RESUMO
The regulation of emotions is a crucial facet of well-being and social adaptability, with explicit strategies receiving primary attention in prior research. Recent studies, however, emphasize the role of implicit emotion regulation, particularly implicating the ventromedial prefrontal cortex (VMPFC) in association with its implementation. This study delves into the nuanced role of the VMPFC through focality-optimized multichannel transcranial direct current stimulation (tDCS), shedding light on its causal involvement in implicit reappraisal. The primary goal was to evaluate the effectiveness of VMFPC-targeted tDCS and elucidate its role in individuals with high trait anxiety. Participants engaged in implicit and explicit emotion regulation tasks during multichannel tDCS targeting the VMPFC. The outcome measures encompassed negative emotion ratings, pupillary diameter, and saccade count, providing a comprehensive evaluation of emotion regulation efficiency. The intervention exhibited a notable impact, resulting in significant reductions in negative emotion ratings and pupillary reactions during implicit reappraisal, highlighting the indispensable role of the VMPFC in modulating emotional responses. Notably, these effects demonstrated sustained efficacy up to 1 day postintervention. This study underscores the potency of VMPFC-targeted multichannel tDCS in augmenting implicit emotion regulation. This not only contributes insights into the neural mechanisms of emotion regulation but also suggests innovative therapeutic avenues for anxiety disorders. The findings present a promising trajectory for future mood disorder interventions, bridging the gap between implicit emotion regulation and neural stimulation techniques.
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Regulação Emocional , Córtex Pré-Frontal , Estimulação Transcraniana por Corrente Contínua , Humanos , Córtex Pré-Frontal/fisiologia , Córtex Pré-Frontal/diagnóstico por imagem , Regulação Emocional/fisiologia , Masculino , Feminino , Adulto , Adulto Jovem , Ansiedade/fisiopatologia , Ansiedade/terapia , Movimentos Sacádicos/fisiologia , Emoções/fisiologiaRESUMO
Fusarium wilt of Momordica charantia in the greenhouse is one of the most severe crop diseases in Shandong Province, P.R. China. This study aimed to investigate the mechanisms of accumulation and long-term survival of the pathogen in naturally pathogenic soils. Soil physicochemical properties were tested after applying a highly virulent strain of Fusarium wilt to M. charantia in an artificial disease nursery. The functional structure of soil microorganisms was analyzed through amplicon sequencing. The highly virulent strain SG-15 of F. oxysporum f. sp. momordicae was found to cause Fusarium wilt in M. charantia in Shandong Province. The strain SG-15 could not infect 14 non-host crops, including Solanum melongena and Lycopersicon esculentum, but it had varying degrees of pathogenicity towards 11 M. charantia varieties. In the artificial disease nursery for Fusarium wilt of M. charantia, the F. oxysporum was distributed in the soil to a depth of 0-40 cm and was mainly distributed in crop residues at 0-10 cm depth. During crop growth, F. oxysporum primarily grows and reproduces in susceptible host plants, rather than disease-resistant hosts and non-host crops. The colonization of the pathogen of Fusarium wilt significantly changed the soil physicochemical properties, the functional structure of soil microorganisms and the circulation of soil elements such as carbon, nitrogen, phosphorus and sulfur. Soil pH value, organic matter content, available iron content, available manganese content, FDA hydrolase activity and polyphenol oxidase activity were significantly correlated with the relative abundance of Fusarium wilt pathogens in the soil. In general, this study suggests that susceptible host plants facilitate the accumulation of Fusarium wilt pathogens in the soil. These pathogens can mediate the decomposition process of plant residues, particularly those of diseased plants, and indirectly or directly affect soil's chemical properties.
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Evidence suggests that explicit reappraisal has limited regulatory effects on high-intensity emotions, mainly due to the depletion of cognitive resources occupied by the high-intensity emotional stimulus itself. The implicit form of reappraisal has proved to be resource-saving and therefore might be an ideal strategy to achieve the desired regulatory effect in high-intensity situations. In this study, we explored the regulatory effect of explicit and implicit reappraisal when participants encountered low- and high-intensity negative images. The subjective emotional rating indicated that both explicit and implicit reappraisal down-regulated negative experiences, irrespective of intensity. However, the amplitude of the parietal late positive potential (LPP; a neural index of experienced emotional intensity) showed that only implicit reappraisal had significant regulatory effects in the high-intensity context, though both explicit and implicit reappraisal successfully reduced the emotional neural responses elicited by low-intensity negative images. Meanwhile, implicit reappraisal led to a smaller frontal LPP amplitude (an index of cognitive cost) compared to explicit reappraisal, indicating that the implementation of implicit reappraisal consumes limited cognitive control resources. Furthermore, we found a prolonged effect of implicit emotion regulation introduced by training procedures. Taken together, these findings not only reveal that implicit reappraisal is suitable to relieve high-intensity negative experiences as well as neural responses, but also highlight the potential benefit of trained implicit regulation in clinical populations whose frontal control resources are limited.
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Regulação Emocional , Humanos , Eletroencefalografia , Potenciais Evocados/fisiologia , Cognição/fisiologia , Emoções/fisiologiaRESUMO
BACKGROUND: While implicit distraction could ameliorate negative feelings in patients with major depressive disorders (MDD), it remains unclear whether patients could benefit from explicit, voluntary distraction. Meanwhile, though the dorsolateral prefrontal cortex (DLPFC) is established as a crucial brain region involved in attentional control, the causal relationship between the DLPFC and voluntary distraction is unexplored in patients. METHODS: Combing explicit distraction and transcranial magnetic stimulation (TMS), this study investigated whether TMS-activated DLPFC facilitates voluntary distraction in MDD patients. Eighty patients diagnosed with current MDD underwent either active (n = 40) or sham (n = 40) TMS sessions, followed by receiving negative social feedback from other patients, during which they were requied to use distraction strategy to down-regulate their painful feelings. Electroencephalogram was recorded during the task. RESULTS: Both the subjective emotional rating and the amplitude of late positive potential showed that depressed patients successfully down-regulate their negative emotions via voluntary distraction, and the TMS-activated left DLPFC produced a larger benefit of emotion regulation compared to the sham TMS group. Results also revealed that while emotion regulation effect was negatively associated with depressive symptoms in the sham TMS group, this correlation was largely diminished when patients' left DLPFC was activated by TMS during the voluntary distraction. CONCLUSIONS: These findings demonstrated that distraction is valuable for emotion regulation in MDD patients and they could be beneficial in voluntary distraction by activating their left DLPFC using neural modulation techniques. This study has valuable implications for clinical treatement of emotional dysregulation in MDD patients.
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Major depressive disorder (MDD) is characterized by emotion dysregulation. Whether implicit emotion regulation can compensate for this deficit remains unknown. In this study, we recruited 159 subjects who were healthy controls, had subclinical depression, or had MDD, and examined them under baseline, implicit, and explicit reappraisal conditions. Explicit reappraisal led to the most negative feelings and the largest parietal late positive potential (parietal LPP, an index of emotion intensity) in the MDD group compared to the other two groups; the group difference was absent under the other two conditions. MDD patients showed larger regulatory effects in the LPP during implicit than explicit reappraisal, whereas healthy controls showed a reversed pattern. Furthermore, the frontal P3, an index of voluntary cognitive control, showed larger amplitudes in explicit reappraisal compared to baseline in the healthy and subclinical groups, but not in the MDD group, while implicit reappraisal did not increase P3 across groups. These findings suggest that implicit reappraisal is beneficial for clinical depression.
Assuntos
Transtorno Depressivo Maior , Regulação Emocional , Humanos , Transtorno Depressivo Maior/psicologia , Depressão , Emoções/fisiologia , Cognição/fisiologiaRESUMO
Endophytic fungi are effective in plant growth and development by secreting various kinds of plant hormones and nutrients. However, the cellular and molecular interactions between the endophytic fungi and plant growth-promoting have remained less explored. The present study was designed to explore the effects of the infection and colonization events of Chaetomium globosum strain ND35 on cucumber growth and the expression pattern of some metabolically important genes in development of the cucumber radicle. The results demonstrated that strain ND35 can infect and colonize the outer layers (cortical cells) of cucumber root and form a symbiotic structure with the host cell, similar to a periarbuscular membrane and establish chemical communication with the plant. Through transcriptome analysis, we found the differentially expressed genes (DEGs) caused by strain ND35 were mainly enriched in phenylpropanoid biosynthesis, plant hormone signal transduction, plant-pathogen interaction and photosynthesis. Correspondingly, the contents of reactive oxygen species (ROS), hydrogen peroxide (H2O2), indole-3-acetic acid (IAA), gibberellin (GA), zeatin (ZT), salicylic acid (SA), jasmonic acid (JA) and the activity of phenylalanine ammonia lyase (PAL), 4-coumarate-CoA ligase (4CL), cinnamyl alcohol dehydrogenase (CAD), and peroxidase (POD) in ND35-colonized seedlings were generally higher than those of non-inoculated seedlings. Overall, the infection and colonization events of C. globosum strain ND35 increased cucumber growth through complex regulation of plant hormones biosynthesis and metabolism. Furthermore, although the endophytic fungus strain ND35 produced IAA, GA, ZT, and ergosterol in the fermentation broth, and there are enabled to promote growth of cucumber, it is uncertain whether there are ND35-derived microbial hormones in plants. This study of the interaction between cucumber and strain ND35 contributes to a better understanding of the plant-endophytic fungi interactions, and may help to develop new strategies for crop production.
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Fusarium wilt is an important disease of many food crops and often causes serious damages to yield and food quality. Consequently, numerous studies mainly focused on exploring the control strategy for Fusarium oxysporum as well as the mechanism of interaction between the F. oxysporum and other beneficial soil microorganisms. In this study, we have screened and identified an efficient biocontrol strain from the soil with infection of F. oxysporum f. sp. momordica (referred to as Fom), Talaromyces purpurogenus Q2 (referred to as TpQ2), which could be effective to reduce relative abundance of the rhizospheric Fom, leading to a significant decrease of Fusarium wilt disease incidence in bitter gourd during the greenhouse and field trails. TpQ2 can reduce the relative abundance of rhizospheric Fom through inhibition of growth and development of Fom. During the co-cultivation of TpQ2 and Fom, we confirmed that TpQ2 could significantly suppress the growth and development of Fom through disturbing the normal hyphae shape and function of the cell walls of Fom via secreting cell wall-degrading enzymes and suppression of the expression of cell wall biosynthesis genes, such as FomCFEM. In the meantime, TpQ2 showed a strong negative correlation with F. oxysporum in soil and positive correlation with beneficial indigenous microorganisms that had significant negative correlation with Fusarium populations, such as Streptomycetes, Lysobacter, and Sphingobium. To summarize, TpQ2 has a good biocontrol efficacy on Fusarium wilt of bitter gourd. The biocontrol mechanisms of TpQ2 on Fusarium wilt are complex and diverse.
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Socially engaging robots have been increasingly applied to alleviate depressive symptoms and to improve the quality of social life among different populations. Seeing that depression negatively influences social reward processing in everyday interaction, we investigate this influence during simulated interactions with humans or robots. In this study, 35 participants with mild depression and 35 controls (all from nonclinical populations) finished the social incentive delay task with event-related potential recording, in which they received performance feedback from other persons or from a robot. Compared to the controls, the mild depressive symptom (MDS) group represented abnormalities of social reward processing in the human feedback condition: first, the MDS group showed a lower hit rate and a smaller contingent-negative variation (correlated with each other) during reward anticipation; second, depression level modulated both the early phase (indexed by the feedback-related negativity (FRN)) and the late phase (indexed by the P3) of reward consumption. In contrast, the effect of depression was evident only on FRN amplitude in the robot feedback condition. We suggest that compared to human-human interaction, the rewarding properties of human-robot interaction are less likely to be affected by depression. These findings have implications for the utilization of robot-assisted intervention in clinical practice.
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Robótica , Depressão , Eletroencefalografia , Potenciais Evocados , Humanos , RecompensaRESUMO
Talaromyces purpurogenus strain Q2 is a kind of beneficial microbe originally separated from the rhizosphere of healthy cucumber. In this study, we evaluated the biocontrol potential of strain Q2 against four soil-borne diseases by plate confrontation culture, potting in the greenhouse. We further estimated the control efficacy of strain Q2 combined metam-sodium fumigation against Fusarium wilt of bitter gourd in the field. The mechanism of strain Q2 controlling bitter gourd wilt and regulating soil microbial community was examined by plate dilution culture, high throughput sequencing and quantitative PCR. The results showed that strain Q2 could efficiently reduce disease incidence of Fusarium wilt of bitter gourd, potato stem canker, black shank of tobacco and black root rot of tobacco in the green house. Its biocontrol efficacy on black shank of tobacco and bitter gourd wilt was 75.3% and 63.4%, respectively. Biocontrol efficacy of strain Q2 on bitter gourd wilt was 51.0% in the artificial disease nursery inoculated pathogen of bitter gourd wilt, while the control efficacy of strain Q2 combined with soil fumigation technology was more than 80% in the same experiment condition. Strain Q2 application and soil fumigation altered soil microbial community composition and recovery trend. Metam-sodium fumigation significantly reduced the abundances of Fusarium oxysporum and causal agent of bitter gourd wilt. Strain Q2 further suppressed the efficient recovery trend of the pathogen. After application of strain Q2, Penicillium was enriched in soil, as well as the beneficial microbes involved in the suppression of F. oxysporum, such as Bacillus and Gaiella. Overall, after soil fumigation, biocontrol efficacy of strain Q2 on soil-borne diseases such as Fusarium wilt could attribute to the formation of beneficial microbial communities in soil and inhibition of strain Q2 on growth and development of F. oxysporum.
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Fusarium , Microbiota , Doenças das Plantas , Solo , Microbiologia do Solo , TalaromycesRESUMO
Fusarium oxysporum f. sp. momordica was used as the target pathogenic fungus to screen actinomycetes that were isolated from rhizosphere soil of Momordica charantia by confrontation culture and antifungal tests of fermentation filtrate. The candidate strain 0250 had broad antifungal activity. According to cultural characteristics, physiological and biochemical properties, as well as average nucleotide identity analysis of the strains with similar homology, the strain 0250 was identified as Streptomyces rhizosphaericus. Its effects on growth promotion and control of bitter gourd wilt were evaluated in both greenhouse and field. The results showed that the plate inhibition percentage of S. rhizosphaericus strain 0250 against F. oxysporum f. sp. momordica was 69.2%, while the plate inhibition percentage against 17 plant pathogenic fungi reached 64.3%-85.6%. The suspension treatment of the strain could promote the growth and development of roots and stems and improve production of bitter gourd in pots and field. The control efficacy of Fusarium wilt of bitter gourd was 66.9% and 61.5%, respectively. When soils were treated with the strain 0250 suspension in advance and inoculated with the fungal pathogen, the inhibition percentage on the soil F. oxysporum reached 62.1%. The activity of phenylalanine ammonia-lyase, peroxidase and ß-1,3-glucanase as well as root activity were significantly improved in bitter gourd seedlings. In summary, strain 0250 is an actinomycetes resource with biocontrol potential to Fusarium wilt of bitter gourd.
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Actinobacteria , Fusarium , Momordica charantia , Actinomyces , Programas de Rastreamento , Doenças das Plantas/prevenção & controle , StreptomycesRESUMO
The antibiotic pyrrolnitrin (PRN) is a tryptophan-derived secondary metabolite that plays an important role in the biocontrol of plant diseases due to its broad-spectrum of antimicrobial activities. The PRN biosynthetic gene cluster remains to be characterised in Serratia plymuthica, though it is highly conserved in PRN-producing bacteria. To better understand PRN biosynthesis and its regulation in Serratia, the prnABCD operon from S. plymuthica G3 was cloned, sequenced and expressed in Escherichia coli DH5α. Furthermore, an engineered strain prnind which is a conditional mutant of G3 prnABCD under the control of the Ptac promoter was constructed. This mutant was able to overproduce PRN with isopropylthiogalactoside (IPTG) induction by overexpressing prnABCD, whilst behaving as a conditional mutant of G3 prnABCD in the absence of IPTG. These results confirmed that prnABCD is responsible for PRN biosynthesis in strain G3. Further experiments involving lux-/dsRed-based promoter fusions, combined with site-directed mutagenesis of the putative σS extended -10 region in the prnA promoter, and liquid chromatography-mass spectrometry (LC-MS) analysis extended our previous knowledge about G3, revealing that quorum sensing (QS) regulates PRN biosynthesis through cross talk with RpoS, which may directly activated prnABCD transcription. These findings suggest that PRN in S. plymuthica G3 is produced in a tightly controlled manner, and has diverse functions, such as modulation of cell motility, in addition to antimicrobial activities. Meanwhile, the construction of inducible mutants could be a powerful tool to improve PRN production, beyond its potential use for the investigation of the biological function of PRN.
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Regulação Bacteriana da Expressão Gênica , Óperon/genética , Pirrolnitrina/biossíntese , Serratia/genética , Mutação , Percepção de Quorum/fisiologiaRESUMO
A strain LN07 with high laccase yield was identified as basidiomycete fungus Lepista nuda from which a white laccase without type I copper was purified and characterized. The laccase was a monomeric protein with a molecular mass of 56 kDa. Its N-terminal amino acid sequence was AIGPAADLHIVNKDISPDGF. Besides, eight inner peptide sequences were determined and lac4, lac5 and lac6 sequences were in the Cu(2+) combination and conservation zones of laccases. HIV-1 reverse transcriptase was inhibited by the laccase with a half-inhibitory concentration of 0.65 µM. Cu(2+) ions (1.5 mM) enhanced the laccase production and the optimal pH and temperature of the laccase were pH 3.0 and 50 °C, respectively. The Km and Vmax of the laccase using ABTS as substrate were respectively 0.19 mM and 195 µM. Several dyes including laboratory dyes and textile dyes used in this study, such as Methyl red, Coomassie brilliant blue, Reactive brilliant blue and so on, were decolorized in different degrees by the purified laccase. By LC-MS analysis, Methyl red was structurally degraded by the laccase. Moreover, the laccase affected the absorbance at the maximum wavelength of many pesticides. Thus, the white laccase had potential commercial value for textile finishing and wastewater treatment.
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Agaricales/enzimologia , Corantes/química , Lacase/química , Cor , Cobre/química , Transcriptase Reversa do HIV/antagonistas & inibidores , Transcriptase Reversa do HIV/química , HIV-1/efeitos dos fármacos , HIV-1/patogenicidade , Humanos , Lacase/isolamento & purificação , Lacase/farmacologia , Peso Molecular , Especificidade por Substrato , Águas Residuárias/químicaRESUMO
The σ(S) subunit RpoS of RNA polymerase functions as a master regulator of the general stress response in Escherichia coli and related bacteria. RpoS has been reported to modulate biocontrol properties in the rhizobacterium Serratia plymuthica IC1270. However, the role of RpoS in the stress response and biofilm formation in S. plymuthica remains largely unknown. Here we studied the role of RpoS from an endophytic S. plymuthica G3 in regulating these phenotypes. Mutational analysis demonstrated that RpoS positively regulates the global stress response to acid or alkaline stresses, oxidative stress, hyperosmolarity, heat shock and carbon starvation, in addition to proteolytic and chitinolytic activities. Interestingly, rpoS mutations resulted in significantly enhanced swimming motility, biofilm formation and production of the plant auxin indole-3-acetic acid (IAA), which may contribute to competitive colonization and environmental fitness for survival. These findings provide further insight into the strain-specific role of RpoS in the endophytic strain G3 of S. plymuthica, where it confers resistance to general stresses encountered within the plant environment. The heterogeneous functionality of RpoS in rhizosphere and endophytic S. plymuthica populations may provide a selective advantage for better adaptation to various physiological and environmental stresses.
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Proteínas de Bactérias/metabolismo , Biofilmes/crescimento & desenvolvimento , Endófitos/enzimologia , Endófitos/fisiologia , Serratia/enzimologia , Serratia/fisiologia , Fator sigma/metabolismo , Estresse Fisiológico , Proteínas de Bactérias/genética , Análise Mutacional de DNA , Endófitos/genética , Locomoção , Reguladores de Crescimento de Plantas/metabolismo , Serratia/genética , Fator sigma/genéticaRESUMO
The GacS/GacA two-component system plays a central role in the regulation of a broad range of biological functions in many bacteria. In the biocontrol organism Pseudomonas chlororaphis, the Gac system has been shown to positively control quorum sensing, biofilm formation, and phenazine production, but has an overall negative impact on motility. These studies have been performed with strains originated from the rhizosphere predominantly. To investigate the level of conservation between the GacA regulation of biocontrol-related traits in P. chlororaphis isolates from different habitats, the studies presented here focused on the endophytic isolate G5 of P. chlororaphis subsp. aurantiaca. A gacA mutant deficient in the production of N-acylhomoserine lactones (AHLs) and phenazine was isolated through transposon mutagenesis. Further phenotypic characterization revealed that in strain G5, similar to other P. chlororaphis strains, a gacA mutation caused inability to produce biocontrol factors such as phenazine, HCN and proteases responsible for antifungal activity, but overproduced siderophores. LC-MS/MS analysis revealed that AHL production was also practically abolished in this mutant. However, the wild type exhibited an extremely diverse AHL pattern which has never been identified in P. chlororaphis. In contrast to other isolates of this organism, GacA in strain G5 was shown to negatively regulate biofilm formation and oxidative stress response whilst positively regulating cell motility and biosynthesis of indole-3-acetic acid (IAA). To gain a better understanding of the overall impact of GacA in G5, a comparative proteomic analysis was performed revealing that, in addition to some of the traits like phenazine mentioned above, GacA also negatively regulated lipopolysaccharide (LPS) and trehalose biosynthesis whilst having a positive impact on energy metabolism, an effect not previously described in P. chlororaphis. Consequently, GacA regulation shows a differential strain dependency which is likely to be in line with their niche of origin.
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Acil-Butirolactonas/metabolismo , Proteínas de Bactérias/genética , Regulação Bacteriana da Expressão Gênica/genética , Fenazinas/metabolismo , Pseudomonas/genética , Antifúngicos/metabolismo , Biofilmes/crescimento & desenvolvimento , Movimento Celular/genética , Metabolismo Energético/genética , Cianeto de Hidrogênio/metabolismo , Ácidos Indolacéticos/metabolismo , Lipopolissacarídeos/biossíntese , Estresse Oxidativo/genética , Proteômica , Pseudomonas/classificação , Pseudomonas/metabolismo , Percepção de Quorum/genética , Espectrometria de Massas em Tandem , Trealose/biossínteseRESUMO
The RNA-binding protein Hfq has been well studied as a global post-transcriptional regulator which controls diverse cellular processes in bacteria. However, the function in the genus of Serratia has remained unexplored. Here we show that beyond mutation in Hfq resulting in their growth defects, Hfq has global effects on a variety of biocontrol-related phenotypes in the endophytic strain G3 of Serratia plymuthica, including antifungal activity, production of exoenzymes, as well as motility and biofilm formation. Especially for the first time, Hfq is observed to control biosynthesis of auxin indole-3-acetic acid (IAA) and antibiotic pyrrolnitrin (PRN), which are key determinants responsible for plant growth promotion and suppression of phytopathogens, respectively by G3. Additionally, Hfq is also required for the production of RpoS, a major stress sigma factor in G3. In contrast to E. coli, translation of hfq in G3 is positively autoregulated. Further investigation of the molecular mechanisms involved in regulation of IAA and pyrrolnitrin production by Hfq and its role in the regulatory networks of G3 will help to optimize the beneficial plant-microbe interactions.
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Proteínas de Bactérias/metabolismo , Fator Proteico 1 do Hospedeiro/metabolismo , Serratia/metabolismo , Proteínas de Bactérias/genética , Clonagem Molecular , Fator Proteico 1 do Hospedeiro/genética , FilogeniaRESUMO
BACKGROUND: N-acylhomoserine lactone (AHL)-based quorum sensing (QS) systems have been described in many plant-associated Gram-negative bacteria to control certain beneficial phenotypic traits, such as production of biocontrol factors and plant growth promotion. However, the role of AHL-mediated signalling in the endophytic strains of plant-associated Serratia is still poorly understood. An endophytic Serratia sp. G3 with biocontrol potential and high levels of AHL signal production was isolated from the stems of wheat and the role of QS in this isolate was determined. RESULTS: Strain G3 classified as Serratia plymuthica based on 16S rRNA was subjected to phylogenetic analysis. Using primers to conserved sequences of luxIR homologues from the Serratia genus, splIR and spsIR from the chromosome of strain G3 were cloned and sequenced. AHL profiles from strain G3 and Escherichia coli DH5α expressing splI or spsI from recombinant plasmids were identified by liquid chromatography-tandem mass spectrometry. This revealed that the most abundant AHL signals produced by SplI in E. coli were N-3-oxo-hexanoylhomoserine lactone (3-oxo-C6-HSL), N-3-oxo-heptanoylhomoserine lactone (3-oxo-C7-HSL), N-3-hydroxy-hexanoylhomoserine lactone (3-hydroxy-C6-HSL), N-hexanoylhomoserine lactone (C6-HSL), and N-heptanoyl homoserine lactone (C7-HSL); whereas SpsI was primarily responsible for the synthesis of N-butyrylhomoserine lactone (C4-HSL) and N-pentanoylhomoserine lactone (C5-HSL). Furthermore, a quorum quenching analysis by heterologous expression of the Bacillus A24 AiiA lactonase in strain G3 enabled the identification of the AHL-regulated biocontrol-related traits. Depletion of AHLs with this lactonase resulted in altered adhesion and biofilm formation using a microtiter plate assay and flow cells coupled with confocal laser scanning microscopy respectively. This was different from the closely related S. plymuthica strains HRO-C48 and RVH1, where biofilm formation for both strains is AHL-independent. In addition, QS in G3 positively regulated antifungal activity, production of exoenzymes, but negatively regulated production of indol-3-acetic acid (IAA), which is in agreement with previous reports in strain HRO-C48. However, in contrast to HRO-C48, swimming motility was not controlled by AHL-mediated QS. CONCLUSIONS: This is the first report of the characterisation of two AHL-based quorum sensing systems in the same isolate of the genus Serratia. Our results show that the QS network is involved in the global regulation of biocontrol-related traits in the endophytic strain G3. However, although free-living and endophytic S. plymuthica share some conservation on QS phenotypic regulation, the control of motility and biofilm formation seems to be strain-specific and possible linked to the life-style of this organism.
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Biofilmes/crescimento & desenvolvimento , Percepção de Quorum/genética , Serratia/genética , Serratia/fisiologia , Filogenia , RNA Ribossômico 16S/genética , Serratia/classificaçãoRESUMO
We isolated a new strain of endophytic Pseudomonas G5 from the stems of Chinese parsley (Coriandrum sativum L.), and it is tentatively identified as Pseudomonas aurantiaca according to analysis of the entire substrate utilization profiles using BIOLOG Microstation system (BIOLOG, Inc, Hayward CA). An array of evidence established that many Gram-negative bacteria employ Quorum sensing (QS) system to regulate gene expression in response to cell density using small diffusible signal molecules, N-acyl homoserine lactones (AHLs), and control diverse phenotypic traits in plant-associated bacteria. In this study, we showed that Pseudomonas sp. strain G5 can produce several types of AHLs at a detectable level using Thin Layer Chromatography (TLC) analysis combined with bioreporter Chromobacterium violaceum CV026 bioassay, and N-hexanoyl-homoserine lactone (HHL, C6-HSL) with Rf value 0.4 is the major signal molecule. Furthermore, we have identified its quorum sensing system composed of PhzI and PhzR by cloning and sequencing of phzI-phzR. PhzI is responsible for synthesis of AHLs signal molecules, and PhzR is a transcriptional regulator. Finally, we heterologously expressed the recombinant plasmid pMD-phzIR in Escherichia coli JM109 and verified it using C. violaceum CV026 bioassay. The phylogenetic analysis using MEGA4 revealed highly similarities exist among the phzIR homologs, suggesting it is evolutionary well conserved in the genus Pseudomonas.
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Proteínas de Bactérias/genética , Escherichia coli/metabolismo , Pseudomonas/genética , Transativadores/genética , 4-Butirolactona/análogos & derivados , 4-Butirolactona/metabolismo , Acil-Butirolactonas/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/biossíntese , Clonagem Molecular , Escherichia coli/genética , Regulação Bacteriana da Expressão Gênica , Vetores Genéticos/genética , Dados de Sequência Molecular , Filogenia , Pseudomonas/classificação , Pseudomonas/isolamento & purificação , Transativadores/biossínteseRESUMO
Antagonism of three endophytic fungi isolated from common reed (Phragmites australis) against eight soilborne pathogenic fungi was investigated on potato dextrose agar by light microscopy, scanning electron microscopy, and transmission electron microscopy. Inhibitory zones were not observed. The microscopical studies suggested that the endophytes inhibit growth of soilborne pathogens by means of coiling around hyphae and, after penetration, the degradation of hyphal cytoplasm. Since penetration of hyphae seems to play a major role in parasitism, we studied the production of cell wall degrading enzymes by the three endophytes. Choiromyces aboriginum produced higher activities of beta-1,3-glucanases compared to Stachybotrys elegans and Cylindrocarpon sp. For C. aboriginum and S. elegans, colloidal chitin was the best substrate for the induction of beta-1,3-glucanases and chitinases, respectively. This result suggests that mycoparasitism by endophytes on soilborne plant pathogens can be explained by their mycoparasitic activity.
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Antibiose , Ascomicetos/fisiologia , Parede Celular/metabolismo , Fungos/fisiologia , Doenças das Plantas/microbiologia , Poaceae/microbiologia , Microbiologia do Solo , Stachybotrys/fisiologia , Ascomicetos/enzimologia , Ascomicetos/ultraestrutura , Celulases/metabolismo , Quitinases/metabolismo , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Controle Biológico de Vetores , Stachybotrys/enzimologia , Stachybotrys/ultraestruturaRESUMO
Microbial biocontrol agents (BCAs) are generally used for controlling plant diseases via antagonistic mechanisms including competition, antibiosis, parasitism, and cross-protection. Some BCAs can even promote plant growth, and provide induced systemic resistance (ISR), i. e., induce the plants to have resistance against pathogens including phytopathogenic fungi, bacteria and virus, and in some cases, pest insects and nematodes. ISR is characterized by non-specific, wide spectrum and systemic. It is phenotypically similar to the systemic acquired resistance (SAR) induced by the infection of pathogens, and with the same efficiency but without hypersensitive response (HR) and visible symptoms in plant as SAR, which is helpful to open a new way to develop and improve safer and environmentally friendly strategies for plant protection. In this paper, the research advances on ISR mediated by biocontrol fungi and bacteria, elicitors or determinants, and signaling transduction pathways were summarized, with more emphasis on the biochemical and cytological mechanisms of host plant defense reaction induced by free-living and endophytic BCAs. The potential application of ISR in biocontrol of plant diseases was also discussed.