Design, Synthesis, Antibacterial Activity, and Mechanisms of Novel Benzofuran Derivatives Containing Disulfide Moieties.

The unsatisfactory effects of conventional bactericides and antimicrobial resistance have increased the challenges in managing plant diseases caused by bacterial pests. Here, we report the successful design and synthesis of benzofuran derivatives using benzofuran as the core skeleton and splicing the disulfide moieties commonly seen in natural substances with antibacterial properties. Most of our developed benzofurans displayed remarkable antibacterial activities to frequently encountered pathogens, including Xanthomonas oryzae pv oryzae (Xoo), Xanthomonas oryzae pv oryzicola (Xoc), and Xanthomonas axonopodis pv citri (Xac). With the assistance of the three-dimensional quantitative constitutive relationship (3D-QSAR) model, the optimal compound V40 was obtained, which has better in vitro antibacterial activity with EC50 values of 0.28, 0.56, and 10.43 µg/mL against Xoo, Xoc, and Xac, respectively, than those of positive control, TC (66.41, 78.49, and 120.36 µg/mL) and allicin (8.40, 28.22, and 88.04 µg/mL). Combining the results of proteomic analysis and enzyme activity assay allows the antibacterial mechanism of V40 to be preliminarily revealed, suggesting its potential as a versatile bactericide in combating bacterial pests in the future.

Diversity and biopotential of Bacillus velezensis strains A6 and P42 against rice blast and bacterial blight of pomegranate.

Bacillus velezensis is widely known for its inherent biosynthetic potential to produce a wide range of bio-macromolecules and secondary metabolites, including polyketides (PKs) and siderophores, as well as ribosomally and non-ribosomally synthesized peptides. In the present study, we aimed to investigate the bio-macromolecules, such as proteins and peptides of Bacillus velezensis strains, namely A6 and P42 by whole-cell sequencing and highlighted the potential application in controlling phytopathogens. The bioactive compounds, specifically secondary metabolites, were characterized by whole-cell protein profiling, Thin-Layer Chromatography, Infra-Red Spectroscopy, Nuclear Magnetic Resonance, Gas Chromatograph and Electro Spray Liquid Chromatography. Gas Chromatography analysis revealed that the A6 and P42 strains exert different functional groups of compounds, such as aromatic ring, aliphatic, alkene, ketone, amine groups and carboxylic acid. Whole-cell protein profiling of A6 and P42 strains of B. velezensis by nano-ESI LC-MS/MS revealed the presence of 945 and 5303 proteins, respectively. The in vitro evaluation of crude extracts (10%) of A6 and P42 significantly inhibited the rice pathogen, Magnaporthe oryzae (MG01), whereas the cell-free culture filtrate (75%) of strain P42 showed 58.97% inhibition. Similarly, in vitro evaluation of crude extract (10%) of P42 strain inhibited bacterial blight of pomegranate pathogen, Xanthomonas axonopodis pv. punicae, which eventually resulted in a higher inhibition zone of 3 cm, whereas the cell-free extract (75%) of the same strain significantly suppressed the growth of the pathogen with an inhibition zone of 1.48 cm. From the results obtained, the crude secondary metabolites and cell-free filtrates (containing bio-macromolecules) of the strains A6 and P42 of B. velezensis can be employed for controlling the bacterial and fungal pathogens of crop plants.

Phytochemical Screening and Evaluation of Antioxidant Properties and Antimicrobial Activity against Xanthomonas axonopodis of Euphorbia tirucalli Extracts in Binh Thuan Province, Vietnam.

Euphorbia tirucalli is a medicine plant possessing many bioactive properties. This paper focused on phytochemical screening (alkaloid, flavonoid, saponin, tannin, and anthraquinone), quantification of polyphenol and flavonoids, and activating evaluation of antioxidants and antimicrobial properties against Xanthomonas axonopodis of different extracts from Euphorbia tirucalli grown in Binh Thuan, Vietnam. The best activity fraction was used for purification and determining bioactive ingredients. The results showed that the phytochemical study revealed the presence of alkaloids, flavonoids, tannins, and terpenoids in the ethyl acetate fraction. Saponin and anthraquinone did not present in all extracts. The content of polyphenol and flavonoid of Euphorbia tirucalli stem was in the range of 16.65-106.32 mg EqAG/g and 97.97-450.83 µg QE/g. The ethyl acetate fraction showed higher amounts of polyphenol and flavonoids and antimicrobial activity against X. axonopodis than other fractions. The antioxidant (SC50) activity of Euphorbia tirucalli stem was in the range of 12.91 ± 0.70 and 528.33 ± 25.15 µg/mL. At concentrations of 5.0 and 7.5 mg/mL, the diameter of inhibition of the ethyl acetate fraction was 14.33 ± 0.76 mm and 17.87 ± 0.57 mm, respectively. The MIC (minimum inhibitory concentration) was 0.156 mg/mL. Scopoletin, gallic acid, and piperic acid got MICs corresponding to 78, 312, and 312 µg/mL, respectively. Scopoletin, gallic acid, and piperic acid were found in the ethyl acetate fraction of Euphorbia tirucalli and exhibited the treatment of citrus bacteria canker and plant diseases.

Synthesis and Antibacterial Evaluation of N-phenylacetamide Derivatives Containing 4-arylthiazole Moieties.

A series of new N-phenylacetamide derivatives containing 4-arylthiazole moieties was designed and synthesized by introducing the thiazole moiety into the amide scaffold. The structures of the target compounds were confirmed by 1H-NMR, 13C-NMR and HRMS. Their in vitro antibacterial activities were evaluated against three kinds of bacteria-Xanthomonas oryzae pv. Oryzae (Xoo), Xanthomonas axonopodis pv. Citri (Xac) and X.oryzae pv. oryzicola (Xoc)-showing promising results. The minimum 50% effective concentration (EC50) value of N-(4-((4-(4-fluoro-phenyl)thiazol-2-yl)amino)phenyl)acetamide (A1) is 156.7 µM, which is superior to bismerthiazol (230.5 µM) and thiodiazole copper (545.2 µM). A scanning electron microscopy (SEM) investigation has confirmed that compound A1 could cause cell membrane rupture of Xoo. In addition, the nematicidal activity of the compounds against Meloidogyne incognita (M. incognita) was also tested, and compound A23 displayed excellent nematicidal activity, with mortality of 100% and 53.2% at 500 µg/mL and 100 µg/mL after 24 h of treatment, respectively. The preliminary structure-activity relationship (SAR) studies of these compounds are also briefly described. These results demonstrated that phenylacetamide derivatives may be considered as potential leads in the design of antibacterial agents.

Synthesis and Antibacterial Evaluation of Novel 1,3,4-Oxadiazole Derivatives Containing Sulfonate/Carboxylate Moiety.

Abstract: In order to discover new lead compounds with high antibacterial activity, a series of new derivatives were designed and synthesized by introducing a sulfonate or carboxylate moiety into the 1,3,4-oxadiazole structure. Antibacterial activity against two phytopathogens, Xanthomonas oryzae pv. oryzae (Xoo) and Xanthomonas axonopodis pv. citri (Xac), was assayed in vitro. The preliminary results indicated that ten compounds including 4a-1-4a-4 and 4a-11-4a-16 had good antibacterial activity against Xoo, with EC50 values ranging from 50.1-112.5 µM, which was better than those of Bismerthiazol (253.5 µM) and Thiodiazole copper (467.4 µM). Meanwhile, 4a-1, 4a-2, 4a-3 and 4a-4 demonstrated good inhibitory effect against Xanthomonas axonopodis pv. citri with EC50 values around 95.8-155.2 µM which were better than those of bismerthiazol (274.3 µM) and thiodiazole copper (406.3 µM). In addition, in vivo protection activity of compound 4a-2 and 4a-3 against rice bacterial leaf blight was 68.6% and 62.3%, respectively, which were better than bismerthiazol (49.6%) and thiodiazole copper (42.2%). Curative activity of compound 4a-2 and 4a-3 against rice bacterial leaf blight was 62.3% and 56.0%, which were better than bismerthiazol (42.9%) and thiodiazole copper (36.1%). Through scanning electron microscopy (SEM) analysis, it was observed that compound 4a-2 caused the cell membrane of Xanthomonas oryzae pv. oryzae ruptured or deformed. The present results indicated novel derivatives of 5-phenyl sulfonate methyl 1,3,4-oxadiazole might be potential antibacterial agents.

Synthesis and antibacterial activity of solanum torvum mediated silver nanoparticle against Xxanthomonas axonopodis pv.punicae and Ralstonia solanacearum.

To meet the food demand of growing population, agricultural productivity needs to be increased by employing safe strategies without harming ecosystem. Silver nanoparticle (AgNP) using a green approach has become a promising substitute to the synthetic pesticides to overcome pest menace. In this study, AgNPs were synthesized from Solanum torvum fruit extract and their bactericidal property against phyto bacteria was shown. UV-vis spectroscopic observation revealed a surface resonance peak of 440 nm corresponding to the formation of AgNPs. Microscopic and particle-size analyses showed a nearly spherical size, with an average diameter of 27 nm. Surface charge and polydispersity index of the synthesized AgNPs were -11.8 mV and 0.29, respectively. Powder X-ray diffraction, energy-dispersive X-ray and Infrared spectroscopy techniques were used to explore phase formation, composition and possible biological molecules involved in AgNP formation. AgNPs exhibited minimum inhibitory concentrations of 6.25 µg mL-1 and 12.5 µg mL-1 against bacterial plant pathogens Xanthomonas axonopodis pv. punicae and Ralstonia solanacearum. In-vitro disk-diffusion assay showed inhibition zones of 11.4 ± 1 mm for R. solanacearum and 18.1 ± 1 mm for X. axonopodis pv. punicae treated with 50 µg mL-1 AgNPs. The AgNPs generated intracellular reactive oxygen species in the pathogens. DNA damage and DNA replication inhibition studies showed genotoxicity of AgNPs to the bacterial cells. A plant toxicity study demonstrated a nontoxic effect of the synthesized NPs. Overall; the results show that AgNPs can be used as an economically feasible, ecologically safe and effective approach to overcome bacterial diseases.

Antibacterial activity of various chitosan forms against Xanthomonas axonopodis pv. glycines.

In this study, the antibacterial activities of colloidal chitosan, chitosan solution, and chitooligosaccharide solution were evaluated against Xanthomonas axonopodis pv. glycines grown in peptone sucrose broth (PSB) medium. Treatment with colloidal chitosan (0.01, 0.025, and 0.05%) inhibited X. axonopodis pv. glycines growth only until 36 h. Thin-layer chromatography analysis detected some metabolites, consistently with the cell growth pattern. Two chitooligosaccharides (1-3 kDa and 5-10 kDa) dissolved in distilled water and acetic acid did not exhibit antibacterial activity against X. axonopodis pv. glycines at all tested concentrations (0, 0.001, 0.005, 0.01, 0.015, 0.02, 0.025, and 0.05%). Compared to the control, the chitosan solution decreased X. axonopodis pv. glycines cell growth by 58.7% and 99.0% at concentrations of 0.015% and 0.02%, respectively, after 3 d of incubation. The chitosan solution exhibited the highest antibacterial activity at pH 6.5. However, the antibacterial activity of chitosan decreased in the presence of NaCl and MgCl2.

Bactericidal In-Vitro Effect of Zinc Ferrite Nanoparticles and the Orange Wax Extracts on Three Phytopathogen Microorganisms.

Phytopathogenic bacteria affect a wide variety of crops, causing significant economic losses. Natural biocides are the alternative to chemical methods of phytopathogens control. The goal of the present study is the evaluation of the biocidal activity of the following: 1) the extract of orange wax (EOW); 2) zinc ferrite nanoparticles (ZF-NPs); 3) the EOW adsorbed on the ZF-NPs; and 4) the EOW/ZF-NPs washed with 40% ethanol. For the biocidal activity, three phytopathogenic bacteria were used, namely, Xanthomonas axonopodis pv. Vesicatoria (Xav) Erwinia amylovora (Ew), and Pseudomonas syringae pv. Phaseolicola (Psph). For the ZF-NPs, an inhibitory effect higher than 50% ( ) was observed for Xav respect to the antibiotic used as positive control. On the other hand, the ZF-NPs did not show inhibitory effects on both Ew and Psph. In addition, the EOW in dimethyl sulfoxide (DMSO) at 100% caused growth inhibition on Xav, bacteriostatic activity on Ew, and had not biological activity on Psph. To the best of our knowledge, the control of Xav by zinc ferrites and orange wax, and the bacteriostatic effect produced by orange wax extract on Ew have not been reported elsewhere. Orange wax and zinc ferrite nanoparticles show potential in control of phytopathogenic bacteria. However, the bactericidal effect depends on the bacterium, the concentration of treatments, and the method of preparation.

Save the crop: Photodynamic Inactivation of plant pathogens I: bacteria.

The ever growing world-population poses challenges concerning the need for more food free of pesticide residues. The most common means to control plant pathogens is through the application of pesticides, which raises concerns over safety for humans and the environment. Recently, Photodynamic Inactivation (PDI) of microorganisms using natural photosensitizers has shown itself to be a powerful tool to combat bacteria and fungi. This study investigates the efficacy of PDI against the Gram(+) bacterial plant pathogen Rhodococcus fascians and Gram(-) Xanthomonas axonopodis and Erwinia amylovora using two chlorin e6 derivatives as photosensitizers: anionic sodium magnesium chlorophyllin (Chl, approved as food additive E140) in combination with cell wall permeabilizing agents (Na2EDTA or Polyaspartic acid sodium salt (PA)) and B17-0024, a mixture of chlorin e6 derivatives with cationic moieties at physiological pH. Both photosensitizers show excellent efficacy against R. fascians, whereby B17-0024 is phototoxic at a one order of magnitude lower concentration than Chl (10 µM B17-0024: relative inactivation (r.i.) >7.5 × 106, 100 µM Chl: r.i. 2.2 × 106, illumination with 26.6 J cm-2, 395 nm). The phototreatment of Gram(-) bacteria with Chl requires the obligatory use of cell wall permeabilizing agents like Na2EDTA (X. axonopodis) or PA (E. amylovora) to induce significant killing (more than 7 log units at 100 µM). On the other hand, B17-0024 proves to be a highly effective photosensitizer inducing bacterial inactivation at very low concentrations (10 µM for R. fascians and X. axonopodis, 100 µM for E. amylovora) without additives. In summary, PDI using both the natural photosensitizer Chl in combination with cell wall permeabilizing agents is effective and environmentally friendly. As an alternative, B17-0024 is highly photoactive against all model strains tested - even without cell wall permeabilizing agents. The photodynamic approach based on chlorin e6 derivatives should add to the growers' toolbox as a preferred alternative for the control of phytopathogens.

Synthesis and three-dimensional quantitative structure-activity relationship study of quinazoline derivatives containing a 1,3,4-oxadiazole moiety as efficient inhibitors against Xanthomonas axonopodis pv. citri.

A series of quinazoline derivatives containing a 1,3,4-oxadiazole moiety were synthesized and evaluated for their antibacterial activities against Xanthomonas axonopodis pv. citri (Xac) and Ralstonia solanacearum (Rs). Antibacterial bioassays indicated that most of target compounds exhibited significant antibacterial activities against Xac and Rs in vitro. Strikingly, compounds 6d-6i, 6m-6r and 6u-6x showed antibacterial activity against Xac, with [Formula: see text] values ranging from 14.42 to 38.91 [Formula: see text]g/mL, which are better than that of bismerthiazol (39.86 [Formula: see text]g/mL). Based on the antibacterial activity against Xac, comparative molecular filed analysis and comparative molecular similarity index analysis models were generated to investigate the structure-activity relationship of title compounds against Xac. The analytical results indicated that the above models exhibited good predictive accuracy and could be used as practical tools for guiding the design and synthesis of more potent quinazoline derivatives containing a 1,3,4-oxadiazole moiety.

Thymol nanoemulsion exhibits potential antibacterial activity against bacterial pustule disease and growth promotory effect on soybean.

An antibacterial and plant growth promoting nanoemulsion was formulated using thymol, an essential oil component of plant and Quillaja saponin, a glycoside surfactant of Quillaja tree. The emulsion was prepared by a sonication method. Fifty minutes of sonication delivered a long term stable thymol nanoemulsion which was characterized by dynamic light scattering (DLS), transmission electron microscopy (TEM), cryogenic-field emission scanning electron microscopy (Cryo-FESEM) and fourier transform infra-red (FTIR) spectroscopy. Creaming index, pH and dilution stability were also studied for deliberation of its practical applications. The nanoemulsion (0.01-0.06%, v/v) showed substantial in vitro growth inhibition of Xanthomonas axonopodis pv. glycine of soybean (6.7-0.0 log CFU/ml). In pot experiments, seed treatment and foliar application of the nanoemulsion (0.03-0.06%, v/v) significantly lowered the disease severity (DS) (33.3-3.3%) and increased percent efficacy of disease control (PEDC) (54.9-95.4%) of bacterial pustule in soybean caused by X. axonopodis pv. glycine. Subsequently, significant enhancements of plant growth were also recorded in plants treated with thymol nanoemulsion. This is the first report of a thymol based nanoemulsion obtained using Quillaja saponin as a surfactant. Our study claims that nano scale thymol could be a potential antimicrobial and plant growth promoting agent for agriculture.

Activity of Antarctic fungi extracts against phytopathogenic bacteria.

This study aims to obtain secondary metabolites extracts from filamentous fungi isolated from soil and marine sediments from Antarctic ecosystems and to assess its potential antibacterial activity on Xanthomonas euvesicatoria and Xanthomonas axonopodis pv. passiflorae (phytopathogenic bacteria causing diseases in pepper and tomato and passionfruit, respectively). Among the 66 crude intracellular and extracellular extracts obtained from fungi recovered from soil and 79 obtained from marine sediment samples, 25 showed the ability to prevent the growth of X. euvesicatoria in vitro and 28 showed the ability to prevent the growth of X. axonopodis pv. passiflorae in vitro. Intracellular and extracellular extracts from soil fungi inhibited around 97% of X. euvesicatoria and 98% of X. axonopodis pv. passiflorae at 2·1 mg ml-1 . The average inhibition rates against X. euvesicatoria and X. axonopodis pv. passiflorae for intracellular and extracellular extracts from marine sediments fungi were around 96 and 97%, respectively, at 3·0 mg ml-1 . Extracts containing secondary metabolites with antimicrobial activity against X. euvesicatoria and X. axonopodis pv. passiflorae were obtained, containing possible substitutes for the products currently used to control these phytopathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: Micro-organisms from extreme ecosystems, such as the Antarctic ecosystem, need to survive in harsh conditions with low temperatures, low nutrients and high UV radiation. Micro-organisms adapt to these conditions evolving diverse biochemical and physiological adaptations essential for survival. All this makes these micro-organisms a rich source of novel natural products based on unique chemical scaffolds. Discovering novel bioactive compounds is essential because of the rise in antibiotic-resistant micro-organisms and the emergence of new infections. Fungi from Antarctic environments have been proven to produce bioactive secondary metabolites against various micro-organisms, but few studies have shown activity against Xanthomonas phytopathogens.

Rational Design of Cyclic Antimicrobial Peptides Based on BPC194 and BPC198.

A strategy for the design of antimicrobial cyclic peptides derived from the lead compounds c(KKLKKFKKLQ) (BPC194) and c(KLKKKFKKLQ) (BPC198) is reported. First, the secondary ß-structure of BPC194 and BPC198 was analyzed by carrying out molecular dynamics (MD) simulations. Then, based on the sequence pattern and the ß-structure of BPC194 or BPC198, fifteen analogues were designed and synthesized on solid-phase. The best peptides (BPC490, BPC918, and BPC924) showed minimum inhibitory concentration (MIC) values <6.2 µM against Pseudomonas syringae pv. syringae and Xanthomonas axonopodis pv. vesicatoria, and an MIC value of 12.5 to 25 µM against Erwinia amylovora, being as active as BPC194 and BPC198. Interestingly, these three analogues followed the structural pattern defined from the MD simulations of the parent peptides. Thus, BPC490 maintained the parallel alignment of the hydrophilic pairs K¹-K8, K²-K7, and K4-K5, whereas BPC918 and BPC924 included the two hydrophilic interactions K³-Q10 and K5-K8. In short, MD simulations have proved to be very useful for ascertaining the structural features of cyclic peptides that are crucial for their biological activity. Such approaches could be further employed for the development of new antibacterial cyclic peptides.

Label-free quantitative proteomic analysis of inhibition of Xanthomonas axonopodis pv. citri by the novel bactericide Fubianezuofeng.

To understand the antibacterial mechanism of the new bactericide 2-(methylsulfonyl)-5- (4-fluorobenzyl)-1, 3, 4-oxadiazole (Generic name: Fubianezuofeng), we performed label-free quantitative proteomics analysis of the response of Xanthomonas axonopodis pv. citri (Xac) strain 29-1 to Fubianezuofeng. A total of 1133 proteins were identified in the treatment and control groups. Upon treatment with the 1/2 minimum inhibitory concentration (MIC), 339 proteins were found to be differentially expressed (fold changes>1.5, p<0.05) with 99 upregulated and 240 down-regulated. In comparison, 314 proteins were differentially expressed (125 up-regulated, 189 down-regulated) at MIC. The differentially expressed proteins were enriched for those involved in the pyrimidine metabolic pathway. The results offer a complete view of the proteome changes in bacteria in response to Fubianezuofeng.

Effect of cellulose and lignin on disintegration, antimicrobial and antioxidant properties of PLA active films.

This study reports the effects on antimicrobial, antioxidant, migration and disintegrability activities of ternary nanocomposite films based on poly(lactic acid) incorporating two biobased nanofillers, (cellulose nanocrystals (CNC) and lignin nanoparticles (LNP)), in two different amounts (1 and 3% wt.). Results from antimicrobial tests revealed a capacity to inhibit the Gram negative bacterial growth of Xanthomonas axonopodis pv. vesicatoria and Xanthomonas arboricola pv. pruni along the time, offering innovative opportunities against dangerous bacterial plant pathogens. LNP proved to be highly efficient in antioxidation activity, based on the disappearance of the absorption band at 517nm of the free radical, 2,2-diphenyl-1-picrylhydrazyl (DPPH) upon reduction by an antiradical compound; moreover the combination of LNP and CNC generates a synergistic positive effect in the antioxidation response of PLA ternary films. Furthermore, all the studied formulations showed a disintegrability value up to 90% after 15days of incubation in composting conditions. Migration results showed that the films can be considered suitable for application in food packaging field.

Sunroot mediated synthesis and characterization of silver nanoparticles and evaluation of its antibacterial and rat splenocyte cytotoxic effects.

A rapid, green phytosynthesis of silver nanoparticles (AgNPs) using the aqueous extract of Helianthus tuberosus (sunroot tuber) was reported in this study. The morphology of the AgNPs was determined by transmission electron microscopy (TEM). Scanning electron microscopy-energy-dispersive spectroscopy (SEM-EDS) and X-ray powder diffraction (XRD) analysis confirmed the presence of AgNPs. Fourier transform infrared spectroscopy (FTIR) analysis revealed that biomolecules in the tuber extract were involved in the reduction and capping of AgNPs. The energy-dispersive spectroscopy (EDS) analysis of the AgNPs, using an energy range of 2-4 keV, confirmed the presence of elemental silver without any contamination. Further, the synthesized AgNPs were evaluated against phytopathogens such as Ralstonia solanacearum and Xanthomonas axonopodis. The AgNPs (1-4 mM) extensively reduced the growth rate of the phytopathogens. In addition, the cytotoxic effect of the synthesized AgNPs was analyzed using rat splenocytes. The cell viability was decreased according to the increasing concentration of AgNPs and 67% of cell death was observed at 100 µg/mL.

Field evaluation of the bacterial volatile derivative 3-pentanol in priming for induced resistance in pepper.

Plants are defended from attack by emission of volatile organic compounds (VOCs) that can act directly against pathogens and herbivores or indirectly by recruiting natural enemies of herbivores. However, microbial VOC have been less investigated as potential triggers of plant systemic defense responses against pathogens in the field. Bacillus amyloliquefaciens strain IN937a, a plant growth-promoting rhizobacterium that colonizes plant tissues, stimulates induced systemic resistance (ISR) via its emission of VOCs. We investigated the ISR capacity of VOCs and derivatives collected from strain IN937a against bacterial spot disease caused by Xanthomonas axonopodis pv. vesicatoria in pepper. Of 15 bacterial VOCs and their derivatives, 3-pentanol, which is a C8 amyl alcohol reported to be a component of sex pheromones in insects, was selected for further investigation. Pathogens were infiltrated into pepper leaves 10, 20, 30, and 40 days after treatment and transplantation to the field. Disease severity was assessed 7 days after transplantation. Treatment with 3-pentanol significantly reduced disease severity caused by X. axonopodis and naturally occurring Cucumber mosaic virus in field trials over 2 years. We used quantitative real-time polymerase chain analysis to examine Pathogenesis-Related genes associated with salicylic acid (SA), jasmonic acid (JA), and ethylene defense signaling. The expression of Capsicum annuum Pathogenesis-Related protein 1 (CaPR1), CaPR2, and Ca protease inhibitor2 (CaPIN2) increased in field-grown pepper plants treated with 3-pentanol. Taken together, our results show that 3-pentanol triggers induced resistance by priming SA and JA signaling in pepper under field conditions.

The folate precursor para-aminobenzoic acid elicits induced resistance against Cucumber mosaic virus and Xanthomonas axonopodis.

BACKGROUND AND AIMS: The use of vitamins including vitamin B1, B2 and K3 for the induction of systemic acquired resistance (SAR) to protect crops against plant pathogens has been evaluated previously. The use of vitamins is beneficial because it is cost effective and safe for the environment. The use of folate precursors, including ortho-aminobenzoic acid, to induce SAR against a soft-rot pathogen in tobacco has been reported previously. METHODS: In the present study, para-aminobenzoic acid (PABA, also referred to as vitamin Bx) was selected owing to its effect on the induction of SAR against Xanthomonas axonopodis pv. vesicatoria in pepper plants through greenhouse screening. KEY RESULTS: Dipping of pepper seedlings in a 1 mm PABA solution in field trials induced SAR against artificially infiltrated X. axonopodis pv. vesicatoria and naturally occurring cucumber mosaic virus. Expression of the Capsicum annuum pathogenesis-related 4 gene was primed in response to pathogen infection as assessed by quantitative real-time PCR. The accumulation of cucumber mosaic virus RNA was reduced in PABA-treated pepper plants at 40 and 105 d post-treatment. Unexpectedly, fruit yield was increased in PABA-treated plants, indicating that PABA-mediated SAR successfully protected pepper plants from infection by bacterial and viral pathogens without significant fitness allocation costs. CONCLUSIONS: The present study is the first to demonstrate the effective elicitation of SAR by a folate precursor under field conditions.

Exogenous treatment with salicylic acid attenuates occurrence of citrus canker in susceptible navel orange (Citrus sinensis Osbeck).

Citrus canker caused by Xanthomonas axonopodis pv. citri (Xac) is a devastating bacterial disease threatening the citrus industry. Salicylic acid (SA) plays a key role in plant defense response to biotic stress, but information is scarce concerning the application of SA to enhancing Xac resistance. In the present research attempts were made to investigate how exogenous application of SA influenced canker disease outbreak in navel orange (Citrus sinensis). Exogenously applied SA at 0.25 mM significantly enhanced the endogenous free and bound SA, particularly the latter. Upon exposure to Xac, lower disease incidence rate and smaller lesion sites were observed in the samples pre-treated with SA, accompanied by repression of bacterial growth at the lesion sites. Concurrent with the augmented disease resistance, SA-treated leaves had higher H2O2 level and smaller stomata apertures before or after Xac infection when compared with their counterparts pre-treated with water (control). SA treatment elevated the activities of phenylalanine ammonia-lyase and ß-1,3-glucanase, but only the latter was higher in the SA-treated samples after Xac infection. In addition, mRNA levels of two pathogenesis-related genes, CsCHI and CsPR4A, were higher in the SA-treated samples relative to the control. Taken together, our results strongly suggest that the exogenously applied SA has evoked a cascade of physiological and molecular events that function singly or in concert to confer resistance to Xac invasion.

Peptidotriazoles with antimicrobial activity against bacterial and fungal plant pathogens.

We designed and prepared peptidotriazoles based on the antimicrobial peptide BP100 (LysLysLeuPheLysLysIleLeuLysTyrLeu-NH(2)) by introducing a triazole ring in the peptide backbone or onto the side chain of a selected residue. These compounds were screened for their in vitro growth inhibition of bacterial and fungal phytopathogens, and for their cytotoxic effects on eukaryotic cells and tobacco leaves. Their proteolytic susceptibility was also analyzed. The antibacterial activity and the hemolysis were influenced by the amino acid that was modified with the triazole as well as by the absence of presence of a substituent in this heterocyclic ring. We identified sequences active against the bacteria Xanthomonas axonopodis pv. vesicatoria, Erwinia amylovora, Pseudomonas syringae pv. syringae (MIC of 1.6-12.5 µM), and against the fungi Fusarium oxysporum (MIC<6.2-12.5 µM) with low hemolytic activity (0-23% at 50 µM), high stability to protease digestion and no phytotoxicity. These peptidotriazoles constitute good candidates to design new antimicrobial agents.