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.

Transcriptome analysis reveals the regulation of cyclic nucleotide-gated ion channels in response to exogenous abscisic acid and calcium treatment under drought stress in tomato.

Background: Drought stress can limit the growth and development of tomato seedlings and cause considerable loss of tomato yield. Exogenous abscisic acid (ABA) and calcium (Ca2+) can effectively alleviate the damage of drought stress to plants in part because Ca2+ acts as a second messenger in the drought resistance pathway. Although cyclic nucleotide-gated ion channels (CNGCs) are common non-specific Ca2+ osmotic channels in cell membranes, a thorough understanding of the transcriptome characteristics of tomato treated with exogenous ABA and Ca2+ under drought stress is necessary to characterize the molecular mechanism of CNGC involved in tomato drought resistance. Results: There were 12,896 differentially expressed genes in tomato under drought stress, as well as 11,406 and 12,502 differentially expressed genes after exogenous ABA and Ca2+ application, respectively. According to functional annotations and reports, the 19 SlCNGC genes related to Ca2+ transport were initially screened, with 11 SlCNGC genes that were upregulated under drought stress and downregulated after exogenous ABA application. After exogenous Ca2+ application, the data showed that two of these genes were upregulated, while nine genes were downregulated. Based on these expression patterns, we predicted the role of SlCNGC genes in the drought resistance pathway and their regulation by exogenous ABA and Ca2+ in tomato. Conclusion: The results of this study provide foundational data for further study of the function of SlCNGC genes and a more comprehensive understanding of drought resistance mechanisms in tomato.

Calcium nutrition nanoagent rescues tomatoes from mosaic virus disease by accelerating calcium transport and activating antiviral immunity.

As an essential structural, metabolic and signaling element, calcium shows low remobilization from old to young tissues in plants, restricting the nutrient-use efficiency and control efficacy against mosaic virus disease. Nanotechnology has been applied to prevent/minimize nutrient losses and improve the accessibility of poorly-available nutrients. Herein, the current study applied a star polycation (SPc) to prepare a calcium nutrition nanoagent. The SPc could assemble with calcium glycinate through hydrogen bond and Van der Waals force, forming stable spherical particles with nanoscale size (17.72 nm). Transcriptomic results revealed that the calcium glycinate/SPc complex could activate the expression of many transport-related genes and disease resistance genes in tomatoes, suggesting the enhanced transport and antiviral immunity of SPc-loaded calcium glycinate. Reasonably, the calcium transport was accelerated by 3.17 times into tomato leaves with the help of SPc, and the protective effect of calcium glycinate was remarkably improved to 77.40% and 67.31% toward tomato mosaic virus with the help of SPc after the third and fifth applications. Furthermore, SPc-loaded calcium glycinate could be applied to increase the leaf photosynthetic rate and control the unusual fast growth of tomatoes. The current study is the first success to apply nano-delivery system for enhanced calcium transport and antiviral immunity, which is beneficial for increasing nutrient-use efficiency and shows good prospects for field application.

A Preparation Method of Nano-Pesticide Improves the Selective Toxicity toward Natural Enemies.

Various nano-delivery systems have been designed to deliver synthetic/botanical pesticides for improved bioactivity. However, the enhanced toxicity of nanocarrier-loaded pesticides may injure the natural enemies, and their selective toxicity should be evaluated before the large-scale application. In this context, a star polymer (SPc)-based cyantraniliprole (CNAP) nano-delivery system was constructed, and its selective toxicity was evaluated using pest Frankliniella occidentalis (WFT) and predator Orius sauteri. The amide NH of CNAP could assemble with carbonyl groups or tertiary amines of SPc through hydrogen bonds to form CNAP/SPc complex spontaneously. The above self-assembly decreased the particle size of CNAP from 808 to 299 nm. With the help of SPc, the lethal concentration 50 (LC50) values of CNAP decreased from 99 to 54 mg/L and 230 to 173 mg/L toward WFTs and O. sauteri due to the enhancement of broad-spectrum bioactivity. Interestingly, the toxicity selective ratio (TSR) of CNAP increased from 2.33 to 3.23 with the help of SPc, revealing the higher selectivity of SPc-loaded CNAP. To our knowledge, it was the first successful exploration of the selective toxicity of nanocarrier-loaded pesticides, and the higher selective toxicity of SPc-loaded CNAP was beneficial for alleviating the negative impacts on predators.

A Star Polyamine-Based Nanocarrier Delivery System for Enhanced Avermectin Contact and Stomach Toxicity against Green Peach Aphids.

The unscientific application of synthesized/botanical pesticides has not only brought the resistance of plant pathogens and pests, but also led to serious environmental pollution. In recent years, various nano-delivery systems have been used for the development of environmental-friendly pesticides with improved efficacy. Herein, the current study constructed an efficient avermectin B1a (AVM) nano-delivery system based on a star polyamine (SPc) and focused on the characterization and bioactivity of SPc-loaded AVM at various mass ratios. The hydroxyl groups of AVM could assemble with carbonyl groups of SPc through hydrogen bond and van der Waals forces, and the self-assembly of AVM and SPc formed nearly spherical particles of AVM/SPc complex with nanoscale size. The contact angle of SPc-loaded AVM decreased with the increasing mass ratio of SPc, revealing the easier distribution and spreading of the AVM/SPc complex. Furthermore, the stomach and contact toxicity of AVM/SPc complex also increased along with the increasing SPc mass ratio, which could be attributed to the enhanced systemic transportation in plants, enlarged contact area to insect pests and stronger permeability across the insect cuticle. The current study provides an efficient nano-delivery system for increasing stomach and contact toxicity of pesticides with wide applications in the agricultural field.

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.

Simple Osthole/Nanocarrier Pesticide Efficiently Controls Both Pests and Diseases Fulfilling the Need of Green Production of Strawberry.

The application of botanical pesticides is a good choice in organic agriculture. However, most botanical pesticides have limitations of slow action and short persistence for pest and disease management, which constrain their further application. With the objective of exploring a green pesticide for controlling strawberry pests and diseases simultaneously, a star polymer (SPc) with a low production cost was synthesized as a pesticide nanocarrier through simple reactions. The SPc complexed with osthole quickly through electrostatic interaction and hydrophobic association, which decreased the particle size of osthole down to the nanoscale (17.66 nm). With the help of SPc, more nano-sized osthole was delivered into cytoplasm through endocytosis, leading to the enhanced cytotoxicity against insect cells. As a green botanical pesticide, the control efficacy of the osthole/SPc complex was improved against main strawberry pests (green peach aphid and two-spotted spider mite) and disease (powdery mildew), which fulfilled the need of both pest and disease management in sustainable production of strawberry. Meanwhile, the introduction of SPc not only improved plant-uptake but also decreased the residue of osthole due to the higher degradation rate. Furthermore, the application of the osthole/SPc complex exhibited no influence on the strawberry fruit quality and nontarget predators. To our knowledge, it is the first success to control plant pests and diseases simultaneously for sustainable agriculture by only one pesticidal formulation based on nanoparticle-delivered botanical pesticides.

Nanometerization of thiamethoxam by a cationic star polymer nanocarrier efficiently enhances the contact and plant-uptake dependent stomach toxicity against green peach aphids.

BACKGROUND: The utilization efficiency of conventional insecticides is comparatively low in agricultural production, which leads to their excessive application and environmental pollution. Insecticide nanometerization by polymers and polymeric materials has advantages, particularly increased utilization efficiency and reduced insecticide application. RESULTS: To increase the utilization efficiency of insecticides, a star polycation (SPc) was selected as a drug carrier that could be complexed with thiamethoxam through electrostatic interaction. Formation of the complex decreased the particle size of thiamethoxam from 575.77 to 116.16 nm in aqueous solution. Plant uptake of SPc-delivered thiamethoxam was increased 1.69-1.84 times compared with thiamethoxam alone. Nano-sized thiamethoxam/SPc complexes showed enhanced contact and stomach toxicity against green peach aphids. CONCLUSION: SPc is a promising insecticide adjuvant for insecticide nanometerization, and is beneficial in improving insecticidal activity and decreasing the application amounts and application rates of conventional insecticides. © 2020 Society of Chemical Industry.

Identification, characterization and expression analysis of calmodulin and calmodulin-like proteins in Solanum pennellii.

In plants, the calmodulin (CaM) proteins is an important calcium-binding protein, which play a crucial role in both regulating plant growth and development, as well as in the resistance mechanisms to various biotic and abiotic stresses. However, there is limited knowledge available on the CaM family functions in Solanum pennellii, a wild tomato species utilized as a genetic resource for cultivated tomatoes. In this study, 6 CaM (SpCaM) and 45 CaM-like (SpCML) genes from Solanum pennellii were selected for bioinformatics analysis to obtain insights into their phylogenetic relationships, gene structures, conserved motifs, chromosomal locations, and promoters. The results showed that the 6 SpCaM proteins contained 4 EF-hand domains each, and the 45 SpCML proteins had 2-4 EF-hand domains. The 51 CaM and CaM-like genes contained different intron/exon patterns and they were unevenly distributed across the 12 chromosomes of S. pennellii. The results of the analysis of the conserved motifs and promoter cis-regulatory elements also indicated that these proteins were involved in the responses to biotic and abiotic stresses. qRT-PCR analysis indicated that the SpCaM and SpCML genes had broad expression patterns in abiotic stress conditions and with hormone treatments, in different tissues. The findings of this study will be important for further investigations of the calcium signal transduction mechanisms under stress conditions and lay a theoretical foundation for further exploration of the molecular mechanisms of plant resistance.

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.

CsNIP2;1 is a Plasma Membrane Transporter from Cucumis sativus that Facilitates Urea Uptake When Expressed in Saccharomyces cerevisiae and Arabidopsis thaliana.

Urea is an important source of nitrogen (N) for the growth and development of plants. It occurs naturally in soils, is the major N source in agricultural fertilizers and is an important N metabolite in plants. Therefore, the identification and characterization of urea transporters in higher plants is important for the fundamental understanding of urea-based N nutrition in plants and for designing novel strategies for improving the N-use efficiency of urea based-fertilizers. Progress in this area, however, is hampered due to scarce knowledge of plant urea transporters. From what is known, urea uptake from the soil into plant roots is mediated by two types of transporters: the major intrinsic proteins (MIPs) and the DUR3 orthologs, mediating low- and high-affinity urea transport, respectively. Here we characterized a MIP family member from Cucumis sativus, CsNIP2;1, with regard to its contribution to urea transport. We show that CsNIP2;1 is a plasma membrane transporter that mediates pH-dependent urea uptake when expressed in yeast. We also found that ectopic expression of CsNIP2;1 improves growth of wild-type Arabidopsis thaliana and rescues growth and development of the atdur3-3 mutant on medium with urea as the sole N source. In addition, CsNIP2;1 is transcriptionally up-regulated by N deficiency, urea and NO3 (-). These data and results from the analyses of the pattern of CsNIP2;1 expression in A. thaliana and cucumber suggest that CsNIP2;1 might be involved in multiple steps of urea-based N nutrition, including urea uptake and internal transport during N remobilization throughout seed germination and N delivery to developing tissues.

[Effects of organic manure on the profile distribution of soil nitrate-N in a peach orchard].

In 2004 and 2005, a field experiment was conducted in a peach orchard in Pinggu District of Beijing to study the effects of organic manure on the profile distribution of nitrate-N in soil. Four treatments were installed, i.e., applying 6.75 x 10(4) kg x hm(-2) of organic manure both in 2004 and in 2005 (T1), no fertilization in 2004 but applying 13.5 x 10(4) kg x hm(-2) of organic manure in 2005 (T2), no fertilization in 2004 but applying 6.75 x 10(4) kg x hm(-2) of organic manure in 2005 (CK), and no fertilization both in 2004 and in 2005 (CK). In 2006, soil samples at the depth of 0-120 cm were collected from the treatments and analyzed. The results showed that soil nitrate-N had a relatively uniform distribution in the profile in CK, accumulated more at the depth of 0-60 cm and decreased gradually at 60-120 cm in T1 and T3, and increased with increasing depth, with the peak at the depth of 100-120 cm in T2. The soil nitrate-N content below 60 cm in T2 was the highest among all the treatments, indicating that applying excessive amount of organic manure could induce nitrate-N leaching. The profile distribution of soil nitrate-N had significant correlations with the total amount of applied organic manure, the amount of organic manure applied in the nearest year, and the distance of sampling sites from peach tree. A correlation model of organic manure treatments and soil nitrate-N distribution was established.

[Investigation on occurrence of lycium pests and their natural enemies].

OBJECTIVE: To Provide the scientific basis for controlling the lycium pests. METHOD: The kinds and number of pests and their natural enemies were investigated at regular intervals and on the fixed trees. RESULT: There are 13 kinds of lycium pests and 17 kinds of natural enemies, the occurrence of which are closely related. CONCLUSION: Natural enemies can control aphids and psyllidae effectively, but ten-spotted lema and Epithrix sp. must be controlled by pesticides.