Non-Coding Transcriptome Provides Novel Insights into the Escherichia coli F17 Susceptibility of Sheep Lamb

SIMPLE SUMMARY: Diarrhea and vomiting caused by Escherichia coli (E. coli) F17 are considered significant threats to animal farming. In the present study, RNA-Seq was performed to investigate the potential circRNA and miRNA biomarkers for E. coli F17-antagonism (AN) and -sensitive (SE) lambs. The results indicated that circRNA and miRNA expression is closely associated with the susceptibility of E. coli F17 in lambs. Numbers of circRNAs and miRNAs may serve as potential biomarkers for intestinal inflammatory response against E. coli F17 infection. Our study can provide a preliminary understanding of the underlying mechanisms of intestinal immunity. ABSTRACT: It has long been recognized that enterotoxigenic Escherichia coli (ETEC) is the major pathogen responsible for vomiting and diarrhea. E. coli F17, a main subtype of ETEC, is characterized by high morbidity and mortality in young livestock. However, the transcriptomic basis underlying E. coli F17 infection has not been fully understood. In this study, RNA sequencing was performed to explore the expression profiles of circRNAs and miRNAs in the jejunum of E. coli F17-antagonism (AN) and -sensitive (SE) lambs. A total of 16,534 circRNAs and 271 miRNAs (125 novel miRNAs and 146 annotated miRNAs) were screened, and 214 differentially expressed (DE) circRNAs and 53 DE miRNAs were detected between the AN and SE lambs (i.e., novel_circ_0025840, novel_circ_0022779, novel_miR_107, miR-10b). Functional enrichment analyses showed that source genes of DE circRNAs were mainly involved in metabolic-related pathways, while target genes of DE miRNAs were mainly enriched in the immune response pathways. Then, a two-step machine learning approach combining Random Forest (RF) and XGBoost (candidates were first selected by RF and further assessed by XGBoost) was performed, which identified 44 circRNAs and 39 miRNAs as potential biomarkers (i.e., novel_circ_0000180, novel_circ_0000365, novel_miR_192, oar-miR-496-3p) for E. coli infection. Furthermore, circRNA-related and lncRNA-related ceRNA networks were constructed, containing 46 circRNA-miRNA-mRNA competing triplets and 630 lncRNA-miRNA-mRNA competing triplets, respectively. By conducting a serious of bioinformatic analyses, our results revealed important circRNAs and miRNAs that could be potentially developed as candidate biomarkers for intestinal inflammatory response against E. coli F17 infection; our study can provide novel insights into the underlying mechanisms of intestinal immunity

Defining ovine dermal papilla cell markers and identifying key signaling pathways regulating its intrinsic properties

Dermal papilla cell (DPC), one of the key cell types during hair follicle development and regeneration, specifies hair size, shape and cycling. It is also an important in vitro screening model for hair growth. Although some characteristics of DPCs, such as agglutinative growth and marker genes, have been studied in mice and humans, the intrinsic properties of ovine DPCs and the regulatory mechanism of the intrinsic properties during continued culture in vitro remained unknown. In this study, based on our previous single-cell transcriptome sequencing on sheep lambskin, we verified SOX18 and PDGFRA as the novel marker genes of ovine DPCs through immunofluorescence staining on skin sections and cultured DPCs. Using continued cell culture and alkaline phosphatase staining, we found that different from mice and humans, ovine DPCs exhibit particularly robust and stable aggregation with unbated alkaline phosphatase activity till 30 passages during continued culture in vitro. Also, we found that the expression of some marker genes and the activity of Wnt/β-catenin signaling differ between early passaged DPCs and multiple passaged DPCs. Further, using Wnt/β-catenin agonist and antagonist, we demonstrated that Wnt/β-catenin signaling could regulate cell aggregation and alkaline phosphatase activity of ovine DPCs through regulating FGF and IGF signaling. This study provides the basis for isolating ovine DPCs and defines their intrinsic properties, which contribute to improving wool performance and medicine of hair regeneration

Silencing of a Cotton Actin-Binding Protein GhWLIM1C Decreases Resistance against Verticillium dahliae Infection

LIM proteins are widely spread in various types of plant cells and play diversely crucial cellular roles through actin cytoskeleton assembly and gene expression regulation. Till now, it has not been clear whether LIM proteins function in plant pathogen defense. In this study, we characterized a LIM protein, GhWLIM1C, in upland cotton (Gossypium hirsutum). We found that GhWLIM1C could bind and bundle the actin cytoskeleton, and it contains two LIM domains (LIM1 and LIM2). Both the two domains could bind directly to the actin filaments. Moreover, the LIM2 domain additionally bundles the actin cytoskeleton, indicating that it possesses a different biochemical activity than LIM1. The expression of GhWLIM1C responds to the infection of the cotton fungal pathogen Verticillium dahliae (V. dahliae). Silencing of GhWLIM1C decreased cotton resistance to V. dahliae. These may be associated with the down regulated plant defense response, including the PR genes expression and ROS accumulation in the infected cotton plants. In all, these results provide new evidence that a plant LIM protein functions in plant pathogen resistance and the assembly of the actin cytoskeleton are closely related to the triggering of the plant defense response

Effects of Heavy Metals on Phyllosphere and Rhizosphere Microbial Community of Bothriochloa ischaemum

Copper mining has resulted in severe damage to the ecological environment of mining areas. This study investigated heavy metal distribution in plants and compared the driving factors between aboveground and subsurface microorganisms, as well as the diversity in rhizosphere and non-rhizosphere soil microbial community response to heavy metal transfer factors in a copper tailings dam. We analyzed phyllosphere and soil microbial community using high-throughput sequencing and denaturing gradient gel electrophoresis, respectively. Although we detected chromium in aboveground and subsurface of Bothriochloa ischaemum specimens, no chromium was detected in soil. Total nitrogen was negatively correlated to the carbon and nitrogen ratios of plants and soil, respectively, while the total sulfur was negatively correlated to cadmium in roots. On the contrary, soil sulfur was positively correlated to cadmium in soil. Moreover, soil sulphur was the main influencing factor on the soil bacterial community, while ammonium nitrogen, total nitrogen, and zinc were the driving factors of fungi diversity in non-rhizosphere soil. Fungi diversity in the rhizosphere was significantly correlated to phosphatase, and fungi diversity in the non-rhizosphere was significantly correlated to sucrose enzymes. The transfer factor of lead was negatively correlated to rhizosphere fungi diversity, and the transfer factor of copper was significantly correlated to non-rhizosphere bacterial diversity. Results from this study may offer some scientific reference for the improvement of plant-microbe remediation efficiency. At the same time, this study could provide an ecological basis for further studies on soil ecosystem restoration and degradation mechanisms that are associated with copper tailings dams

Silencing of a Cotton Actin-Binding Protein GhWLIM1C Decreases Resistance against <i>Verticillium dahliae</i> Infection

LIM proteins are widely spread in various types of plant cells and play diversely crucial cellular roles through actin cytoskeleton assembly and gene expression regulation. Till now, it has not been clear whether LIM proteins function in plant pathogen defense. In this study, we characterized a LIM protein, GhWLIM1C, in upland cotton (Gossypium hirsutum). We found that GhWLIM1C could bind and bundle the actin cytoskeleton, and it contains two LIM domains (LIM1 and LIM2). Both the two domains could bind directly to the actin filaments. Moreover, the LIM2 domain additionally bundles the actin cytoskeleton, indicating that it possesses a different biochemical activity than LIM1. The expression of GhWLIM1C responds to the infection of the cotton fungal pathogen Verticillium dahliae (V. dahliae). Silencing of GhWLIM1C decreased cotton resistance to V. dahliae. These may be associated with the down regulated plant defense response, including the PR genes expression and ROS accumulation in the infected cotton plants. In all, these results provide new evidence that a plant LIM protein functions in plant pathogen resistance and the assembly of the actin cytoskeleton are closely related to the triggering of the plant defense response

Microstructure Evolution of 7075 Aluminum Alloy by Rotary

This study proposed a rotary back extrusion (RBE) process for an open punch, which is used to produce high-performance 7075 aluminum alloy cup-shaped piece. The RBE experiment was carried out on the Gleeble-3500 testing machine at 400 &deg;C and compared with the conventional back extrusion (CBE). The microstructure was analyzed by optical microscope, scanning electron microscope and DEFORM-3D simulation software. The results shown that compared with CBE, RBE can significantly increase the equivalent strain value and deformation uniformity of 7075 aluminum alloy cup-shape pieces. RBE deformation increases the accumulated strain of the piece, and the rotation of the die causes the piece to produce shear strain, which increases the overall strain of the cup-shape piece. The proportion of dynamic recrystallization increases, and the grain refinement was obvious. The micro-hardness value of the RBE sample is higher than that of the CBE sample, which could be the result of grain refinement strengthening. What is more, RBE and CBE have different metal flow laws

Magnaporthe oryzae Transcription Factor MoBZIP3 Regulates Appressorium Turgor Pressure Formation during Pathogenesis

The devastating fungus Magnaporthe oryzae (M. oryzae) forms a specialized infection structure known as appressorium, which generates enormous turgor, to penetrate the plant cells. However, how M. oryzae regulates the appressorium turgor formation, is not well understood. In this study, we identified MoBZIP3, a bZIP transcription factor that functioned in pathogenesis in M. oryzae. We found that the pathogenicity of the MoBZIP3 knockout strain (&Delta;mobzip3) was significantly reduced, and the defect was restored after re-expression of MoBZIP3, indicating that MoBZIP3 is required for M. oryzae virulence. Further analysis showed that MoBZIP3 functions in utilization of glycogen and lipid droplets for generation of glycerol in appressorium. MoBZIP3 localized in the nucleus and could bind directly to the promoters of the glycerol synthesis-related genes, MoPTH2, MoTGL1 and MoPEX6, and regulate their expression which is critical for glycerol synthesis in the appressorium turgor pressure generation. Furthermore, the critical turgor sensor gene MoSln1 was also down regulated and its subcellular localization was aberrant in &Delta;mobzip3, which leads to a disordered actin assembly in the &Delta;mobzip3 appressorium. Taken together, these results revealed new regulatory functions of the bZIP transcription factor MoBZIP3, in regulating M. oryzae appressorium turgor formation and infection

Anisotropic Low Cycle Behavior of the Extruded 7075 Al Alloy

The quasi-static and low cycle fatigue tests of extruded 7075 Al alloy (Φ200 mm) were investigated in three directions: the extrusion direction (ED), the radial direction (RD), and 45° with ED (45°). Grain morphology analysis, texture measurement, and fatigue fracture characterization were conducted to discuss the relationship between microstructure and mechanical properties. The experimental results showed that the ED specimen had higher ultimate tensile strength (UTS) and low cycle fatigue (LCF) properties, which were mainly attributed to the following three causes. First, the grain boundaries (GBs) had an obvious effect on the crack growth. The number of GBs in the three directions was different due to the shape of the grains elongated along the ED. Second, the sharp &lt;111&gt; texture and the small Schmidt factor along the ED explained the higher ultimate tensile strength (UTS) of the ED specimens. Third, fatigue fracture observation showed that the ED specimen had a narrow fatigue striation spacing, which indicated that the plastic deformation of the ED specimen was the smallest in each cycle. In addition, two fatigue prediction models were established to predict the LCF life of extruded 7075 Al alloy, based on the life response behavior of the three directions under different strains

Effects of ageing on low-cycle fatigue (LCF) properties of AZ80 magnesium alloy wheel hub

A hollow billet Expanding-Reducing Extrusion (ERE) wheel hub forming process was carried out with AZ80 magnesium alloy. The effects of ageing on Low-cycle fatigue (LCF) properties were investigated, and the microstructure was characterized with scanning electronic microscopy (SEM). The LCF testing results exhibited that the stress amplitude increased with increasing ageing time, while the plastic strain decreased with increasing ageing time. Cyclic softening followed by hardening was observed for 0 h and 16 h aged samples; however, there was no obvious cyclic softening or hardening for 36 h aged sample. The tension-compression asymmetry were not obvious due to the low strain, precipitation and the non typical basal texture. Fracture observations indicated that the fatigue cracks were initiated on the surface, and the crack propagation area decreased with increasing ageing time. Thus, it can be concluded that ageing has strong effects on the fatigue life for the studied magnesium alloy