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Temporal transcriptional variation is a major contributor to functional evolution and the developmental process. Parthenogenetic water fleas of the genus Daphnia (Cladocera) provide an ideal model to characterize gene expression patterns across distinct developmental stages. Herein, we report RNA-seq data for female Daphnia mitsukuri at three developmental stages: the embryo, juvenile (three timepoints) and adult. Comparisons of gene expression patterns among these three developmental stages and weighted gene co-expression network analysis based on expression data across developmental stages identified sets of genes underpinning each of the developmental stages of D. mitsukuri. Specifically, highly expressed genes (HEGs) at the embryonic developmental stage were associated with cell proliferation, ensuring the necessary foundation for subsequent development; HEGs at the juvenile stages were associated with chemosensory perception, visual perception and neurotransmission, allowing individuals to enhance detection of potential environmental risks; HEGs at the adult stage were associated with antioxidative defensive systems, enabling adults to mount an efficient response to perceived environmental risks. Additionally, we found a significant overlap between expanded gene families of Daphnia species and HEGs at the juvenile stages, and these genes were associated with visual perception and neurotransmission. Our work provides a resource of developmental transcriptomes, and comparative analyses that characterize gene expression dynamics throughout development of Daphnia.
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Daphnia , Perfilación de la Expresión Génica , Humanos , Animales , Femenino , Daphnia/metabolismo , RNA-Seq , Transcriptoma , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
Ethylene response factors have been shown to be involved in the effects of plant developmental processes and to regulate stress tolerance. The aim of this study was to recognize the regulatory mechanisms of ethylene response factors on tobacco plant height. In this study, a gene-edited mutant (ERF10-KO) and wild type (WT) were utilized as experimental materials. Transcriptome and metabolome analyses were used to investigate the regulatory mechanism of NtERF10 gene editing on plant height in tobacco. Here, through the analysis of differentially expressed genes (DEGs), 2051 genes were upregulated and 1965 genes were downregulated. We characterized the different ERF10-KO and WT plant heights and identified key genes for photosynthesis, the plant hormone signal transduction pathway and the terpene biosynthesis pathway. NtERF10 was found to affect the growth and development of tobacco by regulating the expression levels of the PSAA, PSBA, GLY17 and GGP3 genes. Amino acid metabolism was analyzed by combining analyses of differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs). In addition, we found that members of the bHLH, NAC, MYB, and WRKY transcription factor families have vital roles in regulating plant height. This study not only provides important insights into the positive regulation of the ethylene response factor NtERF10 on plant height during plant growth and development but also provides new research ideas for tobacco molecular breeding.
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Regulación de la Expresión Génica de las Plantas , Nicotiana , Proteínas de Plantas , Factores de Transcripción , Nicotiana/genética , Nicotiana/crecimiento & desarrollo , Nicotiana/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Reguladores del Crecimiento de las Plantas/metabolismo , Reguladores del Crecimiento de las Plantas/genética , Etilenos/metabolismo , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , TranscriptomaRESUMEN
BACKGROUND: DFR is a crucial structural gene in plant flavonoid and polyphenol metabolism, and DFR knockout (DFR-KO) plants may have increased biomass accumulation. It is uncertain whether DFR-KO has comparable effects in tobacco and what the molecular mechanism is. We employed the CRISPR/Cas9 method to generate a knockout homozygous construct and collected samples from various developmental phases for transcriptome and metabolome detection and analysis. RESULTS: DFR-KO turned tobacco blossoms white on homozygous tobacco (Nicotiana tabacum) plants with both NtDFR1 and NtDFR2 knockout. RNA-seq investigation of anthesis leaf (LF), anthesis flower (FF), mature leaf (LM), and mature root (RM) variations in wild-type (CK) and DFR-KO lines revealed 2898, 276, 311, and 101 differentially expressed genes (DEGs), respectively. DFR-KO primarily affected leaves during anthesis. According to KEGG and GSEA studies, DFR-KO lines upregulated photosynthetic pathway carbon fixation and downregulated photosystem I and II genes. DFR-KO may diminish tobacco anthesis leaf photosynthetic light reaction but boost dark reaction carbon fixation. DFR-KO lowered the expression of pathway-related genes in LF, such as oxidative phosphorylation and proteasome, while boosting those in the plant-pathogen interaction and MAPK signaling pathways, indicating that it may increase biological stress resistance. DFR-KO greatly boosted the expression of other structural genes involved in phenylpropanoid production in FF, which may account for metabolite accumulation. The metabolome showed that LF overexpressed 8 flavonoid metabolites and FF downregulated 24 flavone metabolites. In DFR-KO LF, proteasome-related genes downregulated 16 amino acid metabolites and reduced free amino acids. Furthermore, the DEG analysis on LM revealed that the impact of DFR-KO on tobacco growth may progressively diminish with time. CONCLUSION: The broad impact of DFR-KO on different phases and organs of tobacco development was thoroughly and methodically investigated in this research. DFR-KO decreased catabolism and photosynthetic light reactions in leaves during the flowering stage while increasing carbon fixation and disease resistance pathways. However, the impact of DFR-KO on tobacco growth steadily declined as it grew and matured, and transcriptional and metabolic modifications were consistent. This work offers a fresh insight and theoretical foundation for tobacco breeding and the development of gene-edited strains.
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Nicotiana , Complejo de la Endopetidasa Proteasomal , Nicotiana/metabolismo , Complejo de la Endopetidasa Proteasomal/metabolismo , Fitomejoramiento , Flores , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Flavonoides/metabolismo , Regulación de la Expresión Génica de las PlantasRESUMEN
RATIONALE: Laser ablation multicollector inductively coupled plasma mass spectrometry (LA-MC-ICP-MS) has become a powerful technique for in situ Cu isotopic analysis in natural geological samples. Cu isotopic compositions in natural chalcopyrites have been used to reveal aspects of the mineralization processes directly. However, internationally or commercially available matrix-matched chalcopyrite reference materials for mass fractionation correction or quality control purposes are still lacking for in situ Cu isotopic analysis using LA-MC-ICP-MS. METHODS: Three natural chalcopyrites 14ZJ12-1, JGZ-29 and JGZ-78, and one copper metal GBW02141 with different Cu isotopic compositions were investigated as potential microanalytical reference materials by LA-MC-ICP-MS. Ga element was used as an internal standard to correct the mass fractionation of Cu isotopes during LA-MC-ICP-MS analysis. RESULTS: A large number of Cu isotope ratio measurements using femtosecond LA-MC-ICP-MS were conducted and produced good intermediate precision of δ65 CuNIST976 (0.07-0.08, 2 standard deviations), demonstrating the homogeneous Cu isotopic distribution in the recommended samples. The mean δ65 CuNIST976 values of -0.21 ± 0.04, 0.46 ± 0.04, -0.06 ± 0.04 and 0.11 ± 0.05 (2 standard deviations) in 14ZJ12-1, JGZ-29, JGZ-78 and GBW02141, respectively, were obtained using solution-MC-ICP-MS in the four recommended samples. CONCLUSIONS: Here, we describe three natural chalcopyrites 14ZJ12-1, JGZ-29, JGZ-78, and one copper metal GBW02141 as the potential Cu isotopic reference materials for LA-MC-ICP-MS analysis. Our analyses demonstrate that these recommended materials have a high degree of elemental and isotopic homogeneity, indicating that they are suitable for microanalysis techniques for data quality assurance or interlaboratory calibration.
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The biogeography and molecular phylogeny of invertebrate zooplankton populations from inland saline waters remains under-explored in the Eastern Palearctic, especially the Qinghai-Tibetan Plateau. Here, we surveyed the diversity of the Brachionus plicatilis Müller, 1786 species complex from inland saline waters across China. We compared morphometrics with DNA taxonomy (using two genetic markers: the mitochondrial cytochrome c oxidase subunit I (COI) gene and the nuclear internal transcribed spacer (ITS-1)). Our phylogenies based on the sequences of ITS-1 recognized two distinct clades (i.e. two species: B. plicatilis sensu stricto (s.s.) and B. asplanchnoidis) in China. We detected two mitochondrial clades within B. plicatilis s.s and one within B. asplanchnoidis across China, consistent with the three morphogroups present. One of these three clades was novel and restricted to the Qinghai-Tibetan Plateau, where it exhibited evidence of recent expansion across the region. The new mitochondrial clade fell within B. plicatilis s.s. but was sister to all other mitochondrial sequences of that species, suggesting a period of isolation from other populations. Moreover, significant morphological differences were identified: B. plicatilis s.s. from the Qinghai-Tibetan Plateau had a larger lorica length and width than did members of this species from lowland China. Our data demonstrate the successful adaptation of this species complex to the harsh environment of the Qinghai-Tibetan Plateau.
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Rotíferos , Animales , China , ADN Mitocondrial/genética , Variación Genética , Filogenia , Rotíferos/genética , Aguas Salinas , TibetRESUMEN
Copper is a micronutrient essential for plant growth and development. However, Cu is also a heavy metal element that has deleterious impacts on plants when excessively accumulated in the environment. To understand the molecular mechanism underlying tobacco in response to Cu stress, iTRAQ based technology was used to identify differentially expressed proteins (DEPs) and important metabolic pathways in tobacco plants treated with excessive CuSO4. The results showed that 180 DEPs were detected between the treatment and control, among which 78 were upregulated and 102 were downregulated. These DEPs can be functionally divided into 65 categories and are closely related to metabolic pathways, carbon metabolism, secondary metabolite biosynthesis, biosynthesis of antibiotics, glyoxylate and dicarboxylate metabolism, and glycolysis/gluconeogenesis. Peroxidase7 was significantly upregulated and was selected and overexpressed in tobacco. Then, positive transgenic lines and wild type plants were exposed to a Cu stress environment. The results showed that Peroxidase7 transgenic tobacco plants exhibited enhanced Cu stress resistance with decreased malondialdehyde and Cu contents, and increased shoot dry weight, root length, secondary root number, SOD, POD and CAT activity. The present study suggests that the ROS scavenging mechanism is essential for tobacco plants in response to Cu stress and that Peroxidase7 functions in tobacco plant resistance to excessive Cu environment.
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Metales Pesados , Nicotiana , Antibacterianos/metabolismo , Carbono/metabolismo , Cobre/metabolismo , Cobre/toxicidad , Regulación de la Expresión Génica de las Plantas , Glioxilatos/metabolismo , Malondialdehído/metabolismo , Metales Pesados/metabolismo , Micronutrientes/metabolismo , Peroxidasa/metabolismo , Peroxidasas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Proteoma/genética , Proteoma/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Estrés Fisiológico/genética , Superóxido Dismutasa/genética , Nicotiana/genética , Nicotiana/metabolismoRESUMEN
Predation has been a major driver of the evolution of prey species, which consequently develop antipredator adaptations. However, little is known about the genetic basis underpinning the adaptation of prey to intensive predation. Here, we describe a high-quality chromosome-level genome assembly (approx. 145 Mb, scaffold N50 11.45 Mb) of Daphnia mitsukuri, a primary forage for many fish species. Transcriptional profiling of D. mitsukuri exposed to fish kairomone revealed that this cladoceran responds to predation risk through regulating activities of Wnt signalling, cuticle pattern formation, cell cycle regulation and anti-apoptosis pathways. Genes differentially expressed in response to predation risk are more likely to be members of expanded families. Our results suggest that expansions of multiple gene families associated with chemoreception and vision allow Daphnia to enhance detection of predation risk, and that expansions of those associated with detoxification and cuticle formation allow Daphnia to mount an efficient response to perceived predation risk. This study increases our understanding of the molecular basis of prey defences, being important evolutionary adaptations playing a stabilizing role in community dynamics.
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Daphnia , Conducta Predatoria , Adaptación Fisiológica , Animales , Daphnia/genética , Genoma , Genómica , HumanosRESUMEN
Cr(VI) is a common heavy metal ion, which will seriously harm human body and environment. Therefore, the removal of Cr(VI) has become an attractive topic. In this work, cinder was used as a raw material to synthesize a nanoneedle material: γ-(AlOOH@FeOOH) (γ-Al@Fe). The physicochemical properties of γ-Al@Fe were thoroughly characterized, and its effectiveness as a catalyst for photocatalytic reduction of Cr(VI) was evaluated. The results showed that Cr(VI) could be efficiently reduced by γ-Al@Fe in the presence of tartaric acid (TA) under visible light. The variable factors on the reaction were investigated in detail, and the results showed that under optimal conditions (γ-Al@Fe 0.4 g/L, TA 0.6 g/L, pH 2), Cr(VI) was completely reduced within 7 min. Besides, scavenger experiments and EPR proved that O2⢠- and CO2⢠- played a significant role in the photocatalytic reduction of Cr(VI). TA acts as a sacrificial agent to trap the holes and generate strong reducing free radicals: CO2⢠-. Dissolving O2 could react with electrons to generate O2⢠-. This work discussed the performance and mechanism of photocatalytic reduction of Cr(VI) in detail, which provided a new idea for the resource utilization of solid waste and the treatment of heavy metal sewage.
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Cromo , Tartratos , Humanos , Oxidación-ReducciónRESUMEN
This study aims to figure out the methylation of long non-coding RNA GAS5 promoter in cervical cancer and the mechanism of GAS5 on the progression of cervical cancer cells. The expression of GAS5 and methylation state of GAS5 in cervical cancer tissues and cells were determined. With the aim to to explore the ability of GAS5 in the proliferation, cell cycle progression, apoptosis, invasion, migration as well as the tumor growth, and metastasis in nude mice were determined. The expression of GAS5 was decreased and methylation state of GAS5 was elevated in cervical cancer. Overexpression of GAS5 inhibited proliferation, cell cycle progression, invasion, migration while inducing apoptosis of cervical cancer cells as well as suppressed tumor growth and metastasis in nude mice. Our study demonstrates that abnormal methylation of GAS5 contributes to poor expression of GAS5 in cervical cancer. In addition, upregulation of GAS5 inhibits the cervical cancer development.
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Proliferación Celular , ARN Largo no Codificante/metabolismo , Neoplasias del Cuello Uterino/metabolismo , Animales , Apoptosis , Puntos de Control del Ciclo Celular , Movimiento Celular , Metilación de ADN , Transición Epitelial-Mesenquimal , Femenino , Regulación Neoplásica de la Expresión Génica , Células HeLa , Humanos , Ratones Endogámicos BALB C , Ratones Desnudos , Persona de Mediana Edad , Invasividad Neoplásica , ARN Largo no Codificante/genética , Transducción de Señal , Carga Tumoral , Regulación hacia Arriba , Neoplasias del Cuello Uterino/genética , Neoplasias del Cuello Uterino/patologíaRESUMEN
We describe a stoichiometric approach to the synthesis of molecularly imprinted polymers specific for auramine O. Using the stoichiometric interaction in molecular imprinting, no excess of binding sites is necessary and binding sites are only located inside the imprinted cavities. The free base of the template was obtained to facilitate the interaction with the monomers. Itaconic acid was selected as the functional monomer, and stoichiometric ratio of the interaction with the free base was investigated. The molecularly imprinted polymer preparation conditions such as cross-linker, molar ratio, porogen were optimized as divinylbenzene, 1:2:20 and chloroform/N,N-dimethylformamide, respectively. Under the optimum conditions, a good imprinting effect and very high selectivity were achieved. A solid-phase extraction method was developed using the molecularly imprinted polymers as a sorbent and extraction procedure was optimized. The solid-phase extraction method showed a high extraction recovery for auramine O in its hydrochloride form and free form compared to its analogues. The results strongly indicated that stoichiometric imprinting is an efficient method for development of high selectivity molecularly imprinted polymers for auramine O.
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Bacillus velezensis strain S3-1 has a broad range of hosts and is used as a biocontrol agent and biofertilizer. However, the interaction of maize root exudates and colonization of the strain S3-1 has not yet been investigated. In our study, strain S3-1 effectively colonized both rhizosphere soil and root tissue. Collected maize root exudates significantly induced the chemotaxis, cluster movement, and biofilm formation of strain S3-1, showing increases of 1.43, 1.6, and 2.08 times, respectively, compared with the control. In addition, the components of root exudates (organic acids: citric acid, malic acid, and oxalic acid; amino acids: glycine, proline and phenylalanine; sugars: glucose, fructose, and sucrose) were tested. Each of these compounds could induce chemotactic response, swarming motility, and biofilm formation significantly. The strongest chemotactic response and swarming motility were found when malic acid was applied, but maximal ability of biofilm formation was stimulated by proline. Furthermore, we found that these compounds of root exudates stimulated the population of S3-1 adhering to the maize root surface, especially in the presence of malic acid. These results indicate that maize root exudates play an important role in the colonization of S3-1, and provide a deeper understanding of the interaction between plants and microorganisms.
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Bacillus/crecimiento & desarrollo , Exudados de Plantas/metabolismo , Raíces de Plantas/metabolismo , Raíces de Plantas/microbiología , Rizosfera , Microbiología del Suelo , Zea mays , Bacillus/metabolismo , Adhesión Bacteriana/efectos de los fármacos , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Quimiotaxis/efectos de los fármacos , Interacciones Microbiota-Huesped , Locomoción/efectos de los fármacos , Compuestos Orgánicos/análisis , Compuestos Orgánicos/farmacología , Exudados de Plantas/química , Exudados de Plantas/farmacología , Raíces de Plantas/química , Zea mays/metabolismo , Zea mays/microbiologíaRESUMEN
A novel magnetic molecularly imprinted polymer for the selective recognition of auramine O was rationally designed via screening from a library of nonimprinted polymers. A stoichiometric ratio of functional monomer (itaconic acid) and template molecule (auramine O) was found to be 1.5. Meanwhile, the synthesized SiO2 @Fe3 O4 was modified by 0.5 mol/L hydrochloric acid to facilitate the preparation of magnetic molecularly imprinted polymer particles. Adsorption experiments showed that the magnetic polymer particles exhibited good selectivity, recoveries, and enrichment performance. The stoichiometric imprinted polymers have been employed for the selective preconcentration of auramine O from lake water sample. The high specificity of the stoichiometric imprinted polymers was proven in the extraction of mixture solution of auramine O, auramine O hydrochloride, and chrysoidine, and the recoveries ranged between 99.66 and 108.75% (RSD 2.6-3.7%, n = 3) for lake water. These results suggest that this method is effective and can be successfully applied to the analysis of auramine O in environmental water samples.
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Growth arrest special 5 (GAS5) is a long non-coding RNA reported to function as an inhibitor in various tumors including cervical cancer. However, the molecular mechanism of GAS5 involved in cervical cancer progression remains far from being elucidated. The expression of GAS5, forkhead box protein O1 and phosphatase and tensin homolog was examined by quantitative reverse transcription polymerase chain reaction qRT-PCR. cell growth, invasion, and apoptosis were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, colony formation assay, transwell invasion assay, and flow cytometry analysis, respectively. The interaction between GAS5 and miR-196a or miR-205 was confirmed by luciferase reporter assay, RNA immunoprecipitation assay, and qRT-PCR. Xenograft tumor experiments were performed to validate the biological role of GAS5 and its molecular mechanism in cervical cancer in vivo. GAS5 expression was decreased in cervical cancer tissues and cells. GAS5 overexpression suppressed cervical cancer cell proliferation, invasion, and apoptosis. GAS5 was able to directly bind to miR-196a and miR-205 to downregulate their expression. Moreover, GAS5 induced forkhead box protein O1 and phosphatase and tensin homolog expression by repressing miR-196a and miR-205, respectively. Exogenous expression of GAS5 hindered tumor growth in vivo by downregulating miR-196a and miR-205. Upregulation of GAS5 suppressed cell proliferation, invasion, and apoptosis of cervical cancer cells by downregulating miR-196a and miR-205, contributing to our understanding the pathogenesis of cervical cancer and development of long non-coding RNA-mediated clinical therapy against this disease.
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Carcinogénesis/genética , MicroARNs/biosíntesis , ARN Largo no Codificante/genética , Neoplasias del Cuello Uterino/genética , Animales , Apoptosis/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Femenino , Proteína Forkhead Box O1/biosíntesis , Regulación Neoplásica de la Expresión Génica , Humanos , Ratones , MicroARNs/genética , Fosfohidrolasa PTEN/biosíntesis , Neoplasias del Cuello Uterino/patología , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Background: Babesia is a unique apicomplexan parasite that specifically invades and proliferates in red blood cells and can be transmitted via blood transfusion, resulting in transfusion-transmitted babesiosis. However, detecting Babesia in blood before transfusion has not received enough attention, and the risk of transfusing blood containing a low density of Babesia microti (B. microti) is unclear, possibly threatening public health and wellness. Purpose: This study aimed to determine the lower detection limit of B. microti in blood and to evaluate the transmission risk of blood transfusion containing low-density B. microti. Methods: Infected BALB/c mouse models were established by transfusing infected whole blood with different infection rates and densities of B. microti. Microscopic examination, nested Polymerase Chain Reaction (nested PCR), and an enzyme-linked immunosorbent assay (ELISA) were used to evaluate the infection status of the mouse models. Meanwhile, the nested PCR detection limit of B. microti was obtained using pure B. microti DNA samples with serial concentrations and whole blood samples with different densities of B. microti-infected red blood cells. Thereafter, whole mouse blood with a B. microti density lower than that of the nested PCR detection limit and human blood samples infected with B. microti were transfused into healthy mice to assess the transmission risk in mouse models. The infection status of these mice was evaluated through microscopic examination, nested PCR tests, and ELISA. Results: The mice inoculated with different densities of B. microti reached the peak infection rate on different days. Overall, the higher the blood B. microti density was, the earlier the peak infection rate was reached. The levels of specific antibodies against B. microti in the blood of the infected mice increased sharply during the first 30 days of infection, reaching a peak level at 60 days post-infection, and maintaining a high level thereafter. The nested PCR detection limits of B. microti DNA and parasite density were 3 fg and 5.48 parasites/µL, respectively. The whole blood containing an extremely low density of B. microti and human blood samples infected with B. microti could infect mice, confirming the transmission risk of transfusing blood with low-density B. microti. Conclusion: Whole blood containing extremely low density of B. microti poses a high transmission risk when transfused between mice and mice or human and mice, suggesting that Babesia detection should be considered by governments, hospitals, and disease prevention and control centers as a mandatory test before blood donation or transfusion.
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Babesia microti , Babesia , Babesiosis , Humanos , Animales , Ratones , Babesia microti/genética , Babesia/genética , Transfusión Sanguínea , Babesiosis/diagnóstico , Babesiosis/parasitología , ADN Protozoario , Ratones Endogámicos BALB C , Modelos Animales de EnfermedadRESUMEN
MYC2 is a class of bHLH family transcription factors and a major regulatory factor in the JA signaling pathway, and its molecular function in tobacco has not been reported. In this study, CRISPR/Cas9-mediated MYC2 gene NtMYC2a knockout mutants at tobacco was obtained and its agronomic traits, disease resistance, and chemical composition were identified. Comparing with the WT, the leaf width of the KO-NtMYC2a was narrowed, the nornicotine content and mecamylamine content increased significantly and the resistance to Ralstonia solanacearum significantly decreased. The transcriptome sequencing results showed that DEGs related to immunity, signal transduction and growth and development were enriched between KO-NtMYC2a and WT. NtJAR1 and NtCOI1 in KO-NtMYC2a were down-regulated to regulating the JA signaling pathway, result in a significant decrease in tobacco's resistance to R. solanacearum. Our research provides theoretical support for the functional research of MYC2 and the study of the mechanism of tobacco bacterial wilt resistance.
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Sistemas CRISPR-Cas , Resistencia a la Enfermedad , Regulación de la Expresión Génica de las Plantas , Nicotiana , Enfermedades de las Plantas , Proteínas de Plantas , Ralstonia solanacearum , Nicotiana/genética , Nicotiana/microbiología , Resistencia a la Enfermedad/genética , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/genética , Ralstonia solanacearum/patogenicidad , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Técnicas de Inactivación de Genes , Ciclopentanos/metabolismo , Transducción de Señal , Oxilipinas/metabolismo , Plantas Modificadas Genéticamente/genéticaRESUMEN
Ethylene responsive factor (ERF) is a plant-specific transcription factor that plays a pivotal regulatory role in various stress responses. Although the genome of tobacco harbors 375 ER F genes, the functional roles of the majority of these genes remain unknown. Expression pattern analysis revealed that NtERF283 was induced by water deficit and salt stresses and mainly expressed in the roots and leaves. Subcellular localization and transcriptional activity assays confirmed that NtERF283 was localized in the nucleus and exhibited transcriptional activity. In comparison to the wild-type (WT), the NtERF283-overexpressing transgenic plants (OE) exhibited enhanced water deficit tolerance, whereas the knockout mutant erf283 displayed contrasting phenotypes. Transcriptional analysis demonstrated that several oxidative stress response genes were significantly altered in OE plants under water deficit conditions. 3,3'-diaminobenzidine (DAB) and nitroblue tetrazolium (NBT) staining showed that erf283 accumulated a higher level of reactive oxygen species (ROS) compared to the WT under water deficit conditions. Conversely, OE plants displayed the least amount of ROS accumulation. Furthermore, the activities of POD and SOD were higher in OE plants and lower in erf283, suggesting that NtERF283 enhanced the capacity to effectively eliminate ROS, consequently enhancing water deficit tolerance in tobacco. These findings strongly indicate the significance of NtERF283 in promoting tobacco water deficit tolerance through the activation of the antioxidant system.
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Antioxidantes , Agua , Antioxidantes/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Agua/metabolismo , Estrés Oxidativo , Plantas Modificadas Genéticamente/metabolismo , Nicotiana/genética , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/metabolismo , Estrés Fisiológico/genéticaRESUMEN
LYSINE HISTIDINE TRANSPORTER1 (LHT1) is a crucial broad-specificity and high-affinity amino acid transporter affecting the uptake of nitrogen and probably the tolerance to abiotic stress in plants. However, little is known about the phenotypic functions of LHT1 in plant growth and development and abiotic stress tolerance. In this study, we identified the NtLHT1 gene from the tobacco variety Honghuadajinyuan (HD) and determined its important roles in leaf morphological development and plant resistance to abiotic stress. Comprehensive functional analyses using knockout and overexpression transgenic lines (ntlht1 and OE) revealed overexpression of NtLHT1 accelerated leave senescence and increased plant height, leaf number and plant tolerance under cold, salt and drought stresses. In addition, NtLHT1 overexpression significantly decreased the leaf elongation of HD, causing the leaves to change from a long-elliptical shape to an elliptical shape. However silencing NtLHT1 decreased the seed germination rate under NaCl and PEG stresses. Moreover, NtLHT1 significantly affected the contents of various amino acids, such as the neutral, acidic, non-polar and aromatic amino acids, ethylene precursor (ACC), GA3 and IAA in tobacco. These results suggested that the amino acid and ethylene precursor ACC transport activities of NtLHT1 provide fine regulatory function for plant growth and development and plant tolerance to abiotic stress.
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Etilenos , Estrés Fisiológico , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Etilenos/metabolismo , Estrés Fisiológico/genética , Cloruro de Sodio/metabolismo , Aminoácidos/metabolismo , Nicotiana/genética , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , SequíasRESUMEN
This study was designed to investigate the effects of dietary betaine supplementation on growth performance, meat quality and muscle lipid metabolism of growing-finishing pigs. Thirty-six crossbred pigs weighing 24.68 ± 0.97 kg were randomly allotted into two treatments consisting of a basal diet supplemented with 0 or 1200 mg/kg betaine. Each treatment included six replications of three pigs per pen. Following 119 days of feeding trial, dietary betaine supplementation significantly enhanced average daily gain (ADG) (p < 0.05) and tended to improve average daily feed intake (ADFI) (p = 0.08) and decreased the feed intake to gain ratio (F/G) (p = 0.09) in pigs during 100~125 kg. Furthermore, a tendency to increase ADG (p = 0.09) and finial body weight (p = 0.09) of pigs over the whole period was observed in the betaine diet group. Betaine supplementation significantly increased a*45 min and marbling and decreased b*24 h and cooking loss in longissimus lumborum (p < 0.05), tended to increase intramuscular fat (IMF) content (p = 0.08), however had no significant influence on carcass characteristics (p > 0.05). Betaine supplementation influenced the lipid metabolism of pigs, evidenced by a lower serum concentration of low-density lipoprotein cholesterol (p < 0.05), an up-regulation of mRNA abundance of fatty acid synthase and acetyl-CoA carboxylase (p < 0.05), and a down-regulation of mRNA abundance of lipolysis-related genes, including the silent information regulators of transcription 1 (p = 0.08), peroxisome proliferator-activated receptorα (p < 0.05), peroxisome proliferator-activated receptor gamma coactivator-1α (p = 0.07) and carnitine palmitoyl transferase 1 (p < 0.05) in longissimus lumborum. Moreover, betaine markedly improved the expression of microRNA-181a (miR-181a) (p < 0.05) and tended to enhance miR-370 (p = 0.08). Overall, betaine supplementation at 1200 mg/kg could increase the growth performance of growing-finishing pigs. Furthermore, betaine had a trend to improve meat quality and IMF content via increasing lipogenesis and down-regulating the abundance of genes associated with lipolysis, respectively, which was associated with the regulation of miR-181a and miR-370 expression by betaine.
RESUMEN
Two experiments were carried out to evaluate the effects of betaine (BET) supplementation in diets with reduced net energy (NE) levels on growth performance, nutrient digestibility, and serum metabolomic profiles in growing pigs. In experiment 1, 24 growing pigs (initial body weight, BW, 30.83â ±â 2.50 kg) were allotted to one of the four treatments (six replications with 1 pig per pen) in a 2â ×â 2 factorial arrangement, including two dietary NE levels (2475 [N-NE] or 2395 [R80-NE] kcal/kg) and two BET doses (0 or 1500 mg/kg). In experiment 2, 72 growing pigs were used in a 2â ×â 3 factorial arrangement, including three dietary NE levels (2475 [N-NE], 2415 [R60-NE], or 2355 [R120-NE] kcal/kg) and two BET doses (0 or 1500 mg/kg). Pigs with initial BW of 31.44â ±â 1.65 kg were divided to one of the six treatments (six replications with 2 pigs per pen). In experiment 1, lowing NE concentrations increased average daily feed intake (ADFI) by 10.69% in pigs fed the diet without BET (Pâ >â 0.05). BET significantly increased ADFI in N-NE diet (Pâ <â 0.05) but had no influence on ADFI in R80-NE diet (Pâ >â 0.05). BET enhanced the apparent digestibility of crude protein (CP), dry matter (DM), organic matter (OM), gross energy (GE), and ether extract (EE) in R80-NE diet (Pâ <â 0.05). In experiment 2, lowing NE concentrations enhanced ADFI (Pâ >â 0.05) and decreased average daily gain (ADG; Pâ <â 0.05). The reduction in feed intake by BET was further enhanced as NE concentrations decreased from 2415 to 2355 kcal/kg (Pâ <â 0.10). BET reversed the elevation of serum triglyceride, alkaline phosphatase, aspartate aminotransferase, and alanine aminotransferase levels caused by R120-NE diet (Pâ <â 0.05). The concentrations of cholecystokinin and glucagon-like peptide 1 were increased by BET in pigs fed the R120-NE diet (Pâ <â 0.05). Serum metabolomics reveals that lowing dietary NE concentrations affected mainly amino acid biosynthetic pathways (Pâ <â 0.05). BET supplementation in R120-NE diet up-regulated serum BET levels and down-regulated homocysteine, DL-carnitine, and four amino acid secondary metabolites (Pâ <â 0.05). In conclusion, lowing dietary NE contents reduced the growth performance and caused metabolic abnormalities in growing pigs. However, BET decreased feed intake to a certain extent and improved the metabolic health of pigs fed the low-NE diets, which may be related to the dual regulation of amino acid metabolism and the secretion of appetite related hormones by BET.
Energy is an important factor in affecting the production efficiency and feed cost in animal husbandry. For pigs, the reduction of dietary energy will lead to a decreased growth performance. Therefore, additional researches towards ameliorating the negative effects caused by low energy diets are necessary to conduct, so as to develop appropriate nutritional strategies. Betaine, a trimethyl derivative of glycine, is considered to affect energy partitioning. Betaine may influence the growth performance and healthy status of pigs under low-energy conditions. Herein, two experiments were carried out to evaluate the effects of betaine supplementation in diets with reduced net energy levels on growth performance, nutrient digestibility, and serum metabolomic profiles in growing pigs. Results indicated that lowering dietary energy reduced growth performance and caused metabolic abnormalities in growing pigs, however, betaine supplementation in low-energy diets improved metabolic homeostasis and the utilization of energy despite reduced feed intake to a certain extent.
Asunto(s)
Betaína , Suplementos Dietéticos , Porcinos , Animales , Betaína/farmacología , Dieta/veterinaria , Aminoácidos/metabolismo , Nutrientes , Alimentación Animal/análisis , Digestión , Fenómenos Fisiológicos Nutricionales de los AnimalesRESUMEN
The widespread use of chemical herbicides has jeopardized concerns about food safety and ecological consequences. To address these issues and reduce reliance on chemical herbicides, a physical weed control device was developed for the tillering stage in paddy fields. This device features a biomimetic duckbill-like vibration chain that effectively controls weed outbreaks. The chain penetrates the soft surface soil of the paddy field under gravity and rapidly stirs the soil through vibration, leading to the detachment of the weed roots anchored in the surface layer. Simultaneously, the device avoids mechanical damage to rice seedlings rooted in deeper soil. This study aimed to investigate the effects of chain structural parameters (the number of chain rows, vibration amplitude, and length of chains) and operational parameters (vibration frequency and working velocity) on weed control efficiency and rice seedling damage. Through a central composite regression field test, the optimal device structure and operational parameters were determined. The optimization results demonstrated that a vibration amplitude of 78.8 mm, a chain length of 93.47 cm, and 3.4 rows of chains, along with a vibration frequency and working velocity ranging from 0.5 to 1.25 m/s, achieved an optimal weeding effect. Under the optimal parameter combination, field test results demonstrated that approximately 80% of the weeds in the field were effectively cleared. This indicates that the design of the biomimetic duckbill-like vibration chain weeding device exhibits a relatively superior weeding performance, offering a practical solution for the management of weeds in rice fields.