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1.
Science ; 385(6714): 1211-1217, 2024 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-39265009

RESUMO

In plants, a local infection can lead to systemic acquired resistance (SAR) through increased production of salicylic acid (SA). For many years, the identity of the mobile signal and its direct transduction mechanism for systemic SA synthesis in initiating SAR have been debated. We found that in Arabidopsis thaliana, after a local infection, the conserved cysteine residue of the transcription factor CCA1 HIKING EXPEDITION (CHE) undergoes sulfenylation in systemic tissues, which enhances its binding to the promoter of the SA-synthesis gene ISOCHORISMATE SYNTHASE1 (ICS1) and increases SA production. Furthermore, hydrogen peroxide (H2O2) produced through NADPH oxidases is the mobile signal that sulfenylates CHE in a concentration-dependent manner. Accumulation of SA and the previously reported signal molecules, such as N-hydroxypipecolic acid (NHP), then form a signal amplification loop to establish SAR.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Peróxido de Hidrogênio , Doenças das Plantas , Ácido Salicílico , Fatores de Transcrição , Arabidopsis/genética , Arabidopsis/microbiologia , Arabidopsis/imunologia , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Peróxido de Hidrogênio/metabolismo , Ácido Salicílico/metabolismo , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Doenças das Plantas/microbiologia , Resistência à Doença/genética , Regiões Promotoras Genéticas , Transferases Intramoleculares/metabolismo , Transferases Intramoleculares/genética , NADPH Oxidases/metabolismo , NADPH Oxidases/genética , Regulação da Expressão Gênica de Plantas , Cisteína/metabolismo , Transdução de Sinais , Pseudomonas syringae
2.
J Agric Food Chem ; 72(37): 20658-20669, 2024 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-39226125

RESUMO

Sulfur-containing compounds have diverse biological functions and are crucial in crop protection chemistry. In this study, a series of novel 1-methyl-1H-pyrazol-5-amine derivatives incorporating disulfide moieties were synthesized and evaluated for their antimicrobial properties. In vitro bioassays demonstrated that compound 7f displayed potent antifungal activity against Valsa mali, with an EC50 value of 0.64 mg/L, outperforming allicin (EC50 = 26.0 mg/L) but lower than tebuconazole (EC50 = 0.33 mg/L). In vivo experiments confirmed that compound 7f could effectively inhibit V. mali infection on apples at a concentration of 100 mg/L, similar to the positive control tebuconazole. Mechanistic studies revealed that compound 7f could induce hyphal shrinkage and collapse, trigger intracellular reactive oxygen species accumulation, modulate antioxidant enzyme activities, initiate lipid peroxidation, and ultimately cause irreversible oxidative damage to the cells of V. mali. Additionally, compound 7b exhibited notable antibacterial activity, particularly against Pseudomonas syringae pv. actinidiae, with a MIC90 value of 1.56 mg/L, surpassing the positive controls allicin, bismerthiazol, and streptomycin sulfate. These findings suggest that 1-methyl-1H-pyrazol-5-amine derivatives containing disulfide moieties hold promise as potent candidates for the development of novel antimicrobial agents.


Assuntos
Dissulfetos , Testes de Sensibilidade Microbiana , Pirazóis , Dissulfetos/química , Dissulfetos/farmacologia , Pirazóis/farmacologia , Pirazóis/química , Pirazóis/síntese química , Doenças das Plantas/microbiologia , Relação Estrutura-Atividade , Pseudomonas syringae/efeitos dos fármacos , Antibacterianos/farmacologia , Antibacterianos/química , Antibacterianos/síntese química , Anti-Infecciosos/farmacologia , Anti-Infecciosos/química , Anti-Infecciosos/síntese química , Estrutura Molecular , Fungicidas Industriais/farmacologia , Fungicidas Industriais/química , Fungicidas Industriais/síntese química
3.
Mol Plant Pathol ; 25(9): e70005, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39235143

RESUMO

Plant pathogens pose a high risk of yield losses and threaten food security. Technological and scientific advances have improved our understanding of the molecular processes underlying host-pathogen interactions, which paves the way for new strategies in crop disease management beyond the limits of conventional breeding. Cross-family transfer of immune receptor genes is one such strategy that takes advantage of common plant immune signalling pathways to improve disease resistance in crops. Sensing of microbe- or host damage-associated molecular patterns (MAMPs/DAMPs) by plasma membrane-resident pattern recognition receptors (PRR) activates pattern-triggered immunity (PTI) and restricts the spread of a broad spectrum of pathogens in the host plant. In the model plant Arabidopsis thaliana, the S-domain receptor-like kinase LIPOOLIGOSACCHARIDE-SPECIFIC REDUCED ELICITATION (AtLORE, SD1-29) functions as a PRR, which senses medium-chain-length 3-hydroxylated fatty acids (mc-3-OH-FAs), such as 3-OH-C10:0, and 3-hydroxyalkanoates (HAAs) of microbial origin to activate PTI. In this study, we show that ectopic expression of the Brassicaceae-specific PRR AtLORE in the solanaceous crop species Solanum lycopersicum leads to the gain of 3-OH-C10:0 immune sensing without altering plant development. AtLORE-transgenic tomato shows enhanced resistance against Pseudomonas syringae pv. tomato DC3000 and Alternaria solani NL03003. Applying 3-OH-C10:0 to the soil before infection induces resistance against the oomycete pathogen Phytophthora infestans Pi100 and further enhances resistance to A. solani NL03003. Our study proposes a potential application of AtLORE-transgenic crop plants and mc-3-OH-FAs as resistance-inducing biostimulants in disease management.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Resistência à Doença , Ácidos Graxos , Doenças das Plantas , Solanum lycopersicum , Solanum lycopersicum/microbiologia , Solanum lycopersicum/imunologia , Solanum lycopersicum/genética , Arabidopsis/imunologia , Arabidopsis/microbiologia , Arabidopsis/genética , Resistência à Doença/genética , Doenças das Plantas/microbiologia , Doenças das Plantas/imunologia , Ácidos Graxos/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Pseudomonas syringae/patogenicidade , Imunidade Vegetal , Plantas Geneticamente Modificadas
4.
Nat Commun ; 15(1): 7048, 2024 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-39147739

RESUMO

Plants possess cell surface-localized immune receptors that detect microbe-associated molecular patterns (MAMPs) and initiate defenses that provide effective resistance against microbial pathogens. Many MAMP-induced signaling pathways and cellular responses are known, yet how pattern-triggered immunity (PTI) limits pathogen growth in plants is poorly understood. Through a combined metabolomics and genetics approach, we discovered that plant-exuded proline is a virulence-inducing signal and nutrient for the bacterial pathogen Pseudomonas syringae, and that MAMP-induced depletion of proline from the extracellular spaces of Arabidopsis leaves directly contributes to PTI against P. syringae. We further show that MAMP-induced depletion of extracellular proline requires the amino acid transporter Lysine Histidine Transporter 1 (LHT1). This study demonstrates that depletion of a single extracellular metabolite is an effective component of plant induced immunity. Given the important role for amino acids as nutrients for microbial growth, their depletion at sites of infection may be a broadly effective means for defense against many pathogens.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Reconhecimento da Imunidade Inata , Doenças das Plantas , Imunidade Vegetal , Pseudomonas syringae , Sistemas de Transporte de Aminoácidos Básicos/metabolismo , Sistemas de Transporte de Aminoácidos Básicos/genética , Arabidopsis/imunologia , Arabidopsis/microbiologia , Arabidopsis/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/imunologia , Regulação da Expressão Gênica de Plantas/imunologia , Reconhecimento da Imunidade Inata/genética , Metabolômica , Moléculas com Motivos Associados a Patógenos/metabolismo , Moléculas com Motivos Associados a Patógenos/imunologia , Doenças das Plantas/microbiologia , Doenças das Plantas/imunologia , Imunidade Vegetal/genética , Folhas de Planta/microbiologia , Folhas de Planta/metabolismo , Folhas de Planta/imunologia , Prolina/metabolismo , Pseudomonas syringae/imunologia , Pseudomonas syringae/patogenicidade , Transdução de Sinais , Virulência
5.
Nat Commun ; 15(1): 6853, 2024 Aug 10.
Artigo em Inglês | MEDLINE | ID: mdl-39127720

RESUMO

Phytochromes (Phys) are a divergent cohort of bili-proteins that detect light through reversible interconversion between dark-adapted Pr and photoactivated Pfr states. While our understandings of downstream events are emerging, it remains unclear how Phys translate light into an interpretable conformational signal. Here, we present models of both states for a dimeric Phy with histidine kinase (HK) activity from the proteobacterium Pseudomonas syringae, which were built from high-resolution cryo-EM maps (2.8-3.4-Å) of the photosensory module (PSM) and its following signaling (S) helix together with lower resolution maps for the downstream output region augmented by RoseTTAFold and AlphaFold structural predictions. The head-to-head models reveal the PSM and its photointerconversion mechanism with strong clarity, while the HK region is interpretable but relatively mobile. Pr/Pfr comparisons show that bilin phototransformation alters PSM architecture culminating in a scissoring motion of the paired S-helices linking the PSMs to the HK bidomains that ends in reorientation of the paired catalytic ATPase modules relative to the phosphoacceptor histidines. This action apparently primes autophosphorylation enroute to phosphotransfer to the cognate DNA-binding response regulator AlgB which drives quorum-sensing behavior through transient association with the photoreceptor. Collectively, these models illustrate how light absorption conformationally translates into accelerated signaling by Phy-type kinases.


Assuntos
Proteínas de Bactérias , Histidina Quinase , Fitocromo , Pseudomonas syringae , Transdução de Sinais , Histidina Quinase/metabolismo , Histidina Quinase/química , Histidina Quinase/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Fitocromo/metabolismo , Fitocromo/química , Pseudomonas syringae/metabolismo , Modelos Moleculares , Microscopia Crioeletrônica , Conformação Proteica , Multimerização Proteica , Fotorreceptores Microbianos/metabolismo , Fotorreceptores Microbianos/química , Fotorreceptores Microbianos/genética , Luz
6.
Cell Host Microbe ; 32(9): 1552-1565.e8, 2024 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-39111320

RESUMO

Disease tolerance is an essential defense strategy against pathogens, alleviating tissue damage regardless of pathogen multiplication. However, its genetic and molecular basis remains largely unknown. Here, we discovered that protein condensation at the endoplasmic reticulum (ER) regulates disease tolerance in Arabidopsis against Pseudomonas syringae. During infection, Hematopoietic protein-1 (HEM1) and Bax-inhibitor 1 (BI-1) coalesce into ER-associated condensates facilitated by their phase-separation behaviors. While BI-1 aids in clearing these condensates via autophagy, it also sequesters lipid-metabolic enzymes within condensates, likely disturbing lipid homeostasis. Consequently, mutations in hem1, which hinder condensate formation, or in bi-1, which prevent enzyme entrapment, enhance tissue-damage resilience, and preserve overall plant health during infection. These findings suggest that the ER is a crucial hub for maintaining cellular homeostasis and establishing disease tolerance. They also highlight the potential of engineering disease tolerance as a defense strategy to complement established resistance mechanisms in combating plant diseases.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Resistência à Doença , Retículo Endoplasmático , Doenças das Plantas , Pseudomonas syringae , Arabidopsis/microbiologia , Arabidopsis/imunologia , Arabidopsis/genética , Arabidopsis/metabolismo , Retículo Endoplasmático/metabolismo , Doenças das Plantas/microbiologia , Doenças das Plantas/imunologia , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Pseudomonas syringae/patogenicidade , Autofagia
7.
Sci Adv ; 10(33): eado6229, 2024 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-39141726

RESUMO

The choline-glycine betaine pathway plays an important role in bacterial survival in hyperosmotic environments. Osmotic activation of the choline transporter BetT promotes the uptake of external choline for synthesizing the osmoprotective glycine betaine. Here, we report the cryo-electron microscopy structures of Pseudomonas syringae BetT in the apo and choline-bound states. Our structure shows that BetT forms a domain-swapped trimer with the C-terminal domain (CTD) of one protomer interacting with the transmembrane domain (TMD) of a neighboring protomer. The substrate choline is bound within a tryptophan prism at the central part of TMD. Together with functional characterization, our results suggest that in Pseudomonas species, including the plant pathogen P. syringae and the human pathogen Pseudomonas aeruginosa, BetT is locked at a low-activity state through CTD-mediated autoinhibition in the absence of osmotic stress, and its hyperosmotic activation involves the release of this autoinhibition.


Assuntos
Proteínas de Bactérias , Colina , Microscopia Crioeletrônica , Proteínas de Membrana Transportadoras , Proteínas de Membrana Transportadoras/metabolismo , Proteínas de Membrana Transportadoras/química , Colina/metabolismo , Colina/química , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/química , Pseudomonas syringae/metabolismo , Modelos Moleculares , Osmorregulação , Pressão Osmótica , Betaína/metabolismo , Conformação Proteica , Ligação Proteica , Relação Estrutura-Atividade , Domínios Proteicos
8.
BMC Plant Biol ; 24(1): 670, 2024 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-39004723

RESUMO

The most effective strategy for managing wheat bacterial blight caused by Pseudomonas syringae pv. syringae is believed to be the use of resistant cultivars. Researching the correlation between molecular markers and stress resistance can expedite the plant breeding process. The current study aims to evaluate the response of 27 bread wheat cultivars to bacterial blight disease in order to identify resistant and susceptible cultivars and to pinpoint ISSR molecular markers associated with bacterial blight resistance genes. ISSR markers are recommended for assessing a plant's disease resistance. This experiment is focused on identifying ISSR molecular markers linked to bacterial blight resistance. After applying the bacterial solution to the leaves, we performed sampling to determine the infection percentage in the leaves at different intervals (7, 14, and 18 days after spraying). In most cultivars, the average leaf infection percentage decreased 18 days after spraying on young leaves. However, in some cultivars such as Niknegad, Darab2, and Zarin, leaf infection increased in older leaves and reached up to 100% necrosis. In our study, 12 ISSR primers generated a total of 170 bands, with 156 being polymorphic. The primers F10 and F5 showed the highest polymorphism, while the F7 primer exhibited the lowest polymorphism. Cluster analysis grouped these cultivars into four categories. The resistant group included Qods, Omid, and Atrak cultivars, while the semi-resistant and susceptible groups comprised the rest of the cultivars. Through binary logistic analysis, we identified three Super oxide dismutase-related genes that contribute to plant resistance to bacterial blight. These genes were linked to the F3, F5, and F12 primers in regions I (1500 bp), T (1000 bp), and G (850 bp), respectively. We also identified seven susceptibility-associated genes. Atrak, Omid, and Qods cultivars exhibited resistance against bacterial blight, and three genes associated with this resistance were linked to the F3, F5, and F12 primers. These markers can be used for screening or transferring tolerance to other wheat cultivars in breeding programs.


Assuntos
Resistência à Doença , Doenças das Plantas , Pseudomonas syringae , Triticum , Doenças das Plantas/microbiologia , Doenças das Plantas/genética , Triticum/genética , Triticum/microbiologia , Resistência à Doença/genética , Pseudomonas syringae/fisiologia , Marcadores Genéticos , Folhas de Planta/microbiologia , Folhas de Planta/genética , Modelos Logísticos
9.
Plant Physiol Biochem ; 214: 108933, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39033650

RESUMO

WRKY transcription factors are essential for coping with various biotic stresses. Pseudomonas syringae pv. actinidiae (Psa)-induced kiwifruit canker is a major problem restricting kiwifruit yield. Nevertheless, it's unclear how the kiwifruit WRKY genes respond to Psa. Through genome-wide identification, 112 WRKY members were found in 'Hongyang' genome in this work. Promoter analysis revealed that there were many cis-acting elements associated with stress responses in the AcWRKY gene's promoter region. According to transcriptomic analysis, 90 of the AcWRKY genes were differently expressed following Psa, salicylic acid (SA), or methyl jasmonate (MeJA) treatments. Almost all group III WRKYs were responsive to at least one of these treatments, with tissue-specific expression patterns. Quantitative RT-PCR study provided more evidence that Psa and SA treatments significantly induced the expression of the group III WRKY gene AcWRKY94, whereas MeJA treatment repressed it. AcWRKY94 was a transcriptionally active protein localized in the nucleus. Transient overexpression of AcWRKY94 in the leaves of 'Hongyang' enhanced the resistance of kiwifruit to Psa. Overexpression of AcWRKY94 in kiwifruit callus remarkably promoted the expression of PR and JAZ genes associated with SA and JA signals, respectively. These data imply that AcWRKY94 controls the signaling pathway dependent on SA and JA, thereby enhancing resistance to Psa. Taken together, this study establishes the basis for functional research on WRKY genes and provides important information for elucidating the resistance mechanism of kiwifruit canker disease.


Assuntos
Actinidia , Regulação da Expressão Gênica de Plantas , Doenças das Plantas , Proteínas de Plantas , Pseudomonas syringae , Fatores de Transcrição , Actinidia/microbiologia , Actinidia/genética , Pseudomonas syringae/patogenicidade , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Doenças das Plantas/microbiologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Ciclopentanos/metabolismo , Ciclopentanos/farmacologia , Oxilipinas/farmacologia , Oxilipinas/metabolismo , Acetatos/farmacologia , Ácido Salicílico/metabolismo , Ácido Salicílico/farmacologia , Frutas/microbiologia , Frutas/genética , Resistência à Doença/genética , Regiões Promotoras Genéticas/genética
10.
Physiol Plant ; 176(4): e14456, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39072778

RESUMO

Receptor-like cytoplasmic kinases (RLCKs) represent a distinct class of receptor-like kinases crucial for various aspects of plant biology, including growth, development, and stress responses. This study delves into the characterization of RLCK VII-8 members within cucurbits, particularly in melon, examining both structural features and the phylogenetic relationships of these genes/proteins. The investigation extends to their potential involvement in disease resistance by employing ectopic overexpression in Arabidopsis. The promoters of CmRLCK VII-8 genes harbor multiple phytohormone- and stress-responsive cis-acting elements, with the majority (excluding CmRLCK39) displaying upregulated expression in response to defense hormones and fungal infection. Subcellular localization studies reveal that CmRLCK VII-8 proteins predominantly reside on the plasma membrane, with CmRLCK29 and CmRLCK30 exhibiting additional nuclear distribution. Notably, Arabidopsis plants overexpressing CmRLCK30 manifest dwarfing and delayed flowering phenotypes. Overexpression of CmRLCK27, CmRLCK30, and CmRLCK34 in Arabidopsis imparts enhanced resistance against Botrytis cinerea and Pseudomonas syringae pv. tomato DC3000, concomitant with the strengthened expression of defense genes and reactive oxygen species accumulation. The CmRLCK VII-8 members actively participate in chitin- and flg22-triggered immune responses. Furthermore, CmRLCK30 interacts with CmMAPKKK1 and CmARFGAP, adding a layer of complexity to the regulatory network. In summary, this functional characterization underscores the regulatory roles of CmRLCK27, CmRLCK30, and CmRLCK34 in immune responses by influencing pathogen-induced defense gene expression and ROS accumulation.


Assuntos
Arabidopsis , Botrytis , Resistência à Doença , Regulação da Expressão Gênica de Plantas , Doenças das Plantas , Proteínas de Plantas , Pseudomonas syringae , Arabidopsis/genética , Arabidopsis/microbiologia , Doenças das Plantas/microbiologia , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Resistência à Doença/genética , Botrytis/fisiologia , Botrytis/patogenicidade , Pseudomonas syringae/fisiologia , Pseudomonas syringae/patogenicidade , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Cucurbitaceae/microbiologia , Cucurbitaceae/genética , Filogenia , Plantas Geneticamente Modificadas
11.
J Agric Food Chem ; 72(26): 14581-14591, 2024 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-38957087

RESUMO

Plants withstand pathogen attacks by recruiting beneficial bacteria to the rhizosphere and passing their legacy on to the next generation. However, the underlying mechanisms involved in this process remain unclear. In our study, we combined microbiomic and transcriptomic analyses to reveal how the rhizosphere microbiome assembled through multiple generations and defense-related genes expressed in Arabidopsis thaliana under pathogen attack stress. Our results showed that continuous exposure to the pathogen Pseudomonas syringae pv tomato DC3000 led to improved growth and increased disease resistance in a third generation of rps2 mutant Arabidopsis thaliana. It could be attributed to the enrichment of specific rhizosphere bacteria, such as Bacillus and Bacteroides. Pathways associated with plant immunity and growth in A. thaliana, such as MAPK signaling pathways, phytohormone signal transduction, ABC transporter proteins, and flavonoid biosynthesis, were activated under the influence of rhizosphere bacterial communities. Our findings provide a scientific basis for explaining the relationship between beneficial microbes and defense-related gene expression. Understanding microbial communities and the mechanisms involved in plant responses to disease can contribute to better plant management and reduction of pesticide use.


Assuntos
Arabidopsis , Resistência à Doença , Doenças das Plantas , Pseudomonas syringae , Rizosfera , Arabidopsis/microbiologia , Arabidopsis/genética , Arabidopsis/imunologia , Doenças das Plantas/microbiologia , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Resistência à Doença/genética , Microbiota , Bactérias/genética , Bactérias/classificação , Bactérias/metabolismo , Bactérias/isolamento & purificação , Microbiologia do Solo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Adaptação Fisiológica , Raízes de Plantas/microbiologia , Raízes de Plantas/genética , Raízes de Plantas/imunologia , Raízes de Plantas/metabolismo , Regulação da Expressão Gênica de Plantas
12.
Cell Rep ; 43(8): 114544, 2024 Aug 27.
Artigo em Inglês | MEDLINE | ID: mdl-39052478

RESUMO

Although RNA structures play important roles in regulating gene expression, the mechanism and function of mRNA folding in plant bacterial pathogens remain elusive. Therefore, we perform dimethyl sulfate sequencing (DMS-seq) on the Pseudomonas syringae under nutrition-rich and -deficient conditions, revealing that the mRNA structure changes substantially in the minimal medium (MM) that tunes global translation efficiency (TE), thereby inducing virulence. This process is led by the increased expression of hfq, which is directly activated by transcription regulators RpoS and CysB. The co-occurrence of Hfq and RpoS in diverse bacteria and the deep conservation of Hfq Y25 is critical for RNA-mediated regulation and implicates the wider biological importance of mRNA structure and feedback loops in the control of global gene expression.


Assuntos
Regulação Bacteriana da Expressão Gênica , Fator Proteico 1 do Hospedeiro , Pseudomonas syringae , Transcriptoma , Fator Proteico 1 do Hospedeiro/metabolismo , Fator Proteico 1 do Hospedeiro/genética , Virulência/genética , Transcriptoma/genética , Pseudomonas syringae/patogenicidade , Pseudomonas syringae/genética , Pseudomonas syringae/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Fator sigma/metabolismo , Fator sigma/genética , RNA Mensageiro/metabolismo , RNA Mensageiro/genética , RNA Bacteriano/genética , RNA Bacteriano/metabolismo , Doenças das Plantas/microbiologia
13.
J Biol Chem ; 300(8): 107600, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-39059490

RESUMO

RNase R (encoded by the rnr gene) is a highly processive 3' → 5' exoribonuclease essential for the growth of the psychrotrophic bacterium Pseudomonas syringae Lz4W at low temperature. The cell death of a rnr deletion mutant at low temperature has been previously attributed to processing defects in 16S rRNA, defective ribosomal assembly, and inefficient protein synthesis. We recently showed that RNase R is required to protect P. syringae Lz4W from DNA damage and oxidative stress, independent of its exoribonuclease activity. Here, we show that the processing defect in 16S rRNA does not cause cell death of the rnr mutant of P. syringae at low temperature. Our results demonstrate that the rnr mutant of P. syringae Lz4W, complemented with a RNase R deficient in exoribonuclease function (RNase RD284A), is defective in 16S rRNA processing but can grow at 4 °C. This suggested that the processing defect in ribosomal RNAs is not a cause of the cold sensitivity of the rnr mutant. We further show that the rnr mutant accumulates copies of the indigenous plasmid pLz4W that bears a type II toxin-antitoxin (TA) system (P. syringae antitoxin-P. syringae toxin). This phenotype was rescued by overexpressing antitoxin psA in the rnr mutant, suggesting that activation of the type II TA system leads to cold sensitivity of the rnr mutant of P. syringae Lz4W. Here, we report a previously unknown functional relationship between the cold sensitivity of the rnr mutant and a type II TA system in P. syringae Lz4W.


Assuntos
Proteínas de Bactérias , Pseudomonas syringae , RNA Ribossômico 16S , Sistemas Toxina-Antitoxina , Pseudomonas syringae/metabolismo , Pseudomonas syringae/genética , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo , Sistemas Toxina-Antitoxina/genética , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Temperatura Baixa , Exorribonucleases/metabolismo , Exorribonucleases/genética , Mutação , Toxinas Bacterianas/metabolismo , Toxinas Bacterianas/genética
14.
J Nanobiotechnology ; 22(1): 446, 2024 Jul 29.
Artigo em Inglês | MEDLINE | ID: mdl-39075467

RESUMO

Bacterial diseases are one of the most common issues that result in crop loss worldwide, and the increasing usage of chemical pesticides has caused the occurrence of resistance in pathogenic bacteria and environmental pollution problems. Nanomaterial mediated gene silencing is starting to display powerful efficiency and environmental friendliness for improving plant disease resistance. However, the internalization of nanomaterials and the physiological mechanisms behind nano-improved plant disease resistance are still rarely understood. We engineered the polyethyleneimine (PEI) functionalized gold nanoparticles (PEI-AuNPs) with fluorescent properties and ROS scavenging activity to act as siRNA delivery platforms. Besides the loading, protection, and delivery of nucleic acid molecules in plant mature leaf cells by PEI-AuNPs, its fluorescent property further enables the traceability of the distribution of the loaded nucleic acid molecules in cells. Additionally, the PEI-AuNPs-based RNAi delivery system successfully mediated the silencing of defense-regulated gene AtWRKY1. Compared to control plants, the silenced plants performed better resistance to Pseudomonas syringae, showing a reduced bacterial number, decreased ROS content, increased antioxidant enzyme activities, and improved chlorophyll fluorescence performance. Our results showed the advantages of AuNP-based RNAi technology in improving plant disease resistance, as well as the potential of plant nanobiotechnology to protect agricultural production.


Assuntos
Resistência à Doença , Ouro , Nanopartículas Metálicas , Doenças das Plantas , Pseudomonas syringae , RNA Interferente Pequeno , Espécies Reativas de Oxigênio , Ouro/química , Nanopartículas Metálicas/química , Espécies Reativas de Oxigênio/metabolismo , Doenças das Plantas/microbiologia , Polietilenoimina/química , Inativação Gênica , Arabidopsis/genética
15.
Sci Adv ; 10(27): eadn6606, 2024 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-38959312

RESUMO

Ice-nucleating proteins (INpro) trigger the freezing of supercooled water droplets relevant to atmospheric, biological, and technological applications. The high ice nucleation activity of INpro isolated from the bacteria Pseudomonas syringae could be linked to the aggregation of proteins at the bacterial membrane or at the air-water interface (AWI) of droplets. Here, we imaged freezing onsets, providing direct evidence of these proposed mechanisms. High-speed cryo-microscopy identified the onset location of freezing in droplets between two protein-repellent glass slides. INpro from sterilized P. syringae (Snomax) statistically favored nucleation at the AWI of the droplets. Removing cellular fragments by filtration or adding surfactants increased the frequency of nucleation events at the AWI. On the other hand, cultivated intact bacteria cells or lipid-free droplets nucleated ice without an affinity to the AWI. Overall, we provide visual evidence that INpro from P. syringae trigger freezing at hydrophobic interfaces, such as the AWI or the bacterial membrane, with important mechanistic implications for applications of INpro.


Assuntos
Congelamento , Interações Hidrofóbicas e Hidrofílicas , Pseudomonas syringae , Pseudomonas syringae/metabolismo , Pseudomonas syringae/química , Proteínas da Membrana Bacteriana Externa/química , Proteínas da Membrana Bacteriana Externa/metabolismo , Gelo , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo
16.
Antonie Van Leeuwenhoek ; 117(1): 92, 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38949726

RESUMO

Biological control is a promising approach to enhance pathogen and pest control to ensure high productivity in cash crop production. Therefore, PGPR biofertilizers are very suitable for application in the cultivation of tea plants (Camellia sinensis) and tobacco, but it is rarely reported so far. In this study, production of a consortium of three strains of PGPR were applied to tobacco and tea plants. The results demonstrated that plants treated with PGPR exhibited enhanced resistance against the bacterial pathogen Pseudomonas syringae (PstDC3000). The significant effect in improving the plant's ability to resist pathogen invasion was verified through measurements of oxygen activity, bacterial colony counts, and expression levels of resistance-related genes (NPR1, PR1, JAZ1, POD etc.). Moreover, the application of PGPR in the tea plantation showed significantly reduced population occurrences of tea green leafhoppers (Empoasca onukii Matsuda), tea thrips (Thysanoptera:Thripidae), Aleurocanthus spiniferus (Quaintanca) and alleviated anthracnose disease in tea seedlings. Therefore, PGPR biofertilizers may serve as a viable biological control method to improve tobacco and tea plant yield and quality. Our findings revealed part of the mechanism by which PGPR helped improve plant biostresses resistance, enabling better application in agricultural production.


Assuntos
Nicotiana , Controle Biológico de Vetores , Doenças das Plantas , Pseudomonas syringae , Animais , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Nicotiana/microbiologia , Pseudomonas syringae/fisiologia , Controle Biológico de Vetores/métodos , Camellia sinensis/microbiologia , Camellia sinensis/crescimento & desenvolvimento , Insetos/microbiologia , Tisanópteros/microbiologia , Resistência à Doença , Desenvolvimento Vegetal , Agentes de Controle Biológico , Hemípteros/microbiologia
17.
Methods Mol Biol ; 2832: 205-212, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38869797

RESUMO

One of the major plant stress level indicators is reactive oxygen species (ROS). They have been known to play a central role in regulating plant responses to various environmental stresses. This book chapter specifically covers abiotic stress induced by a drought hormone abscisic acid and biotic stress induced by Pseudomonas syringe DC3000 on single cell-type guard cells. We describe in detail the measurement of ROS production starting from sample preparation to data analysis by fluorescence intensity acquisition using ImageJ software. We discussed the problems faced while performing the experiment and addressed how to overcome them by providing specific guidelines to ensure high quality repeatable data.


Assuntos
Arabidopsis , Espécies Reativas de Oxigênio , Estresse Fisiológico , Espécies Reativas de Oxigênio/metabolismo , Arabidopsis/metabolismo , Arabidopsis/genética , Ácido Abscísico/metabolismo , Pseudomonas syringae
18.
Microb Biotechnol ; 17(6): e14489, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38864499

RESUMO

Treating plant bacterial diseases is notoriously difficult because of the lack of available antimicrobials. Pseudomonas syringae pathovar syringae (Pss) is a major pathogen of cherry (Prunus avium) causing bacterial canker of the stem, leaf and fruit, impacting productivity and leading to a loss of trees. In an attempt to find a treatment for this disease, naturally occurring bacteriophage (phage) that specifically target Pss is being investigated as a biocontrol strategy. However, before using them as a biocontrol treatment, it is important to both understand their efficacy in reducing the bacterial population and determine if the bacterial pathogens can evolve resistance to evade phage infection. To investigate this, killing curve assays of five MR phages targeting Pss showed that phage resistance rapidly emerges in vitro, even when using a cocktail of the five phages together. To gain insight to the changes occurring, Pss colonies were collected three times during a 66-h killing curve assay and separately, Pss and phage were also coevolved over 10 generations, enabling the measurement of genomic and fitness changes in bacterial populations. Pss evolved resistance to phages through modifications in lipopolysaccharide (LPS) synthesis pathways. Bacterial fitness (growth) and virulence were affected in only a few mutants. Deletion of LPS-associated genes suggested that LPS was the main target receptor for all five MR phages. Later generations of coevolved phages from the coevolution experiment were more potent at reducing the bacterial density and when used with wild-type phages could reduce the emergence of phage-resistant mutants. This study shows that understanding the genetic mechanisms of bacterial pathogen resistance to phages is important for helping to design a more effective approach to kill the bacteria while minimizing the opportunity for phage resistance to manifest.


Assuntos
Doenças das Plantas , Pseudomonas syringae , Pseudomonas syringae/virologia , Pseudomonas syringae/genética , Doenças das Plantas/microbiologia , Fagos de Pseudomonas/genética , Fagos de Pseudomonas/fisiologia , Bacteriófagos/genética , Bacteriófagos/fisiologia
19.
mBio ; 15(7): e0087124, 2024 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-38899869

RESUMO

Chemosensory systems allow bacteria to respond and adapt to environmental conditions. Many bacteria contain more than one chemosensory system, but knowledge of their specific roles in regulating different functions remains scarce. Here, we address this issue by analyzing the function of the F6, F8, and alternative (non-motility) cellular functions (ACF) chemosensory systems of the model plant pathogen Pseudomonas syringae pv. tomato. In this work, we assign PsPto chemoreceptors to each chemosensory system, and we visualize for the first time the F6 and F8 chemosensory systems of PsPto using cryo-electron tomography. We confirm that chemotaxis and swimming motility are controlled by the F6 system, and we demonstrate how different components from the F8 and ACF systems also modulate swimming motility. We also determine how the kinase and response regulators from the F6 and F8 chemosensory systems do not work together in the regulation of biofilm, whereas both components from the ACF system contribute together to regulate these traits. Furthermore, we show how the F6, F8, and ACF kinases interact with the ACF response regulator WspR, supporting crosstalk among chemosensory systems. Finally, we reveal how all chemosensory systems play a role in regulating virulence. IMPORTANCE: Chemoperception through chemosensory systems is an essential feature for bacterial survival, as it allows bacterial interaction with its surrounding environment. In the case of plant pathogens, it is especially relevant to enter the host and achieve full virulence. Multiple chemosensory systems allow bacteria to display a wider plasticity in their response to external signals. Here, we perform a deep characterization of the F6, F8, and alternative (non-motility) cellular functions chemosensory systems in the model plant pathogen Pseudomonas syringae pv. tomato DC3000. These chemosensory systems regulate key virulence-related traits, like motility and biofilm formation. Furthermore, we unveil an unexpected crosstalk among these chemosensory systems at the level of the interaction between kinases and response regulators. This work shows novel results that contribute to the knowledge of chemosensory systems and their role in functions alternative to chemotaxis.


Assuntos
Proteínas de Bactérias , Biofilmes , Quimiotaxia , Pseudomonas syringae , Solanum lycopersicum , Pseudomonas syringae/genética , Pseudomonas syringae/patogenicidade , Pseudomonas syringae/metabolismo , Pseudomonas syringae/fisiologia , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Biofilmes/crescimento & desenvolvimento , Solanum lycopersicum/microbiologia , Virulência , Doenças das Plantas/microbiologia , Regulação Bacteriana da Expressão Gênica
20.
J Gen Virol ; 105(6)2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38833289

RESUMO

Relatively few phages that infect plant pathogens have been isolated and investigated. The Pseudomonas syringae species complex is present in various environments, including plants. It can cause major crop diseases, such as bacterial canker on apricot trees. This study presents a collection of 25 unique phage genomes that infect P. syringae. These phages were isolated from apricot orchards with bacterial canker symptoms after enrichment with 21 strains of P. syringae. This collection comprises mostly virulent phages, with only three being temperate. They belong to 14 genera, 11 of which are newly discovered, and 18 new species, revealing great genetic diversity within this collection. Novel DNA packaging systems have been identified bioinformatically in one of the new phage species, but experimental confirmation is required to define the precise mechanism. Additionally, many phage genomes contain numerous potential auxiliary metabolic genes with diversified putative functions. At least three phages encode genes involved in bacterial tellurite resistance, a toxic metalloid. This suggests that viruses could play a role in bacterial stress tolerance. This research emphasizes the significance of continuing the search for new phages in the agricultural ecosystem to unravel novel ecological diversity and new gene functions. This work contributes to the foundation for future fundamental and applied research on phages infecting phytopathogenic bacteria.


Assuntos
Genoma Viral , Doenças das Plantas , Fagos de Pseudomonas , Pseudomonas syringae , Pseudomonas syringae/virologia , Pseudomonas syringae/genética , Doenças das Plantas/microbiologia , Doenças das Plantas/virologia , Fagos de Pseudomonas/genética , Filogenia , Variação Genética
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