RESUMEN
By screening the compounding combination of Wuyiencin and chemical agents, this study aims to delay the emergence of chemical agent resistance, and provide a technical reference for scientific and rational fungicides technology. This study investigated the impacts of the antibiotic wuyiencin derived from Streptomyces albulus var. wuyiensis and its combination with pyrimethanil on the inhibition of Botrytis cinerea. Treatment with wuyiencin (≥80 µg mL-1) strongly inhibited the pathogenicity of B. cinerea and activated the plant defense response against B. cinerea. Application of 80-100 µg mL-1 wuyiencin effectively controlled grape gray mold (by 57.6-88.1% on leaves and 46.7-96.6% on fruits). Consequently, the application of 80-100 µg mL-1 wuyiencin effectively mitigated grape gray mold incidence, leading to a substantial reduction in disease symptoms to nearly imperceptible levels. When wuyiencin (at the median effective concentration [EC50]) was combined with pyrimethanil (EC50) at a ratio of 7:3, it exhibited the highest efficacy in inhibiting B. cinerea growth. This combination was significantly more potent (p < 0.05) than using wuyiencin or pyrimethanil alone in controlling gray mold on grape leaves and fruits. Furthermore, the combination effectively delayed resistance development in gray mold. The experimental results show that wuyiencin can delay resistance development by affecting the expression of methionine biosynthesis genes and reducing the activity of the cell wall-degrading enzyme activity.
RESUMEN
Grey mould caused by Botrytis cinerea is a devastating disease responsible for large losses to agricultural production, and B. cinerea is a necrotrophic model fungal plant pathogen. Membrane proteins are important targets of fungicides and hotspots in the research and development of fungicide products. Wuyiencin affects the permeability and pathogenicity of B. cinerea, parallel reaction monitoring revealed the association of membrane protein Bcsdr2, and the bacteriostatic mechanism of wuyiencin was elucidated. In the present work, we generated and characterised ΔBcsdr2 deletion and complemented mutant B. cinerea strains. The ΔBcsdr2 deletion mutants exhibited biofilm loss and dissolution, and their functional activity was illustrated by reduced necrotic colonisation on strawberry and grape fruits. Targeted deletion of Bcsdr2 also blocked several phenotypic defects in aspects of mycelial growth, conidiation and virulence. All phenotypic defects were restored by targeted gene complementation. The roles of Bcsdr2 in biofilms and pathogenicity were also supported by quantitative real-time RT-PCR results showing that phosphatidylserine decarboxylase synthesis gene Bcpsd and chitin synthase gene BcCHSV II were downregulated in the early stages of infection for the ΔBcsdr2 strain. The results suggest that Bcsdr2 plays important roles in regulating various cellular processes in B. cinerea. KEY POINTS: ⢠The mechanism of wuyiencin inhibits B. cinerea is closely associated with membrane proteins. ⢠Wuyiencin can downregulate the expression of the membrane protein Bcsdr2 in B. cinerea. ⢠Bcsdr2 is involved in regulating B. cinerea virulence, growth and development.
Asunto(s)
Biopelículas , Botrytis , Fragaria , Proteínas Fúngicas , Hifa , Proteínas de la Membrana , Enfermedades de las Plantas , Botrytis/patogenicidad , Botrytis/genética , Botrytis/crecimiento & desarrollo , Botrytis/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Biopelículas/efectos de los fármacos , Virulencia , Hifa/crecimiento & desarrollo , Hifa/efectos de los fármacos , Enfermedades de las Plantas/microbiología , Fragaria/microbiología , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Vitis/microbiología , Esporas Fúngicas/crecimiento & desarrollo , Esporas Fúngicas/efectos de los fármacos , Esporas Fúngicas/genética , Eliminación de GenRESUMEN
OBJECTIVE: The aim of this study was to investigate the main factors influencing the survival of patients with advanced gastric cancer. METHODS: The clinicopathological data of 120 patients with advanced gastric cancer were analyzed retrospectively, and clinical and pathological data were collected. Tumor tissue staging and grading were re-evaluated, and 5-year overall survival was followed up. The classified data were described by percentages, and the continuous data were described by standard deviations or medians. Univariate analysis was performed using the χ2 test or rank-sum test, followed by Kaplan-Meier survival analysis to calculate the median survival time and 5-year cumulative survival. A multivariate Cox regression model was used to evaluate the independent risk factors affecting survival. The test level was α = 0.05. RESULTS: Patients were followed up for 0 to 60 months, the 5-year overall survival rate was 36.2%, and the median survival time was 53.0 ± 1.461 months. K-M and log-rank test results revealed that tumor location, degree of differentiation, depth of invasion, regional lymph node involvement, and postoperative tumor stage were correlated with a decreased 5-year survival rate (P < 0.05). A multivariate Cox risk regression model was used to analyze the degree of histological differentiation (HR = 1.441; 95% CI = 1.049-1.979; P = 0.024), regional lymph node (HR = 1.626; 95% CI = 1.160-2.279; P = 0.005), and pTNM stage (HR = 2.266; 95% CI = 1.335-3.847; P = 0.002), which are independent risk factors for poor survival. Tumor location (P = 0.191), invasion depth (P = 0.579) and tumor size (P = 0.324) were not found to be independent risk factors. CONCLUSION: The degree of tumor differentiation, regional lymph node metastasis and postoperative pathological stage were found to be independent risk factors for 5-year overall survival in patients with advanced gastric cancer. Standardized and reasonable lymph node dissection and accurate postoperative pathological staging were very important.
Asunto(s)
Estadificación de Neoplasias , Neoplasias Gástricas , Humanos , Neoplasias Gástricas/mortalidad , Neoplasias Gástricas/patología , Masculino , Femenino , Persona de Mediana Edad , Estudios Retrospectivos , Anciano , Factores de Riesgo , Adulto , Estimación de Kaplan-Meier , Modelos de Riesgos Proporcionales , Pronóstico , Tasa de Supervivencia , Metástasis Linfática/patologíaRESUMEN
We present a unique case of left atrial (LA) dissection in a 46-year-old man following aortic dissection surgery. The LA dissection was attributed to coronary sinus catheter-related injury. This report highlights the importance of recognizing this rare complication and the crucial role of transesophageal echocardiography in its diagnosis. We discuss the pathogenesis, diagnostic criteria, and management strategies for LA dissection.
RESUMEN
BACKGROUND: Left bundle branch block (LBBB) represents a frequently encountered conduction system disorder. Despite its widespread occurrence, a continual dilemma persists regarding its intricate association with underlying cardiomyopathy and its pivotal role in the initiation of dilated cardiomyopathy. The pathologic alterations linked to LBBB-induced cardiomyopathy (LBBB-CM) have remained elusive. OBJECTIVE: This study sought to investigate the chronologic dynamics of LBBB to left ventricular dysfunction and the pathologic mechanism of LBBB-CM. METHODS: LBBB model was established through main left bundle branch trunk ablation in 14 canines. All LBBB dogs underwent transesophageal echocardiography and electrocardiography before ablation and at 1 month, 3 months, 6 months, and 12 months after LBBB induction. Single-photon emission computed tomography imaging was performed at 12 months. We then harvested the heart from all LBBB dogs and 14 healthy adult dogs as normal controls for anatomic observation, Purkinje fiber staining, histologic staining, and connexin43 protein expression quantitation. RESULTS: LBBB induction caused significant fibrotic changes in the endocardium and mid-myocardium. Purkinje fibers exhibited fatty degeneration, vacuolization, and fibrosis along with downregulated connexin43 protein expression. During a 12-month follow-up, left ventricular dysfunction progressively worsened, peaking at the end of the observation period. The association between myocardial dysfunction, hypoperfusion, and fibrosis was observed in the LBBB-afflicted canines. CONCLUSION: LBBB may lead to profound myocardial injury beyond its conduction impairment effects. The temporal progression of left ventricular dysfunction and the pathologic alterations observed shed light on the complex relationship between LBBB and cardiomyopathy. These findings offer insights into potential mechanisms and clinical implications of LBBB-CM.
Asunto(s)
Bloqueo de Rama , Modelos Animales de Enfermedad , Electrocardiografía , Perros , Animales , Bloqueo de Rama/fisiopatología , Bloqueo de Rama/etiología , Sistema de Conducción Cardíaco/fisiopatología , Tomografía Computarizada de Emisión de Fotón Único/métodos , Ramos Subendocárdicos/fisiopatología , Miocardio/patología , Disfunción Ventricular Izquierda/fisiopatología , Disfunción Ventricular Izquierda/etiologíaRESUMEN
The plant microbiota can substantially contribute to various functions related to host health, fitness, and productivity. Therefore, maintaining the integrity of the microbiota is beginning to be seen as a crucial factor in modern agriculture. Here, we evaluated the effects of two chemical pesticides (azoxystrobin and carbendazim) and an antibiotic-based biopesticide (wuyiencin) on the rhizosphere microbiome of tomato plants. It was found that all treatments resulted in changes in the bacterial community structure to varying degrees. The most pronounced changes were observed with the biopesticide, which resulted in an enrichment of Streptomyces in the microbiome. In contrast, the relative abundance of Actinobacteria decreased in samples that were treated with low and high dosages of carbendazim. Clostridia were enriched after the applications of azoxystrobin and wuyiencin. When functioning of the microbiome was assessed, it was shown that genes encoding multidrug efflux pumps and ABC transporters related to nutrient uptake were enriched. This enrichment is likely to overcome potentially negative effects linked to the exposure to the employed substances. The study provides new insights into the potential of different pesticides to modulate native plant microbiomes, and thus highlights the importance to include such evaluations when new active agents are developed.
Asunto(s)
Agentes de Control Biológico , Plaguicidas , Microbiología del Suelo , Rizosfera , Antibacterianos/farmacología , Bacterias/genética , Raíces de Plantas/microbiologíaRESUMEN
Botrytis cinerea is a necrotrophic model fungal plant pathogen that causes grey mould, a devastating disease responsible for large losses in the agriculture sector. As important targets of fungicides, membrane proteins are hot spots in the research and development of fungicide products. We previously found that membrane protein Bcest may be closely related to the pathogenicity of Botrytis cinerea. Herein, we further explored its function. We generated and characterised ΔBcest deletion mutants of B. cinerea and constructed complemented strains. The ΔBcest deletion mutants exhibited reduced conidia germination and germ tube elongation. The functional activity of ΔBcest deletion mutants was investigated by reduced necrotic colonisation of B. cinerea on grapevine fruits and leaves. Targeted deletion of Bcest also blocked several phenotypic defects in aspects of mycelial growth, conidiation and virulence. All phenotypic defects were restored by targeted-gene complementation. The role of Bcest in pathogenicity was also supported by reverse-transcriptase real-time quantitative PCR results indicating that melanin synthesis gene Bcpks13 and virulence factor Bccdc14 were significantly downregulated in the early infection stage of the ΔBcest strain. Taken together, these results suggest that Bcest plays important roles in the regulation of various cellular processes in B. cinerea.
RESUMEN
Streptomyces species are known for their ability to efficiently produce secondary metabolites, including various antibiotics. Wuyiencin, an antibiotic produced by Streptomyces albulus CK15, is commonly used in agriculture to control fungal diseases in crops and vegetables. In this study, we utilized atmospheric and room temperature plasma (ARTP) mutagenesis to generate mutant S. albulus strains with improved fermentation capabilities for wuyiencin production. After mutagenizing the wild-type S. albulus CK15 strain once and conducting two rounds of antimicrobial screening, three genetically stable mutants (M19, M26, and M28) were identified. These mutants showed increased wuyiencin production by 17.4%, 13.6%, and 18.5% in comparison to the CK15 strain in flask culture, respectively. The M28 mutant exhibited the highest wuyiencin activity, producing 1443.0 ± 134.6 U/mL in flask culture and 1673.8 ± 127.4 U/mL in a 5 L fermenter. These results demonstrate that ARTP is an efficient tool for microbial mutation breeding and improving wuyiencin production.
Asunto(s)
Fitomejoramiento , Streptomyces , Temperatura , Mutagénesis , Streptomyces/genética , Streptomyces/metabolismo , Fermentación , Antibacterianos/farmacología , Antibacterianos/metabolismoRESUMEN
Sclerotinia sclerotiorum (Lib) de Bary, a destructive fungal pathogen with an extensive host range, causes major economic losses to crop production activities globally. Streptomyces spp. produce secondary metabolites with diverse structures and biological activities with potential applications in the control of crop disease. This study explored the potential application of wuyiencin, a secondary metabolite of Streptomyces albulus CK-15, to induce defense responses in soybean against S. sclerotiorum. Lesion size was reduced by nearly 60% in wuyiencin-treated soybean plants compared with plants infected with S. sclerotiorum only in greenhouse experiments. Wuyiencin induced callose deposition at 6 h postinoculation and increased reactive-oxygen-scavenging enzyme activities, including superoxide dismutase, catalase, and peroxidase. Moreover, wuyiencin inoculated before S. sclerotiorum infection significantly increased polyphenol oxidase, phenylalanine ammonia lyase, chitinase, and ß-1,3-glucanase activity, suggesting their involvement in soybean defense responses to S. sclerotiorum. Further, qRT-PCR results showed expression levels of the hormone signaling markers CO11, MYC2, PR4, PR1, NPR1, and ERF1 were upregulated in infected leaves treated with wuyiencin.
Asunto(s)
Ascomicetos , Streptomyces , Glycine max , Ascomicetos/fisiología , Streptomyces/genética , Streptomyces/metabolismoRESUMEN
As a neuronal transmembrane protein, leucine-rich repeat and fibronectin type-III domain-containing protein 2 (LRFN2) can recruit and combine with N-methyl-d-aspartate receptors (NMDARs) to promote nerve growth. Genetic studies suggest that mutations in LRFN2 are associated with various cancers. However, the role and mechanism of LRFN2 in the progression of ESCC have not been elucidated. In this study, we demonstrated that LRFN2 was significantly downregulated in ESCC tissues by qRT-PCR and immunohistochemistry. Low LRFN2 expression was an adverse prognostic factor in patients with ESCC. Overexpression of LRFN2 effectively suppressed the proliferation, migration, invasion, and epithelial-to-mesenchymal transition in vitro and tumor growth in vivo. Bioinformatics analysis indicated that Wnt/ß-catenin signaling regulation was one of the most potential mechanisms and studies confirmed that overexpression of LFRN2 obviously downregulated the expression of ß-catenin, c-Myc, and cyclin D1 in ESCC cells and tumor tissues. Further studies revealed that LRFN2 plays an anti-ESCC role by binding with NMDAR-GRIN2B and this effect can be weakened by NR2B-selective NMDA antagonist-NMDA-IN-1. Moreover, the bioinformatics analysis showed that the interaction of GRIN2B and GSK3ß affects the NF-κB pathway, which was demonstrated by western blot experiments. Collectively, our results indicate that LRFN2 binding to NMDARs inhibits the progression of ESCC by regulating the Wnt/ß-catenin and NF-κB pathway, which provides a new therapeutic target for improving the prognosis of patients with ESCC.
Asunto(s)
Neoplasias Esofágicas , beta Catenina , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Ciclina D1/metabolismo , Neoplasias Esofágicas/patología , Fibronectinas/metabolismo , Regulación Neoplásica de la Expresión Génica , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Humanos , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , N-Metilaspartato/genética , N-Metilaspartato/metabolismo , FN-kappa B/metabolismo , Proteínas del Tejido Nervioso , Receptores de N-Metil-D-Aspartato/genética , Receptores de N-Metil-D-Aspartato/metabolismo , Vía de Señalización Wnt , beta Catenina/metabolismoRESUMEN
This study mainly evaluated the broad-spectrum fungicidal activity of ε-poly L lysine (ε-PL) against 12 pathogenic fungi. We further demonstrated synergistic antifungal activity of ε-PL combined with wuyiencin against Botryris cinerea. The combined bio-fungicide achieved an inhibition rate of 100% for mycelial growth using ε-PL at 500 µg/mL + wuyiencin at 50 µg/mL and for spore germination using ε-PL at 200 µg/mL + wuyiencin at 80 µg/mL in vitro. This synergistic spore and mycelia-damaging effect of the combination was confirmed using scanning electron microscopy. In vivo assays with combined bio-fungicide (1500 µg/mL ε-PL + 60 µg/mL wuyiencin) on detached leaves showed depressed growth and development of the spores of B. cinerea. The synergistic effect was further tested in combinations of ε-PL with wuyiencin by measuring the fractional inhibition concentration index (FICI) value below 0.5. Moreover, ε-PL and wuyiencin inoculation before B. cinerea infection significantly increased the superoxide dismutase, peroxidase, catalase, and phenylalanine ammonia-lyase activities, which suggested their involvement in tomato defense responses to disease to minimize damage to B. cinerea. These findings revealed that a combined bio-fungicide comprising ε-PL and wuyiencin had a good prospect for controlling plant fungal disease.
RESUMEN
Streptomyces albulus CK-15 produces various secondary metabolites, including the antibiotics wuyiencin and toyocamycin, which can reportedly control a broad range of plant fungal diseases. The production of these nucleoside antibiotics in CK-15 is regulated by two biosynthesis gene clusters. To investigate the potential effect of toyocamycin biosynthesis on wuyiencin production, we herein generated S. albulus strains in which a key gene in the toyocamycin biosynthesis gene cluster, namely toyF, was either deleted or overexpressed. The toyF deletion mutant ∆toyF did not produce toyocamycin, while the production of wuyiencin increased by 23.06% in comparison with that in the wild-type (WT) strain. In addition, ΔtoyF reached the highest production level of wuyiencin 4 h faster than the WT strain (60 h vs. and 64 h). Further, toyocamycin production by the toyF overexpression strain was two-fold higher than by the WT strain, while wuyiencin production was reduced by 29.10%. qRT-PCR showed that most genes in the toyocamycin biosynthesis gene cluster were expressed at lower levels in ∆toyF as compared with those in the WT strain, while the expression levels of genes in the wuyiencin biosynthesis gene cluster were upregulated. Finally, the growth rate of ∆toyF was much faster than that of the WT strain when cultured on solid or liquid medium. Based on our findings, we report that in industrial fermentation processes, ∆toyF has the potential to increase the production of wuyiencin and reduce the timeframe of fermentation.
Asunto(s)
Streptomyces , Toyocamicina , Antibacterianos/metabolismo , Familia de Multigenes , Streptomyces/metabolismoRESUMEN
The study aimed to investigate left ventricular (LV) motion pattern in patients with LBBB patterns including patients with pacemaker rhythm (PM), type B Wolff-Parkinson-White syndrome (B-WPW), premature ventricular complexes originating from the right ventricular outflow tract (RVOT-PVC), and complete left bundle branch block (CLBBB). Two-dimensional speckle tracking was used to evaluate peak value and time to peak value of the LV twist, LV apex rotation, and LV base rotation in patients with PM, B-WPW, RVOT-PVC, and CLBBB with normal LV ejection fraction, and in age-matched control subjects. The LV motion patterns were altered in all patients compared to the control groups. Patients with PM and CLBBB had a similar LV motion pattern with a reduced peak value of LV apex rotation and LV twist. Patients with B-WPW demonstrated the opposite trend in the reduction of LV rotation peak value, which was more dominant in the basal layer. The most impairment in the LV twist/rotation peak value was identified in patients with RVOT-PVC. Compared to the control group, the apical-basal rotation delay was prolonged in patients with CLBBB, followed by those with B-WPW, PM, and RVOT-PVC. The LV motion patterns were different among patients with different patterns of LBBB. CLBBB and PM demonstrated a reduction in LV twist/rotation that was pronounced in the apical layer, B-WPW showed a reduction in the basal layer, and RVOT-PVC in both layers. CLBBB had the most pronounced LV apical-basal rotation dyssynchrony.
RESUMEN
Sclerotinia sclerotiorum (Lib.) de Bary, a destructive fungal pathogen with an extensive host range, causes various diseases with the potential to cause huge economic losses to crops worldwide. Streptomyces species produce secondary metabolites with variable structures and biological activities that offer possible control methods for crop diseases. Herein, we evaluated the inhibitory effects of wuyiencin, a secondary metabolite of Streptomyces albulus CK-15, against S. sclerotiorum. The results showed that wuyiencin markedly inhibited mycelial growth and germination and the formation of sclerotia. It also increased cell membrane permeability, resulting in leakage of intracellular substances in pathogen mycelia. Wuyiencin markedly decreased oxalic acid content and the activities of polygalacturonase and pectin methyl-galacturonic enzymes. Moreover, it downregulated Nox1, ITL, pph1, Caf1, and sca1, all genes related to growth and infection. Lesions were smaller and less pronounced on soybean (Glycine max [L.] Merr.) leaves pretreated with wuyiencin in vitro, and the inhibition rate reached 78.36%. The results suggest that wuyiencin holds promise for the management of diseases caused by S. sclerotiorum, and the findings provide clues on the mechanism of action.
Asunto(s)
Ascomicetos , Streptomyces , MicelioRESUMEN
BACKGROUND: Knee osteoarthritis (KOA) is a common chronic degenerative disorder with an increasingly prevalence among the older individuals and the leading cause of pain in the elderly. Baduanjin, one of the ancient traditional Chinese mind-body exercise routine, has been recognized to have clinical benefits for KOA. We aim to evaluate the efficacy and safety of Baduanjin for patients with KOA through this systematic review and meta-analysis. METHODS: Four English databases (Cochrane Central Register of Controlled Trials, PubMed, EMBASE, and Web of Science), and 4 Chinese databases (China National Knowledge Infrastructure, Chinese Biomedical Literature Database, Chinese Scientific Journal Database, and Wanfang Database), will be searched from establishment of the database until October 2020. The reference lists and the citation lists of studies meeting the inclusion criteria will also be searched to identify further studies for inclusion. The search languages are English and Chinese. The randomized controlled trials of Baduanjin training for patients with KOA will be included. The primary outcome will be assessed according to the Western Ontario and McMaster Universities Osteoarthritis Index. Meta-analysis will be conducted with the use of RevMan 5.3. RESULTS: The results of this research will be submitted to a peer-reviewed publications. CONCLUSION: This systematic review aims to present evidence for whether Baduanjin training is an effective intervention which can improve both physical condition and life quality in patients suffering KOA. INPLASY REGISTRATION NUMBER: INPLASY202090051.
Asunto(s)
Terapia por Ejercicio/métodos , Medicina Tradicional China/métodos , Terapias Mente-Cuerpo/métodos , Osteoartritis de la Rodilla/terapia , Humanos , Metaanálisis como AsuntoRESUMEN
Grey mould is caused by the ascomycetes Botrytis cinerea in a range of crop hosts. As a biological control agent, the nucleoside antibiotic wuyiencin has been industrially produced and widely used as an effective fungicide. To elucidate the effects of wuyiencin on the transcriptional regulation in B. cinerea, we, for the first time, report a genome-wide transcriptomic analysis of B. cinerea treated with wuyiencin. 2067 genes were differentially expressed, of them, 886 and 1181 genes were significantly upregulated and downregulated, respectively. Functional categorization indicated that transcript levels of genes involved in amino acid metabolism and those encoding putative secreted proteins were altered in response to wuyiencin treatment. Moreover, the expression of genes involved in protein synthesis and energy metabolism (oxidative phosphorylation) and of those encoding ATP-binding cassette transporters was markedly upregulated, whereas that of genes participating in DNA replication, cell cycle, and stress response was downregulated. Furthermore, wuyiencin resulted in mycelial malformation and negatively influenced cell growth rate and conidial yield in B. cinerea. Our results suggest that this nucleoside antibiotic regulates all aspects of cell growth and differentiation in B. cinerea. To summarize, some new candidate pathways and target genes that may related to the protective and antagonistic mechanisms in B. cinerea were identified underlying the action of biological control agents.
Asunto(s)
Antifúngicos/farmacología , Botrytis/genética , Farmacorresistencia Fúngica , Transcriptoma , Transportadoras de Casetes de Unión a ATP/genética , Transportadoras de Casetes de Unión a ATP/metabolismo , Botrytis/efectos de los fármacos , Botrytis/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulación Fúngica de la Expresión Génica , Regulación hacia ArribaRESUMEN
BACKGROUND: Grey mould is an important plant disease worldwide, caused by Botrytis cinerea, resulting in serious economic loss. Wuyiencin, a low toxicity, high efficiency, and broad-spectrum agricultural antibiotic, has been demonstrated effectiveness against B. cinerea. RESULTS: Wuyiencin treatment inhibited growth and sporulation of B. cinerea, specifically altering hypha morphology and intracellular structures. These changes were accompanied by differential expression (fold change > 2.0) of 316 proteins identified by iTRAQ-labelling LC-MS/MS analysis (P < 0.05). Up-regulation of 14 proteins, including carbohydrate metabolism proteins and cell wall stabilization proteins, was validated by parallel reaction monitoring (PRM). Down-regulation of 13 proteins was validated by PRM, including regulators of energy metabolism, nucleotide/protein synthesis, and the biosynthesis of mediators of plant stress and decay. CONCLUSION: Our results confirm the inhibitory biological effects of wuyiencin on B. cinereal and elaborate on the differentially expressed proteins and associated pathways implicated in the capacity of wuyiencin to debilitate the growth and pathogenicity of grey mould. This study provides validated candidates for further targeted exploration with the goal of optimizing wuyiencin as a safe, low-toxicity agent for biological control.
Asunto(s)
Botrytis/efectos de los fármacos , Fungicidas Industriales/farmacología , Proteómica , Agricultura , Botrytis/crecimiento & desarrollo , Pared Celular/efectos de los fármacos , Cromatografía Liquida , Hifa/efectos de los fármacos , Enfermedades de las Plantas/microbiología , Espectrometría de Masas en TándemRESUMEN
BACKGROUND: Indole-3-acetic acid (IAA) is produced by microorganisms and plants via either tryptophan-dependent or tryptophan-independent pathways. Herein, we investigated the optimisation of IAA production by Streptomyces fradiae NKZ-259 and its formulation as a plant growth promoter to improve economic and agricultural development. RESULTS: The maximum IAA yield achieved using optimal conditions was 82.363 µg/mL in the presence of 2 g/L tryptophan after 6 days of incubation. Thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) analysis of putative IAA revealed an RF value of 0.69 and a retention time of 11.842 min, comparable with the IAA standard. Regarding product formulation, kaolin-based powder achieved a suspension rate of 73.74% and a wetting time of 80 s. This carrier exhibited good shelf life stability for NKZ-259, and the cell population did not decrease obviously over 4 months of storage at 4 °C. In vivo analysis of plant growth promotion showed that tomato seedlings treated with kaolin powder containing NKZ-259 cells displayed a significant increase in root and shoot length of 7.97 cm and 32.77 cm, respectively, and an increase in fresh weight and dry weight of 6.72 g and 1.34 g. Compared to controls, plant growth parameters were increased almost it two-fold. CONCLUSION: Optimising the culture conditions resulted in an almost four-fold increase in IAA secretion by NKZ-259 cells. The results clearly demonstrate that S. fradiae NKZ-259 holds great potential for plant growth promotion and IAA production. Furthermore, kaolin-based powder is an effective carrier for NKZ-259 cells and may be useful for commercial applications.
Asunto(s)
Ácidos Indolacéticos/metabolismo , Desarrollo de la Planta/efectos de los fármacos , Reguladores del Crecimiento de las Plantas/biosíntesis , Streptomyces/metabolismo , Técnicas de Cultivo de Célula/métodos , Cromatografía Líquida de Alta Presión , Cromatografía en Capa Delgada , Caolín , Solanum lycopersicum/efectos de los fármacos , Solanum lycopersicum/crecimiento & desarrollo , Streptomyces/crecimiento & desarrollo , Triptófano/metabolismoRESUMEN
Antibiotic resistance genes (ARGs) have raised numerous concerns in recent years as emerging environmental contaminants. At present, research on environmental contamination by antibiotics focuses on medical, animal husbandry, and aquaculture fields, with few studies on environmental contamination by agricultural antibiotics in the field of plant protection. Wuyiencin is a low toxicity, high efficiency, and broad-spectrum agricultural antibiotic. It has been widely used in agricultural production and it effectively controls crop fungal diseases. In the present study, pot experiments with four soil treatments (A, B, C and D) were set up in a greenhouse to investigate the effect of the application of wuyiencin on the fate of typical ARGs and microbial community. Eight typical ARGs were detected by real-time PCR and the microbial communities were analyzed using high-throughput sequencing. The results showed that wuyiencin neither significantly influenced ARG abundance and absolute gene copy numbers, nor significantly varied microbial community among treatments. Since it only was short-term results, and the detection number of ARGs was limited, whether wuyiencin is safe or not to ecological environment when using for long-term will need further deep research.
Asunto(s)
Antibacterianos/farmacología , Microbiota/efectos de los fármacos , Agricultura , Farmacorresistencia Microbiana/genética , Dosificación de Gen/efectos de los fármacos , Dosificación de Gen/genética , Genes Bacterianos/genética , Microbiota/genética , Suelo , Microbiología del SueloRESUMEN
Wuyiencin is produced by Streptomyces albulus var. wuyiensis and used widely in agriculture to control a variety of fungal diseases, such as cucumber downy mildew, strawberry powdery mildew, and tomato gray mold. As an industrially-produced biopesticide, reducing production costs is very important for popularization of this approach. To obtain a rapidly growing strain that effectively shortens the fermentation time, we investigated the effects of knockout and overexpression of the wysPII gene, a member of the LuxR regulatory gene family, in S. albulus strain CK-15. The ΔwysPII mutant exhibited a reduced rate of growth and sporulation. The time taken to reach the greatest mycelial biomass was approximately 18 h shorter in the ooPII (wysPII overexpressing) strain compared with that of the wild-type (WT) strain. In addition, the time to reach the greatest wuyiencin production was 56 h in the ooPII strain compared with 62 h in the WT strain. Furthermore, wysPII was shown to act as an activator of morphological development without affecting wuyiencin production. Thus, the ooPII strain can be used to reduce costs and increase efficiency in industrial fermentation processes for wuyiencin production.