RESUMO
The L-type voltage-gated Ca2+ channel gene CACNA1C is a risk gene for various psychiatric conditions, including schizophrenia and bipolar disorder. However, the cellular mechanism by which CACNA1C contributes to psychiatric disorders has not been elucidated. Here, we report that the embryonic deletion of Cacna1c in neurons destined for the cerebral cortex using an Emx1-Cre strategy disturbs spontaneous Ca2+ activity and causes abnormal brain development and anxiety. By combining computational modeling with electrophysiological membrane potential manipulation, we found that neural network activity was driven by intrinsic spontaneous Ca2+ activity in distinct progenitor cells expressing marginally increased levels of voltage-gated Ca2+ channels. MRI examination of the Cacna1c knockout mouse brains revealed volumetric differences in the neocortex, hippocampus, and periaqueductal gray. These results suggest that Cacna1c acts as a molecular switch and that its disruption during embryogenesis can perturb Ca2+ handling and neural development, which may increase susceptibility to psychiatric disease.
Assuntos
Transtornos de Ansiedade/metabolismo , Encéfalo/crescimento & desenvolvimento , Encéfalo/metabolismo , Canais de Cálcio Tipo L/metabolismo , Cálcio/metabolismo , Animais , Relógios Biológicos , Canais de Cálcio Tipo L/genética , Regulação da Expressão Gênica no Desenvolvimento , Predisposição Genética para Doença , Camundongos , Camundongos Knockout , Células-Tronco NeuraisRESUMO
Passionfruit (Passiflora edulis) is a significant fruit crop in the commercial sector, owing to its high nutritional and medicinal value. The advent of high-throughput genomics sequencing technology has led to the publication of a vast amount of passionfruit omics data, encompassing complete genome sequences and transcriptome data under diverse stress conditions. To facilitate the efficient integration, storage, and analysis of these large-scale datasets, and to enable researchers to effectively utilize these omics data, we developed the first passionfruit genome database (PGD). The PGD platform comprises a diverse range of functional modules, including a genome browser, search function, heatmap, gene expression patterns, various tools, sequence alignment, and batch download, thereby providing a user-friendly interface. Additionally, supplementary practical tools have been developed for the PGD, such as gene family analysis tools, gene ontology (GO) terms, a pathway enrichment analysis, and other data analysis and mining tools, which enhance the data's utilization value. By leveraging the database's robust scalability, the intention is to continue to collect and integrate passionfruit omics data in the PGD, providing comprehensive and in-depth support for passionfruit research. The PGD is freely accessible via http://passionfruit.com.cn .
Assuntos
Passiflora , Diagnóstico Pré-Implantação , Feminino , Gravidez , Humanos , Passiflora/genética , Genômica , Genoma , Análise de Sequência , Bases de Dados GenéticasRESUMO
BACKGROUND: Mechano-growth factor (MGF), which is a growth factor produced specifically in response to mechanical stimuli, with potential of tissue repair and regeneration. Our previous research has shown that MGF plays a crucial role in repair of damaged periodontal ligaments by promoting differentiation of periodontal ligament stem cells (PDLSCs). However, the molecular mechanism is not fully understood. This study aimed to investigated the regulatory effect of MGF on differentiation of PDLSCs and its molecular mechanism. METHODS: Initially, we investigated how MGF impacts cell growth and differentiation, and the relationship with the activation of Fyn-p-YAPY357 and LATS1-p-YAPS127. Then, inhibitors were used to interfere Fyn phosphorylation to verify the role of Fyn-p-YAP Y357 signal after MGF stimulation; moreover, siRNA was used to downregulate YAP expression to clarify the function of YAP in PDLSCs proliferation and differentiation. Finally, after C3 was used to inhibit the RhoA expression, we explored the role of RhoA in the Fyn-p-YAP Y357 signaling pathway in PDLSCs proliferation and differentiation. RESULTS: Our study revealed that MGF plays a regulatory role in promoting PDLSCs proliferation and fibrogenic differentiation by inducing Fyn-YAPY357 phosphorylation but not LATS1-YAP S127 phosphorylation. Moreover, the results indicated that Fyn could not activate YAP directly but rather activated YAP through RhoA in response to MGF stimulation. CONCLUSION: The research findings indicated that the Fyn-RhoA-p-YAPY357 pathway is significant in facilitating the proliferation and fibrogenic differentiation of PDLSCs by MGF. Providing new ideas for the study of MGF in promoting periodontal regenerative repair.
RESUMO
BACKGROUND: Rhizoctonia solani is an important plant pathogen worldwide, and causes serious tobacco target spot in tobacco in the last five years. This research studied the biological characteristics of four different anastomosis groups strains (AG-3, AG-5, AG-6, AG-1-IB) of R. solani from tobacco. Using metabolic phenotype technology analyzed the metabolic phenotype differences of these strains. RESULTS: The results showed that the suitable temperature for mycelial growth of four anastomosis group strains were from 20 to 30oC, and for sclerotia formation were from 20 to 25oC. Under different lighting conditions, R. solani AG-6 strains produced the most sclerotium, followed by R. solani AG-3, R. solani AG-5 and R. solani AG-1-IB. All strains had strong oligotrophic survivability, and can grow on water agar medium without any nitrutions. They exhibited three types of sclerotia distribution form, including dispersed type (R. solani AG-5 and AG-6), peripheral type (R. solani AG-1-IB), and central type (R. solani AG-3). They all presented different pathogenicities in tobacco leaves, with the most virulent was noted by R. solani AG-6, followed by R. solani AG-5 and AG-1-IB, finally was R. solani AG-3. R. solani AG-1-IB strains firstly present symptom after inoculation. Metabolic fingerprints of four anastomosis groups were different to each other. R. solani AG-3, AG-6, AG-5 and AG-1-IB strains efficiently metabolized 88, 94, 71 and 92 carbon substrates, respectively. Nitrogen substrates of amino acids and peptides were the significant utilization patterns for R. solani AG-3. R. solani AG-3 and AG-6 showed a large range of adaptabilities and were still able to metabolize substrates in the presence of the osmolytes, including up to 8% sodium lactate. Four anastomosis groups all showed active metabolism in environments with pH values from 4 to 6 and exhibited decarboxylase activities. CONCLUSIONS: The biological characteristics of different anastomosis group strains varies, and there were significant differences in the metabolic phenotype characteristics of different anastomosis group strains towards carbon source, nitrogen source, pH, and osmotic pressure.
Assuntos
Nicotiana , Fenótipo , Doenças das Plantas , Rhizoctonia , Nicotiana/microbiologia , Doenças das Plantas/microbiologia , Temperatura , Micélio/metabolismo , Micélio/crescimento & desenvolvimento , Folhas de Planta/microbiologia , VirulênciaRESUMO
Plant viral diseases compromise the growth and yield of the crop globally, and they tend to be more serious under extreme temperatures and drought climate changes. Currently, regulatory dynamics during plant development and in response to virus infection at the plant cell level remain largely unknown. In this study, single-cell RNA sequencing on 23 226 individual cells from healthy and tomato chlorosis virus-infected leaves was established. The specific expression and epigenetic landscape of each cell type during the viral infection stage were depicted. Notably, the mesophyll cells showed a rapid function transition in virus-infected leaves, which is consistent with the pathological changes such as thinner leaves and decreased chloroplast lamella in virus-infected samples. Interestingly, the F-box protein SKIP2 was identified to play a pivotal role in chlorophyll maintenance during virus infection in tomato plants. Knockout of the SlSKIP2 showed a greener leaf state before and after virus infection. Moreover, we further demonstrated that SlSKIP2 was located in the cytomembrane and nucleus and directly regulated by ERF4. In conclusion, with detailed insights into the plant responses to viral infections at the cellular level, our study provides a genetic framework and gene reference in plant-virus interaction and breeding in the future research.
Assuntos
Folhas de Planta , Solanum lycopersicum , Transcriptoma , Solanum lycopersicum/virologia , Solanum lycopersicum/genética , Solanum lycopersicum/crescimento & desenvolvimento , Folhas de Planta/virologia , Folhas de Planta/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Análise de Célula Única , Doenças das Plantas/virologia , Doenças das Plantas/genética , Regulação da Expressão Gênica de Plantas , Crinivirus/genética , Crinivirus/fisiologiaRESUMO
A novel negative-sense single-stranded RNA mycovirus, designated as "Magnaporthe oryzae mymonavirus 1" (MoMNV1), was identified in the rice blast fungus Magnaporthe oryzae isolate NJ39. MoMNV1 has a single genomic RNA segment consisting of 10,515 nucleotides, which contains six open reading frames. The largest open reading frame contains 5837 bases and encodes an RNA replicase. The six open reading frames have no overlap and are arranged linearly on the genome, but the spacing of the genes is small, with a maximum of 315 bases and a minimum of 80 bases. Genome comparison and phylogenetic analysis indicated that MoMNV1 is a new member of the genus Penicillimonavirus of the family Mymonaviridae.
Assuntos
Micovírus , Genoma Viral , Fases de Leitura Aberta , Oryza , Filogenia , Doenças das Plantas , Vírus de RNA , RNA Viral , Vírus de RNA/genética , Vírus de RNA/isolamento & purificação , Vírus de RNA/classificação , Micovírus/genética , Micovírus/isolamento & purificação , Micovírus/classificação , Oryza/microbiologia , Oryza/virologia , Doenças das Plantas/microbiologia , Doenças das Plantas/virologia , RNA Viral/genética , Ascomicetos/virologia , Ascomicetos/genética , Proteínas Virais/genética , Magnaporthe/virologia , Magnaporthe/genéticaRESUMO
P0 proteins encoded by the pepper vein yellow virus (PeVYV) are pathogenic factors that cause hypersensitive response (HR). However, the host gene expression related to PeVYV P0-induced HR has not been thoroughly studied. Transcriptomic technology was used to investigate the host pathways mediated by the PeVYV P0 protein to explore the molecular mechanisms underlying its function. We found 12,638 differentially expressed genes (DEGs); 6784 and 5854 genes were significantly upregulated and downregulated, respectively. Transcriptomic and reverse-transcription quantitative polymerase chain reaction (RT-qPCR) analyses revealed that salicylic acid (SA) and jasmonic acid (JA) synthesis-related gene expression was upregulated, and ethylene synthesis-related gene expression was downregulated. Ultrahigh performance liquid chromatography-tandem mass spectrometry was used to quantify SA and JA concentrations in Nicotiana benthamiana, and the P0 protein induced SA and JA biosynthesis. We then hypothesized that the pathogenic activity of the P0 protein might be owing to proteins related to host hormones in the SA and JA pathways, modulating host resistance at different times. Viral gene silencing suppression technology was used in N. benthamiana to characterize candidate proteins, and downregulating NbHERC3 (Homologous to E6-AP carboxy-terminus domain and regulator of choromosome condensation-1 dmain protein 3) accelerated cell necrosis in the host. The downregulation of NbCRR reduced cell death, while that of NbBax induced necrosis and curled heart leaves. Our findings indicate that NbHERC3, NbBax, and NbCRR are involved in P0 protein-driven cell necrosis.
Assuntos
Ciclopentanos , Regulação da Expressão Gênica de Plantas , Nicotiana , Oxilipinas , Doenças das Plantas , Proteínas de Plantas , Ácido Salicílico , Proteínas Virais , Ciclopentanos/metabolismo , Oxilipinas/metabolismo , Doenças das Plantas/virologia , Ácido Salicílico/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas Virais/genética , Proteínas Virais/metabolismo , Nicotiana/virologia , Nicotiana/genética , Potyvirus/patogenicidade , Potyvirus/genética , Folhas de Planta/virologia , Folhas de Planta/metabolismo , Resistência à Doença/genética , Interações Hospedeiro-Patógeno , Perfilação da Expressão Gênica , Capsicum/virologia , Capsicum/genética , Capsicum/metabolismo , Reguladores de Crescimento de Plantas/metabolismoRESUMO
Strigolactones (SLs) are plant hormones that regulate diverse developmental processes and environmental responses in plants. It has been discovered that SLs play an important role in regulating plant immune resistance to pathogens but there are currently no reports on their role in the interaction between Nicotiana benthamiana and the tobacco mosaic virus (TMV). In this study, the exogenous application of SLs weakened the resistance of N. benthamiana to TMV, promoting TMV infection, whereas the exogenous application of Tis108, a SL inhibitor, resulted in the opposite effect. Virus-induced gene silencing (VIGS) inhibition of two key SL synthesis enzyme genes, NtCCD7 and NtCCD8, enhanced the resistance of N. benthamiana to TMV. Additionally, we conducted a screening of N. benthamiana related to TMV infection. TMV-infected plants treated with SLs were compared to the control by using RNA-seq. The KEGG enrichment analysis and weighted gene co-expression network analysis (WGCNA) of differentially expressed genes (DEGs) suggested that plant hormone signaling transduction may play a significant role in the SL-TMV-N. benthamiana interactions. This study reveals new functions of SLs in regulating plant immunity and provides a reference for controlling TMV diseases in production.
Assuntos
Resistência à Doença , Regulação da Expressão Gênica de Plantas , Lactonas , Nicotiana , Doenças das Plantas , Vírus do Mosaico do Tabaco , Nicotiana/virologia , Nicotiana/genética , Nicotiana/imunologia , Vírus do Mosaico do Tabaco/fisiologia , Lactonas/farmacologia , Resistência à Doença/genética , Doenças das Plantas/virologia , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Imunidade Vegetal/genética , Imunidade Vegetal/efeitos dos fármacos , Inativação GênicaRESUMO
Isolation and analysis of double-stranded RNA (dsRNA) from the phytopathogenic fungus Setosphaeria turcica f. sp. zeae revealed the presence of a new double-stranded RNA (dsRNA) virus, tentatively named "Setosphaeria turcica polymycovirus 2" (StPmV2). The genome of StPmV2 consists of five segments (dsRNA1-5), ranging in size from 965 bp to 2462 bp. Each dsRNA contains one open reading frame (ORF) flanked by 5' and 3' untranslated regions (UTRs) with conserved terminal sequences. The putative protein encoded by dsRNA1 shows 64.52% amino acid sequence identity to the RNA-dependent RNA polymerase (RdRp) of the most closely related virus, Cladosporium cladosporioides virus 1, which belongs to the family Polymycoviridae. dsRNAs 2-4 encode the putative coat protein, methyltransferase (MTR), and proline-alanine-serine-rich protein (PASrp), respectively, and dsRNA5 encodes a protein of unknown function. Phylogenetic analysis based on the RdRp protein indicated that StPmV2 clustered with members of the family Polymycoviridae and is therefore a new mycovirus belonging to the genus Polymycovirus in the family Polymycoviridae. In addition, three other distinct isolates of StPmV2 were identified: one isolated from S. turcica f. sp. zeae and two from S. turcica f. sp. sorghi. To our knowledge, this is the first report of a polymycovirus infecting both S. turcica f. sp. zeae and S. turcica f. sp. sorghi.
Assuntos
Micovírus , Vírus de RNA , RNA Viral , RNA de Cadeia Dupla/genética , Filogenia , Genoma Viral , Vírus de RNA/genética , RNA Polimerase Dependente de RNA/genética , RNA Polimerase Dependente de RNA/química , Fases de Leitura AbertaRESUMO
BACKGROUND: Coronavirus papain-like proteases (PLpros) play a crucial role in virus replication and the evasion of the host immune response. Infectious bronchitis virus (IBV) encodes a proteolytically defective remnant of PL1pro and an active PL2pro. However, the function of PL1pro in IBV remains largely unknown. This study aims to explore the effect of PL1pro on virus replication and underlying mechanisms. RESULTS: The recombinant viruses rIBV-ΔPL1pro and rIBV-ΔPL1pro-N were obtained using reverse genetic techniques through the deletion of the IBV PL1pro domain and the N-terminal conserved sequence of PL1pro (PL1pro-N). We observed significantly lower replication of rIBV-ΔPL1pro and rIBV-ΔPL1pro-N than wild-type IBV. Further investigation revealed that the lack of PL1pro-N in IBV decreased virus resistance to interferon (IFN) while also inducing host immune response by enhancing the production of IFN-ß and activating the downstream STAT1 signaling pathway of IFNs. In addition, the overexpression of PL1pro-N significantly suppressed type I IFN response by down-regulating the expressions of genes in the IFN pathway. CONCLUSIONS: Our data demonstrated that IBV PL1pro plays a crucial role in IBV replication and the suppression of host innate immune responses, suggesting that IBV PL1pro could serve as a promising molecular target for antiviral therapy.
Assuntos
Infecções por Coronavirus , Vírus da Bronquite Infecciosa , Animais , Vírus da Bronquite Infecciosa/genética , Imunidade Inata , Interferons , Replicação Viral , Transdução de Sinais , Infecções por Coronavirus/veterinária , GalinhasRESUMO
Nanoparticles (NPs) derived from RNA interference (RNAi) are considered a potentially revolutionary technique in the field of plant protection in the future. However, the application of NPs in RNAi is hindered by the conflict between the high cost of RNA production and the large quantity of materials required for field application. This study aimed to evaluate the antiviral efficacy of commercially available nanomaterials, such as chitosan quaternary ammonium salt (CQAS), amine functionalized silica nano powder (ASNP), and carbon quantum dots (CQD), that carried double-stranded RNA (dsRNA) via various delivery methods, including infiltration, spraying, and root soaking. ASNP-dsRNA NPs are recommended for root soaking, which is considered the most effective method of antiviral compound application. The most effective antiviral compound tested was CQAS-dsRNA NPs delivered by root soaking. Using fluorescence, FITC-CQAS-dsCP-Cy3, and CQD-dsCP-Cy3 NPs demonstrated the uptake and transport pathways of dsRNA NPs in plants when applied to plants in different modes. The duration of protection with NPs applied in various modes was then compared, providing references for evaluating the retention period of various types of NPs. All three types of NPs effectively silenced genes in plants and afforded at least 14 days of protection against viral infection. Particularly, CQD-dsRNA NPs could protect systemic leaves for 21 days following spraying.
Assuntos
Nanopartículas , Potyvirus , RNA de Cadeia Dupla , Potyvirus/genética , Antivirais/farmacologia , Interferência de RNARESUMO
Heterostructured TiO2@MXene rich in oxygen vacancies defects (VO-TiO2@MXene) has been developed to construct an electrochemical sensing platform for imidacloprid (IMI) determination. For the material design, TiO2 nanoparticles were firstly in situ grown on MXene and used as a scaffolding to prevent the stack of MXene nanosheets. The obtained TiO2@MXene heterostructure displays excellent layered structure and large specific surface area. After that, electrochemical activation is utilized to treat TiO2@MXene, which greatly increases the concentration of surface oxygen vacancies (VOs), thereby remarkably enhancing the conductivity and adsorption capacity of the composite. Accordingly, the prepared VO-TiO2@MXene displays excellent electrocatalytic activity toward the reduction of IMI. Under optimum conditions, cyclic voltammetry and linear sweep voltammetry techniques were utilized to investigate the electrochemical behavior of IMI at the VO-TiO2@MXene/GCE. The proposed sensor based on VO-TiO2@MXene presents an obvious reduction peak at -1.05 V(vs. Hg|Hg2Cl2) with two linear ranges from 0.07 - 10.0 µM and 10.0 - 70.0 µM with a detection limit of 23.3 nM (S/N= 3). Furthermore, the sensor provides a reliable result for detecting IMI in fruit and vegetable samples with a recovery of 97.9-103% and RSD≤ 4.3%. A sensitive electrochemical sensing platform was reported for imidacloprid (IMI) determination based on heterostructured TiO2@MXene rich in oxygen vacancy defects.
Assuntos
Oxigênio , Verduras , Frutas , Técnicas Eletroquímicas/métodosRESUMO
Simultaneously being a nonradiative and noninvasive technique makes magnetic resonance imaging (MRI) one of the highly required imaging approaches for the early diagnosis and follow-up of tumors, specifically for brain cancer. Paramagnetic gadolinium (Gd)-based contrast agents (CAs) are the most widely used ones in brain MRI acquisitions with special interest when assessing blood-brain barrier (BBB) integrity, a characteristic of high-grade tumors. However, alternatives to Gd-based contrast agents (CAs) are highly required to overcome their established toxicity. Organic radicals anchored on a dendrimer macromolecule surface (radical dendrimers) are promising alternatives since they also exhibit paramagnetic properties and can act as T1 CAs like Gd-based CAs while being organic species (mitigating concerns about toxic metal accumulation). Here, we studied the third generation of a water-soluble family of poly(phosphorhydrazone) radical dendrimers, with 48 PROXYL radical units anchored on their branches, exploring their potential of ex vivo and in vivo contrast enhancement in brain tumors (in particular, of immunocompetent, orthotopic GL261 murine glioblastoma (GB)). Remarkably, this radical species provides suitable contrast enhancement on murine GL261 GB tumors, which was comparable to that of commercial Gd-based CAs (at standard dose 0.1 mmol/kg), even at its 4 times lower administered dose (0.025 mmol/kg). Importantly, no signs of toxicity were detected in vivo. In addition, it showed a selective accumulation in brain tumor tissues, exhibiting longer retention within the tumor, which allows performing imaging acquisition over longer time frames (≥2.5 h) as opposed to Gd chelates. Finally, we observed high stability of the radicals in biological media, on the order of hours instead of minutes, characteristic of the isolated radicals. All of these features allow us to suggest that the G3-Tyr-PROXYL-ONa radical dendrimer could be a viable alternative to metal-based MRI contrast agents, particularly on MRI analysis of GB, representing, to the best of our knowledge, the first case of organic radical species used for this purpose and one of the very few examples of these types of radical species working as MRI CAs in vivo.
Assuntos
Neoplasias Encefálicas , Dendrímeros , Glioblastoma , Animais , Neoplasias Encefálicas/diagnóstico por imagem , Meios de Contraste , Radicais Livres , Glioblastoma/diagnóstico por imagem , Humanos , Imageamento por Ressonância Magnética/métodos , Metais , CamundongosRESUMO
A novel positive-sense single-stranded RNA mycovirus, designated as "Magnaporthe oryzae botourmiavirus 10" (MoBV10), was identified in the rice blast fungus Magnaporthe oryzae isolate HF04. MoBV10 has a single genomic RNA segment consisting of 2,448 nucleotides, which contains a single open reading frame encoding an RNA-dependent RNA polymerase. Genome comparison and phylogenetic analysis indicated that MoBV10 is a new member of the genus Betascleroulivirus in the family Botourmiaviridae. The 5'- and 3'-terminal sequences of the genomic RNA of MoBV10 have inverted complementarity and potentially form a panhandle structure, which is very rare in RNA viruses.
Assuntos
Magnaporthe , Oryza , Vírus de RNA , Ascomicetos , Genoma Viral , Magnaporthe/genética , Oryza/microbiologia , Filogenia , Doenças das Plantas/microbiologia , RNA Viral/genéticaRESUMO
Two double stranded RNAs (dsRNAs) that likely represent the genome of an alphapartitivirus, tentatively named "impatiens cryptic virus 1" (ICV1), were recovered from Impatiens balsamina L. RNA1 (2008 bp) codes for the RNA-dependent RNA polymerase (RdRp) of ICV1, which shares <83% amino acid sequence identity with the RdRps of other alphapartitiviruses. RNA2 (1906 bp) codes for the coat protein (CP) of ICV1, which shares <60% amino acid sequence identity with the CPs of other alphapartitiviruses. Phylogenetic analysis suggested that ICV1 is closely related to plant alphapartitiviruses, including vicia cryptic virus, beet cryptic virus 1, carrot cryptic virus, and white clover cryptic virus 1. Using primers specific for RNA1 or RNA2, ICV1 could be detected in I. balsamina from various parts of China.
Assuntos
Impatiens , Vírus de RNA , Genoma Viral , Impatiens/genética , Filogenia , Doenças das Plantas , Vírus de RNA/genética , RNA de Cadeia Dupla/genética , RNA Viral/genética , RNA Polimerase Dependente de RNA/genéticaRESUMO
Fusarium pseudograminearum is a phytopathogen that causes wheat crown rot disease worldwide. Fusarium pseudograminearum megabirnavirus 1 (FpgMBV1) was isolated from the hypovirulent strain FC136-2A of F. pseudograminearum as a novel double-stranded RNA mycovirus belonging to the family Megabirnaviridae. Here we examined the effects of FpgMBV1 on colony morphology and pathogenicity of F. pseudograminearum. Through hyphal tip culture, we obtained virus-free progeny of strain FC136-2A, referred to as FC136-2A-V-. FpgMBV1 was transferred horizontally to another virus-free strain, WZ-8A-HygR-V-. The progeny obtained through horizontal transfer was referred to as WZ-8A-HygR-V+. Colony morphology was similar between the FpgMBV1-positive and -negative strains. The ability to penetrate cellophane in vitro was lost, and pathogenicity on wheat plants was reduced significantly in the FpgMBV1-positive strains relative to the FpgMBV1-negative strains. Microscopic observations showed a 6-h delay in the formation of appressoria-like structures in FC136-2A relative to FC136-2A-V-. Mycelium extension was significantly longer in wheat coleoptiles infected by WZ-8A-HygR-V- than in that infected by WZ-8A-HygR-V+ at 12 and 20 h after inoculation (hai). In addition, expression of five genes that encode cell wall-degrading enzymes differed significantly between FpgMBV1-positive and -negative strains at 12 and 20 hai during early infection of wheat cells by conidia. This study provides evidence for the hypovirulence effect of FpgMBV1 on F. pseudograminearum and suggests that the underlying mechanism involves unsuccessful early infection and perhaps cell wall degradation.
Assuntos
Fusarium , Vírus de RNA , Doenças das Plantas/genética , Triticum/genética , VirulênciaRESUMO
The structural DNA nanotechnology holds great potential application in bioimaging, drug delivery and cancer therapy. Herein, an intelligent aptamer-incorporated DNA nanonetwork (Apt-Nnes) is demonstrated for cancer cell imaging and targeted drug delivery, which essentially is a micron-scale pattern with the thickness of double-stranded monolayer. Cancer cell-surface receptors can make it perform magical transformation into small size of nanosheet intermediates and specifically enter target cells. The binding affinity of Apt-Nnes is increased by 3-fold due to multivalent binding effect of aptamers and it can maintain the structural integrity in fetal bovine serum (FBS) for 8â¯h. More interestingly, target cancer cells can cause the structural disassembly, and each resulting unit transports 4963 doxorubicin (Dox) into target cells, causing the specific cellular cytotoxicity. The cell surface receptor-mediated disassembly of large size of DNA nanostructures into small size of fractions provides a valuable insight into developing intelligent DNA nanostructure suitable for biomedical applications.
Assuntos
Aptâmeros de Nucleotídeos , Neoplasias , Aptâmeros de Nucleotídeos/química , Linhagem Celular Tumoral , DNA/química , Doxorrubicina , Sistemas de Liberação de Medicamentos/métodos , Neoplasias/tratamento farmacológicoRESUMO
A facile and sensitive electrochemical aptamer sensor (aptasensor) based on Au nanoparticles-decorated porous carbon (AuNPs/PC) composite was developed for the efficient determination of the antibiotic drug chloramphenicol (CAP). AuNPs modified metal-organic framework (AuNPs/ZIF-8) is applied as a precursor to synthesize the porous carbon with homogeneous AuNPs distribution through a direct carbonization step under nitrogen atmosphere. The as-synthesized AuNPs/PC exhibits high surface area and improved conductivity. Moreover, the loading AuNPs could enhance the attachment of the aptamers on the surface of electrode through the Au-S bond. When added to CAP, poorly conductive aptamer-CAP complexes are formed on the sensor surface, which increases the hindrance to electron transfer resulting in a decrease in electrochemical signal. Based on this mechanism, the developed CAP aptasensor represents a wide linear detection range of 0.1 pM to 100 nM with a low detection limit of 0.03 pM (S/N = 3). In addition, the proposed aptasensor was employed for the analysis of CAP in honey samples and provided satisfactory recovery.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Grafite , Nanopartículas Metálicas , Estruturas Metalorgânicas , Estruturas Metalorgânicas/química , Ouro/química , Cloranfenicol , Carbono/química , Aptâmeros de Nucleotídeos/química , Técnicas Eletroquímicas/métodos , Técnicas Biossensoriais/métodos , Porosidade , Nanopartículas Metálicas/química , Limite de Detecção , Grafite/química , Nitrogênio/química , AntibacterianosRESUMO
A simple and label-free electrochemical aptasensor was developed for ultra-sensitive determination of chloramphenicol (CAP) based on a 2D transition of metal carbides (MXene) loaded with gold nanoparticles (AuNPs). The embedded AuNPs not only inhibit the aggregation of MXene sheets, but also improve the quantity of active sites and electronic conductivity. The aptamers (Apts) were able to immobilize on the MXene-AuNP modified electrode surface through Au-S interaction. Upon specifically binding with CAP with high affinity, the CAP-Apt complexes produced low conductivity on the aptasensor surface, leading to a decreased electrochemical signal. The resulting current change was quantitatively correlated with CAP concentration. Under optimized experimental conditions, the constructed aptasensor exhibited a good linear relationship within a wide range of 0.0001-10 nM and with a low detection limit of 0.03 pM for CAP. Moreover, the developed aptasensor has been applied to the determination of CAP concentration in honey samples with satisfactory results.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Mel , Nanopartículas Metálicas , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Cloranfenicol/análise , Técnicas Eletroquímicas/métodos , Ouro/química , Nanopartículas Metálicas/químicaRESUMO
Beauveria bassiana, an entomopathogenic fungus, is used for arthropod pest control worldwide. Here, we report the discovery and characterization of a novel double-stranded RNA (dsRNA) mycovirus, Beauveria bassiana chrysovirus 2 (BbCV-2), isolated from a Chinese B. bassiana strain. The genome sequence of the virus was determined by metagenomic sequencing, RT-PCR, and RACE cloning and was found to consist of four dsRNA segments that are 3441 bp, 2779 bp, 2925 bp, and 2688 bp long, respectively. Each dsRNA segment contains a single ORF. The ORF of dsRNA1 encodes a 1114-amino-acid (aa) protein (123.4 kDa) with a conserved RNA-dependent RNA polymerase (RdRp) motif, the sequence of which showed the highest identity of only 16.13% to that of Beauveria bassiana chrysovirus-1 (BbCV-1). The ORF of dsRNA2 encodes an 805-aa coat protein (CP) (84.7 kDa). The ORFs of dsRNAs 3 and 4 encodes proteins of undetermined function. The virus is a new member of the family Chrysoviridae from B. bassiana.