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1.
Plant Cell ; 36(4): 987-1006, 2024 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-37831412

RESUMO

Plant immunity is fine-tuned to balance growth and defense. However, little is yet known about molecular mechanisms underlying immune homeostasis in rice (Oryza sativa). In this study, we reveal that a rice calcium-dependent protein kinase (CDPK), OsCPK17, interacts with and stabilizes the receptor-like cytoplasmic kinase (RLCK) OsRLCK176, a close homolog of Arabidopsis thaliana BOTRYTIS-INDUCED KINASE 1 (AtBIK1). Oxidative burst and pathogenesis-related gene expression triggered by pathogen-associated molecular patterns are significantly attenuated in the oscpk17 mutant. The oscpk17 mutant and OsCPK17-silenced lines are more susceptible to bacterial diseases than the wild-type plants, indicating that OsCPK17 positively regulates rice immunity. Furthermore, the plant U-box (PUB) protein OsPUB12 ubiquitinates and degrades OsRLCK176. OsCPK17 phosphorylates OsRLCK176 at Ser83, which prevents the ubiquitination of OsRLCK176 by OsPUB12 and thereby enhances the stability and immune function of OsRLCK176. The phenotypes of the ospub12 mutant in defense responses and disease resistance show that OsPUB12 negatively regulates rice immunity. Therefore, OsCPK17 and OsPUB12 reciprocally maintain OsRLCK176 homeostasis and function as positive and negative immune regulators, respectively. This study uncovers positive cross talk between CDPK- and RLCK-mediated immune signaling in plants and reveals that OsCPK17, OsPUB12, and OsRLCK176 maintain rice immune homeostasis.


Assuntos
Oryza , Oryza/metabolismo , Resistência à Doença , Imunidade Vegetal/genética , Transdução de Sinais/fisiologia , Homeostase , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Doenças das Plantas/microbiologia , Regulação da Expressão Gênica de Plantas
2.
BMC Biol ; 22(1): 68, 2024 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-38520013

RESUMO

BACKGROUND: The brown planthopper (BPH) is a kind of piercing-sucking insect specific to rice, with the damage tops the list of pathogens and insects in recent years. microRNAs (miRNAs) are pivotal regulators of plant-environment interactions, while the mechanism underlying their function against insects is largely unknown. RESULTS: Here, we confirmed that OsmiR319, an ancient and conserved miRNA, negatively regulated resistance to BPHs, with overexpression of OsmiR319 susceptible to BPH, while suppression of OsmiR319 resistant to BPH in comparison with wild type. Meanwhile, we identified several targets of OsmiR319 that may mediate BPH resistance. Among them, OsPCF5 was the most obviously induced by BPH feeding, and over expression of OsPCF5 was resistance to BPH. In addition, various biochemical assays verified that OsPCF5 interacted with several MYB proteins, such as OsMYB22, OsMYB30, and OsMYB30C.Genetically, we revealed that both OsMYB22 and OsMYB30C positively regulated BPH resistance. Genetic interaction analyses confirmed that OsMYB22 and OsMYB30C both function in the same genetic pathway with OsmiR319b to mediate BPH resistance. CONCLUSIONS: Altogether, we revealed that OsPCF5 regulates BPH resistance via association with several MYB proteins downstream of OsmiR319, these MYB proteins might function as regulators of BPH resistance through regulating the phenylpropane synthesis.


Assuntos
Hemípteros , MicroRNAs , Oryza , Animais , Oryza/fisiologia , Hemípteros/genética , MicroRNAs/genética , MicroRNAs/metabolismo
3.
Anal Chem ; 96(4): 1717-1724, 2024 01 30.
Artigo em Inglês | MEDLINE | ID: mdl-38217876

RESUMO

In this work, we demonstrate for the first time the application of the phosphorothioated-terminal hairpin formation and self-priming extension (PS-THSP) reaction for miRNA assays. A self-priming amplification accelerating CRISPR sensor was well-established for sensitive and specific miRNA detection by integrating the PS-THSP reaction and CRISPR/Cas12a system. The sensor consists of three steps: (1) the formation of a complete PS-THSP template in the presence of target miRNA and ligase; (2) the exponential isothermal amplification of the PS-THSP reaction under the action of DNA polymerase; (3) the activation of the CRISPR/Cas12a fluorescence system to generate signals. We used miR-21 as a model target. The sensor can achieve sensitive detection of miR-21 without the involvement of any primers, and the special design of the CRISPR proto-spacer neighbor motif (PAM) sequence effectively avoids the interference of the background signal. In addition, the sensor can not only identify single-base mutant homologous sequences but also show stable performance in complex biological matrices. We have successfully used this sensor to accurately analyze miR-21 in different cell lines and real clinical samples, demonstrating its great potential in clinical diagnosis.


Assuntos
Técnicas Biossensoriais , MicroRNAs , Bioensaio , Linhagem Celular , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Primers do DNA , MicroRNAs/genética , Técnicas de Amplificação de Ácido Nucleico
4.
New Phytol ; 243(6): 2332-2350, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39056291

RESUMO

Protein posttranslational modifications play crucial roles in plant immunity through modulating a complicated signaling network mediated by different hormones. We previously demonstrated that OsATL32, an ATL-type E3 ligase, negatively contributes to rice immunity against Magnaporthe oryzae. Here, we show that OsATL32 forms a loop with OsPPKL2 and OsGSK2 through distinct protein posttranslational modifications to modulate rice immunity. OsATL32 ubiquitinates OsPPKL2, a protein phosphatase with Kelch-like repeat domains that exerts positive roles in regulating rice immunity against M. oryzae and chitin-triggered immune responses, for degradation. The glycogen synthase kinase 2 (OsGSK2), which acts as a negative regulator of rice immunity against M. oryzae and chitin-triggered immune responses, phosphorylates OsATL32 to elevate its protein stability and E3 ligase activity on OsPPKL2. Moreover, OsPPKL2 directly dephosphorylates OsGSK2, affecting its kinase activity on substrates including OsATL32 for phosphorylation. Like OsGSK2 as a BR signaling repressor, OsATL32 negatively regulates BR signaling; conversely, OsPPKL2 plays a positive role in BR signaling. These findings provide a molecular mechanism in which OsATL32 serves as a node connecting BR signaling and immunity by associating with OsPPKL2 and OsGSK2, assembling into a distinct protein posttranslational modifications-linked loop that functions in rice BR signaling and immunity.


Assuntos
Oryza , Doenças das Plantas , Imunidade Vegetal , Proteínas de Plantas , Processamento de Proteína Pós-Traducional , Oryza/genética , Oryza/imunologia , Oryza/microbiologia , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Doenças das Plantas/microbiologia , Doenças das Plantas/imunologia , Fosforilação , Ubiquitinação , Transdução de Sinais , Magnaporthe/fisiologia , Brassinosteroides/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitina-Proteína Ligases/genética , Regulação da Expressão Gênica de Plantas , Quitina/metabolismo , Quinases da Glicogênio Sintase/metabolismo , Fosfoproteínas Fosfatases/metabolismo , Fosfoproteínas Fosfatases/genética , Ascomicetos
5.
Langmuir ; 40(16): 8721-8729, 2024 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-38598618

RESUMO

Experimental studies have demonstrated that the gas phase contact angle (CA) of a surface nanobubble (SNB) is much smaller than that of a macroscopic gas bubble. This reduced CA plays a crucial role in prolonging the lifetime of SNBs by lowering the bubble pressure and preventing gas molecules from dissolving in the surrounding liquids. Despite extensive efforts to explain the anomalously small CA, a consensus about the underlying reasons is yet to be reached. In this study, we conducted experimental investigations to explore the influence of gas molecules adsorbed at the solid-liquid interface on the CA of SNBs created through the solvent exchange (SE) method and temperature difference (TD). Interestingly, no significant change is observed in the CA of SNBs on highly oriented pyrolytic graphite (HOPG) surfaces. Even for nanobubbles on micro/nano pancakes, the CA only exhibited a slight reduction compared to SNBs on bare HOPG surfaces. These findings suggest that gas adsorption at the immersed solid surface may not be the primary factor contributing to the small CA of the SNBs. Furthermore, the CA of SNBs formed on polystyrene (PS) and octadecyltrichlorosilane (OTS) substrates was also investigated, and a considerable increase in CA was observed. In addition, the effects of other factors including impurity, electric double layer (EDL) line tension, and pinning force upon the CA of SNBs were discussed, and a comprehensive model about multiple factors affecting the CA of SNBs was proposed, which is helpful for understanding the abnormally small CA and the stability of SNBs.

6.
Physiol Plant ; 176(4): e14456, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39072778

RESUMO

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


Assuntos
Arabidopsis , Botrytis , Resistência à Doença , Regulação da Expressão Gênica de Plantas , Doenças das Plantas , Proteínas de Plantas , Pseudomonas syringae , Arabidopsis/genética , Arabidopsis/microbiologia , Doenças das Plantas/microbiologia , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Resistência à Doença/genética , Botrytis/fisiologia , Botrytis/patogenicidade , Pseudomonas syringae/fisiologia , Pseudomonas syringae/patogenicidade , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Cucurbitaceae/microbiologia , Cucurbitaceae/genética , Filogenia , Plantas Geneticamente Modificadas
7.
Neurol Sci ; 45(7): 3093-3105, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38381393

RESUMO

Post-sepsis psychiatric disorder, encompassing anxiety, depression, post-traumatic stress disorder and delirium, is a highly prevalent complication secondary to sepsis, resulting in a marked increase in long-term mortality among affected patients. Regrettably, psychiatric impairment associated with sepsis is frequently disregarded by clinicians. This review aims to summarize recent advancements in the understanding of the pathophysiology, prevention, and treatment of post-sepsis mental disorder, including coronavirus disease 2019-related psychiatric impairment. The pathophysiology of post-sepsis psychiatric disorder is complex and is known to involve blood-brain barrier disruption, overactivation of the hypothalamic-pituitary-adrenal axis, neuroinflammation, oxidative stress, neurotransmitter dysfunction, programmed cell death, and impaired neuroplasticity. No unified diagnostic criteria for this disorder are currently available; however, screening scales are often applied in its assessment. Modifiable risk factors for psychiatric impairment post-sepsis include the number of experienced traumatic memories, the length of ICU stay, level of albumin, the use of vasopressors or inotropes, daily activity function after sepsis, and the cumulative dose of dobutamine. To contribute to the prevention of post-sepsis psychiatric disorder, it may be beneficial to implement targeted interventions for these modifiable risk factors. Specific therapies for this condition remain scarce. Nevertheless, non-pharmacological approaches, such as comprehensive nursing care, may provide a promising avenue for treating psychiatric disorder following sepsis. In addition, although several therapeutic drugs have shown preliminary efficacy in animal models, further confirmation of their potential is required through follow-up clinical studies.


Assuntos
Transtornos Mentais , Sepse , Humanos , COVID-19/complicações , Delírio/etiologia , Delírio/terapia , Delírio/prevenção & controle , Delírio/fisiopatologia , Transtornos Mentais/etiologia , Transtornos Mentais/terapia , SARS-CoV-2 , Sepse/complicações , Sepse/fisiopatologia , Sepse/terapia , Transtornos de Estresse Pós-Traumáticos/terapia , Transtornos de Estresse Pós-Traumáticos/fisiopatologia , Transtornos de Estresse Pós-Traumáticos/etiologia
8.
Ecotoxicol Environ Saf ; 285: 117117, 2024 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-39342753

RESUMO

Not all antibiotic resistance genes (ARGs) pose an ecological risk to their host animals. A standard should be developed to study which types of ARGs posed an ecological risk to wild animals under human disturbances (HDs). In this study, the golden snub-nosed monkeys (Rhinopithecus roxellana) were used as sentinel species. According to the animals-associated enrichment, mobility, and pathogenicity, the ARGs in habitat of sentinel species were divided into four levels. If the mobile and pathogenic ARGs that could be collinear with the metagenome-assembled genome (MAGs) in the gut of the sentinel species, the ARGs were defined as Rank I ARGs and they were considered to have ecological risk to sentinel species. Functional genes in the MAGs that collinear with the Rank I ARGs were used to predict the health risks of sentinel species. The ecological risk to sentinel species was present in 0.158 % of the ARGs-contigs in the habitat. Cultivation and villages, but not grazing, agriculture and ecotourism, increased the ecological risk of the ARGs to wild animals, The ability of gut microbiome to acquire mobile and pathogenic ARGs increased, as did the collinear functional genes, and the health risks of the wild animals also enhanced by the disturbances of cultivation and villages. Cultivation and villages increased the nutrient content of the soil, and they had a positive effect on the ecological risk of Rank I ARGs by affecting the mobile genetic elements (MGEs), microbiome and the resistant group in the habitat, which was why the cultivation and villages increased the health risks of wild animals. We proposed that cultivation and living should be controlled, while grazing, agriculture and ecotourism could be developed in nature reserves of wild animals, but the nutrients in the wild animals' habitat should be monitored.


Assuntos
Animais Selvagens , Resistência Microbiana a Medicamentos , Ecossistema , Microbioma Gastrointestinal , Animais , Microbioma Gastrointestinal/efeitos dos fármacos , Resistência Microbiana a Medicamentos/genética , Humanos , Animais Selvagens/microbiologia , Medição de Risco , Colobinae/microbiologia , Colobinae/genética , Metagenômica , Metagenoma , Monitoramento Ambiental , Antibacterianos/farmacologia , Antibacterianos/toxicidade
9.
Plant Dis ; 108(6): 1461-1469, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38240714

RESUMO

Northern corn leaf blight (NCLB), caused by Exserohilum turcicum, is one of the most devastating foliar diseases of maize. Rapid and accurate diagnosis for this disease is urgently needed but still limited. Here, we establish a field-deployable diagnostic method to detect E. turcicum based on loop-mediated isothermal amplification (LAMP) assays. A software application called K-mer Elimination by Cross-reference was used to search for the specific sequences belonging to E. turcicum by comparing the whole genome sequence between E. turcicum and other known maize pathogens. Five LAMP primer sets were designed based on specific and single-copy fragments of E. turcicum. Post-LAMP analyses indicated that only the primer set, Et9468_set1, was the most suitable, producing a ladder-like amplification pattern in the agarose gel electrophoresis and a strong fluorescence signal in the presence of SYBR Green I. The LAMP assay using Et9468_set1 primers demonstrated a high level of specificity in distinguishing E. turcicum from six other common fungal pathogens of maize, as well as 12 more fungal and oomycete strains including the epiphytic fungi from maize leaves and other crop pathogens. Moreover, it exhibited remarkable sensitivity by detecting five copies per reaction, which was approximately 104 times more sensitive compared with conventional PCR. The LAMP assay successfully detected E. turcicum in field maize leaves without DNA extraction, demonstrating its suitability for rapid on-spot detection of NCLB. Our study provides a direct LAMP diagnostic method to detect E. turcicum, which enables on-site pathogen detection in the field and the development of preventive strategies for NCLB management.


Assuntos
Ascomicetos , Primers do DNA , Técnicas de Amplificação de Ácido Nucleico , Doenças das Plantas , Zea mays , Doenças das Plantas/microbiologia , Técnicas de Amplificação de Ácido Nucleico/métodos , Zea mays/microbiologia , Ascomicetos/genética , Ascomicetos/isolamento & purificação , Primers do DNA/genética , Folhas de Planta/microbiologia , Sensibilidade e Especificidade , DNA Fúngico/genética , Técnicas de Diagnóstico Molecular/métodos
10.
J Integr Plant Biol ; 66(7): 1459-1480, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38629772

RESUMO

Ubiquitination-mediated protein degradation is integral to plant immunity, with E3 ubiquitin ligases acting as key factors in this process. Here, we report the functions of OsATL32, a plasma membrane-localized Arabidopsis Tóxicos En Levadura (ATL)-type E3 ubiquitin ligase, in rice (Oryza sativa) immunity and its associated regulatory network. We found that the expression of OsATL32 is downregulated in both compatible and incompatible interactions between rice and the rice blast fungus Magnaporthe oryzae. The OsATL32 protein level declines in response to infection by a compatible M. oryzae strain or to chitin treatment. OsATL32 negatively regulates rice resistance to blast and bacterial leaf blight diseases, as well as chitin-triggered immunity. Biochemical and genetic studies revealed that OsATL32 suppresses pathogen-induced reactive oxygen species (ROS) accumulation by mediating ubiquitination and degradation of the ROS-producing OsRac5-OsRbohB module, which enhances rice immunity against M. oryzae. The protein phosphatase PHOSPHATASE AND TENSIN HOMOLOG enhances rice blast resistance by dephosphorylating OsATL32 and promoting its degradation, preventing its negative effect on rice immunity. This study provides insights into the molecular mechanism by which the E3 ligase OsATL32 targets a ROS-producing module to undermine rice immunity.


Assuntos
Oryza , Doenças das Plantas , Imunidade Vegetal , Proteínas de Plantas , Espécies Reativas de Oxigênio , Ubiquitinação , Oryza/microbiologia , Oryza/genética , Oryza/imunologia , Oryza/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Imunidade Vegetal/genética , Doenças das Plantas/microbiologia , Doenças das Plantas/imunologia , Ubiquitina-Proteína Ligases/metabolismo , Ubiquitina-Proteína Ligases/genética , Regulação da Expressão Gênica de Plantas , Resistência à Doença/genética , Ascomicetos
11.
J Integr Plant Biol ; 66(9): 2017-2041, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38953747

RESUMO

NAC transcription factors (TFs) are pivotal in plant immunity against diverse pathogens. Here, we report the functional and regulatory network of MNAC3, a novel NAC TF, in rice immunity. MNAC3, a transcriptional activator, negatively modulates rice immunity against blast and bacterial leaf blight diseases and pathogen-associated molecular pattern (PAMP)-triggered immune responses. MNAC3 binds to a CACG cis-element and activates the transcription of immune-negative target genes OsINO80, OsJAZ10, and OsJAZ11. The negative function of MNAC3 in rice immunity depends on its transcription of downstream genes such as OsINO80 and OsJAZ10. MNAC3 interacts with immunity-related OsPP2C41 (a protein phosphatase), ONAC066 (a NAC TF), and OsDjA6 (a DnaJ chaperone). ONAC066 and OsPP2C41 attenuate MNAC3 transcriptional activity, while OsDjA6 promotes it. Phosphorylation of MNAC3 at S163 is critical for its negative functions in rice immunity. OsPP2C41, which plays positive roles in rice blast resistance and chitin-triggered immune responses, dephosphorylates MNAC3, suppressing its transcriptional activity on the target genes OsINO80, OsJAZ10, and OsJAZ11 and promoting the translocation of MNAC3 from nucleus to cytoplasm. These results establish a MNAC3-centered regulatory network in which OsPP2C41 dephosphorylates MNAC3, attenuating its transcriptional activity on downstream immune-negative target genes in rice. Together, these findings deepen our understanding of molecular mechanisms in rice immunity and offer a novel strategy for genetic improvement of rice disease resistance.


Assuntos
Resistência à Doença , Regulação da Expressão Gênica de Plantas , Oryza , Doenças das Plantas , Imunidade Vegetal , Proteínas de Plantas , Fatores de Transcrição , Oryza/genética , Oryza/microbiologia , Oryza/imunologia , Oryza/metabolismo , Doenças das Plantas/microbiologia , Doenças das Plantas/imunologia , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Imunidade Vegetal/genética , Fatores de Transcrição/metabolismo , Fatores de Transcrição/genética , Resistência à Doença/genética , Redes Reguladoras de Genes , Fosforilação
12.
Zhongguo Zhong Yao Za Zhi ; 49(2): 361-369, 2024 Jan.
Artigo em Zh | MEDLINE | ID: mdl-38403312

RESUMO

The 4-coumarate: CoA ligase(4CL) is a key enzyme in the upstream pathway of phenylpropanoids such as flavonoids, soluble phenolic esters, lignans, and lignins in plants. In this study, 13 4CL family members of Arabidopsis thaliana were used as reference sequences to identify the 4CL gene family candidate members of Isatis indigotica from the reported I. indigotica genome. Further bioinformatics analysis and analysis of the expression pattern of 4CL genes and the accumulation pattern of flavonoids were carried out. Thirteen 4CL genes were obtained, named Ii4CL1-Ii4CL13, which were distributed on chromosomes 1, 2, 3, 4, and 6. The analysis of the gene structure and conserved structural domains revealed the intron number of I. indigotica 4CL genes was between 1 and 12 and the protein structural domains were highly conserved. Cis-acting element analysis showed that there were multiple response elements in the promoter sequence of I. indigotica 4CL gene family, and jasmonic acid had the largest number of reaction elements. The collinearity analysis showed that there was a close relationship between the 4CL gene family members of I. indigotica and A. thaliana. As revealed by qPCR results, the expression analysis of the 4CL gene family showed that 10 4CL genes had higher expression levels in the aboveground part of I. indigotica. The content assay of flavonoids in different parts of I. indigotica showed that flavonoids were mainly accumulated in the aboveground part of plants. This study provides a basis for further investigating the roles of the 4CL gene family involved in the biosynthesis of flavonoids in I. indigotica.


Assuntos
Isatis , Ligases , Ligases/genética , Isatis/genética , Regiões Promotoras Genéticas , Plantas/metabolismo , Flavonoides , Coenzima A Ligases/genética , Coenzima A Ligases/química , Coenzima A Ligases/metabolismo
13.
Anal Chem ; 95(7): 3606-3612, 2023 02 21.
Artigo em Inglês | MEDLINE | ID: mdl-36565296

RESUMO

Exosome-based liquid biopsy technologies play an increasingly prominent role in tumor diagnosis. However, the simple and sensitive method for counting exosomes still faces considerable challenges. In this work, the CD63 aptamer-modified DNA tetrahedrons on the gold electrode were used as recognition elements for the specific capture of exosomes. Partially complementary DNA probes act as bridges linking trapped exosomes and three AuNP-DNA signal probes. This clover-like structure can tackle the recognition and sensitivity issues arising from the undesired AuNP aggregation event. When cancerous exosomes are present in the system, the high accumulation of methylene blue molecules from DNA-AuNP nanocomposites on the surface of the electrode leads to an intense current signal. According to the results, the aptasensor responds to MCF-7 cell-derived exosomes in the concentration range from 1.0 × 103 to 1.0 × 108 particles·µL-1, with the detection limit of 158 particles·µL-1. Furthermore, the aptasensor has been extended to serum samples from breast cancer patients and exhibited excellent specificity. To sum it up, the aptasensor is sensitive, straightforward, less expensive, and fully capable of receiving widespread application in clinics for tumor monitoring.


Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Neoplasias da Mama , Exossomos , Nanopartículas Metálicas , Humanos , Feminino , Exossomos/química , Ouro/química , Neoplasias da Mama/diagnóstico , DNA/análise , Células MCF-7 , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Nanopartículas Metálicas/química
14.
Anal Chem ; 95(2): 1549-1555, 2023 01 17.
Artigo em Inglês | MEDLINE | ID: mdl-36598887

RESUMO

Long non-coding RNAs (lncRNAs) played vital roles in physiological and pathological conditions. Consistent results from cell experiments, animal experiments, and clinical studies suggested that lncRNA HULC was an oncogenic lncRNA serving as a potential diagnostic and prognostic marker of hepatocellular carcinoma. In this study, we developed a fluorescent biosensor for lncRNA HULC detection based on rolling circle amplification (RCA) induced by multi-primer probes. Multiple primer probes can not only combine with lncRNA to break its secondary structure, which was conducive to lncRNA captured by Y-shaped probes, but also trigger multiple RCA reactions to achieve signal amplification and the goal of sensitive detection of lncRNA. Compared to previous detection methods, in this scheme, we took advantage of the long sequence characteristics of lncRNA to make it a carrier that can bind multiple primers to initiate RCA. This newly designed biosensor provided a linear range from 1 pM to 100 nM with a detection limit of 0.06 pM. This method can provide a new idea for the application of isothermal amplification in detecting lncRNA. Furthermore, the application of the biosensor in liver cancer cell lines and whole blood samples from hepatocellular carcinomatosis patients also confirmed that the method had good selectivity and sensitivity to lncRNA HULC. This method offered a new way for transforming specific lncRNA into clinical application for diagnosis, prognosis, or predicting treatment response.


Assuntos
Técnicas Biossensoriais , Carcinoma Hepatocelular , Neoplasias Hepáticas , RNA Longo não Codificante , Animais , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/genética , RNA Longo não Codificante/genética , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/genética , Detecção Precoce de Câncer , Primers do DNA/metabolismo , Técnicas Biossensoriais/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Limite de Detecção
15.
Anal Chem ; 95(23): 9006-9013, 2023 06 13.
Artigo em Inglês | MEDLINE | ID: mdl-37261725

RESUMO

Due to its high efficiency and selectivity, cell-free biosynthesis has found broad utility in the fields of bioproduction, environment monitoring, and disease diagnostics. However, the practical application is limited by its low productivity. Here, we introduce the entropy-driven assembly of transcription templates as dynamic amplifying modules to accelerate the cell-free transcription process. The catalytic DNA circuit with high sensitivity and enzyme-free format contributes to the production of large amounts of transcription templates, drastically accelerating the as-designed cell-free transcription system without interference from multiple enzymes. The proposed approach was successfully applied to the ultrasensitive detection of SARS-CoV-2, improving the sensitivity by 3 orders of magnitude. Thanks to the high programmability and diverse light-up RNA pairs, the method can be adapted to multiplexing detection, successfully demonstrated by the analysis of two different sites of the SARS-CoV-2 gene in parallel. Further, the flexibility of the entropy-driven circuit enables a dynamic responding range by tuning the circuit layers, which is beneficial for responding to targets with different concentration ranges. The strategy was also applied to the analysis of clinical samples, providing an alternative for sensitively detecting the current SARS-CoV-2 RNA that quickly mutates.


Assuntos
Técnicas Biossensoriais , COVID-19 , Humanos , DNA/análise , Entropia , RNA Viral , SARS-CoV-2/genética , Técnicas Biossensoriais/métodos
16.
Small ; 19(15): e2207136, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36599658

RESUMO

The nano-enabled crop protecting agents have been emerging as a cost-effective, eco-friendly, and sustainable alternative to conventional chemical pesticides. Here, the antibacterial activity and disease-suppressive potential of biogenic copper nanoparticles (bio-CuNPs) against bacterial fruit blotch (BFB), caused by Acidovorax citrulli (Ac), in watermelon (Citrullus lanatus L.) is discussed. CuNPs are extracellularly biosynthesized using a locally isolated bacterial strain Bacillus altitudinis WM-2/2, and have spherical shapes of 29.11-78.56 nm. Various metabolites, such as alcoholic compounds, carboxylic acids, alkenes, aromatic amines, and halo compounds, stabilize bio-CuNPs. Foliar application of bio-CuNPs increases the Cu accumulation in shoots/roots (66%/27%), and promotes the growth performance of watermelon plants by improving fresh/dry weight (36%/39%), through triggering various imperative physiological and biochemical processes. Importantly, bio-CuNPs at 100 µg mL-1 significantly suppress watermelon BFB through balancing reactive oxygen species system, improving photosynthesis capacity, and modulating stomatal immunity. Bio-CuNPs show obvious antibacterial activity against Ac by inducing oxidative stress, biofilm inhibition, and cellular integrity disruption. These findings demonstrate that bio-CuNPs can suppress watermelon BFB through direct antibacterial activity and induction of active immune response in watermelon plants, and highlight the value of this approach as a powerful tool to increase agricultural production and alleviate food insecurity.


Assuntos
Citrullus , Citrullus/microbiologia , Frutas/microbiologia , Cobre , Doenças das Plantas/prevenção & controle , Doenças das Plantas/microbiologia , Antibacterianos/farmacologia
17.
Small ; 19(2): e2205687, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36382544

RESUMO

The use of nanofabricated materials is being explored for the potential in crop disease management. Chemically synthesized micronutrient nanoparticles (NPs) have been shown to reduce crop diseases; however, the potential of biogenic manganese NPs (bio-MnNPs) in disease control is unknown. Here, the potential and mechanism of bio-MnNPs in suppression of watermelon Fusarium wilt, caused by Fusarium oxysporum f. sp. niveum (Fon) are reported. Bio-MnNPs are synthesized by cell-free cultural filtrate of a waterrmelon rhizosphere bacterial strain Bacillus megaterium NOM14, and are found spherical in shape with a size range of 27.0-65.7 nm. Application of bio-MnNPs at 100 µg mL-1 increases Mn content in watermelon roots/shoots and improves growth performance through enhancing multiple physiological processes, including antioxidative capacity. Bio-MnNPs at 100 µg mL-1 suppress Fusarium wilt through inhibiting colonization and invasive growth of Fon in watermelon roots/stems, and inhibit Fon vegetative growth, conidiation, conidial morphology, and cellular integrity. Bio-MnNPs potentiate watermelon systemic acquired resistance by triggering the salicylic acid signaling upon Fon infection, and reshape the soil microbial community by improving fungal diversity. These findings demonstrate that bio-MnNPs suppress watermelon Fusarium wilt by multiple ex planta and in planta mechanisms, and offer a promising nano-enabled strategy for the sustainable management of crop diseases.


Assuntos
Citrullus , Fusarium , Citrullus/microbiologia , Solo , Fusarium/fisiologia , Manganês , Doenças das Plantas/prevenção & controle , Doenças das Plantas/microbiologia
18.
Plant Biotechnol J ; 21(10): 2125-2139, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37402218

RESUMO

The plant cell wall is the first line of defence against physical damage and pathogen attack. Wall-associated kinase (WAK) has the ability to perceive the changes in the cell wall matrix and transform signals into the cytoplasm, being involved in plant development and the defence response. Downy mildew, caused by Hyaloperonospora brassicae, can result in a massive loss in Chinese cabbage (Brassica rapa L. ssp. pekinensis) production. Herein, we identified a candidate resistant WAK gene, BrWAK1, in a major resistant quantitative trait locus, using a double haploid population derived from resistant inbred line T12-19 and the susceptible line 91-112. The expression of BrWAK1 could be induced by salicylic acid and pathogen inoculation. Expression of BrWAK1 in 91-112 could significantly enhance resistance to the pathogen, while truncating BrWAK1 in T12-19 increased disease susceptibility. Variation in the extracellular galacturonan binding (GUB) domain of BrWAK1 was found to mainly confer resistance to downy mildew in T12-19. Moreover, BrWAK1 was proved to interact with BrBAK1 (brassinosteroid insensitive 1 associated kinase), resulting in the activation of the downstream mitogen-activated protein kinase (MAPK) cascade to trigger the defence response. BrWAK1 is the first identified and thoroughly characterized WAK gene conferring disease resistance in Chinese cabbage, and the plant biomass is not significantly influenced by BrWAK1, which will greatly accelerate Chinese cabbage breeding for downy mildew resistance.


Assuntos
Brassica rapa , Brassica , Oomicetos , Brassica rapa/genética , Melhoramento Vegetal , Oomicetos/genética , Locos de Características Quantitativas , Resistência à Doença/genética , Brassica/genética , Doenças das Plantas/genética
19.
Crit Rev Food Sci Nutr ; 63(25): 7489-7509, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-35254111

RESUMO

Biotic and abiotic environmental stresses affect the production and quality of agricultural products worldwide. The extensive use of traditional preventive measures comprising toxic chemicals has become more problematic due to severe ecotoxicological challenges. To address this issue, engineered nanoparticles (NPs) with their distinct physical and chemical properties has gained scientific attention and can help plants to confront environmental challenges. Despite their ameliorative and beneficial effects, toxicological concerns have been raised about NPs. The recent development of biogenic NPs (bio-NPs) is getting attention in agriculture due to their diverse biocompatibility, better functional efficacy, and eco-friendly nature compared to the recalcitrant NPs, providing a promising strategy for increased crop protection against biotic and abiotic environmental stresses, with the ultimate goal of ensuring global food security. This review summarizes the recent advances in the engineering of bio-NPs with particular emphasis on the functions of bio-NPs in protecting plants from biotic and abiotic environmental stresses, delivery and entry routes of NPs to plant systems, nanotoxicity, and plant physiological/biochemical responses to nanotoxicity. Future perspectives of bio-NP-enabled strategies, remaining pitfalls, and possible solutions to combat environmental challenges via advanced nanotechnology to achieve global food security and a sustainable agricultural system are also discussed.


Assuntos
Nanopartículas , Plantas , Agricultura , Nanotecnologia , Segurança Alimentar
20.
Conserv Biol ; 37(4): e14069, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-36751969

RESUMO

Over the past 40 years, the climate has been changing and human disturbance has increased in the vast Qinghai-Tibet Plateau (QTP). These 2 factors are expected to affect the distribution of a large number of endemic vertebrate species. However, quantitative relationships between range shifts and climate change and human disturbance of these species in the QTP have rarely been evaluated. We used occurrence records of 19 terrestrial vertebrate species (birds, mammals, amphibians, and reptiles) occurring in the QTP from 1980 to 2020 to quantify the effects of climate change and anthropogenic impacts on the distribution of these 4 taxonomic groups and estimated species range changes in each species. The trend in distribution changes differed among the taxonomic groups, although, generally, ranges shifted to central QTP. Climate change contributed more to range variation than human disturbance (the sum of the 4 climatic variables contributed more than the sum of the 4 human disturbance variables for all 4 taxonomic groups). Suitable geographic range increased for most mammals, amphibians, and reptiles (+27.6%, +18.4%, and +27.8% on average, respectively), whereas for birds range decreased on average by 0.9%. Quantitative evidence for climate change and human disturbance associations with range changes for endemic vertebrate species in the QTP can provide useful insights into biodiversity conservation under changing environments.


En los últimos 40 años, el clima ha cambiado y las perturbaciones humanas han aumentado en la vasta meseta Qinghai-Tíbet (MQT). Se espera que estos dos factores afecten la distribución de un gran número de especies de vertebrados endémicos. Sin embargo, las relaciones cuantitativas entre los cambios del área de distribución y el cambio climático y las perturbaciones humanas en estas especies de la MQT han sido poco evaluadas. Utilizamos los registros de presencia de 19 especies de vertebrados terrestres (aves, mamíferos, anfibios y reptiles) de la MQT tomados entre 1980 y 2020 para cuantificar los efectos del cambio climático y los impactos antropogénicos sobre la distribución de estos cuatro grupos taxonómicos y estimar los cambios en el área de distribución de cada especie. La tendencia en los cambios de distribución difirió entre los grupos taxonómicos, aunque, en general, las áreas de distribución se desplazaron hacia el centro de la MQT. El cambio climático contribuyó más a la variación del área de distribución que las perturbaciones humanas (la suma de las cuatro variables climáticas contribuyó más que la suma de las cuatro variables de perturbaciones humanas para los cuatro grupos taxonómicos). El área de distribución geográfica adecuada aumentó para la mayoría de los mamíferos, anfibios y reptiles (+27.6%, +18.4% y +27.8% en promedio, respectivamente), mientras que para las aves disminuyó en promedio un 0.9%. Las pruebas cuantitativas de la asociación del cambio climático y las perturbaciones humanas con los cambios en el área de distribución de las especies vertebradas endémicas de la MQT pueden aportar información útil para la conservación de la biodiversidad en entornos cambiantes.


Assuntos
Efeitos Antropogênicos , Mudança Climática , Animais , Humanos , Tibet , Conservação dos Recursos Naturais , Vertebrados , Mamíferos
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