RESUMO
Strategies to reduce crop losses due to drought are needed as climate variability affects agricultural productivity. Wheat (Triticum aestivum var. Juniper) growth in a nutrient-sufficient, solid growth matrix containing varied doses of CuO, ZnO, and SiO2 nanoparticles (NPs) was used to evaluate NP mitigation of drought stress. NP amendments were at fertilizer levels, with maxima of 30 Cu, 20 Zn, and 200 Si (mg metal/kg matrix). Seeds of this drought-tolerant cultivar were inoculated with Pseudomonas chlororaphis O6 (PcO6) to provide a protective root microbiome. An 8 day drought imposed on 14 day-old wheat seedlings decreased shoot and root mass, shoot water content, and the quantum yield of photosystem II when compared to watered plants. PcO6 root colonization was not impaired by drought or NPs. A dose-dependent increase in the Cu, Zn, and Si from the NPs was observed from analysis of the rhizosphere solution, and this process was not affected by drought. Consequently, fertilizer concentrations of the NPs did not further improve drought tolerance in wheat seedlings under the growth conditions of adequate mineral nutrition and the presence of a beneficial microbiome. These findings suggest that potential NP benefits in promoting plant drought tolerance occur only under certain environmental conditions.
Assuntos
Nanopartículas , Óxido de Zinco , Secas , Nutrientes , Raízes de Plantas , Plântula , Dióxido de Silício , TriticumRESUMO
A biocontrol bacterium, Pseudomonas chlororaphis O6 promotes plant health through multifaceted mechanisms. In this study, we used P. chlororaphis O6 mutants to examine metabolites with aphicidal activity. Direct application of intact P. chlororaphis cells to the surface of second-instar nymphs of the green peach aphid resulted in no mortality. However, nymphs died when exposed only to the volatiles produced by the P. chlororaphis O6 wild-type strain grown on rich media. Mutants lacking the production of two antibiotics, phenazine and pyrrolnitrin, or the insect toxin FitD retained the aphicidal potential of the wild-type strain. However, the volatiles produced by mutants deficient in the production of hydrogen cyanide (HCN) or defective in the synthesis of the global regulator GacS, which regulates HCN synthesis, showed no aphicidal activity. Direct application of potassium cyanide caused mortality of green peach aphid nymphs. These results indicate that HCN production by a plant probiotic is involved in preventing insect growth.
Assuntos
Afídeos/efeitos dos fármacos , Cianeto de Hidrogênio/farmacologia , Inseticidas/farmacologia , Pseudomonas chlororaphis/metabolismo , Animais , Cianeto de Hidrogênio/metabolismo , Inseticidas/metabolismoRESUMO
Cost-effective "green" methods of producing Ag nanoparticles (NPs) are being examined because of the potential of these NPs as antimicrobials. Ag NPs were generated from Ag ions using extracellular metabolites from a soil-borne Pythium species. The NPs were variable in size, but had one dimension less than 50 nm and were biocoated; aggregation and coating changed with acetone precipitation. They had dose-dependent lethal effects on a soil pseudomonad, Pseudomonas chlororaphis O6, and were about 30-fold more effective than Ag(+) ions. A role of reactive oxygen species in cell death was demonstrated by use of fluorescent dyes responsive to superoxide anion and peroxide accumulation. Also mutants of the pseudomonad, defective in enzymes that protect against oxidative stress, were more sensitive than the wild type strain; mutant sensitivity differed between exposure to Ag NPs and Ag(+) ions demonstrating a nano-effect. Imaging of bacterial cells treated with the biocoated Ag NPs revealed no cell lysis, but there were changes in surface properties and cell height. These findings support that biocoating the NPs results in limited Ag release and yet they retained potent antimicrobial activity.
Assuntos
Anti-Infecciosos/farmacologia , Nanopartículas Metálicas/química , Prata/farmacologia , Anti-Infecciosos/química , Reatores Biológicos , Peróxido de Hidrogênio/farmacologia , Testes de Sensibilidade Microbiana , Tamanho da Partícula , Pseudomonas chlororaphis/efeitos dos fármacos , Pseudomonas chlororaphis/ultraestrutura , Pythium/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Prata/químicaRESUMO
Zn is an essential element for plants yet some soils are Zn-deficient and/or have low Zn-bioavailability. This paper addresses the feasibility of using ZnO nanoparticles (NPs) as soil amendments to improve Zn levels in the plant. The effects of soil properties on phytotoxicity and Zn bioavailability from the NPs were studied by using an acidic and a calcareous alkaline soil. In the acid soil, the ZnO NPs caused dose-dependent phytotoxicity, observed as inhibition of elongation of roots of wheat, Triticum aestivum. Phytotoxicity was mitigated in the calcareous alkaline soil although uptake of Zn from the ZnO NPs occurred doubling the Zn level compared to control plants. This increase occurred with a low level of Zn in the soil solution as expected from the interactions of Zn with the soil components at the alkaline pH. Soluble Zn in the acid soil was 200-fold higher and shoot levels were tenfold higher than from the alkaline soil correlating with phytotoxicity. Mitigation of toxicity was not observed in plants grown in sand amended with a commercial preparation of humic acid: growth, shoot uptake and solubility of Zn from the NPs was not altered by the humic acid. Thus, variation in humic acid between soils may not be a major factor influencing plant responses to the NPs. These findings illustrate that formulations of ZnO NPs to be used as a soil amendment would need to be tuned to soil properties to avoid phytotoxicity yet provide increased Zn accumulations in the plant.
Assuntos
Nanopartículas/química , Nanopartículas/toxicidade , Triticum/efeitos dos fármacos , Óxido de Zinco/química , Óxido de Zinco/toxicidade , Raízes de Plantas/efeitos dos fármacos , Brotos de Planta/efeitos dos fármacos , SoloRESUMO
We provide here a comparative genome analysis of ten strains within the Pseudomonas fluorescens group including seven new genomic sequences. These strains exhibit a diverse spectrum of traits involved in biological control and other multitrophic interactions with plants, microbes, and insects. Multilocus sequence analysis placed the strains in three sub-clades, which was reinforced by high levels of synteny, size of core genomes, and relatedness of orthologous genes between strains within a sub-clade. The heterogeneity of the P. fluorescens group was reflected in the large size of its pan-genome, which makes up approximately 54% of the pan-genome of the genus as a whole, and a core genome representing only 45-52% of the genome of any individual strain. We discovered genes for traits that were not known previously in the strains, including genes for the biosynthesis of the siderophores achromobactin and pseudomonine and the antibiotic 2-hexyl-5-propyl-alkylresorcinol; novel bacteriocins; type II, III, and VI secretion systems; and insect toxins. Certain gene clusters, such as those for two type III secretion systems, are present only in specific sub-clades, suggesting vertical inheritance. Almost all of the genes associated with multitrophic interactions map to genomic regions present in only a subset of the strains or unique to a specific strain. To explore the evolutionary origin of these genes, we mapped their distributions relative to the locations of mobile genetic elements and repetitive extragenic palindromic (REP) elements in each genome. The mobile genetic elements and many strain-specific genes fall into regions devoid of REP elements (i.e., REP deserts) and regions displaying atypical tri-nucleotide composition, possibly indicating relatively recent acquisition of these loci. Collectively, the results of this study highlight the enormous heterogeneity of the P. fluorescens group and the importance of the variable genome in tailoring individual strains to their specific lifestyles and functional repertoire.
Assuntos
Genoma Bacteriano , Plantas , Pseudomonas fluorescens/genética , Pseudomonas fluorescens/metabolismo , Análise de Sequência de DNA , Animais , Proteínas de Bactérias/genética , Toxinas Bacterianas/genética , Bacteriocinas/genética , Heterogeneidade Genética , Variação Genética , Interações Hospedeiro-Patógeno/genética , Insetos/genética , Família Multigênica , Filogenia , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Plantas/genética , Plantas/microbiologia , Sequências Repetitivas de Ácido Nucleico/genética , Resorcinóis/metabolismoRESUMO
The expansion of nanotechnology raises concerns about the consequences of nanomaterials in plants. Here, the effects of nanoparticles (NPs; 100-500 mg/kg) on processes related to micronutrient accumulation were evaluated in bean (Phaseolus vulgaris) exposed to CuO NPs, a mixture of CuO and ZnO (CuO:ZnO) NPs, and in CuO NP-exposed plants colonized by a root bacterium, Pseudomonas chlororaphis O6 (PcO6) in a sand matrix for 7 days. Depending on exposure levels, the inhibition of growth by CuO NPs was more apparent in roots (10-66 %) than shoots (9-25 %). In contrast, CuO:ZnO NPs or root colonization with PcO6 partially mitigated growth inhibition. At 500 mg/kg exposure, CuO NPs increased soluble Cu in the growth matrix by 23-fold, relative to the control, while CuO:ZnO NPs increased soluble Cu (26-fold), Zn (127-fold) and Ca (4.5-fold), but reduced levels of Fe (0.8-fold) and Mn (0.75-fold). Shoot accumulations of Cu (3.8-fold) and Na (1-fold) increased, while those of Fe (0.4-fold), Mn (0.2-fold), Zn (0.5-fold) and Ca (0.5-fold) were reduced with CuO NP (500 mg/kg) exposure. CuO:ZnO NPs also increased shoot Cu, Zn and Na levels, while decreasing that of Fe, Mn, Ca and Mg. Root colonization reduced shoot uptake of Cu and Na, 15 and 24 %, respectively. CuO NPs inhibited ferric reductase (up to 49 %) but stimulated cupric (up to 273 %) reductase activity; while CuO:ZnO NPs or root colonization by PcO6 altered levels of ferric, but not copper reductase activity, relative to CuO NPs. Cu ions at the level released from the NPs did not duplicate these effects. Our findings demonstrate that in addition to the apparent phytotoxic effects of NPs, NP exposure may also have subtle impacts on secondary processes such as metal nutrition.
Assuntos
Cobre/toxicidade , Nanopartículas Metálicas/toxicidade , Phaseolus/efeitos dos fármacos , Microbiologia do Solo , Óxido de Zinco/toxicidade , Metais/metabolismo , Phaseolus/metabolismo , Phaseolus/microbiologia , Pseudomonas/crescimento & desenvolvimentoRESUMO
An aggressive root colonizer, Pseudomonas chlororaphis O6 produces various secondary metabolites that impact plant health. The sensor kinase GacS is a key regulator of the expression of biocontrol-related traits. Biofilm formation is one such trait because of its role in root surface colonization. This paper focuses on the effects of carbon source on biofilm formation. In comparison with the wild type, a gacS mutant formed biofilms at a reduced level with sucrose as the major carbon source but at much higher level with mannitol in the defined medium. Biofilm formation by the gacS mutant occurred without phenazine production and in the absence of normal levels of acyl homoserine lactones, which promote biofilms with other pseudomonads. Colonization of tomato roots was similar for the wild type and gacS mutant, showing that any differences in biofilm formation in the rhizosphere were not of consequence under the tested conditions. The reduced ability of the gacS mutant to induce systemic resistance against tomato leaf mold and tomato gray mold was consistent with a lack of production of effectors, such as phenazines. These results demonstrated plasticity in biofilm formation and root colonization in the rhizosphere by a beneficial pseudomonad.
Assuntos
Proteínas de Bactérias/metabolismo , Biofilmes/crescimento & desenvolvimento , Carbono/metabolismo , Manitol/metabolismo , Pseudomonas/fisiologia , Solanum lycopersicum/microbiologia , Acil-Butirolactonas/metabolismo , Proteínas de Bactérias/genética , Fenazinas/metabolismo , Raízes de Plantas/microbiologia , Pseudomonas/metabolismo , Plântula/microbiologia , Sacarose/metabolismoRESUMO
Hydroponic plant growth studies indicate that silver nanoparticles (Ag NPs) are phytotoxic. In this work, the phytotoxicity of commercial Ag NPs (10 nm) was evaluated in a sand growth matrix. Both NPs and soluble Ag were recovered from water extracts of the sand after growth of plants challenged with the commercial product; the surface charge of the Ag NPs in this extract was slightly reduced compared to the stock NPs. The Ag NPs reduced the length of shoots and roots of wheat in a dose-dependent manner. Furthermore, 2.5 mg/kg of the NPs increased branching in the roots of wheat (Triticum aestivum L.), thereby affecting plant biomass. Micron-sized (bulk) Ag particles (2.5 mg/kg) as well as Ag ions (63 µg Ag/kg) equivalent to the amount of soluble Ag in planted sand with Ag NPs (2.5 mg/kg) did not affect plant growth compared to control. In contrast, higher levels of Ag ions (2.5 mg/kg) reduced plant growth to a similar extent as the Ag NPs. Accumulation of Ag was detected in the shoots, indicating an uptake and transport of the metal from the Ag NPs in the sand. Transmision electron microscopy indicated that Ag NPs were present in shoots of plants with roots exposed to the Ag NPs or high levels of Ag ions. Both of these treatments caused oxidative stress in roots, as indicated by accumulation of oxidized glutathione, and induced expression of a gene encoding a metallothionein involved in detoxification by metal ion sequestration. Our findings demonstrate the potential effects of environmental contamination by Ag NPs on the metabolism and growth of food crops in a solid matrix.
Assuntos
Nanopartículas Metálicas/toxicidade , Prata/toxicidade , Triticum/efeitos dos fármacos , Triticum/crescimento & desenvolvimento , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Dióxido de Silício/químicaRESUMO
The environmental fate of metal oxide particles as a function of size was assessed by comparing the behavior of CuO or ZnO nanoparticles (NPs) to that of the corresponding microparticles (MPs) in a sand matrix, with and without wheat (Triticum aestivum L.) growth. After 14 days of incubation in the planted sand, the CuO and ZnO NPs were increased from their nominal sizes of <50 nm and <100 nm, to ~317 nm and ~483 nm, respectively. Accordingly, the negative surface charge of colloids present in aqueous extracts from the sand amended with CuO (-27.0 mV) and ZnO (-10.0 mV) NPs was reduced by the presence of plants, to -19.8 mV and -6.0 mV, respectively. The surface charge of the MPs was not influenced by plants. Plant growth increased dissolution of NPs and MPs of both metal oxides in the sand from <0.3 mg/kg to about 1.0 mg/kg for the CuO products, and from ≤0.6 mg/kg to between 1.0 and 2.2 mg/kg for the Zn products. The NP or MP products reduced wheat root length by ~60% or ~50% from control levels; CuO was more toxic than ZnO. X-ray absorption spectroscopy (XAS) analysis showed that treatments with MPs or NPs of ZnO led to similar accumulations of Zn-phosphate species in the shoots, likely from dissolution of ZnO. Exposure to CuO NPs or MPs resulted in similar XAS spectra for Cu in the shoots explained by plant accumulation of both CuO and Cu(I)-sulfur complexes. These findings demonstrate the similarities between commercial NPs and MPs of CuO or ZnO in wheat plants, with greater root toxicity correlating with smaller particle size. Factors from the sand and the plant modified the aggregation or dissolution of both types of particles, thus, influencing their environmental fates.
Assuntos
Cobre/química , Nanopartículas , Plantas/química , Óxido de Zinco/química , Microscopia de Força Atômica , Solubilidade , Espectroscopia por Absorção de Raios XRESUMO
Fungal plant pathogens such as Fusarium graminearum cause severe global economic losses in cereals crops, and current control measures are limited. This work addresses the potential for ZnO nanoparticles (NPs) and biocontrol bacteria to be used in plant fungal control strategies. Growth of F. graminearum was significantly (p = 0.05) inhibited by inclusion of the NPs in a mung bean broth agar and in sand. Suspension in mung bean broth medium modified the surface charge, dissolution, and aggregation state of the ZnO NPs, in comparison to processes occurring in water suspension. The ZnO NPs were significantly more inhibitory to fungal growth than micro-sized particles of ZnO, although both types of particles released similar levels of soluble Zn, indicating size-dependent toxicity of the particles. Zn ions produced dose-dependent inhibition, noticeable at the level of soluble Zn released from NPs after seven-day suspension in medium; inhibitory levels caused acidification of the growth medium. Transfer of fungal inoculum after exposure to the ZnO NPs to fresh medium did not indicate adaptation to the stress because growth was still inhibited by the NPs. The ZnO NPs did not prevent metabolites from a biocontrol bacterium, Pseudomonas chlororaphis O6, from inhibiting Fusarium growth: no synergism was observed in the mung bean agar. Because other studies find that soil amendment with ZnO NPs required high doses for inhibition of plant growth, the findings of pathogen growth control reported in this paper open the possibility of using ZnO NP-based formulations to complement existing strategies for improving crop health in field settings.
Assuntos
Antifúngicos/farmacologia , Fusarium/efeitos dos fármacos , Nanopartículas/toxicidade , Controle Biológico de Vetores , Pseudomonas/fisiologia , Óxido de Zinco/farmacologia , Zinco/farmacologia , Ágar , Antibiose , Cátions Bivalentes , Meios de Cultura/química , Grão Comestível/microbiologia , Fabaceae/química , Fusarium/crescimento & desenvolvimento , Tamanho da PartículaRESUMO
Pseudomonas chlororaphis O6 possesses many beneficial traits involved in biocontrol of plant diseases. In this paper, we examined the effect of a mutation in rpoS encoding a stress-related alternative sigma factor to better understand the regulation of these traits. Biochemical studies indicated that production of acyl homoserine lactones was altered and phenazine was increased in the P. chlororaphis O6 rpoS mutant. The rpoS mutation reduced hydrogen cyanide levels, but the rpoS mutant still displayed a level of in vitro antifungal activity against Fusarium graminearum and Alternaria alternata. Tomato root colonization by the rpoS mutant was lower than that by the wild type at 5, 7, and 13 days after inoculation. The rpoS mutant was less effective than the wild type in induction of systemic resistance to two foliar pathogens after root inoculation of the tomato plants. Our findings demonstrate that the stationary-phase sigma factor RpoS regulates production of several key factors involved in the biocontrol potential of P. chlororaphis O6, some independently of the global regulator GacS.
Assuntos
Proteínas de Bactérias/metabolismo , Pseudomonas/genética , Pseudomonas/metabolismo , Fator sigma/metabolismo , Acil-Butirolactonas/metabolismo , Antifúngicos/metabolismo , Proteínas de Bactérias/genética , Fusarium/crescimento & desenvolvimento , Cianeto de Hidrogênio/metabolismo , Solanum lycopersicum/microbiologia , Interações Microbianas , Mutação/genética , Fenazinas/metabolismo , Raízes de Plantas/microbiologia , Fator sigma/genética , Fatores de Transcrição/metabolismoRESUMO
Mycobacterium sp. strain KMS was isolated from soils where remediation of polycyclic aromatic hydrocarbons was active. This isolate is a competent plant root colonizer through utilization of an array of carbon substrates available in the root exudates. Bioinformatic analyses based on the KMS genome propose pathways for C4- and C3-intermediate conversions during growth of the isolate on substrates requiring gluconeogenesis. Expression of candidate genes for these pathways was compared using semi-quantitative RT-PCR from cells grown on acetate, succinate, benzoate, or pyrene as sole carbon sources requiring gluconeogenesis during growth. Expression was examined for cells grown on fructose and mannitol, where gluconeogenesis would not be essential. Transcript accumulation in cells grown on all the carbon sources confirmed expression from genes involved in the glyoxylate shunt and a gene encoding a novel enzyme to complete the tricarboxylic acid cycle, a membrane-associated malate:quinone oxidoreductase (MQO). Transcript accumulations for genes encoding phosphoenolpyruvate carboxykinase, malic enzyme, and phosphoenolpyruvate synthase were weak for mannitol growth but were detected for the other carbon sources. Activities for PEP synthase and the membrane-associated MQO were confirmed in cell extracts at different levels indicating feasibility of their function in production of PEP for gluconeogenesis in this soil Mycobacterium.
Assuntos
Gluconeogênese , Redes e Vias Metabólicas/genética , Mycobacterium/genética , Mycobacterium/metabolismo , Microbiologia do Solo , Biodegradação Ambiental , Meios de Cultura/química , Perfilação da Expressão Gênica , Mycobacterium/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo RealRESUMO
The soil isolate, Mycobacterium sp. strain KMS, utilizes an array of carbon compounds including the aromatics benzoate and pyrene as sole carbon sources. Growth on pyrene induced both chromosomal and plasmid nidA genes encoding pyrene ring-hydroxylating dioxygenase α-subunits for pyrene oxidation. Diauxic growth occurred when KMS was cultured with pyrene plus either acetate, succinate, fructose, or benzoate and nidA expression only was detected in the second slower log-phase period. Potential cAMP-CRP binding sites exist within the promoter region of both nidA genes indicating that cAMP-CRP may be involved in catabolite repression of pyrene utilization. When cultured with benzoate plus either acetate, succinate, or fructose, there was no diauxic growth. Also there was no diauxic growth on fructose plus succinate or acetate. Expression of a benA gene, encoding a benzoate dioxygenase α-subunit involved in the initiation of benzoate oxidation, was detected in log-phase cells from the benzoate-mixed substrate cultures at the same level as when the cells were cultured on benzoate alone. These findings suggested that catabolite repression of pyrene but not benzoate occurred in isolate KMS. These differences may help the microbe exploit the varied carbon sources available in the soil and rhizosphere environments.
Assuntos
Benzoatos/metabolismo , Mycobacterium/metabolismo , Mycobacterium/fisiologia , Pirenos/metabolismo , Acetatos/metabolismo , Adenilil Ciclases/genética , Adenilil Ciclases/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Sequência de Bases , Sítios de Ligação , Repressão Catabólica , AMP Cíclico/metabolismo , Proteína Receptora de AMP Cíclico/metabolismo , Dioxigenases/genética , Dioxigenases/metabolismo , Frutose/metabolismo , Regulação Bacteriana da Expressão Gênica , Genes Bacterianos/genética , Dados de Sequência Molecular , Mycobacterium/enzimologia , Mycobacterium/genética , Microbiologia do Solo , Ácido Succínico/metabolismoRESUMO
The essential metals Cu, Zn, and Fe are involved in many activities required for normal and stress responses in plants and their microbiomes. This paper focuses on how drought and microbial root colonization influence shoot and rhizosphere metabolites with metal-chelation properties. Wheat seedlings, with and without a pseudomonad microbiome, were grown with normal watering or under water-deficit conditions. At harvest, metal-chelating metabolites (amino acids, low molecular weight organic acids (LMWOAs), phenolic acids, and the wheat siderophore) were assessed in shoots and rhizosphere solutions. Shoots accumulated amino acids with drought, but metabolites changed little due to microbial colonization, whereas the active microbiome generally reduced the metabolites in the rhizosphere solutions, a possible factor in the biocontrol of pathogen growth. Geochemical modeling with the rhizosphere metabolites predicted Fe formed Fe-Ca-gluconates, Zn was mainly present as ions, and Cu was chelated with the siderophore 2'-deoxymugineic acid, LMWOAs, and amino acids. Thus, changes in shoot and rhizosphere metabolites caused by drought and microbial root colonization have potential impacts on plant vigor and metal bioavailability.
RESUMO
The beneficial bacterium Pseudomonas chlororaphis O6 produces indole-3-acetic acid (IAA), a plant growth regulator. However, the pathway involved in IAA production in this bacterium has not been reported. In this paper we describe the involvement of the indole-3-acetamide (IAM) pathway in IAA production in P. chlororaphis O6 and the effects of CuO and ZnO nanoparticles (NPs). Sublethal levels of CuO and ZnO NPs differentially affected the levels of IAA secreted in medium containing tryptophan as the precursor. After 15 h of growth, CuO NP-exposed cells had metabolized more tryptophan than the control and ZnO NP-challenged cells. The CuO NP-treated cells produced higher IAA levels than control cultures lacking NPs. In contrast, ZnO NPs inhibited IAA production. Mixing of CuO and ZnO NPs resulted in an intermediate level of IAA production relative to the levels in the separate CuO and ZnO NP treatments. The effect of CuO NPs on IAA levels could be duplicated by ions at the concentrations released from the NPs. However, ion release did not account for the inhibition caused by the ZnO NPs. The mechanism underlying changes in IAA levels cannot be accounted for by effects on transcript accumulation from genes encoding a tryptophan permease or the IAM hydrolase in 15-h cultures. These findings raise the issue of whether sublethal doses of NPs would modify the beneficial effects of association between plants and bacteria.
Assuntos
Antibacterianos/metabolismo , Cobre/metabolismo , Ácidos Indolacéticos/metabolismo , Nanopartículas , Pseudomonas/efeitos dos fármacos , Óxido de Zinco/metabolismo , Meios de Cultura/química , Plantas/microbiologia , Pseudomonas/metabolismo , Triptofano/metabolismoRESUMO
CuO nanoparticles (NPs) exhibit dose-dependent toxicity to bacteria, whereas sublethal concentrations of these NPs change bacterial metabolism. Siderophores are model metabolites to study the impact of sublethal levels of metallic NPs on bacteria because they are involved in survival and interaction with other organisms and with metals. We report that a sublethal level of CuO NPs modify the production of the fluorescent siderophore pyoverdine (PVD) in a soil beneficial bacterium, Pseudomonas chlororaphis O6. The production of PVD was inhibited by CuO NPs but not by bulk CuO nor Cu ions at concentrations equivalent to those released from the NPs. The cell responses occurred despite the NPs forming near micrometer-sized aggregates. The CuO NPs reduced levels of periplasmic and secreted PVD and impaired expression from genes encoding proteins involved in PVD maturation in the periplasm and export through cell membranes. EDTA restored the fluorescence of PVD quenched by Cu ions but did not generate fluorescence with cultures of NP-challenged cells, confirming the absence of PVD. Consequently, depending on the bacterium, this nanoparticle-specific phenomenon mediating cellular reprogramming through effects on secondary metabolism could have an impact on critical environmental processes including bacterial pathogenicity.
Assuntos
Cobre/toxicidade , Nanopartículas/toxicidade , Oligopeptídeos/metabolismo , Pseudomonas/efeitos dos fármacos , Pseudomonas/metabolismo , Sideróforos/metabolismo , Cobre/química , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Nanopartículas/química , Nanopartículas/ultraestrutura , Oligopeptídeos/genética , Pseudomonas/genética , Sideróforos/genética , Microbiologia do SoloRESUMO
Mycobacterium sp. strain KMS utilizes pyrene, a high-molecular weight polycyclic aromatic hydrocarbon (PAH), as a sole carbon source. Bioinformatic analysis of the genome of isolate KMS predicted 25 genes with the potential to encode 17 pyrene-induced proteins identified by proteomics; these genes were clustered on both the chromosome and a circular plasmid. RT-PCR analysis of total RNA isolated from KMS cells grown with or without pyrene showed that the presence of pyrene increased the transcript accumulation of 20 of the predicted chromosome- and plasmid-located genes encoding pyrene-induced proteins. The transcribed genes from both the chromosome and a circular plasmid were within larger regions containing genes required for PAH degradation constituting PAH-degrading gene islands. Genes encoding integrases and transposases were found within and outside the PAH-degrading gene islands. The lower GC content of the genes within the gene island (61%-64%) compared with the average genome content (68%) suggested that these mycobacteria initially acquired these genes by horizontal gene transfer. Synteny was detected for the PAH-degrading islands in the genomes of two additional Mycobacterium isolates from the same PAH-polluted site and of two other pyrene-degrading Mycobacterium from different sites in the United States of America. Consequently, the gene islands have been conserved from a common ancestral strain.
Assuntos
Genes Bacterianos/genética , Ilhas Genômicas/genética , Mycobacterium/genética , Mycobacterium/metabolismo , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Pirenos/metabolismo , Biodegradação Ambiental , Cromossomos Bacterianos/genética , Duplicação Gênica , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Família Multigênica/genética , Mycobacterium/isolamento & purificação , Plasmídeos/genética , Pirenos/farmacologia , Homologia de Sequência do Ácido Nucleico , Sintenia , Estados Unidos , Poluentes Químicos da Água/farmacologiaRESUMO
Mycobacterium sp. strain KMS has bioremediation potential for polycyclic aromatic hydrocarbons (PAHs), such as pyrene, and smaller ring aromatics, such as benzoate. Degradation of these aromatics involves oxidation catalyzed by aromatic ring-hydroxylating dioxygenases. Multiple genes encoding dioxygenases exist in KMS: ten genes encode large-subunits with homology to phenylpropionate dioxygenase genes, sixteen pairs of adjacent genes encode alpha- and beta-subunits of dioxygenase and two genes encode beta-subunits. These genes include orthologs of nid genes essential for degradation of multi-ring PAHs in M. vanbaalenii isolate PYR-1. The multiplicity of genes in part is explained by block duplication that results in two or three copies of certain genes on the chromosome, a linear plasmid, and a circular plasmid within the KMS genome. Quantitative real-time PCR showed that four dioxygenase beta-subunit nid genes from operons with almost identical promoter sequences otherwise unique in the genome were induced by pyrene to similar extents. No induction occurred with benzoate. Unlike isolate PYR-1, isolate KMS has an operon specifying benzoate catabolism and the expression of the alpha-subunit dioxygenase gene was activated by benzoate but not pyrene. These studies showed that isolate KMS had a genome well adapted to utilization of different aromatic compounds.
Assuntos
Proteínas de Bactérias/genética , Dioxigenases/genética , Regulação Enzimológica da Expressão Gênica , Família Multigênica , Mycobacterium/enzimologia , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Sequência de Bases , Dioxigenases/química , Dioxigenases/metabolismo , Dados de Sequência Molecular , Mycobacterium/classificação , Mycobacterium/genética , Mycobacterium/metabolismo , Filogenia , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Regiões Promotoras Genéticas , Subunidades Proteicas/química , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismoRESUMO
Biological control is an important process for sustainable plant production, and this trait is found in many plant-associated microbes. This study reviews microbes that could be formulated into pesticides active against various microbial plant pathogens as well as damaging insects or nematodes. The focus is on the beneficial microbes that colonize the rhizosphere where, through various mechanisms, they promote healthy plant growth. Although these microbes have adapted to cohabit root tissues without causing disease, they are pathogenic to plant pathogens, including microbes, insects, and nematodes. The cocktail of metabolites released from the beneficial strains inhibits the growth of certain bacterial and fungal plant pathogens and participates in insect and nematode toxicity. There is a reinforcement of plant health through the systemic induction of defenses against pathogen attack and abiotic stress in the plant; metabolites in the beneficial microbial cocktail function in triggering the plant defenses. The review discusses a wide range of metabolites involved in plant protection through biocontrol in the rhizosphere. The focus is on the beneficial firmicutes and pseudomonads, because of the extensive studies with these isolates. The review evaluates how culture conditions can be optimized to provide formulations containing the preformed active metabolites for rapid control, with or without viable microbial cells as plant inocula, to boost plant productivity in field situations.
RESUMO
A plethora of compounds stimulate protective mechanisms in plants against microbial pathogens and abiotic stresses. Some defense activators are synthetic compounds and trigger responses only in certain protective pathways, such as activation of defenses under regulation by the plant regulator, salicylic acid (SA). This review discusses the potential of naturally occurring plant metabolites as primers for defense responses in the plant. The production of the metabolites, hexanoic acid and melatonin, in plants means they are consumed when plants are eaten as foods. Both metabolites prime stronger and more rapid activation of plant defense upon subsequent stress. Because these metabolites trigger protective measures in the plant they can be considered as "vaccines" to promote plant vigor. Hexanoic acid and melatonin instigate systemic changes in plant metabolism associated with both of the major defense pathways, those regulated by SA- and jasmonic acid (JA). These two pathways are well studied because of their induction by different microbial triggers: necrosis-causing microbial pathogens induce the SA pathway whereas colonization by beneficial microbes stimulates the JA pathway. The plant's responses to the two metabolites, however, are not identical with a major difference being a characterized growth response with melatonin but not hexanoic acid. As primers for plant defense, hexanoic acid and melatonin have the potential to be successfully integrated into vaccination-like strategies to protect plants against diseases and abiotic stresses that do not involve man-made chemicals.