Natural product guvermectin inhibits guanosine 5'-monophosphate synthetase and confers broad-spectrum antibacterial activity.

Bacterial diseases caused substantial yield losses worldwide, with the rise of antibiotic resistance, there is a critical need for alternative antibacterial compounds. Natural products (NPs) from microorganisms have emerged as promising candidates due to their potential as cost-effective and environmentally friendly bactericides. However, the precise mechanisms underlying the antibacterial activity of many NPs, including Guvermectin (GV), remain poorly understood. Here, we sought to explore how GV interacts with Guanosine 5'-monophosphate synthetase (GMPs), an enzyme crucial in bacterial guanine synthesis. We employed a combination of biochemical and genetic approaches, enzyme activity assays, site-directed mutagenesis, bio-layer interferometry, and molecular docking assays to assess GV's antibacterial activity and its mechanism targeting GMPs. The results showed that GV effectively inhibits GMPs, disrupting bacterial guanine synthesis. This was confirmed through drug-resistant assays and direct enzyme inhibition studies. Bio-layer interferometry assays demonstrated specific binding of GV to GMPs, with dependency on Xanthosine 5'-monophosphate. Site-directed mutagenesis identified key residues crucial for the GV-GMP interaction. This study elucidates the antibacterial mechanism of GV, highlighting its potential as a biocontrol agent in agriculture. These findings contribute to the development of novel antibacterial agents and underscore the importance of exploring natural products for agricultural disease management.

Efficient Polyamine-Based Nanodelivery System for Proline: Enhanced Uptake Improves the Drought Tolerance of Tobacco.

Drought stress is one of the most unfavorable factors affecting plant growth and productivity among various environmental stresses. Nanotechnology is expected to enhance the effectiveness of conventional biostimulants. Herein, the current study constructed an efficient proline (Pro) nanodelivery system based on a star polyamine (SPc). The hydroxyl groups of Pro could assemble with carbonyl groups of SPc, and the self-assembly of Pro with SPc formed the nanoscale particles of the Pro/SPc complex. Compared to Pro alone, the contact angle of SPc-loaded Pro decreased, and its retentivity and plant uptake increased. Importantly, the tobacco (Nicotiana benthamiana) seeds and seedlings treated with Pro/SPc complex exhibited stronger drought tolerance. RNA-Seq analysis indicated that the SPc-loaded Pro could further upregulate photosynthesis-related genes and endocytosis-related genes. The current study constructed an efficient nanodelivery system for improving the bioactivity of biostimulants, which has broad application prospects in the agricultural field.

2-Bromopalmitate inhibits malignant behaviors of HPSCC cells by hindering the membrane location of Ras protein.

Palmitoylation, which is mediated by protein acyltransferase (PAT) and performs important biological functions, is the only reversible lipid modification in organism. To study the effect of protein palmitoylation on hypopharyngeal squamous cell carcinoma (HPSCC), the expression levels of 23 PATs in tumor tissues of 8 HPSCC patients were determined, and high mRNA and protein levels of DHHC9 and DHHC15 were found. Subsequently, we investigated the effect of 2-bromopalmitate (2BP), a small-molecular inhibitor of protein palmitoylation, on the behavior of Fadu cells in vitro (50 µM) and in nude mouse xenograft models (50 µmol/kg), and found that 2BP suppressed the proliferation, invasion, and migration of Fadu cells without increasing cell apoptosis. Mechanistically, the effect of 2BP on the transduction of BMP, Wnt, Shh, and FGF signaling pathways was tested with qRT-PCR, and its drug target was explored with western blotting and acyl-biotinyl exchange assay. Our results showed that 2BP inhibited the transduction of the FGF/ERK signaling pathway. The palmitoylation level of Ras protein decreased after 2BP treatment, and its distribution in the cell membrane structure was reduced significantly. The findings of this work reveal that protein palmitoylation mediated by DHHC9 and DHHC15 may play important roles in the occurrence and development of HPSCC. 2BP is able to inhibit the malignant biological behaviors of HPSCC cells, possibly via hindering the palmitoylation and membrane location of Ras protein, which might, in turn, offer a low-toxicity anti-cancer drug for targeting the treatment of HPSCC.

Star Polymer-Based Nanodelivery System for Pesticides: Enhanced Broad-Spectrum Toxicity and Selective Toxicity.

The application of nanotechnology in agriculture can break through many traditional problems of synthetic pesticides, especially for increasing bioactivity and reducing application amount. However, the safety and selective toxicity of nanocarrier-loaded pesticides should be clarified toward natural predators. In this context, an efficient spirotetramat nanodelivery system was successfully constructed based on a star polymer (SPc). Spirotetramat could complex with SPc through hydrogen bonding and van der Waals forces spontaneously. The self-assembly of the spirotetramat/SPc complex decreased the particle size of spirotetramat from 1292 to 710 nm. After the complexation with SPc, the lethal concentration 50 (LC50) values of spirotratemat decreased from 252.064 to 108.871 and 332.079 to 189.257 mg/L toward target pest Frankliniella occidentalis and nontarget predator Orius sauteri with the synergic ratios of 2.315 and 1.755, respectively. The possible reason might be due to the enhancement of the broad-spectrum toxicity of SPc-loaded pesticides. Importantly, the selective toxicity ratio (STR) of spirotetramat increased from 1.32 to 1.73 with the help of SPc, indicating the higher selectivity of the spirotratemat/SPc complex toward predators. Meanwhile, the safety coefficient (SC) of spirotratemat was not significantly changed after complexation with SPc, and the spirotratemat/SPc complex belonged to the medium risk pesticide. Overall, the assembly with SPc could not only improve the control efficacy of spirotetramat but also increase its selectivity as well as alleviate its negative effects on predators. The current study is beneficial for understanding the enhancement of broad-spectrum toxicity and the selective toxicity of nanocarrier-loaded pesticides.

Biotoxicity Evaluation of a Cationic Star Polymer on a Predatory Ladybird and Cooperative Pest Control by Polymer-Delivered Pesticides and Ladybird.

Although employing nanocarriers for gene/drug delivery shows great potential in agricultural fields, the biotoxicity of nanocarriers is a major concern for large-scale applications. Herein, we synthesized a cationic star polymer (SPc) as a pesticide nanocarrier/adjuvant to evaluate its safety against a widely used predatory ladybird (Harmonia axyridis). The application of SPc at extremely high concentrations nearly did not influence the hatching of ladybird eggs but it led to the death of ladybird larvae at lethal concentration 50 (LC50) values of 43.96 and 19.85 mg/mL through the soaking and feeding methods, respectively. The oral feeding of SPc downregulated many membrane protein genes and lysosome genes significantly, and the cell membrane and nucleus in gut tissues were remarkably damaged by SPc application, revealing that the lethal mechanism might be SPc-mediated membrane damage. Furthermore, the oral feeding of SPc increased the relative abundance of Serratia bacteria in ladybird guts to result in bacterial infection. Coapplication of ladybird and SPc-loaded thiamethoxam/matrine achieved desired control efficacies of more than 80% against green peach aphids, revealing that the coapplication could overcome the slow-acting property of ladybirds. To our knowledge, this is the first attempt to investigate the polymer-mediated lethal mechanism toward natural enemies and explore the possibility of coapplying SPc-loaded pesticides and natural enemies for pest management.

Overlapping and differential roles of plasma membrane calcium ATPases in Arabidopsis growth and environmental responses.

Plant cells have multiple plasma membrane (PM)-localized calcium ATPases (ACAs) pumping calcium ions out of the cytosol. Although the involvement of some of these ACAs in plant growth and immunity has been reported, their individual and combined functions have not been fully examined. Here, we analysed the effects of single and combined mutations of four ACA genes, ACA8, ACA10, ACA12, and ACA13, in a number of processes. We found that these four genes had both overlapping and differential involvements in vegetative growth, inflorescence growth, seeds setting, disease resistance and stomatal movement. Disruption of any of these four genes reduces seed setting, indicating their contribution to the overall fitness of the plants. While ACA10 and ACA8 play major roles in vegetative growth and immunity, ACA13 and ACA12 are also involved in these processes especially when the function of ACA10 and/or ACA8 is compromised. The loss of ACA13 and ACA10 function in combination with a reduction in function of ACA8 leads to seedling death at bolting, revealing the essential role of their collective function in plant growth. Taken together, this study indicates a highly tuned calcium system involving these PM-localized calcium pumps in plant growth and environmental responses.