The genome of the soybean pathogen Phytophthora sojae contains nearly 400 genes encoding candidate effector proteins carrying the host cell entry motif RXLR-dEER. Here, we report a broad survey of ...the transcription, variation, and functions of a large sample of the P. sojae candidate effectors. Forty-five (12%) effector genes showed high levels of polymorphism among P. sojae isolates and significant evidence for positive selection. Of 169 effectors tested, most could suppress programmed cell death triggered by BAX, effectors, and/or the PAMP INF1, while several triggered cell death themselves. Among the most strongly expressed effectors, one immediate-early class was highly expressed even prior to infection and was further induced 2- to 10-fold following infection. A second early class, including several that triggered cell death, was weakly expressed prior to infection but induced 20- to 120-fold during the first 12 h of infection. The most strongly expressed immediate-early effectors could suppress the cell death triggered by several early effectors, and most early effectors could suppress INF1-triggered cell death, suggesting the two classes of effectors may target different functional branches of the defense response. In support of this hypothesis, misexpression of key immediate-early and early effectors severely reduced the virulence of P. sojae transformants.
This study experimentally investigates the impact of a single piezoelectric (PZT) actuator on a turbulent boundary layer from a statistical viewpoint. The working conditions of the actuator include a ...range of frequencies and amplitudes. The streamwise velocity signals in the turbulent boundary layer flow are measured downstream of the actuator using a hot-wire anemometer. The mean velocity profiles and other basic parameters are reported. Spectra results obtained by discrete wavelet decomposition indicate that the PZT vibration primarily influences the near-wall region. The turbulent intensities at different scales suggest that the actuator redistributes the near-wall turbulent energy. The skewness and flatness distributions show that the actuator effectively alters the sweep events and reduces intermittency at smaller scales. Moreover, under the impact of the PZT actuator, the symmetry of vibration scales’ velocity signals is promoted and the structural composition appears in an orderly manner. Probability distribution function results indicate that perturbation causes the fluctuations in vibration scales and smaller scales with high intensity and low intermittency. Based on the flatness factor, the bursting process is also detected. The vibrations reduce the relative intensities of the burst events, indicating that the streamwise vortices in the buffer layer experience direct interference due to the PZT control.
Soluble N‐ethylmaleimide‐sensitive factor attachment protein receptors (SNAREs) mediate cellular membrane fusion and intracellular vesicle trafficking in eukaryotic cells, and are critical in the ...growth and development of pathogenic fungi such as Magnaporthe oryzae which causes rice blast. Rice blast is thought to involve distinct SNARE‐mediated transport and secretion of fungal effector proteins into the host to modulate rice immunity. We have previously characterized two SNARE proteins, secretory protein (MoSec22) and vesicle‐associated membrane protein (MoVam7), as being important in cellular transport and pathogenicity. Here, we show that syntaxin 8 (MoSyn8), a Qc‐SNARE protein homolog, also plays important roles in growth, conidiation, and pathogenicity. The MoSYN8 deletion mutant (∆Mosyn8) mutant exhibits defects in endocytosis and F‐actin organization, appressorium turgor pressure generation, and host penetration. In addition, the ∆Mosyn8 mutant cannot elaborate biotrophic invasion of the susceptible rice host, or secrete avirulence factors Avr‐Pia (corresponding to the rice resistance gene Pia) and Avrpiz‐t (the cognate Avr gene for the resistance gene Piz‐t) proteins. Our study of MoSyn8 advances our understanding of SNARE proteins in effector secretion which underlies the normal physiology and pathogenicity of M. oryzae, and it sheds new light on the mechanism of the blight disease caused by M. oryzae.
The production of reactive oxygen species (ROS) is a ubiquitous defense response in plants. Adapted pathogens evolved mechanisms to counteract the deleterious effects of host-derived ROS and promote ...infection. How plant pathogens regulate this elaborate response against ROS burst remains unclear. Using the rice blast fungus
, we uncovered a self-balancing circuit controlling response to ROS in planta and virulence. During infection, ROS induces phosphorylation of the high osmolarity glycerol pathway kinase MoOsm1 and its nuclear translocation. There, MoOsm1 phosphorylates transcription factor MoAtf1 and dissociates MoAtf1-MoTup1 complex. This releases MoTup1-mediated transcriptional repression on oxidoreduction-pathway genes and activates the transcription of MoPtp1/2 protein phosphatases. In turn, MoPtp1/2 dephosphorylate MoOsm1, restoring the circuit to its initial state. Balanced interactions among proteins centered on MoOsm1 provide a means to counter host-derived ROS. Our findings thereby reveal new insights into how
utilizes a phosphor-regulatory circuitry to face plant immunity during infection.
Returning straw to soil is an effective way to sustain or improve soil quality and crop yields. However, a robust understanding of the impact of straw return on the composition of the soil microbial ...communities under field conditions has remained elusive. In this study, we characterized the effects of wheat straw return on soil bacterial and fungal communities in a wheat-soybean rotation system over a 3-year period, using Illumina-based 16S rRNA, and internal transcribed region (ITS) amplicon sequencing. Wheat straw return significantly affected the α-diversity of the soil bacterial, but not fungal, community. It enhanced the relative abundance of the bacterial phylum Proteobacteria and the fungal phylum Zygomycota, but reduced that of the bacterial phylum Acidobacteria, and the fungal phylum Ascomycota. Notably, it enriched the relative abundance of nitrogen-cycling bacterial genera such as
and
. Preliminary analysis of soil chemical properties indicated that straw return soils had significantly higher total nitrogen (TN) contents than no straw return soils. In addition, the relative abundance of fungal genera containing pathogens was significantly lower in straw return soils relative to control soils, such as
,
, and
. These results suggested a selection effect from the 3-year continuous straw return treatment and the soil bacterial and fungal communities were moderately changed.
In order to further improve the utility of unmanned aerial vehicle (UAV) remote-sensing for quickly and accurately monitoring the growth of winter wheat under film mulching, this study examined the ...treatments of ridge mulching, ridge–furrow full mulching, and flat cropping full mulching in winter wheat. Based on the fuzzy comprehensive evaluation (FCE) method, four agronomic parameters (leaf area index, above-ground biomass, plant height, and leaf chlorophyll content) were used to calculate the comprehensive growth evaluation index (CGEI) of the winter wheat, and 14 visible and near-infrared spectral indices were calculated using spectral purification technology to process the remote-sensing image data of winter wheat obtained by multispectral UAV. Four machine learning algorithms, partial least squares, support vector machines, random forests, and artificial neural network networks (ANN), were used to build the winter wheat growth monitoring model under film mulching, and accuracy evaluation and mapping of the spatial and temporal distribution of winter wheat growth status were carried out. The results showed that the CGEI of winter wheat under film mulching constructed using the FCE method could objectively and comprehensively evaluate the crop growth status. The accuracy of remote-sensing inversion of the CGEI based on the ANN model was higher than for the individual agronomic parameters, with a coefficient of determination of 0.75, a root mean square error of 8.40, and a mean absolute value error of 6.53. Spectral purification could eliminate the interference of background effects caused by mulching and soil, effectively improving the accuracy of the remote-sensing inversion of winter wheat under film mulching, with the best inversion effect achieved on the ridge–furrow full mulching area after spectral purification. The results of this study provide a theoretical reference for the use of UAV remote-sensing to monitor the growth status of winter wheat with film mulching.
The fungal cell wall plays an essential role in maintaining cell morphology, transmitting external signals, controlling cell growth, and even virulence. Relaxation and irreversible stretching of the ...cell wall are the prerequisites of cell division and development, but they also inevitably cause cell wall stress. Both Mitotic Exit Network (MEN) and Cell Wall Integrity (CWI) are signaling pathways that govern cell division and cell stress response, respectively, how these pathways cross talk to govern and coordinate cellular growth, development, and pathogenicity remains not fully understood. We have identified MoSep1, MoDbf2, and MoMob1 as the conserved components of MEN from the rice blast fungus Magnaporthe oryzae. We have found that blocking cell division results in abnormal CWI signaling. In addition, we discovered that MoSep1 targets MoMkk1, a conserved key MAP kinase of the CWI pathway, through protein phosphorylation that promotes CWI signaling. Moreover, we provided evidence demonstrating that MoSep1-dependent MoMkk1 phosphorylation is essential for balancing cell division with CWI that maintains the dynamic stability required for virulence of the blast fungus.
Abstract
Background
Vascular invasion is a major route for intrahepatic and distant metastasis in hepatocellular carcinoma (HCC) and is a strong negative prognostic factor. Circular RNAs (circRNAs) ...play important roles in tumorigenesis and metastasis. However, the regulatory functions and underlying mechanisms of circRNAs in the development of vascular invasion in HCC are largely unknown.
Methods
High throughput sequencing was used to screen dysregulated circRNAs in portal vein tumor thrombosis (PVTT) tissues. The biological functions of candidate circRNAs in the migration, vascular invasion, and metastasis of HCC cells were examined in vitro and in vivo. To explore the underlying mechanisms, RNA sequencing, MS2-tagged RNA affinity purification, mass spectrometry, and RNA immunoprecipitation assays were performed.
Results
circRNA sequencing followed by quantitative real-time PCR (qRT-PCR) revealed that circRNA pleckstrin and Sect. 7 domain containing 3 (circPSD3) was significantly downregulated in PVTT tissues. Decreased circPSD3 expression in HCC tissues was associated with unfavourable characteristics and predicted poor prognosis in HCC. TAR DNA-binding protein 43 (TDP43) inhibited the biogenesis of circPSD3 by interacting with the downstream intron of pre-PSD3. circPSD3 inhibited the intrahepatic vascular invasion and metastasis of HCC cells in vitro and in vivo. Serpin family B member 2 (SERPINB2), an endogenous
bona fide
inhibitor of the urokinase-type plasminogen activator (uPA) system, is the downstream target of circPSD3. Mechanistically, circPSD3 interacts with histone deacetylase 1 (HDAC1) to sequester it in the cytoplasm, attenuating the inhibitory effect of HDAC1 on the transcription of SERPINB2. In vitro and in vivo studies demonstrated that circPSD3 is a promising inhibitor of the uPA system.
Conclusions
circPSD3 is an essential regulator of vascular invasion and metastasis in HCC and may serve as a prognostic biomarker and therapeutic target.
This study proposes a fatigue life analysis method for long-span CFST arch bridges based on a vehicle–bridge coupled vibration analysis model, which can analyze the structural dynamic effects and the ...excessive fatigue damage caused by the passage of vehicles. In situ test analysis of bridge dynamic characteristics is carried out, and a numerical model considering the vehicle–bridge coupled system is validated according to the measured vibration modes, frequency, and displacement time history. The results indicate that the proposed vehicle–bridge coupled vibration numerical model can be used to simulate the dynamic response of the bridge under various conditions. The factors of vehicle speed, vehicle weight, and road surface condition are further selected to analyze the vehicle–bridge coupled vibration effect, and it is found that the response time history is more sensitive to the vehicle weight factor. In addition, the fatigue life of suspenders at different positions is compared, which is found to decrease significantly with a reduction in suspender length. Due to damage to the suspender caused by environmental erosion, the cross-sectional area decreases and the stress amplitude changes, resulting in a decrease in the fatigue reliability of the suspender under different conditions.
GTP-binding protein (G-protein) and regulator of G-protein signaling (RGS) mediated signal transduction are critical in the growth and virulence of the rice blast pathogen Magnaporthe oryzae. We have ...previously reported that there are eight RGS and RGS-like proteins named MoRgs1 to MoRgs8 playing distinct and shared regulatory functions in M. oryzae and that MoRgs1 has a more prominent role compared to others in the fungus. To further explore the unique regulatory mechanism of MoRgs1, we screened a M. oryzae cDNA library for genes encoding MoRgs1-interacting proteins and identified MoCkb2, one of the two regulatory subunits of the casein kinase (CK) 2 MoCk2. We found that MoCkb2 and the sole catalytic subunit MoCka1 are required for the phosphorylation of MoRgs1 at the plasma membrane (PM) and late endosome (LE). We further found that an endoplasmic reticulum (ER) membrane protein complex (EMC) subunit, MoEmc2, modulates the phosphorylation of MoRgs1 by MoCk2. Interestingly, this phosphorylation is also essential for the GTPase-activating protein (GAP) function of MoRgs1. The balance among MoRgs1, MoCk2, and MoEmc2 ensures normal operation of the G-protein MoMagA-cAMP signaling required for appressorium formation and pathogenicity of the fungus. This has been the first report that an EMC subunit is directly linked to G-protein signaling through modulation of an RGS-casein kinase interaction.
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