the pH-sensitive probe 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS) were employed to monitor pH variations. Furthermore, FM1-43FX dye was utilized to delineate the extracellular calcifying medium (ECM), whereas calcein was applied to visualize paracellular gaps and ECM. These methodologies provide profound insights into the intricate structural and functional dynamics of coral spats calcification. Findings suggest a potential reconsideration of established models of marine calcification and highlight the necessity to reassess the role of marine calcifying organisms in the carbon cycle, particularly their influence on CO2 fluxes.

Integrated roles of nitric oxide and melatonin in enhancing chromium resilience in cotton plants: modulation of thiol metabolism and antioxidant responses.

Chromium (Cr) is a hazardous metal found in various oxidation states, posing significant environmental and health risks. The study focuses on understanding how melatonin (MT), known for its diverse functions, including stress alleviation and antioxidant properties, interacts with nitric oxide (NO) to regulate sulfur metabolism and enhance cotton resilience under Cr toxicity. Cr toxicity negatively affected plant growth and photosynthesis and induced oxidative stress. MT treatment ameliorated these effects by enhancing photosynthetic pigments and gas exchange traits and upregulating the activities of antioxidant enzymes and the expression of FeSOD, CuZnSOD, and APX1. Moreover, MT reduced Cr accumulation in leaves, protecting photosynthetic organs from direct toxicity. Additionally, MT promotes the level of sulfur-based defense substances like glutathione (GSH) and phytochelatins (PC), which arecrucial for detoxifying heavy metals. This is achieved by upregulating genes involved in cysteine metabolism (CYC1, CYC2, CAS1, CAS2, DES1, DES2, and SSCS), increasing cysteine availability for GSH and PC synthesis, and enhancing Cr sequestration in vacuoles. However, when the inhibitor of MT biosynthesis (p-CPA) and NO scavenger (cPTIO) were used along with MT in Cr-stressed plants, they hindered the stimulatory effects of MT. The study highlights the importance of the NO-MT interaction in mediating these protective effects, indicating a potential signaling role for NO in plant defense mechanisms against Cr toxicity. Overall, the findings reveal that MT fertilizing may serve as an effective strategy to enhance Cr resistance in cotton plants, with NO potentially playing a signaling role in this response pathway.

A triplicated wheat-rye chromosome segment including several 12-OXOPHYTODIENOATE REDUCTASE III genes influences magnesium partitioning and impacts wheat performance at low magnesium supply

We previously reported a structural rearrangement between wheat (Triticum aestivum) and rye (Secale cereale) chromosomes 1BS/1RS that increased the dosage of 12-OXOPHYTODIENOATE REDUCTASE III (OPRIII) genes involved in jasmonate biosynthesis (henceforth, 1RW line), and that drastically reduced primary root growth relative to a control line with the intact 1RS chromosome (henceforth, 1RS). In this study, we show that the increased gene-dosage of this region is associated with increases in the shoot-root partitioning of magnesium (Mg). Moreover, both a CRISPR-edited 1RW line with reduced OPRIII dosage and the 1RW line treated with the jasmonate biosynthesis inhibitor ibuprofen showed reduced differences in shoot-root Mg partitioning than 1RW. The observed differences in Mg partitioning between 1RS and 1RW plants occur over a wide range of external Mg supplies and imply opposite trends of Mg accumulation in roots and shoots. Furthermore, we show an association between the increase of shoot-root Mg partitioning and increased tolerance of the 1RW line to low levels of Mg supply. In summary, our results provide evidence of the role of the jasmonate pathway on the dynamics of Mg accumulation in roots and shoots, which correlates with the performance of wheat plants under conditions of Mg scarcity.

Enhancing Mycoprotein Yield: Metabolic Modulation of Chitin Synthase in Fusarium venenatum.

Fusarium venenatum is being extensively utilized for microbial protein production. However, its high dietary fiber content results in substantial carbon loss. Inhibition of chitin biosynthesis presents a promising strategy to improve the mycoprotein yield. Through transcriptomic and bioinformatics analyses, chitin synthase gene FvChs3 was identified as crucial for chitin synthesis in F. venenatum. Knockout of the FvChs3 gene resulted in mycelial expansion and a 26% reduction in the chitin content of strain ΔFvChs3. Ethanol production from fermentation decreased by 47%, while the carbon conversion efficiency and protein conversion increased by 16% and 36%, respectively. Transcriptomic analysis revealed an upregulation of nitrogen metabolism in ΔFvChs3, while genes related to the glycolysis pathway for ethanol synthesis were downregulated. Further knockout of pyruvate decarboxylase gene FvPDC6 in ΔFvChs3 accelerated growth, leading to improvements in carbon and protein conversion of 29% and 40%, respectively. This research lays the foundation for enhancing fungal protein production.

Characterization of SUPPRESSOR OF MAX2 1-LIKE (SMXL) Genes in ‘duli’ (Pyrus betulifolia L.) and Expression Analysis of PbSMXLs in Response to Plant Growth Regulators and Salt Stress

SUPPRESSOR OF MAX2 1-LIKE (SMXL) proteins are negative regulators of strigolactone (SL) signal transduction that play an important role in regulating plant branching and responses to abiotic stress. Here, we studied the role of SMXL proteins in pear growth, development, and stress resistance. A total of 18 SMXL members were characterized in ‘duli’. All SMXL members were localized to chloroplasts. Chromosome mapping analysis showed that the members of this family were unevenly distributed on 14 chromosomes. Gene fragment replication analysis showed that there were no tandem repeat genes in PbSMXLs, and 12 pairs of homologous genes were fragment duplications. There were 30 pairs of homologous genes between ‘duli’ and apples, and 17 between ‘duli’ and Arabidopsis thaliana. Analysis of cis-acting elements showed that there was a large number of photo-effector elements, short-effector elements, hormone-responsive elements, and abiotic stress-responsive elements in the promoter sequences of this family. Analysis of enzyme activity and endogenous SL showed that β-carotenoid isomerase (D27), carotenoid cleavage dioxygenase 7 (CCD7), lateral branch oxidoreductase (LBO) levels, and SL content were higher in ‘duli’ roots and leaves compared in the control under exogenous GA3 (gibberellin 3), IAA (indole-3-acetic acid), GR24 (synthetic SL analog), and NaCl. Most SMXL genes in ‘duli’ were highly expressed in branches and axillary lobes, but their expression was low in fruits. qRT-PCR analysis revealed that eight PbSMXL genes were responsive to GA3, PAC (Paclobutrazol), IAA, ABA (abscisic acid), GR24, and Tis108 (SL biosynthesis inhibitor). PbSMXLs responded positively to salt stress. The expression of PbSMXL6 and PbSMXL15 was significantly induced under salt stress. The expression of PbSMXL7, PbSMXL10, and PbSMXL15 was significantly induced by Tis108 treatment. The results of this study enhance our understanding of the role of SMXL genes in the responses to plant growth regulators and salt stress. Our findings will also aid future studies of the functions of SMXL genes in ‘duli’.

Engineering an Escherichia coli strain for enhanced production of flavonoids derived from pinocembrin.

Flavonoids are a structurally diverse group of secondary metabolites, predominantly produced by plants, which include a range of compounds with pharmacological importance. Pinocembrin is a key branch point intermediate in the biosynthesis of a wide range of flavonoid subclasses. However, replicating the biosynthesis of these structurally diverse molecules in heterologous microbial cell factories has encountered challenges, in particular the modest pinocembrin titres achieved to date. In this study, we combined genome engineering and enzyme candidate screening to significantly enhance the production of pinocembrin and its derivatives, including chrysin, pinostrobin, pinobanksin, and galangin, in Escherichia coli. By implementing a combination of established strain engineering strategies aimed at enhancing the supply of the building blocks phenylalanine and malonyl-CoA, we constructed an E. coli chassis capable of accumulating 353 ± 19mg/L pinocembrin from glycerol, without the need for precursor supplementation or the fatty acid biosynthesis inhibitor cerulenin. This chassis was subsequently employed for the production of chrysin, pinostrobin, pinobanksin, and galangin. Through an enzyme candidate screening process involving eight type-1 and five type-2 flavone synthases (FNS), we identified Petroselinum crispum FNSI as the top candidate, producing 82 ± 5mg/L chrysin. Similarly, from a panel of five flavonoid 7-O-methyltransferases (7-OMT), we found pinocembrin 7-OMT from Eucalyptus nitida to yield 153 ± 10mg/L pinostrobin. To produce pinobanksin, we screened seven enzyme candidates exhibiting flavanone 3-hydroxylase (F3H) or F3H/flavonol synthase (FLS) activity, with the bifunctional F3H/FLS enzyme from Glycine max being the top performer, achieving a pinobanksin titre of 12.6 ± 1.8mg/L. Lastly, by utilising a combinatorial library of plasmids encoding G. max F3H and Citrus unshiu FLS, we obtained a maximum galangin titre of 18.2 ± 5.3mg/L. Through the integration of microbial chassis engineering and screening of enzyme candidates, we considerably increased the production levels of microbially synthesised pinocembrin, chrysin, pinostrobin, pinobanksin, and galangin. With the introduction of additional chassis modifications geared towards improving cofactor supply and regeneration, as well as alleviating potential toxic effects of intermediates and end products, we anticipate further enhancements in the yields of these pinocembrin derivatives, potentially enabling greater diversification in microbial hosts.

VvFHY3 links auxin and endoplasmic reticulum stress to regulate grape anthocyanin biosynthesis at high temperatures.

Anthocyanins affect quality in fruits such as grape (Vitis vinifera). High temperatures reduce anthocyanin levels by suppressing the expression of anthocyanin biosynthesis genes and decreasing the biosynthetic rate. However, the regulatory mechanisms that coordinate these two processes remain largely unknown. In this study, we demonstrate that high-temperature-mediated inhibition of anthocyanin biosynthesis in grape berries depends on the auxin and endoplasmic reticulum (ER) stress pathways. Inactivation of these pathways restores anthocyanin accumulation under high temperatures. We identified and characterized FAR-RED ELONGATED HYPOCOTYL3 (FHY3), a high-temperature-modulated transcription factor that activates multiple anthocyanin biosynthesis genes by binding to their promoters. The auxin response factor VvARF3 interacts with VvFHY3 and represses its transactivation activity, antagonizing VvFHY3-induced anthocyanin biosynthesis. Additionally, we found that the ER stress sensor VvbZIP17 represses anthocyanin biosynthesis. VvFHY3 suppresses VvbZIP17 activity by directly binding to the VvbZIP17 promoter to repress its transcription and by physically interacting with VvbZIP17 to block its DNA binding ability. Furthermore, AUXIN RESPONSE FACTOR 3 (ARF3) interferes with the VvFHY3-VvbZIP17 interaction, releasing VvbZIP17 to activate the unfolded protein response and further suppress anthocyanin production. Our results unravel the VvARF3-VvFHY3-VvbZIP17 regulatory module, which links the auxin and ER stress pathways to coordinately repress anthocyanin structural gene expression and biosynthesis under high-temperature stress.

Gibberellin action‐based strategies show promise in the control of Striga—A major threat to global agriculture

Societal Impact StatementThe root‐parasitic plant Striga hermonthica can cause severe crop failure, leading to significant agricultural and economic losses in Africa. Despite the promising target offered by the Striga seed germination mechanism for developing control methods, effective control methods have yet to be established. For seed germination, Striga must undergo an incubation under warm and humid conditions, a process termed conditioning, which allows it to germinate. Here, we report that the plant hormone gibberellin stimulates the conditioning process in Striga seeds. Our findings suggest that the quantitative control of gibberellin in plants is a promising approach for controlling Striga infestations and may help alleviate substantial threats to food security.Summary The root‐parasitic plant S. hermonthica is a serious agricultural threat in Africa. Although a number of control methods have been employed to combat Striga, there is still a need to develop more effective control methods. Striga seeds germinate only in the presence of host plants, owing to the need for host‐released stimulants, primarily strigolactone. To detect signals from the host, Striga seeds must be exposed to a specific warm, dark, and moist environment, which is known as conditioning. Gibberellin directly promotes germination in many nonparasitic plants, but Striga seeds are insensitive to exogenous gibberellin. Consequently, the role of gibberellin in the germination of Striga seeds is unclear. We demonstrated that the effect of the gibberellin biosynthesis inhibitor paclobutrazol was higher in seeds during the conditioning, compared with those already conditioned, suggesting that gibberellin primarily regulates conditioning rather than germination. The content of an active form gibberellin, gibberellin A4 (GA4) exhibited an increase throughout the conditioning period, and we quantified the elevated expression levels of some gibberellin biosynthesis genes. The GA4 treatment shortened the conditioning period required for strigolactone‐induced seed germination. We also discovered that gibberellin upregulated ShACO1, the gene responsible for ethylene synthesis, during conditioning, and that the inhibition of GR24‐induced germination by the ethylene biosynthesis inhibitor α‐aminoisobutyric acid was gradually alleviated by GA4. Our findings demonstrate that gibberellin is critical for conditioning and germination processes but acts as a minor stimulant in Striga seeds. This study demonstrates the potential of quantitative endogenous gibberellin control for Striga management, providing valuable insights for alleviating food security issues.

CNSC-68. SEX DIFFERENCES IN GLYCINE BIOSYNTHESIS DRIVE RESPONSES TO COMBINED NUTRIENT DEPRIVATION AND PHARMACOLOGICAL INHIBITION IN PEDIATRIC HGG

Abstract In children and adults, high-grade gliomas (HGGs) are the most common and deadly primary brain malignancies. Generally, male HGGs occur at higher incidence and demonstrate more rapid proliferation, resulting in shorter overall male survival. Obscurity of molecular mechanisms driving this phenomenon prevents implementation of clinically effective treatments. Recently, a positive correlation between HGG cellular proliferation and metabolic reprogramming toward glycine biosynthesis was identified. By analyzing pediatric and adult HGG patient data, we discovered significantly worse outcomes unique to pediatric males bearing upregulation of serine and glycine biosynthesis transcripts. Previously, we have identified differential flux of glucose into serine and glycine biosynthesis drives sex differences in lung cancer. Therefore, we sought to confirm and target this paradigm in our NF1-/- DNp53 model of HGG. By performing metabolic tracing with [13C]-labeled glucose in male and female cells, we identified higher enrichment of [13C]-labeled serine and glycine in male cells relative to female cells. Through Western blotting, we identify male-specific upregulation of serine and glycine biosynthetic pathway enzymes in transformed cells, supporting our labelling results. Next, we sought to determine if inhibition of de-novo serine and glycine biosynthesis was similarly sex-biased. Utilizing serine-and-glycine-deprived media conditions, we identified a male-biased decrease in proliferation following pharmacological inhibition of de-novo serine and glycine biosynthesis. More precisely, in the absence of exogenous glycine, we discovered that inhibition of de-novo synthesis of glycine from serine drove male-specific decreases in proliferation. Additionally, we identified that transformed female cell robustness against inhibition of glycine biosynthesis is driven by the uptake of exogenous nucleotide precursors. These data confirm that sex differences in de-novo glycine biosynthesis drive sex differences in HGG cellular proliferation. As well, our work suggests that dietary restriction of serine and glycine, in-combination with pharmacological inhibition of de-novo glycine biosynthesis, represents a novel treatment paradigm.

TMIC-49. TUMOR-DERIVED SPERMIDINE DRIVES IMMUNE SUPPRESSION IN THE GLIOBLASTOMA MICROENVIRONMENT VIA CD8 T CELL DEPLETION AND FUNCTIONAL ATTENUATION

Abstract A hallmark of glioblastoma (GBM) is an immunosuppressive microenvironment that is partially driven by tumor cell-derived secreted factors, highlighting the need for a more complete understanding of local immune suppressive mechanism. Altered secreted metabolite levels are observed in the GBM microenvironment; specifically, spermidine (a polyamine), which functions to drive cell proliferation, apoptosis, T cell lineage commitment, and myeloid cell-mediated suppression. However, the direct link between GBM cell-derived spermidine and the tumor microenvironment has yet to be determined. In syngeneic mouse GBM models, spermidine levels were increased in the tumor microenvironment and exogenous administration of spermidine accelerated tumor aggressiveness in an immune dependent-manner. Mechanistically, spermidine administration increased apoptosis of CD8+ T-cells and generated an exhausted phenotype. When ornithine decarboxylase (ODC1, the rate limiting enzyme in spermidine synthesis), was knocked down in tumor cells, no direct effect was observed on tumor cell growth. However, there was an increase in time to tumor succumbence and a concomitant increase in CD8+ T-cells and decrease in CD8+ T cell exhaustion. In GBM patients, there was a negative correlation between ODC1 expression and CD8+ T-cells present near the tumor. Additionally, patients with a favorable prognosis had significantly lower intratumoral levels of spermidine compared to patients with a poor prognosis. Taken together, these results demonstrate that spermidine functions as a tumor cell-derived metabolite that drives tumor progression through reducing CD8+T cell number and altering their phenotype. These findings have clinical implications as clinical trials using a polyamine biosynthesis inhibitor alone have not been very effective in GBM patients, suggesting the need for additional combination immunotherapies. Finally, spermidine is also produced at prominent levels by commensal gut microbes as well as absorbed from consumption of foods such as mushrooms; investigation into how these distal spermidine sources impact GBM growth via the gut-brain axis is actively ongoing.

Metabolic profiling unveils enhanced antibacterial synergy of polymyxin B and teixobactin against multi-drug resistant Acinetobacter baumannii

This untargeted metabolomics study investigated the synergistic antibacterial activity of polymyxin B and Leu10-teixobactin, a depsipeptide inhibitor of cell wall biosynthesis. Checkerboard microdilution assays revealed a significant synergy against polymyxin-susceptible and -resistant A. baumannii, excluding lipopolysaccharide-deficient variants. Time-kill assays confirmed bactericidal synergy, reducing bacterial burden by approximately 4-6-log10CFU/mL. The combination (2xMIC polymyxin B and 0.5xMIC Leu10-teixobactin) prevented bacterial regrowth after 24 h, indicating sustained efficacy against the emergence of resistant mutants. The analysis of A. baumannii ATCC™ 19606 metabolome demonstrated that the polymyxin B–Leu10-teixobactin combination produced more pronounced perturbation compared to the individual antibiotics across all time points (1, 3 and 6 h). Pathway analysis revealed that lipid metabolism, cell envelope biogenesis, and cellular respiration were predominantly impacted by the combination, and to a lesser extent by polymyxin B monotherapy. Leu10-teixobactin treatment alone had only a minor impact on the metabolome, primarily at the 6 h time point. Peptidoglycan assays confirmed the combination’s concerted deleterious effects on bacterial cell envelope integrity. Electron microscopy further substantiated these findings, revealing pronounced cell envelope damage, membrane blebbing, and vacuole formation. These findings highlight the potential of the polymyxin B–Leu10-teixobactin combination as an effective treatment in preventing resistance in A. baumannii.

Single Amine or Guanidine Modification on Norvancomycin and Vancomycin to Overcome Multidrug-Resistance through Augmented Lipid II Binding and Increased Membrane Activity.

Vancomycin and norvancomycin have diminished antibacterial efficacy due to acquired or intrinsic resistance from mutations in the terminal dipeptide of lipid II in Gram-positive bacteria or failure to penetrate into the periplasm in Gram-negative bacteria. Herein, we rationally designed and synthesized a series of vancomycin analogues bearing single amine or guanidine functionality, altering various linkers and modification sites, to combat the resistance. Extensive antibacterial screening was performed to delineate a comprehensive SAR. Many derivatives revitalized the activity in vitro, exhibiting a 4-128-fold or 2-16-fold enhancement against the acquired or intrinsic resistance with lower toxicity. Significantly, the optimal compound 4g demonstrated greater pharmacokinetic and pharmacodynamic profiles. Further studies uncovered additional independent and synergistic mechanisms for 4g, including the enhanced membrane activity and augmented inhibition of peptidoglycan biosynthesis via increased lipid II binding, highlighting its potential as a future lead candidate to replenish the glycopeptide antibiotic arsenal.

Inhibition of cell growth and glycosaminoglycan biosynthesis by xylose analog 2-Az-Xyl

Sugar analogs are versatile chemical tools for probing or inhibiting glycan functions. However, chemical tools are insufficient for glycosaminoglycans (GAGs), which play crucial roles in various biological processes, such as extracellular matrix formation and growth factor signaling. To develop a new compound for detecting GAGs or manipulating GAG functions, we chemically synthesized 2-azide-xylose (2-Az-Xyl), an azide-type analog of Xyl that is a component of the common linkage tetrasaccharide in GAGs, and explored its application to biological experiments. Treatment of cultured cells with 2-Az-Xyl inhibited cell proliferation and reduced the levels of GAGs, particularly heparan sulfate (HS). Although this sugar analog did not perturb the biosynthesis of nucleotide sugars and expression of the key enzymes for HS biosynthesis, 2-Az-Xyl directly inhibited the activity of XYLT2, an initial enzyme for GAG biosynthesis, indicating that 2-Az-Xyl directly inhibits GAG biosynthesis. These findings suggest that 2-Az-Xyl inhibits cell proliferation by blocking GAG biosynthesis through inhibiting XYLT2 activity.

Systemic inhibition of de novo purine biosynthesis prevents weight gain and improves metabolic health by increasing thermogenesis and decreasing food intake.

Obesity is a major health concern, largely because it contributes to type 2 diabetes mellitus (T2DM), cardiovascular disease, and various malignancies. Increase in circulating amino acids and lipids, in part due to adipose dysfunction, have been shown to drive obesity-mediated diseases. Similarly, elevated purines and uric acid, a degradation product of purine metabolism, are found in the bloodstream and in adipose tissue. These metabolic changes are correlated with metabolic syndrome, but little is known about the physiological effects of targeting purine biosynthesis. To determine the effects of purine biosynthesis on organismal health we treated mice with mizoribine, an inhibitor of inosine monophosphate dehydrogenase 1 and 2 (IMPDH1/2), key enzymes in this pathway. Mice were fed either a low-fat (LFD; 13.5% kcal from fat) or a high-fat (HFD; 60% kcal from fat) diet for 30 days during drug or vehicle treatment. We ascertained the effects of mizoribine on weight gain, body composition, food intake and absorption, energy expenditure, and overall metabolic health. Mizoribine treatment prevented mice on a HFD from gaining weight, but had no effect on mice on a LFD. Body composition analysis demonstrated that mizoribine significantly reduced fat mass but did not affect lean mass. Although mizoribine had no effect on lipid absorption, food intake was reduced. Furthermore, mizoribine treatment induced adaptive thermogenesis in skeletal muscle by upregulating sarcolipin, a regulator of muscle thermogenesis. While mizoribine-treated mice exhibited less adipose tissue than controls, we did not observe lipotoxicity. Rather, mizoribine-treated mice displayed improved glucose tolerance and reduced ectopic lipid accumulation. Inhibiting purine biosynthesis prevents mice on a HFD from gaining weight, and improves their metabolic health, to a significant degree. We also demonstrated that the purine biosynthesis pathway plays a previously unknown role in skeletal muscle thermogenesis. A deeper mechanistic understanding of how purine biosynthesis promotes thermogenesis and decreases food intake may pave the way to new anti-obesity therapies. Crucially, given that many purine inhibitors have been FDA-approved for use in treating various conditions, our results indicate that they may benefit overweight or obese patients. A purine biosynthesis inhibitor, mizoribine, protects against diet-induced weight gainMizoribine prevents fat mass gain in high-fat diet-fed male miceMizoribine reduces food intake and increases thermogenesisMizoribine induces expression of sarcolipin, a regulator of thermogenesisMizoribine treatment reduces ectopic lipids and increases glucose tolerance.

A peptide-receptor module links cell wall integrity sensing to pattern-triggered immunity.

Plants employ cell-surface receptors to perceive non- or altered-self, including the integrity of their cell wall. Here we identify a specific ligand-receptor module responsive to cell wall damage that potentiates immunity in Arabidopsis. Disruption of cell wall integrity by inhibition of cellulose biosynthesis promotes pattern-triggered immunity transcriptionally in a manner dependent on the receptor kinase MALE DISCOVERER 1-INTERACTING RECEPTOR-LIKE KINASE 2 (MIK2). Notably, while MIK2 can perceive peptides of the large SERINE RICH ENDOGENOUS PEPTIDE family, a single member of this family, SCOOP18, is transcriptionally induced upon cell wall damage and is required for subsequent responses such as lignification and immunity potentiation. Collectively, our results identify the SCOOP18-MIK2 ligand-receptor module as an important central hub, connecting plant cell wall integrity sensing with immunity.

A Hybrid Alloying Nanozyme-Glutathione Inhibitor Co-Delivery System Initiates a Dual-Disruption on Cancer Redox Homeostasis.

Altered redox homeostasis has long been observed in cancer cells, which can be exploited for therapeutic benefits. However, reactive oxygen species (ROS) pleiotropy coupling with reductive adaptation in cancer cells poses a formidable challenge for redox dyshomeostasis-based cancer therapy. Herein, a AuPd alloying nanozyme-glutathione (GSH) biosynthesis inhibitor co-delivery system (B-BMES) is developed using dendritic SiO2 as a matrix to target cancer redox homeostasis. By optimizing element composition, the alloying nanozyme in B-BMES exhibits a potent peroxidase (POD)-like activity to trigger ROS insults-mediated redox dyshomeostasis. Such a POD functionality is attributed to the optimized electronic structure and catalytic activity. Simultaneously, the B-BMES abrogates the reductive adaptation by exerting its molecule-targeted GSH suppression, thereby achieving a dual-disruption on cancer redox homeostasis. Camouflaging B-BMES with tumor-homologous cytomembrane, a hybrid nanosystem with biological stability and tumor-targeting ability is further fabricated, which initiates a safe, precise redox disruption-based cancer therapy and sensibilizes standard chemotherapy.

The Antifungal Effects of Berberine and Its Proposed Mechanism of Action Through CYP51 Inhibition, as Predicted by Molecular Docking and Binding Analysis.

Fungal infections present a significant health risk, particularly in immunocompromised individuals. Berberine, a natural isoquinoline alkaloid, has demonstrated broad-spectrum antimicrobial activity, though its antifungal potential and underlying mechanisms against both yeast-like and filamentous fungi are not fully understood. This study investigates the antifungal efficacy of berberine against Candida albicans, Cryptococcus neoformans, Trichophyton rubrum, and Trichophyton mentagrophytes in vitro, as well as its therapeutic potential in a murine model of cryptococcal infection. Berberine showed strong antifungal activity, with MIC values ranging from 64 to 128 µg/mL. SEM and TEM analyses revealed that berberine induced notable disruptions to the cell wall and membrane in C. neoformans. No signs of cell necrosis or apoptosis were observed in fungal cells treated with 2 × MIC berberine, and it did not increase intracellular ROS levels or affect mitochondrial membrane potential. Molecular docking and binding affinity assays demonstrated a strong interaction between berberine and the fungal enzyme CYP51, with a dissociation constant (KD) of less than 1 × 10-12 M, suggesting potent inhibition of ergosterol biosynthesis. In vivo studies further showed that berberine promoted healing in guinea pigs infected with T. mentagrophytes, and in a murine cryptococcal infection model, it prolonged survival and reduced lung inflammation, showing comparable efficacy to fluconazole. These findings indicate that berberine exerts broad-spectrum antifungal effects through membrane disruption and CYP51 inhibition, highlighting its potential as a promising therapeutic option for fungal infections.

Functional Characterization of the Sterol-Synthesis-Related Gene CgCYP51 in the Poplar Anthracnose-Causing Fungus Colletotrichum gloeosporioides

Poplar is an economically and ecologically valuable tree species. Anthracnose, which severely affects poplar tree growth, is mainly caused by Colletotrichum gloeosporioides. In the infestation cycle of poplar anthracnose, the entry of C. gloeosporioides into the host tissue depends on the formation of an appressorium. The subsequent development of the appressorium determines the pathogenesis of poplar anthracnose and the degree of damage. Previous studies have found that the transcription factor CgSte12 affects appressorium formation and development by regulating the expression of a series of genes, including the sterol-synthesis-related gene CgCYP51, which influences appressorium formation and development. In this study, knockout and functional analyses of CgCYP51 revealed decreases in differentiation, darkening rate, and turgor pressure of appressoria in mutants. Additionally, compared with the wild-type appressorium, mutant appressoria secreted less mucus and exhibited abnormal penetration pore formation, ultimately leading to decreased pathogenicity. Moreover, CgCyp51 affected the sensitivity of C. gloeosporioides to sterol biosynthesis inhibitors. Considered together, the study findings indicate CgCYP51 is a key CgSte12-regulated gene that affects C. gloeosporioides appressorium formation and development. Furthermore, the study data provide new insights into the molecular basis of C. gloeosporioides appressorium formation and development.

Maize DLR1/NHX7 Is Required for Root Development Under Potassium Deficiency.

Root System Architecture (RSA) is a crucial plant trait that governs a plant's ability to absorb water and nutrients. In this study, we describe a mutant with nutrient-dependent defects in root development, affecting both the primary root and lateral roots (LRs). This mutant, identified through a screen for defects in LR development, has been designated dlr1-1. The dlr1-1 mutant exhibits impaired LR emergence rather than defects in the LR primordium (LRP) formation, particularly under potassium (K+)-deprivation conditions. This impairment likely stems from inhibited cell proliferation caused by the dlr1-1 mutation. K+ deprivation specifically leads to the accumulation of salicylic acid (SA) in the dlr1-1 mutant, consistent with the upregulation of SA biosynthesis genes. Moreover, exogenous application of SA to wild-type plants (B73) mimics the dlr1-1 phenotype. Conversely, treatment of the dlr1-1 mutant with 2-aminoindane-2-phosphonic acid, an SA biosynthesis inhibitor, partially restores LR emergence, indicating that elevated SA levels may be responsible for the mutant's developmental defects. MutMap analysis and allelism tests confirmed that the phenotypes of the dlr1-1 mutant results from the loss of the Na+/H+ antiporter, ZmNHX7. Additionally, the application of NaCl exacerbates the dlr1-1 mutant phenotype, suggesting that the root defects in dlr1-1 mutant depend on ion homoeostasis. In conclusion, our findings demonstrate that maize DLR1/NHX7 is essential for root development under potassium deprivation.

Antifungal Efficacy of Ultrashort β-Peptides against Candida Species: Mechanistic Understanding and Therapeutic Implications.

Candidiasis, a condition spurred by the unchecked proliferation of Candida species, poses a formidable global health threat, particularly in immunocompromised individuals. The emergence of drug-resistant strains complicates management strategies, necessitating novel therapeutic avenues. Antimicrobial peptides (AMPs) have garnered attention for their potent antifungal properties and broad-spectrum activity against Candida species. This study assessed the antifungal effectiveness of ultrashort β-peptides against Candida strains, with a specific focus on peptide P3 (LAU-β3,3-Pip-β2,2-Ac6c-PEA). Our findings showed P3's remarkable fungistatic and fungicidal activities against Candida albicans, exhibiting an MIC of 4 μg/mL, comparable to those of standard antifungal drugs. The MIC value remained unchanged in the presence of ADC and BSA, indicating that serum albumin does not diminish the activity of P3. P3 demonstrates synergistic effects when combined with Fluconazole (FLU), Itraconazole (ITR), and Nystatin (NYS) to the extent that it becomes effective at 0.125, 0.125, and 0.03125 μg/mL, respectively. Concentration versus time-kill kinetics showed its time-dependent activity up to the first 12 h against C. albicans, and later concentration also played a role; indeed, at 24 h the whole culture was sterilized at 8× MIC. Post-antifungal effect assays confirmed prolonged suppression of pathogen growth after the removal of P3 from the media for significant durations. More importantly, P3 inhibits hyphae formation and biofilm development of Candida, outperforming Fluconazole with respect to these properties. Mechanistic insights display P3's potential to disrupt fungal cell membrane integrity and dose-dependent inhibition of ergosterol biosynthesis, essential for fungal cell wall integrity. Using the Bradford assay, it was observed that extracellular protein concentrations increased with higher doses of the compound, thereby validating the effect of P3 on membrane integrity. A comparative gene analysis using RT-PCR showed that P3 downregulates ERG3, ERG11, and HWP1, which are crucial for the survival and pathogenicity of C. albicans. The impact of P3 on ERG11 and ERG3 is more effective than that of Fluconazole. Molecular docking studies revealed strong binding of P3 to various isoforms of lanosterol 14-α-demethylase, a key enzyme in ergosterol synthesis. Furthermore, molecular dynamic simulations validated the stability of the most promising docking complex. Overall, our findings underscore P3's potential as a leading candidate for the development of innovative antifungal therapies, warranting further investigation and optimization.