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At present, in the process of weld induction heat treatment, the common method is to carry out centralized induction heating in the weld area, which will lead to large radial temperature difference of the weld, poor controllability of temperature distribution and easy to cause the defects of residual stress concentration in the weld area. To solve the above problems, this paper adopts the two-sided method to conduct induction heating on both sides of the weld, and at the same time, the auxiliary pulse current is passed into the weld to improve the quality of the weld. ANSYS finite element software is used to establish a multi-field coupling prediction model of electric-magnetic-thermal structure, and explore the distribution law of the auxiliary pulse current and the temperature field of the weld. Finally, an experimental study of pulsed current assisted two-sided induction heating is carried out. Temperature test and metallographic test were carried out respectively to verify the effectiveness of pulsed current assisted induction heating technology.
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During the welding process, temperature filed distribution of weldment is one of the key factors which influence welding quality. In order to improve the mechanical properties of welding joint, a technology for heating process of weldment using auxiliary pulse current was proposed in this article. Firstly, through metallographic experiment, strength experiment and hardness experiment, it was proved that the auxiliary pulse current not only can refine grain size but also improves the mechanical property of the welding joint. Then the paper used ANSYS and its ANSYS Parametric Design Language parametric design language to simulate the welding process with assistant pulse current, and the temperature field, the pulse current density distribution and the time of effective pulse current in the welded joint were obtained. Quantitative analyses were undertaken on the influence laws of auxiliary pulse current parameters on weld temperature field, auxiliary pulse current density distribution and the time of effective pulse current. A new parameter was defined as time-current density (the time integral of current density) to demonstrate the change rule of auxiliary pulse current density distribution with time. Finally, analyzing the reasons for raising the welding quality. There were two reasons for such phenomenon: one was auxiliary pulse current change the state of heat distribution in the weld seam area, the other was that the impulse oscillation of the auxiliary pulse influences the nucleation process of melting region.
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This study investigated the impact of chitosan (CH, 1%) and aloe vera gel (AL, 30%) edible coatings on the preservation of blue honeysuckle quality during a 28-day storage at -1 °C. Coating with CH, AL, and CH+AL led to notable enhancements in several key attributes. These included increased firmness, total soluble solids, acidity, pH, and antioxidant capacity (measured through DPPH, ABTS, and FRAP assays), as well as the preservation of primary (ascorbic acid) and secondary metabolites (TPC, TAC, and TFC). The TAC and TFC levels were approximately increased by 280% and 17%, respectively, in coated blue honeysuckle after 28 d compared to uncoated blue honeysuckle. These coatings also resulted in reduced weight loss, respiration rate, color, abscisic acid, ethylene production, and malondialdehyde content. Notably, the CH+AL treatment excelled in preserving secondary metabolites and elevating FRAP-reducing power, demonstrating a remarkable 1.43-fold increase compared to the control after 28 days. Overall, CH+AL exhibited superior effects compared to CH or AL treatment alone, offering a promising strategy for extending the shelf life and preserving the quality of blue honeysuckle during storage.
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Blue honeysuckle seeds are often overlooked by the processing industry, but they are a good source of healthy oil. The composition, volatiles, and antioxidant capacity of blue honeysuckle seeds and seed oil were investigated for the first time. The fatty acid profile of the seed oil was analysed using GC-MS. The seed oil was particularly rich in polyunsaturated fatty acid, especially linoleic acid (71.24 ± 1.64 %). HS-SPME-GC-MS analysis temporarily detected 34 and 37 volatiles in the seeds and seed oil, respectively. Notably, aldehydes were identified as the major contributors to the aroma. The phytosterols, tocopherols, and triglycerides were identified in the seed oil. Interestingly, the total phenolic content and antioxidant capacity of the seeds were found to be much higher than the seed oil. This study evaluates the nutritional profile and value of blue honeysuckle seed oil, and suggests that it can be used as new functional oil.
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The aim of this study was to investigate a more practical method for obtaining non-extractable polyphenols (NEPPs) from blue honeysuckle fruit pomace. Three methods, namely acid, alkaline, and enzymatic hydrolysis, were utilized to extract NEPPs. The findings indicated that alkaline hydrolysis was the most effective method for releasing NEPPs, which demonstrated higher levels of total flavonoid content (TFC) and total phenolic content (TPC) from blue honeysuckle fruit pomace. Additionally, higher TPC and TFC levels were related to a stronger antioxidant capacity. Qualitative and quantitative analysis using HPLC-HR-TOF-MS/MS revealed that acid hydrolysis resulted in a greater concentration of certain phenolic acids, while alkaline hydrolysis yielded a higher concentration of flavonoids, and enzymatic hydrolysis produced a wider range of phenolic compositions. Despite the fact that enzymatic hydrolysis is considered a gentler method, the researchers concluded that alkaline hydrolysis was the most appropriate method for obtaining NEPPs from blue honeysuckle fruit pomace.
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Lonicera , Polifenóis , Polifenóis/análise , Antioxidantes/análise , Frutas/química , Espectrometria de Massas em Tandem , Fenóis/análise , FlavonoidesRESUMO
Mitochondrial transcription termination factor (mTERF) is a DNA-binding protein that is encoded by nuclear genes, ultimately functions in mitochondria and can affect gene expression. By combining with mitochondrial nucleic acids, mTERF regulates the replication, transcription and translation of mitochondrial genes and plays an important role in the response of plants to abiotic stress. However, there are few studies on mTERF genes in tomato, which limits the in-depth study and utilization of mTERF family genes in tomato stress resistance regulation. In this study, a total of 28 mTERF gene family members were obtained through genome-wide mining and identification of the tomato mTERF gene family. Bioinformatics analysis showed that all members of the family contained environmental stress or hormone response elements. Gene expression pattern analysis showed that the selected genes had different responses to drought, high salt and low temperature stress. Most of the genes played key roles under drought and salt stress, and the response patterns were more similar. The VIGS method was used to silence the SLmTERF13 gene, which was significantly upregulated under drought and salt stress, and it was found that the resistance ability of silenced plants was decreased under both kinds of stress, indicating that the SLmTERF13 gene was involved in the regulation of the tomato abiotic stress response. These results provide important insights for further evolutionary studies and contribute to a better understanding of the role of the mTERF genes in tomato growth and development and abiotic stress response, which will ultimately play a role in future studies of tomato gene function.
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In this paper, the structures of polyphenols and their bioactivity of black mulberry (Morus nigra L.) cv. 'Heisang No. 1' were comprehensively analyzed. The 11 anthocyanins and 20 non-anthocyanin phenolic compounds were identified and quantified by liquid chromatography high-resolution time-of-flight mass spectrometry (LC-HR-TOF/MS2). The cyanidin-3-glucoside and cyanidin-3-rutinoside were the major anthocyanins in the black mulberry. In addition, the black mulberry showed potent antioxidant capacity as assessed by DPPH, ABTS, and FRAP assays. Black mulberry anthocyanins exhibited stronger inhibition activities against α-amylase, α-glucosidase, and lipase compared to non-anthocyanin polyphenols, with IC50 values of 1.10, 4.36, and 9.18 mg/mL, respectively. The total anthocyanin content of black mulberry crude extracts and anthocyanins was 570.10 ± 77.09 and 1278.23 ± 117.60 mg C3GE/100 g DW, respectively. Black mulberry may be a rich source of polyphenols, natural antioxidants, and effective antidiabetic substances with great potential in the food industry.
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Morus , Polifenóis , Polifenóis/análise , Antocianinas/análise , Antioxidantes/química , Morus/química , Fenóis/análise , Frutas/química , Extratos Vegetais/químicaRESUMO
Genes play regulatory roles in plants' response to low-temperature stress. Our understanding of the mechanism of plants' response to low-temperature stress can be expanded by studying the functions of these genes. SfGPX was cloned from Spiraea fritschiana (S. fritschiana) with the highest low-temperature tolerance, to explore the molecular mechanisms of SfGPX in response to low-temperature stress and the physiological mechanisms involved in the regulation of SfGPX to adapt to low temperature, in two species of Spiraea. SfGPX, which was localized in the cytoplasm, was induced by low temperature. The low-temperature tolerance of Spiraea fritschiana was decreased via the interference of SfGPX, and the low-temperature tolerance of Spiraea japonica 'Gold Mound' (S. japonica 'Gold Mound') was elevated via the overexpression of SfGPX. Under low-temperature stress, the photosynthetic capacity of two species of Spiraea was affected by SfGPX; it was higher in the cold-tolerant plants and lower in the cold-intolerant plants. Under low-temperature stress, the transfer intensity of Ca2+ was affected by SfGPX. The transfer intensity of cold-tolerant plants with lower influx level of Ca2+ kinetics was weaker than that of cold-intolerant plants. Under low-temperature stress, the transfer velocity of Ca2+ was affected by SfGPX, and there were slower effluxes of Ca2+ from Ca2+ reservoir in cold-tolerant plants than in cold-intolerant plants. The above results indicate that the response of Spiraea to low temperature is regulated by SfGPX through affecting photosynthetic capacity as well as intensity and velocity of Ca2+ transfer in response to low temperature in Spiraea.
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Spiraea , Temperatura Baixa , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , Spiraea/metabolismo , TemperaturaRESUMO
The structure of proanthocyanidins extracted from bird cherry fruits was characterized by HPLC-ESI/MS2 and MALDI-TOF/MS analyses, and their subunits and mean degree of polymerization (mDP) were investigated by thiolysis reaction, and the inhibition activity against starch hydrolases measured using the high-throughput turbidity assay. This is the first mass spectrometric analysis to thoroughly investigate the structure and mDP of proanthocyanidins in bird cherry fruits. Bird cherry proanthocyanidins were categorized as oligomeric proanthocyanidins (mDP = 5.6), which constituted of (epi)gallocatechins and (epi)catechins. The proanthocyanidins increased from a (epi)gallocatechin-[(epi)catechin]3 tetramer to a (epi)gallocatechin-[(epi)catechin]11 dodecamer through the addition of one (epi)catechin with both A-type and B-type linkages. The proanthocyanidins had potent α-amylase and α-glucosidase inhibition activities with IC50 values of 0.19 ± 0.01 µg/mL and 0.18 ± 0.006 µg/mL, comparing favorably to commercial drug acarbose. Bird cherry oligomeric proanthocyanidins are a promising starch hydrolase inhibitor for the application of potential functional food components.
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Catequina , Proantocianidinas , Prunus , Catequina/química , Polimerização , Proantocianidinas/análise , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Amido , alfa-AmilasesRESUMO
Variations in iodinated aromatic disinfection byproducts (DBPs) in the presence of I- and organic compounds as a function of reaction time in different molar ratios (MRs) of HOCl:NH3-N were investigated. Up to 17 kinds of iodinated aromatic DBPs were identified in the breakpoint chlorination of iodide (I-)/organic (phenol, bisphenol S (BPS) and p-nitrophenol (p-NP)) systems, and the possible pathways for the formation of iodinated aromatic DBPs were proposed. The reaction pathways include HOCl/HOI electrophilic substitution and oxidation, while the dominant iodinated DBPs were quantified. In the I-/phenol system (pH = 7.0), the sum of the concentrations of four iodinated aliphatic DBPs ranged from 0.32 to 1.04 µM (triiodomethane (TIM), dichloroiodomethane (DCIM), diiodochloromethane (DICM) and monoiodoacetic acid (MIAA)), while the concentration of 4-iodophenol ranged from 2.99 to 12.87 µM. The concentration of iodinated aromatic DBPs remained stable with an MR = 1:1. When the MR was 6:1, iodinated aromatic DBPs decreased with increasing reaction time, in which the main disinfectant in the system was active chlorine. This study proposed the formation mechanism of iodinated aromatic DBPs during the breakpoint chlorination of iodide-containing water. These results can be used to control the formation of hazardous iodinated aromatic DBPs in the disinfection of iodine containing water.
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Desinfetantes , Poluentes Químicos da Água , Purificação da Água , Cloro , Desinfetantes/análise , Desinfecção , Halogenação , Iodetos , Nitrogênio , Água , Poluentes Químicos da Água/análiseRESUMO
Membrane fouling has been one of the most important challenges in membrane separation operations. In this study, we report a facile strategy to prepare antifouling polysulfone (PSf) UF membranes by blending amphiphilic zwitterion polysulfone-co-sulfobetaine polysulfone (PSf-co-SBPSf) copolymer. The copolymer chemical structure was characterized by 1HNMR spectroscopy. The PSf/PSf-co-SBPSf blend membranes with various zwitterionic SBPSf segment contents exhibited better surface hydrophilicity and excellent antifouling ability compared to PSf and PSf/PEG membranes. The significant increase of both porosity and water permeance indicates that the PSf-co-SBPSf has a pore-forming effect. The pure water flux and flux recovery ratio of the PSf/PSf-co-SBPSf blend membranes were both remarked to improve 286.43 L/m2h and 92.26%, while bovine serum albumin (BSA) rejection remained at a high level (97.66%). More importantly, the water flux and BSA rejection see minimal variance after heat treatment, indicating excellent thermostability. Overall, the PSf/PSf-co-SBPSf blend membranes achieved a comprehensive performance of sustainable hydrophilic, high permeation flux, and remarkable antifouling ability, thus becoming a promising candidate in high-temperature separation application.
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This study intended to explore the effect and mechanism of total flavonoids of Drynariae Rhizoma in improving scopola-mine-induced learning and memory impairments in model mice. Ninety four-month-old Kunming(KM) mice were randomly divided into six groups. The ones in the model group and blank group were treated with intragastric administration of normal saline, while those in the medication groups separately received the total flavonoids of Drynariae Rhizoma, Kangnaoshuai Capsules, donepezil, as well as total flavonoids of Rhizoma Drynariae plus estrogen receptor(ER) blocker by gavage. The mouse model of learning and memory impairments was established via intraperitoneal injection of scopolamine. Following the measurement of mouse learning and memory abilities in Morris water maze test, the hippocampal ERß expression was detected by immunohistochemistry, and the expression levels of ERß and phosphorylated p38(p-p38) in the hippocampus and B-cell lymphoma 2(Bcl-2), Bcl-2-associated death promoter(Bad), and cysteinyl aspartate-specific protease-3(caspase-3) in the apoptotic system were assayed by Western blot. The contents of malondia-ldehyde(MDA), superoxide dismutase(SOD), and nitric oxide(NO) in the hippocampus were then determined using corresponding kits. Compared with the control group, the model group exhibited significantly prolonged incubation period, reduced frequency of cros-sing the platform, shortened residence time in the target quadrant, lowered ERß, Bcl-2 and SOD activity in the hippocampus, and increased p-p38/p38, Bad, caspase-3, MDA, and NO. Compared with the model group, the total flavonoids of Rhizoma Drynariae increased the expression of ERß and SOD in the hippocampus, down-regulated the expression of neuronal pro-apoptotic proteins, up-re-gulated the expression of anti-apoptotic proteins, and reduced p-p38/p38, MDA, and NO. The effects of total flavonoids of Drynariae Rhizoma on the above indexes were reversed by ER blocker. It has been proved that the total flavonoids of Drynariae Rhizoma obviously alleviate scopolamine-induced learning and memory impairments in mice, which may be achieved by regulating the neuronal apoptotic system and oxidative stress via the ER-p38 mitogen-activated protein kinase(ER-p38 MAPK) signaling pathway.
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Polypodiaceae , Animais , Flavonoides , Hipocampo , Aprendizagem em Labirinto , Camundongos , Receptores de Estrogênio , Escopolamina/toxicidade , Transdução de Sinais , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
Iodinated disinfection by-products (DBPs) have been attracting great attention due to their potential high toxicity to human health. Understanding of formation mechanisms and transformation process of iodinated aromatic DBPs during the chlorination of iodide-containing water is crucial. Phenol was therefore chosen as a representative of phenolic compounds to investigate the generation of iodinated aromatic DBPs in a chlorine/phenol/iodide system. The presence of iodide in water could enhance the removal of phenol by chlorine due to higher second order rate constants of HOI with phenol than that of HOCl with phenol. Fourteen kinds of iodinated aromatic DBPs were identified, which were generated from oxidation and electrophilic substitution of phenol by HOCl and HOI. Iodinated phenolic DBPs were sources of iodinated quinone DBPs and chlorinated/iodinated phenolic DBPs. Alkaline condition favored the formation of iodinated phenolic DBPs, while acid condition favored the production of iodinated quinone DBPs. Neutral condition might be the most suitable pH condition to control the formation of iodinated aromatic DBPs. The relative concentration of almost all iodinated aromatic DBPs first increased and then decreased with time, indicating iodinated aromatic DBPs might be further converted into halogenated aliphatic DBPs during the chlorination. This research provides a research basis for the removal of phenol in water.
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Desinfetantes , Poluentes Químicos da Água , Purificação da Água , Cloro , Desinfecção , Halogenação , Humanos , Iodetos , Fenol , Fenóis , Poluentes Químicos da Água/análiseRESUMO
The electroactive ß-phase in Poly (vinylidene fluoride, PVDF) is the most desirable conformation due to its highest pyro- and piezoelectric properties, which make it feasible to be used as flexible sensors, wearable electronics, and energy harvesters etc. In this study, we successfully developed a method to obtain high-content ß-phase PVDF films and nanofiber meshes by mechanical stretching and electric spinning. The phase transition process and pyro- and piezoelectric effects of stretched films and nanofiber meshes were characterized by monitoring the polarized light microscopy (PLM) images, outputting currents and open-circuit voltages respectively, which were proved to be closely related to stretching ratio (λ) and concentrations. This study could expand a new route for the easy fabrication and wide application of PVDF films or fibers in wearable electronics, sensors, and energy harvesting devices.
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The nucleotide-binding site-leucine-rich repeat (NBS-LRR) gene family is the largest group of plant disease resistance (R) genes widespread in response to viruses, bacteria, and fungi usually involved in effector triggered immunity (ETI). Forty members of the Chinese cabbage CC type NBS-LRR family were investigated in this study. Gene and protein characteristics, such as distributed locations on chromosomes and gene structures, were explored through comprehensive analysis. CC-NBS-LRR proteins were classified according to their conserved domains, and the phylogenetic relationships of CC-NBS-LRR proteins in Brassica rapa, Arabidopsis thaliana, and Oryza sativa were compared. Moreover, the roles of BrCC-NBS-LRR genes involved in pathogenesis-related defense were studied and analyzed. First, the expression profiles of BrCC-NBS-LRR genes were detected by inoculating with downy mildew and black rot pathogens. Second, sensitive and resistant Chinese cabbage inbred lines were screened by downy mildew and black rot. Finally, the differential expression levels of BrCC-NBS-LRR genes were monitored at 0, 1, 3, 6, 12 and 24 h for short and 0, 3, 5, 7, 10 and 14 days for long inoculation time. Our study provides information on BrCC-NBS-LRR genes for the investigation of the functions and mechanisms of CC-NBS-LRR genes in Chinese cabbage.
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Brassica/metabolismo , Brassica/microbiologia , Doenças das Plantas/microbiologia , Arabidopsis/microbiologia , Resistência à Doença , Regulação da Expressão Gênica de Plantas , Oryza/microbiologia , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
An acidic heteropolysaccharide HEP-2 was obtained from blue honeysuckle berry extracts after purification using Sepharose 6B and Sephadex G-200 column chromatography. The molecular weight of HEP-2, determined with high-performance gel permeation chromatography, was 3.01 × 106 Da. Fourier transform infrared spectroscopy, gas chromatography and nuclear magnetic resonance analyses revealed the presence of α-, ß-pyranose ring and six sugar residues in HEP-2. Amorphous aggregates of HEP-2 with irregular shape and lacking the triple helical structure were detected using the Congo red test, scanning electron and atomic force microscopy, and X-ray diffraction studies. Rheological characterization showed typical shear-thinning behavior and viscoelastic property of the polysaccharide. HEP-2 exerted significant inhibitory effects on α-amylase and α-glucosidase and displayed competitive inhibition kinetics. The collective results support the potential utility of HEP-2 as a hypoglycemic agent for enzyme-targeted treatment of diabetes or functional food.
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Polissacarídeos/química , alfa-Amilases/química , alfa-Glucosidases/química , Animais , Bovinos , Cinética , Espectroscopia de Ressonância Magnética , Microscopia de Força Atômica , Peso Molecular , Polissacarídeos/isolamento & purificação , Reologia , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios XRESUMO
BACKGROUND: Members of the cytochrome P450 (CYP450) and UDP-glycosyltransferase (UGT) gene superfamily have been shown to play essential roles in regulating secondary metabolite biosynthesis. However, the systematic identification of CYP450s and UGTs has not been reported in Aralia elata (Miq.) Seem, a highly valued medicinal plant. RESULTS: In the present study, we conducted the RNA-sequencing (RNA-seq) analysis of the leaves, stems, and roots of A. elata, yielding 66,713 total unigenes. Following annotation and KEGG pathway analysis, we were able to identify 64 unigenes related to triterpenoid skeleton biosynthesis, 254 CYP450s and 122 UGTs, respectively. A total of 150 CYP450s and 92 UGTs encoding > 300 amino acid proteins were utilized for phylogenetic and tissue-specific expression analyses. This allowed us to cluster 150 CYP450s into 9 clans and 40 families, and then these CYP450 proteins were further grouped into two primary branches: A-type (53%) and non-A-type (47%). A phylogenetic analysis of 92 UGTs and other plant UGTs led to clustering into 16 groups (A-P). We further assessed the expression patterns of these CYP450 and UGT genes across A. elata tissues, with 23 CYP450 and 16 UGT members being selected for qRT-PCR validation, respectively. From these data, we identified CYP716A295 and CYP716A296 as the candidate genes most likely to be associated with oleanolic acid synthesis, while CYP72A763 and CYP72A776 were identified as being the most likely to play roles in hederagenin biosynthesis. We also selected five unigenes as the best candidates for oleanolic acid 3-O-glucosyltransferase. Finally, we assessed the subcellular localization of three CYP450 proteins within Arabidopsis protoplasts, highlighting the fact that they localize to the endoplasmic reticulum. CONCLUSIONS: This study presents a systematic analysis of the CYP450 and UGT gene family in A. elata and provides a foundation for further functional characterization of these two multigene families.
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Aralia/genética , Família Multigênica , Saponinas/biossíntese , Transcriptoma , Triterpenos/metabolismo , Aralia/metabolismo , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Genes de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Saponinas/genéticaRESUMO
Crop monocropping usually results in an enrichment of soil-borne pathogens in soil. Crop rotation is an environmentally friendly method for controlling soil-borne diseases. Plant rhizosphere microorganisms, especially plant-beneficial microorganisms, play a major role in protecting plants from pathogens, but responses of these microorganisms to crop rotation remain unclear. Here, we evaluated the effects of rotations with Indian mustard (Brassica juncea) and wild rocket (Diplotaxis tenuifolia (L.) DC.) on cucumber Fusarium wilt disease caused by Fusarium oxysporum f.sp. cucumerinum Owen (FOC). Cucumber rhizosphere bacterial community composition was analyzed by high-throughput amplicon sequencing. Bacteria, Pseudomonas spp., 2,4-diacetylphloroglucinol (an antifungal secondary metabolite) producer and FOC abundances were estimated by real-time PCR. Rotations with Indian mustard and wild rocket suppressed cucumber Fusarium wilt disease and cucumber rhizosphere FOC abundance. Crop rotations increased cucumber rhizosphere bacteria, Pseudomonas spp. and 2,4-diacetylphloroglucinol producer abundances. Moreover, crop rotations changed cucumber rhizosphere bacterial community composition and increased bacterial community diversity. However, crop rotations decreased soil inorganic nitrogen content and inhibited cucumber seedling growth. Overall, rotations with Indian mustard and wild rocket suppressed cucumber Fusarium wilt disease, which might be linked to the increased rhizosphere bacterial diversity and abundances of potential plant-beneficial microorganisms (such as Pseudomonas spp. and 2,4-diacetylphloroglucinol producer).
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Plant viruses usually encode one or more movement proteins (MP) to accomplish their intercellular movement. A group of positive-strand RNA plant viruses requires three viral proteins (TGBp1, TGBp2, and TGBp3) that are encoded by an evolutionarily conserved genetic module of three partially overlapping open reading frames (ORFs), termed the triple gene block (TGB). However, how these three viral movement proteins function cooperatively in viral intercellular movement is still elusive. Using a novel in vivo double-stranded RNA (dsRNA) labeling system, we showed that the dsRNAs generated by potato virus X (PVX) RNA-dependent RNA polymerase (RdRp) are colocalized with viral RdRp, which are further tightly covered by "chain mail"-like TGBp2 aggregates and localizes alongside TGBp3 aggregates. We also discovered that TGBp2 interacts with the C-terminal domain of PVX RdRp, and this interaction is required for the localization of TGBp3 and itself to the RdRp/dsRNA bodies. Moreover, we reveal that the central and C-terminal hydrophilic domains of TGBp2 are required to interact with viral RdRp. Finally, we demonstrate that knockout of the entire TGBp2 or the domain involved in interacting with viral RdRp attenuates both PVX replication and movement. Collectively, these findings suggest that TGBp2 plays dual functional roles in PVX replication and intercellular movement.IMPORTANCE Many plant viruses contain three partially overlapping open reading frames (ORFs), termed the triple gene block (TGB), for intercellular movement. However, how the corresponding three proteins coordinate their functions remains obscure. In the present study, we provided multiple lines of evidence supporting the notion that PVX TGBp2 functions as the molecular adaptor bridging the interaction between the RdRp/dsRNA body and TGBp3 by forming "chain mail"-like structures in the RdRp/dsRNA body, which can also enhance viral replication. Taken together, our results provide new insights into the replication and movement of PVX and possibly also other TGB-containing plant viruses.
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Nicotiana/virologia , Proteínas do Movimento Viral em Plantas/metabolismo , Potexvirus/metabolismo , Replicação Viral/fisiologia , Retículo Endoplasmático/metabolismo , Doenças das Plantas/virologia , Domínios Proteicos/genética , RNA Viral/genéticaRESUMO
A microwave assisted extraction technology was used to extract chrysophanol from rhubarb. The present study will focus on the optimum extraction conditions of chrysophanol and discuss the inhibitory effect of chrysophanol on the biofilm formation of Streptococcus suis (S. suis). A Box-Behnken design based on single-factor experiments was applied to optimize the microwave assisted extraction process and to study the factors' relationships with each other. The results showed that a microwave temperature of 56 °C, ethanol concentration of 70%, microwave power of 540 W and liquid to raw material ratio of 55 mL g-1 were the optimal conditions for the microwave method. The yield of chrysophanol was 2.54 ± 0.07% under the optimal conditions, which was in agreement with the predicted value (2.64%). Then, the chemical structure of the extracted chrysophanol was identified by LC-MS. In addition, in vitro experiments showed that chrysophanol has an inhibitory effect on S. suis (minimum inhibitory concentration was 1.98 µg mL-1) and was shown to significantly inhibit the capability of S. suis to form a biofilm using crystal violet staining. Finally, scanning electron microscopy analysis showed that the three-dimensional structure of the biofilm deposited by the S. suis community was destroyed by chrysophanol.