Nonphytotoxic and pH-responsive ZnO-ZIF­8 loaded with honokiol as a "nanoweapon" effectively controls the soil-borne bacterial pathogen Ralstonia solanacearum.

The development of intelligently released and environmentally safe nanocarriers not only aligns with the sustainable agricultural strategy but also offers a potential solution for controlling severe soil-borne bacterial diseases. Herein, the core-shell structured nanocarrier loaded with honokiol bactericide (honokiol@ZnO-ZIF-8) was synthesized via a one-pot method for the targeted control of Ralstonia solanacearum, the causative agent of tobacco bacterial wilt disease. Results indicated that honokiol@ZnO-ZIF-8 nanoparticles induced bacterial cell membrane and DNA damage through the production of excessive reactive oxygen species (ROS), thereby reducing bacterial cell viability and ultimately leading to bacterial death. Additionally, the dissociation mechanism of the nanocarriers was elucidated for the first time through thermodynamic computational simulation. The nanocarriers dissociate primarily due to H+ attacking the N atom on imidazole, causing the rupture of the Zn-N bond under acidic conditions and at room temperature. Furthermore, honokiol@ZnO-ZIF-8 exhibited potent inhibitory effects against other prominent Solanaceae pathogenic bacteria (Pseudomonas syringae pv. tabaci), demonstrating its broad-spectrum antibacterial activity. Biosafety assessment results indicated that honokiol@ZnO-ZIF-8 exhibited non-phytotoxicity towards tobacco and tomato plants, with its predominant accumulation in the roots and no translocation to aboveground tissues within a short period. This study provides potential application value for the intelligent release of green pesticides. ENVIRONMENT IMPLICATION: The indiscriminate use of agrochemicals poses a significant threat to environmental, ecological security, and sustainable development. Slow-release pesticides offer a green and durable strategy for crop disease control. In this study, we developed a non-phytotoxic and pH-responsive honokiol@ZnO-ZIF-8 nano-bactericide based on the pathogenesis of Ralstonia solanacearum. Thermodynamic simulation revealed the dissociation mechanism of ZIF-8, with different acidity controlling the dissociation rate. This provides a theoretical basis for on-demand pesticide release while reducing residue in the. Our findings provide strong evidence for effective soil-borne bacterial disease control and on-demand pesticide release.

Naringenin restricts the colonization and growth of Ralstonia solanacearum in tobacco mutant KCB-1.

Bacterial wilt severely jeopardizes plant growth and causes enormous economic loss in the production of many crops, including tobacco (Nicotiana tabacum). Here, we first demonstrated that the roots of bacterial wilt-resistant tobacco mutant KCB-1 can limit the growth and reproduction of Ralstonia solanacearum. Secondly, we demonstrated that KCB-1 specifically induced an upregulation of naringenin content in root metabolites and root secretions. Further experiments showed that naringenin can disrupt the structure of R. solanacearum, inhibit the growth and reproduction of R. solanacearum, and exert a controlling effect on bacterial wilt. Exogenous naringenin application activated the resistance response in tobacco by inducing the burst of reactive oxygen species and salicylic acid deposition, leading to transcriptional reprogramming in tobacco roots. Additionally, both external application of naringenin in CB-1 and overexpression of the Nicotiana tabacum chalcone isomerase (NtCHI) gene, which regulates naringenin biosynthesis, in CB-1 resulted in a higher complexity of their inter-root bacterial communities than in untreated CB-1. Further analysis showed that naringenin could be used as a marker for resistant tobacco. The present study provides a reference for analyzing the resistance mechanism of bacterial wilt-resistant tobacco and controlling tobacco bacterial wilt.

g-C3N4@CuO electrostatic self-assembly toward Ralstonia solanacearum: Insights from cytomembrane and motility disruption.

BACKGROUND: Ralstonia solanacearum, a notorious and refractory bacterial plant pathogen, threatens multiple vegetable crops and causes significant economic loss in agriculture. Long-term use of traditional medicines not only increases the problem of drug resistance, but also causes great environmental pollution. Therefore, there is an urgent need to develop new agents with high efficacy and low toxicity. RESULTS: In this study, we have synthesized and characterized graphitic carbon nitride incorporated copper oxide composite (g-C3N4@CuO), which showed higher antimicrobial effect than graphitic carbon nitride nanosheets (g-C3N4 nanosheets) and copper oxide nanoparticles (CuONPs). Ralstonia solanacearum exposed to g-C3N4@CuO exhibited higher levels of oxygen toxicity, cell membrane damage, DNA damage, motility disruption and even cell death compared to g-C3N4 nanosheets and CuONPs. In addition, g-C3N4@CuO was more effective in the control of tobacco bacterial wilt than g-C3N4 nanosheets and CuONPs. CONCLUSION: Thus, this study provides a new perspective on g-C3N4@CuO control of bacterial diseases in crops, and the mechanism is related to the destruction of cell membrane damage and motility disruption. © 2024 Society of Chemical Industry.

Synthesis, In-Vitro and In-Silico Evaluation of Silver Nanoparticles with Root Extract of Withania somnifera for Antibacterial Activity via Binding of Penicillin-Binding Protein-4.

BACKGROUND: Metal Nanoparticles (NPs) have been widely used for various applications in biomedical sciences, including in drug delivery, and as therapeutic agents, but limited owing to their toxicity towards the healthy tissue. This warrants an alternative method, which can achieve the desired activity with much reduced or no toxicity. Being a biological product, Withania somnifera (W. somnifera) is environment friendly, besides being less toxic as compared to metal-based NPs. However, the exact mechanism of action of W. somnifera for its antibacterial activities has not been studied so far. OBJECTIVE: To develop "silver nanoparticles with root extract of W. somnifera (AgNPs-REWS)" for antimicrobial and anticancer activities. Furthermore, the analysis of their mechanism of action will be studied. METHODS: Using the in-silico approach, the molecular docking study was performed to evaluate the possible antibacterial mechanism of W. somnifera phytochemicals such as Anaferine, Somniferine, Stigmasterol, Withaferin A, Withanolide- A, G, M, and Withanone by the inhibition of Penicillin- Binding Protein 4 (PBP4). Next, we utilized a bottom-up approach for the green synthesis of AgNPs- REWS, performed an in-detail phytochemical analysis, confirmed the AgNPs-REWS by SEM, UVvisible spectroscopy, XRD, FT-IR, and HPLC. Eventually, we examined their antibacterial activity. RESULTS: The result of molecular docking suggests that WS phytochemicals (Somniferine, Withaferin A, Withanolide A, Withanolide G, Withanolide M, and Withanone) possess the higher binding affinity toward the active site of PBP4 as compared to the Ampicillin (-6.39 kcal/mol) reference molecule. These phytochemicals predicted as potent inhibitors of PBP4. Next, as a proof-of-concept, AgNPs- REWS showed significant antibacterial effect as compared to crude, and control; against Xanthomonas and Ralstonia species. CONCLUSION: The in-silico and molecular docking analysis showed that active constituents of W. somnifera such as Somniferine, Withaferin A, Withanolide A, Withanolide G, Withanolide M, and Withanone possess inhibition potential for PBP4 and are responsible for the anti-bacterial property of W. somnifera extract. This study also establishes that AgNPs via the green synthesis with REWS showed enhanced antibacterial activity towards pathogenic bacteria.

Zinc thiazole enhances defense enzyme activities and increases pathogen resistance to Ralstonia solanacearum in peanut (Arachis hypogaea) under salt stress.

Crop plants always encounter multiple stresses in the natural environment. Here, the effects of the fungicide zinc thiazole (ZT) on propagation of Ralstonia solanacearum, a bacterial pathogen, were investigated in peanut seedlings under salt stress. Compared with water control, salt stress markedly reduced pathogen resistance in peanut seedlings. However, impaired pathogen resistance was alleviated by treatment with dimethylthiourea, a specific ROS scavenger, or ZT. Subsequently, salt stress or combined salt and pathogen treatment resulted in inhibition of photosynthesis, loss of chlorophyll and accumulation of thiobarbituric acid reactive substances, which could be reversed by ZT. In addition, ZT treatment suppressed the salt stress up-regulated Na+ content and Na+/K+ ratios in peanut roots. Furthermore, salt stress or combined salt and pathogen treatment impaired the activities of antioxidant (e.g. superoxide dismutase/SOD and catalase/CAT), and defense-related (e.g. phenylalanine ammonia lyase /PAL and polyphenol oxidase/PPO) enzymes, which could be rescued by addition of ZT. In contrast, only slight changes of SOD and CAT activities were observed in pathogen-infected seedlings. Similarly, activities of PAL and PPO were slightly modified by salt stress in peanut seedlings. These results suggest that the ZT-enhanced pathogen resistance can be partly attributed to the improvement of photosynthetic capacity and defense enzyme activities, and also the inhibition of Na+/K+ ratios, in this salt-stressed crop plant.

Fabrication of pH-Sensitive Tetramycin Releasing Gel and Its Antibacterial Bioactivity against Ralstonia solanacearum.

Ralstonia solanacearum (R. solanacearum)-induced bacterial wilt of the nightshade family causes a great loss in agricultural production annually. Although there has been some efficient pesticides against R. solanacearum, inaccurate pesticide releasing according to the onset time of bacterial wilt during the use of pesticides still hinders the disease management efficiency. Herein, on the basis of the soil pH change during R. solanacearum growth, and pH sensitivity of the Schiff base structure, a pH-sensitive oxidized alginate-based double-crosslinked gel was fabricated as a pesticide carrier. The gel was prepared by crosslinking oxidized sodium alginate (OSA) via adipic dihydrazide (ADH) and Ca2+. After loading tetramycin into the gel, it showed a pH-dependent pesticide releasing behavior and anti-bacterial activity against R. solanacearum. Further study also showed that the inhibition rate of the tetramycin-loaded gel was higher than that of industrial pesticide difenoconazole. This work aimed to reduce the difficulty of pesticide administration in the high incidence period of bacterial wilt and we believe it has a great application potential in nightshade production.

Biogenic synthesis of iron oxide nanoparticles via Skimmia laureola and their antibacterial efficacy against bacterial wilt pathogen Ralstonia solanacearum.

Plant diseases are threat to global food security. The excessive use of agrochemicals is the leading cause of pesticides resistance and toxicity to beneficial life forms. The quest for innocuous and alternate antimicrobial agent is crucial in order to overcome the pathogen resistance and the birth of nanotech offers pledge to combat pathogenic organisms. In this study, a facile benign biogenic approach was adopted for the synthesis of biocompatible iron oxide nanoparticles (Fe2O3-NPs) via Skimmia laureola leaf extract and the synthesized nanoparticles were evaluated for their antibacterial efficacy against bacterial wilt pathogen Ralstonia solanacearum in vitro and in planta. Physico-chemical characterization of the synthesized nanoparticles was performed through UV-Visible spectroscopy, Fourier Transform Infrared Spectroscopy, X-Ray Diffraction, Energy Dispersive X-ray Spectroscopy and Scanning Electron Microscopy. The results revealed polydisperse nanoparticles in the size range of 56 nm to 350 nm. The culture media containing 6 mg/mL of Fe2O3-NPs dramatically inhibited the bacterial growth in vitro. Scanning electron microscopy revealed degenerative characteristics including degraded, shriveled and concentrated cell walls. Diseases severity was effectively reduced with 6% w/v of Fe2O3-NPs treated root zone in planta. Plant shoots, root length and fresh biomass were enhanced with Fe2O3-NPs treatments. The results indicated that the biosynthesized Fe2O3-NPs have the potential to control agriculturally important phytopathogen Ralstonia solanacearum in vitro and in planta.

Efficacy of Leaf Oil from Pimenta racemosa var. racemosa in Controlling Bacterial Wilt of Tomato.

Bacterial wilt, caused by Ralstonia solanacearum, is a major plant disease throughout the Caribbean. The ability of the essential oil from Pimenta racemosa var. racemosa to control bacterial wilt of tomato (R. solanacearum, phylotype IIB/4NPB) was investigated. Lemongrass (chemotype 1)-, aniseed (chemotype 2)-, and clove (chemotype 3)-scented chemotypes of P. racemosa var. racemosa essential oil were tested. Six concentrations of emulsified essential oil (from 0.01 to 0.14% [v/v]) were evaluated by in vitro culture amendment assays and by in vivo experiments in greenhouse. Chemotype 3 displayed remarkable in vitro antibacterial activity against R. solanacearum, because the minimum inhibitory concentration was only 0.03%, compared with 0.14% for chemotypes 1 and 2. In greenhouse experiments, no incidence of bacterial wilt was observed in tomato plants grown in soil treated with chemotype 3 of P. racemosa var. racemosa at a concentration of 0.14%. In the untreated control soil, 62% of plants displayed symptoms of bacterial wilt. Treatment with chemotype 3 significantly increased the growth of tomato plants compared with untreated controls. These results suggest that chemotype 3 of P. racemosa var. racemosa essential oil is a good candidate for further development as a soil biofumigant for the control of tomato bacterial wilt.

CaHDZ27, a Homeodomain-Leucine Zipper I Protein, Positively Regulates the Resistance to Ralstonia solanacearum Infection in Pepper.

Homeodomain-leucine zipper class I (HD-Zip I) transcription factors have been functionally characterized in plant responses to abiotic stresses, but their roles in plant immunity are poorly understood. Here, a HD-Zip I gene, CaHZ27, was isolated from pepper (Capsicum annum) and characterized for its role in pepper immunity. Quantitative real-time polymerase chain reaction showed that CaHDZ27 was transcriptionally induced by Ralstonia solanacearum inoculation and exogenous application of methyl jasmonate, salicylic acid, or ethephon. The CaHDZ27-green fluorescent protein fused protein was targeted exclusively to the nucleus. Chromatin immunoprecipitation demonstrated that CaHDZ27 bound to the 9-bp pseudopalindromic element (CAATAATTG) and triggered ß-glucuronidase expression in a CAATAATTG-dependent manner. Virus-induced gene silencing of CaHDZ27 significantly attenuated the resistance of pepper plants against R. solanacearum and downregulated defense-related marker genes, including CaHIR1, CaACO1, CaPR1, CaPR4, CaPO2, and CaBPR1. By contrast, transient overexpression of CaHDZ27 triggered strong cell death mediated by the hypersensitive response and upregulated the tested immunity-associated marker genes. Ectopic CaHDZ27 expression in tobacco enhances its resistance against R. solanacearum. These results collectively suggest that CaHDZ27 functions as a positive regulator in pepper resistance against R. solanacearum. Bimolecular fluorescence complementation and coimmunoprecipitation assays indicate that CaHDZ27 monomers bind with each other, and this binding is enhanced significantly by R. solanacearum inoculation. We speculate that homodimerization of CaHZ27 might play a role in pepper response to R. solanacearum, further direct evidence is required to confirm it.

Graphene Oxide Induces Toxicity and Alters Energy Metabolism and Gene Expression in Ralstonia solanacearum.

Graphene oxide (GO) is a promising material for development as an antibacterial, phytoprotective agent due to its contact-based antibacterial activity induced by its physical and chemical properties. However, the mechanism underlying the antibacterial effect of GO has yet to be elucidated. In the current study, we investigated the effects of GO on the phytopathogen R. solanacearum at the molecular level with a specific focus on energy metabolism. Under controlled conditions, the bacteriostatic and bactericidal actions of GO were investigated with respect to concentration, treatment time and rotation speed. Transmission electron microscopy (TEM) and destabilization assays revealed that GO caused injury to bacterial cell membrane structures. Furthermore, adenosine triphosphate (ATP) levels decreased after exposure to sheets of GO, while malondialdehyde levels significantly increased, indicating the occurrence of lipid oxidation. A series of genes related to bacterial virulence, motility and oxidative stress were selected to evaluate the molecular mechanism underlying GO's effects on R. solanacearum. Using quantitative reverse transcription polymerase chain reaction (RT-qPCR), we showed that in the presence of GO, the expression levels of genes involved in virulence and motility were down regulated, with the exception of popA. The phcA, hrpB and flgG genes were significantly downregulated by 2.61-, 3.45- and 4.22-fold, respectively. Conversely, the expression levels of sodB, oxyR and dps, three important oxidative stress genes, were upregulated by 1.82-, 2.17-, and 3.79-fold, respectively. These findings confirmed that cell membrane damage and oxidative stress were responsible for the antibacterial actions of GO, in addition to disturbances to energy metabolism processes.

A novel leucine-rich repeat protein, CaLRR51, acts as a positive regulator in the response of pepper to Ralstonia solanacearum infection.

The leucine-rich repeat (LRR) proteins play important roles in the recognition of corresponding ligands and signal transduction networks in plant defence responses. Herein, a novel LRR protein from Capsicum annuum, CaLRR51, was identified and characterized. It was localized to the plasma membrane and transcriptionally up-regulated by Ralstonia solanacearum infection (RSI), as well as the exogenous application of salicylic acid (SA), jasmonic acid (JA) and ethephon (ETH). Virus-induced gene silencing of CaLRR51 significantly increased the susceptibility of pepper to RSI. By contrast, transient overexpression of CaLRR51 in pepper plants activated hypersensitive response (HR)-like cell death, and up-regulated the defence-related marker genes, including PO2, HIR1, PR1, DEF1 and ACO1. Moreover, ectopic overexpression of CaLRR51 in transgenic tobacco plants significantly enhanced the resistance to RSI. Transcriptional expression of the corresponding defence-related marker genes in transgenic tobacco plants was also found to be enhanced by the overexpression of CaLRR51, which was potentiated by RSI. These loss- and gain-of-function assays suggest that CaLRR51 acts as a positive regulator in the response of pepper to RSI. In addition, the putative signal peptide and transmembrane region were found to be required for plasma membrane targeting of CaLRR51, which is indispensable for the role of CaLRR51 in plant immunity.

Extract of Syringa oblata: A new biocontrol agent against tobacco bacterial wilt caused by Ralstonia solanacearum.

Ralstonia solanacearum causes serious wilt disease in tobacco. To effectively control this disease, the antibacterial activity of 95% ethanol extracts from the flower buds of Syringa oblata was examined. Based on GC-MS analysis and an inhibition experiment against R. solanacearum, the main antibacterial component is eugenol. We further determined the effect of eugenol on the physiology, biochemistry, and cellular morphology of R. solanacearum. The results showed that eugenol can destroy wilt bacteria, leading to the disappearance of flagella, the leakage of contents, and the appearance of a cavity. SDS-PAGE showed that eugenol decreased protein content in R. solanacearum, reduced medium carbohydrate utilization, and inhibited CAT and SDH activity. The above results showed that eugenol had a significant inhibitory effect on R. solanacearum and this component has the potential to prevent tobacco bacterial wilt.

Extract from Maize (Zea mays L.): Antibacterial Activity of DIMBOA and Its Derivatives against Ralstonia solanacearum.

Many cereals accumulate hydroxamic acids involved in defense of plant against various fungi, bacteria, and insects. 2,4-dihydroxy-7-methoxy-1,4-benzoxazine-3-one, commonly known as DIMBOA, is one of the principal cyclic hydroxamic acids in aqueous extracts of maize. The aim of this study was to evaluate the antibacterial activity of the isolated DIMBOA and its derivatives 2-benzoxazolinone (BOA), 6-chloro-2-benzoxazolinone (CDHB), and 2-mercaptobenzothiazole (MBT) against Ralstonia solanacearum. MBT showed the strongest antibacterial activity, followed by CDHB and DIMBOA, with minimum inhibitory concentrations (MICs) of 50, 100 and 200 mg/L, respectively, better than the BOA with 300 mg/L. These compounds also significantly affect bacterial growth, reduce biofilm formation, and inhibit swarming motility within 24 h. This paper is the first to report the anti-R. solanacearum activity of DIMBOA from Z. mays. The bioassay and pot experiment results suggested that DIMBOA and its derivatives exhibit potential as a new matrix structure of designing target bactericide or elicitor for controlling tobacco bacterial wilt. Further studies must evaluate the efficacy of DIMBOA and its derivatives in controlling bacterial wilt under natural field conditions where low inoculum concentrations exist.

Bioactive Dibenzo-α-pyrone Derivatives from the Endophytic Fungus Rhizopycnis vagum Nitaf22.

Six new dibenzo-α-pyrones, rhizopycnolides A (1) and B (2) and rhizopycnins A-D (3-6), together with eight known congeners (7-14), were isolated from the endophytic fungus Rhizopycnis vagum Nitaf22 obtained from Nicotiana tabacum. The structures of the new compounds were unambiguously elucidated using NMR, HRESIMS, TDDFT ECD calculation, and X-ray crystallography data. Rhizopycnolides A (1) and B (2) feature an uncommon γ-butyrolactone-fused dibenzo-α-pyrone tetracyclic skeleton (6/6/6/5), while rhizopycnin B (4) was the first amino group containing dibenzo-α-pyrone. Rhizopycnolides A (1) and B (2) are proposed to be biosynthesized from polyketide and tricarboxylic acid cycle pathways. The isolated compounds were tested for their antibacterial, antifungal, and cytotoxic activities. Among them, rhizopycnolide A (1), rhizopycnins C (5) and D (6), TMC-264 (8), penicilliumolide D (11), and alternariol (12) were active against the tested pathogenic bacteria Agrobacterium tumefaciens, Bacillus subtilis, Pseudomonas lachrymans, Ralstonia solanacearum, Staphylococcus hemolyticus, and Xanthomonas vesicatoria with MIC values in the range 25-100 µg/mL. Rhizopycnin D (6) and TMC-264 (8) strongly inhibited the spore germination of Magnaporthe oryzae with IC50 values of 9.9 and 12.0 µg/mL, respectively. TMC-264 (8) showed potent cytotoxicity against five human cancer cell lines (HCT-116, HepG2, BGC-823, NCI-H1650, and A2780) with IC50 values of 3.2-7.8 µM.

Biological Potential of Bioorganic Fertilizer Fortified with Bacterial Antagonist for the Control of Tomato Bacterial Wilt and the Promotion of Crop Yields.

The application of Bacillus sp. in the biological control of plant soilborne diseases has been shown to be an environmentally friendly alternative to the use of chemical fungicides. In this study, the effects of bioorganic fertilizer (BOF) fortified with Bacillus amyloliquefaciens SQY 162 on the suppression of tomato bacterial wilt were investigated in pot experiments. The disease incidence of tomato wilt after the application of BOF was 65.18% and 41.62% lower at 10 and 20 days after transplantation, respectively, than in the control condition. BOF also promoted the plant growth. The SQY 162 populations efficiently colonized the tomato rhizosphere, which directly suppressed the number of Ralstonia solanacearum in the tomato rhizosphere soil. In the presence of BOF, the activities of defense-related enzymes in tomato were lower than in the presence of the control treatment, but the expression levels of the defense-related genes of the plants in the salicylic acid and jasmonic acid pathways were enhanced. It was also found that strain SQY 162 could secrete antibiotic surfactin, but not volatile organic compounds, to suppress Ralstonia. The strain could also produce plant growth promotion compounds such as siderophores and indole-3-acetic acid. Thus, owing to its innate multiple-functional traits and its broad biocontrol activities, we found that this antagonistic strain isolated from the tobacco rhizosphere could establish itself successfully in the tomato rhizosphere to control soilborne diseases.

Evaluation of the Antibacterial Effects and Mechanism of Action of Protocatechualdehyde against Ralstonia solanacearum.

Protocatechualdehyde (PCA) is an important plant-derived natural product that has been associated with a wide variety of biological activities and has been widely used in medicine as an antioxidant, anti-aging and an anti-inflammatory agent. However, fewer reports concerning its antibacterial effects on plant-pathogenic bacteria exist. Therefore, in this study, protocatechualdehyde was evaluated for its antibacterial activity against plant pathogens along with the mechanism of its antibacterial action. PCA at 40 µg/mL was highly active against R. solanacearum and significantly inhibited its growth. The minimum bactericidal concentration and minimum inhibitory concentration values for PCA were 40 µg/mL and 20 µg/mL, respectively. Further investigation of the mechanism of action of PCA via transmission electron microscopy and biological assays indicated that the destruction of the cell structure, the shapes and the inhibition of biofilm formation were important. In addition, the application of PCA effectively reduced the incidence of bacterial wilt on tobacco under greenhouse conditions, and the control efficiency was as high as 92.01% at nine days after inoculation. Taken together, these findings suggest that PCA exhibits strong antibacterial activity against R. solanacearum and has the potential to be applied as an effective antibacterial agent for controlling bacterial wilt caused by R. solanacearum.

Spiculisporic acid analogues of the marine-derived fungus, Aspergillus candidus strain HDf2, and their antibacterial activity.

Two novel antibiotic spiculisporic acid analogues, named as spiculisporic acid F (1) and G (2), and two known compounds, (-)-spiculisporic acid (3) and secospiculisporic acid B (4), were isolated by bioactivity-guided fractionation from the fermentation broth of the sea urchin-derived Aspergillus candidus strain HDf2. Their structures were unambiguously established by comprehensive analysis of 1D and 2D NMR, and high-resolution MS spectra, and by comparison with known compounds. Biological experiments demonstrated that compounds 1 and 2 displayed antibacterial activity against Gram-negative Pseudomonas solanacearum and Gram-positive Staphylococcus aureus, but showed no cytotoxicity against SGC-7901 human gastric adenocarcinoma and SPC-A-1 human lung adenocarcinoma tumor cell lines. This is the first critical evidence identifying spiculisporic acid derivatives as a potential bio-control agent for the soil borne pathogen P. solanacearum (E. F. Smith) Smith. These findings provide further insight into the chemical and biological activity diversity of this class of compounds.

Antibacterial activities against rice bacterial leaf blight and tomato bacterial wilt of 2-mercapto-5-substituted-1,3,4-oxadiazole/thiadiazole derivatives.

In this study, a series of 2-mercapto-5-substituted-1,3,4-oxadiazole/thiadiazole derivatives were synthesized and evaluated for their antibacterial activities against rice bacterial leaf blight and tomato bacterial wilt caused by Xanthomonas oryzae pv. oryzae (Xoo) and Ralstonia solanacearum (R. solanacearum) via the turbidimeter test in vitro. Antibacterial bioassays indicated that most compounds demonstrated appreciable antibacterial bioactivities against Xoo and R. solanacearum. Among the title compounds, compound 4i demonstrated the best inhibitory effect against Xoo and R. solanacearum with half-maximal effective concentration (EC50) values of 14.69 and 15.14µg/mL, respectively, which were even better than those of commercial agents Bismerthiazol and Thiodiazole Copper. In vivo antibacterial activities tests under greenhouse conditions revealed that the control efficiency of compound 4i against rice bacterial leaf blight and tobacco bacterial wilt were better than those of Bismerthiazol and Thiodiazole Copper. Meanwhile, field trials also indicated that compound 4i demonstrated appreciable control efficiency against rice bacterial leaf blight and tomato bacterial wilt.

Effect of seed treatment by cold plasma on the resistance of tomato to Ralstonia solanacearum (Bacterial Wilt).

This study investigated the effect of cold plasma seed treatment on tomato bacterial wilt, caused by Ralstonia solanacearum (R. solanacearum), and the regulation of resistance mechanisms. The effect of cold plasma of 80W on seed germination, plant growth, nutrient uptake, disease severity, hydrogen peroxide (H2O2) concentration and activities of peroxidase (POD; EC 1.11.1.7), polyphenol oxidase (PPO; EC 1.10.3.2) and phenylalanine ammonia lyase (PAL; EC 4.3.1.5) were examined in tomato plants. Plasma treatment increased tomato resistance to R. solanacearum with an efficacy of 25.0%. Plasma treatment significantly increased both germination and plant growth in comparison with the control treatment, and plasma-treated plants absorbed more calcium and boron than the controls. In addition, H2O2 levels in treated plants rose faster and reached a higher peak, at 2.579 µM gFW-1, 140% greater than that of the control. Activities of POD (421.3 U gFW-1), PPO (508.8 U gFW-1) and PAL (707.3 U gFW-1) were also greater in the treated plants than in the controls (103.0 U gFW-1, 166.0 U gFW-1 and 309.4 U gFW-1, respectively). These results suggest that plasma treatment affects the regulation of plant growth, H2O2 concentration, and POD, PPO and PAL activity in tomato, resulting in an improved resistance to R. solanacearum. Consequently, cold plasma seed treatment has the potential to control tomato bacterial wilt caused by R. solanacearum.

Recovery of several cell pellet-associated antibiotics produced by Bacillus amyloliquefaciens NJN-6.

UNLABELLED: In this study, six antibiotics which are antagonistic to phytopathogens were extracted from the water-washed cell pellets of Bacillus amyloliquefaciens NJN-6. They were purified by HPLC and identified using electrospray ionization mass spectroscopy (ESI-MS). Besides three iturin A homologous and macrolactin A, two other kinds of antibiotics were extracted from the cell pellets of B. amyloliquefaciens NJN-6. An antifungal compound with the molecular mass of 1072 Da was identified as bacillomycin D, and the antibacterial compound with the molecular mass of 400 Da was identified as macrolactin E. We also quantified iturin A in both cell pellets and culture media, and the concentrations were 751·12 mg kg(-1) and 21·02 mg l(-1) , respectively. These results show that in addition to the culture medium, the cell pellets could also be a potential resource for antibiotics and the strain B. amyloliquefaciens NJN-6 might be an attractive source for antibiotic production and an antagonist for plant diseases. SIGNIFICANCE AND IMPACT OF THE STUDY: The novelty of this work is the production of several antimicrobial substances associated with cell pellets and the production of antibiotic macrolactin E by B. amyloliquefaciens strain. Here, cell pellet-associated antimicrobial compounds were identified, and strain NJN-6 was able to produce three cell pellet-associated iturin A family homologues, bacillomycin D, macrolactin A and macrolactin E. With the exception of macrolactin E, all other antimicrobial compounds were identified in the liquid culture. The production of macrolactin E by any B. amyloliquefaciens strain has not been previously reported; therefore, this is the first report about macrolactin E production by B. amyloliquefaciens.