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1.
Appl Microbiol Biotechnol ; 108(1): 398, 2024 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-38940906

RESUMO

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.


Assuntos
Biofilmes , Botrytis , Fragaria , Proteínas Fúngicas , Hifas , Proteínas de Membrana , Doenças das Plantas , Botrytis/patogenicidade , Botrytis/genética , Botrytis/crescimento & desenvolvimento , Botrytis/efeitos dos fármacos , Biofilmes/crescimento & desenvolvimento , Biofilmes/efeitos dos fármacos , Virulência , Hifas/crescimento & desenvolvimento , Hifas/efeitos dos fármacos , Doenças das Plantas/microbiologia , Fragaria/microbiologia , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Vitis/microbiologia , Esporos Fúngicos/crescimento & desenvolvimento , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/genética , Deleção de Genes
2.
Plant Dis ; 107(1): 107-115, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-35771107

RESUMO

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.


Assuntos
Ascomicetos , Streptomyces , Glycine max , Ascomicetos/fisiologia , Streptomyces/genética , Streptomyces/metabolismo
3.
World J Microbiol Biotechnol ; 39(8): 202, 2023 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-37209223

RESUMO

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.


Assuntos
Melhoramento Vegetal , Streptomyces , Temperatura , Mutagênese , Streptomyces/genética , Streptomyces/metabolismo , Fermentação , Antibacterianos/farmacologia , Antibacterianos/metabolismo
4.
Cancer Sci ; 113(10): 3566-3578, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35879265

RESUMO

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.


Assuntos
Neoplasias Esofágicas , beta Catenina , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Ciclina D1/metabolismo , Neoplasias Esofágicas/patologia , Fibronectinas/metabolismo , Regulação Neoplásica da Expressão Gênica , Glicogênio Sintase Quinase 3 beta/metabolismo , Humanos , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , N-Metilaspartato/genética , N-Metilaspartato/metabolismo , NF-kappa B/metabolismo , Proteínas do Tecido Nervoso , Receptores de N-Metil-D-Aspartato/genética , Receptores de N-Metil-D-Aspartato/metabolismo , Via de Sinalização Wnt , beta Catenina/metabolismo
5.
Plant Dis ; 106(1): 156-164, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34184553

RESUMO

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.


Assuntos
Ascomicetos , Streptomyces , Micélio
6.
World J Microbiol Biotechnol ; 38(4): 65, 2022 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-35229201

RESUMO

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.


Assuntos
Streptomyces , Toiocamicina , Antibacterianos/metabolismo , Família Multigênica , Streptomyces/metabolismo
7.
BMC Microbiol ; 19(1): 280, 2019 12 11.
Artigo em Inglês | MEDLINE | ID: mdl-31829181

RESUMO

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.


Assuntos
Botrytis/efeitos dos fármacos , Fungicidas Industriais/farmacologia , Proteômica , Agricultura , Botrytis/crescimento & desenvolvimento , Parede Celular/efeitos dos fármacos , Cromatografia Líquida , Hifas/efeitos dos fármacos , Doenças das Plantas/microbiologia , Espectrometria de Massas em Tandem
8.
BMC Microbiol ; 19(1): 155, 2019 07 08.
Artigo em Inglês | MEDLINE | ID: mdl-31286877

RESUMO

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.


Assuntos
Ácidos Indolacéticos/metabolismo , Desenvolvimento Vegetal/efeitos dos fármacos , Reguladores de Crescimento de Plantas/biossíntese , Streptomyces/metabolismo , Técnicas de Cultura de Células/métodos , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , Caulim , Solanum lycopersicum/efeitos dos fármacos , Solanum lycopersicum/crescimento & desenvolvimento , Streptomyces/crescimento & desenvolvimento , Triptofano/metabolismo
9.
Appl Microbiol Biotechnol ; 100(20): 8819-28, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27372076

RESUMO

Wuyiencin is produced by Streptomyces ahygroscopicus var. wuyiensis, which has been widely used in China as an industrially produced biopesticide to control various fungal diseases. Although its mechanism of action, breeding, and fermentation had been extensively characterized, less is known about the regulatory functions that affect its biosynthesis or morphological development. The wysR3 gene of S. ahygroscopicus strain CK-15, a novel member of the DeoR family of regulatory genes, was assessed to determine its function by gene knockdown. Herein, we demonstrate for the first time that DeoR family proteins derived from the same source are likely to be a single branch in a phylogenetic tree and show that wysR3 acts as a repressor for its morphological development without effecting wuyiencin production. We found that the ΔwysR3 strain can grow quickly to reach a plateau stage of maximum biomass at 60 h, which is ∼12 h faster than the wild-type strain. In the industrial fermentation production process, the ΔwysR3 strain can reduce consumption and save both time and money.


Assuntos
Regulação Bacteriana da Expressão Gênica , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Streptomyces/crescimento & desenvolvimento , Streptomyces/genética , Deleção de Genes , Técnicas de Silenciamento de Genes
10.
Arch Virol ; 160(5): 1145-52, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25698104

RESUMO

Pathogens causing significant economic losses in chrysanthemum include tomato aspermy virus (TAV), chrysanthemum virus B (CVB), cucumber mosaic virus (CMV), tobacco mosaic virus (TMV), potato virus Y (PVY), chrysanthemum stunt viroid (CSVd) and chrysanthemum chlorotic mottle viroid (CChMVd). A multiplex reverse transcription polymerase chain reaction (RT-PCR) method, using specific primer sets for each virus or viroid, was developed for simultaneous detection and differentiation of TAV, CVB, CMV, TMV, PVY, CChMVd, and CSVd. The RT-PCR method was validated by testing chrysanthemum samples collected from different regions of China. In this study, CVB, TAV, TMV, PVY, CSVd, CMV, and CChMVd were detected, respectively, in 24.7 %, 17.5 %, 4.4 %, 4.4 %, 2.9 %, 2.5 %, and 1.5 % of the samples tested. These results indicate that CVB and TAV (24.7 % and 17.5 %) are common, whereas CMV, TMV, CChMVd, CSVd, and PVY (all below 5 %) are less frequently encountered. This new multiplex RT-PCR method has potential to be used routinely in large-scale virus and viroid surveys.


Assuntos
Chrysanthemum/virologia , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Vírus de RNA/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Viroides/isolamento & purificação , Virologia/métodos , China , Doenças das Plantas/virologia , Prevalência , Vírus de RNA/classificação , Vírus de RNA/genética , Viroides/classificação , Viroides/genética
11.
Arch Virol ; 159(12): 3197-210, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25233939

RESUMO

Potato virus M (PVM, genus Carlavirus, family Betaflexviridae) is considered to be one of most economically important pathogens of pepino in China. However, the details and the mechanisms underlying PVM evolution are unknown. In this study, we determined and analyzed 40 TGB 1 gene sequences, 67 TGB 2 and TGB 3 gene sequences, and 88 CP and NABP gene sequences from viruses isolated from 19 samples of pepino (Solanum muricatum) and one sample of tomato (S. lycopersicum) collected from different areas of China. Recombination analysis identified only one clear recombinant in the TGB2-TGB3-CP region, but no recombinants were detected for each of the five individual genes. Phylogenetic analysis showed that all PVM isolates could be divided into at least two lineages in trees derived from the TGB 2, CP, and NABP gene sequences, and the lineages seemed to reflect geographical origin. The five PVM genes in this study were found to be under strong negative selection pressure. The PVM isolates examined showed frequent gene flow between the Chinese and European populations, and also within the Chinese population. Clear star phylogenies and the neutral equilibrium model test showed that pepino isolates of PVM appear to be experiencing a new expansion after a recent introduction into China, and these isolates display low levels of genetic diversity. To our knowledge, this study is the first report describing genetic structure, recombination, and gene flow in PVM populations, and it provides strong evolutionary evidence for the virus populations from different geographic regions of China.


Assuntos
Carlavirus/classificação , Carlavirus/genética , Variação Genética , Doenças das Plantas/virologia , Solanum/virologia , Carlavirus/isolamento & purificação , China , Análise por Conglomerados , Evolução Molecular , Fluxo Gênico , Dados de Sequência Molecular , Filogenia , RNA Viral/genética , Análise de Sequência de DNA
12.
Aging (Albany NY) ; 16(10): 8541-8551, 2024 05 13.
Artigo em Inglês | MEDLINE | ID: mdl-38742950

RESUMO

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.


Assuntos
Estadiamento de Neoplasias , Neoplasias Gástricas , Humanos , Neoplasias Gástricas/mortalidade , Neoplasias Gástricas/patologia , Masculino , Feminino , Pessoa de Meia-Idade , Estudos Retrospectivos , Idoso , Fatores de Risco , Adulto , Estimativa de Kaplan-Meier , Modelos de Riscos Proporcionais , Prognóstico , Taxa de Sobrevida , Metástase Linfática/patologia
13.
JACC Case Rep ; 29(6): 102253, 2024 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-38549853

RESUMO

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.

14.
Heart Rhythm ; 2024 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-38490601

RESUMO

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.

15.
Arch Virol ; 158(6): 1181-7, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23338706

RESUMO

Potato virus M (PVM), pepino mosaic virus (PepMV), tomato mosaic virus (ToMV), and potato virus S (PVS) infect pepino and cause serious crop losses. In this study, a multiplex RT-PCR method was developed for simultaneous detection and differentiation of PVS, ToMV, PepMV and PVM. The method was highly reliable and sensitive; validation was accomplished by testing pepino samples collected from different regions of China. In this survey, PVM, ToMV and PVS were detected in 37.0 %, 31.0 % and 5.5 % of samples tested, respectively, confirming the widespread occurrence of these three viruses in China. PepMV was not detected in any of the samples, which indicated that this virus may not be prevalent in China. The results suggest that the new multiplex RT-PCR method has potential to be used routinely for surveys of pepino for virus infection.


Assuntos
Carlavirus/genética , Doenças das Plantas/virologia , Potexvirus/genética , Solanum/virologia , Tobamovirus/genética , China , Ensaio de Imunoadsorção Enzimática , Reação em Cadeia da Polimerase Multiplex , Folhas de Planta/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade
16.
J Hazard Mater ; 458: 132035, 2023 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-37453358

RESUMO

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.


Assuntos
Agentes de Controle Biológico , Praguicidas , Microbiologia do Solo , Rizosfera , Antibacterianos/farmacologia , Bactérias/genética , Raízes de Plantas/microbiologia
17.
Microorganisms ; 11(5)2023 May 06.
Artigo em Inglês | MEDLINE | ID: mdl-37317199

RESUMO

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.

18.
Microorganisms ; 10(5)2022 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-35630418

RESUMO

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.

19.
Artigo em Inglês | MEDLINE | ID: mdl-35137282

RESUMO

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.

20.
PLoS One ; 15(4): e0224643, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32348310

RESUMO

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.


Assuntos
Antifúngicos/farmacologia , Botrytis/genética , Farmacorresistência Fúngica , Transcriptoma , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Botrytis/efeitos dos fármacos , Botrytis/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Regulação para Cima
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