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Rice false smut, caused by Ustilaginoidea virens, is a devastating disease that greatly reduces rice yield and quality. However, controlling rice false smut disease is challenging due to the unique infection mode of U. virens. Therefore, there is a need for early diagnosis and monitoring techniques to prevent the spread of this disease. Lateral flow strip-based recombinase polymerase amplification (LF-RPA) overcomes the limitations of current U. virens detection technologies, which are time-consuming, require delicate equipment, and have a high false-positive rate. In this study, we used a comparative genomics approach to identify Uv_3611, a specific gene of U. virens, as the target for the LF-RPA assay. The designed primers and probe efffectively detected the genomic DNA (gDNA) of U. virens and demonstrated no cross-reactivity with related pathogens. Under optimal conditions, the LF-RPA assay demonstrated a sensitivity of 10 pg of U. virens gDNA. Additionally, by incorporating a simplified PEG-NaOH method for plant DNA extraction, the LF-RPA assay enabled the detection of U. virens in rice spikelets within 30 min, without the need for specialized equipment. Furthermore, the LF-RPA assay successfully detected U. virens in naturally infected rice and seed samples in the field. Therefore, the LF-RPA assay is sensitive, efficient, and convenient, and could be developed as a kit for monitoring rice false smut disease in the field.

Sorafenib (SF), a multi-targeted tyrosine kinase inhibitor, serves as a primary therapeutic modality for advanced liver cancer. Nonetheless, its clinical efficacy is hindered by various obstacles, such as limited bioavailability and inadequate accumulation. This study introduces a novel biomimetic mineralization enzyme, known as BSA@Pt/Ce6/SF@M (PCFM). The PCFM incorporates platinum (Pt) as a catalytic agent, SF as a molecular-targeted therapeutic agent, and Ce6 as a photosensitizer within liver cancer cell membranes. This strategy enables the combination of various anti-tumor treatments, such as photodynamic therapy (PDT) and autophagy induction, leading to increased bioavailability of SF and achieving a multidimensional synergistic anticancer effect. The PDT effect produced by Ce6 in PCFM greatly enhances SF-induced autophagy, effectively promoting autophagic cell death. Furthermore, Pt dissociates from the biomineralization process, acquiring peroxidase properties through chemokinetic reactions. This facilitates the catalysis of significant oxygen generation, addressing the challenge of hypoxia in the tumor microenvironment and improving the efficacy of PDT. Moreover, the SF further enhances therapeutic efficacy by inducing autophagy in response to energy deprivation, as indicated by the reduced levels of HIF-1α, p62, along with increased levels of ROS and LC3-Ⅱ/Ι. This biomineralization-based nanoenzyme exhibits strong anti-tumor characteristics, offering a novel strategy for overcoming challenges in liver cancer treatment.

Mitochondria are highly dynamic organelles that constantly change their morphology to adapt to the cellular environment through fission and fusion, which is critical for a cell to maintain normal cellular functions. Despite the significance of this process in the development and pathogenicity of the rice blast fungus Magnaporthe oryzae, the underlying mechanism remains largely elusive. Here, we identified and characterized a mitochondrial outer membrane translocase, MoTom20, in M. oryzae. Targeted gene deletion revealed that MoTom20 plays an important role in vegetative growth, conidiogenesis, penetration, and infectious growth of M. oryzae. The growth rate, conidial production, appressorium turgor, and pathogenicity are decreased in the ΔMotom20 mutant compared with the wild-type and complemented strains. Further analysis revealed that MoTom20 localizes in mitochondrion and plays a key role in regulating mitochondrial fission and fusion balance, which is critical for infectious growth. Finally, we found that MoTom20 is involved in fatty-acid utilization, and its yeast homolog ScTom20 is able to rescue the defects of ΔMotom20 in mitochondrial morphology and pathogenicity. Overall, our data demonstrate that MoTom20 is a key regulator for mitochondrial morphology maintenance, which is important for infectious growth of the rice blast fungus M. oryzae. [Graphic: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

ABSTRACT Regulator of G-protein signaling (RGS) proteins exhibit GTPase-accelerating protein activities to govern G-protein function. In the rice blast fungus Magnaporthe oryzae, there is a family of at least eight RGS and RGS-like proteins (MoRgs1 to MoRgs8), each exhibiting distinct or shared functions in the growth, appressorium formation, and pathogenicity. MoRgs3 recently emerged as one of the crucial regulators that senses intracellular oxidation during appressorium formation. To explore this unique regulatory mechanism of MoRgs3, we identified the nucleoside diphosphate kinase MoNdk1 that interacts with MoRgs3. MoNdk1 phosphorylates MoRgs3 under induced intracellular reactive oxygen species levels, and MoRgs3 phosphorylation is required for appressorium formation and pathogenicity. In addition, we showed that MoRgs3 phosphorylation determines its interaction with MoCrn1, a coronin-like actin-binding protein homolog, which regulates MoRgs3 internalization. Finally, we provided evidence demonstrating that MoRgs3 functions in MoMagA-mediated cAMP signaling to regulate normal appressorium induction. By revealing a novel signal perception mechanism, our studies highlighted the complexity of regulation during the appressorium function and pathogenicity of the blast fungus.IMPORTANCEWe report that MoRgs3 becomes phosphorylated in an oxidative intracellular environment during the appressorium formation stage. We found that this phosphorylation is carried out by MoNdk1, a nucleoside diphosphate kinase. In addition, this phosphorylation leads to a higher binding affinity between MoRgs3 and MoCrn1, a coronin-like actin-binding protein that was implicated in the endocytic transport of several other RGS proteins of Magnaporthe oryzae. We further found that the internalization of MoRgs3 is indispensable for its GTPase-activating protein function toward the Gα subunit MoMagA. Importantly, we characterized how such cellular regulatory events coincide with cAMP signaling-regulated appressorium formation and pathogenicity in the blast fungus. Our studies uncovered a novel intracellular reactive oxygen species signal-transducing mechanism in a model pathogenic fungus with important basic and applied implications.

Although significant amounts of methane are present in anthracite coal seams, coalbed methane resources cannot be extracted effectively and quickly. This study mainly focused on investigating the pore system and methane adsorption capacity to clarify the storage of coalbed methane. Anthracite coal samples from the Anzenan coalbed methane block in China were collected, and pore characterization methods (low-pressure N2 adsorption, mercury injection experiments, and scanning electron microscopy (SEM) observations) and methane isothermal adsorption experiments were conducted. The results showed that few mesopores and nanoscale macropores were present in the anthracite coal samples. The volume of the 2–300 nm pores in these coal samples obtained from the N2 adsorption experiment was lower than 0.01 cm3/g. SEM observations also revealed that only a small number of mesopores and nanoscale macropores could be seen, and most of these pores were isolated. In terms of the methane isothermal adsorption data, it was found these anthracite coals have a large methane adsorption capacity, and the Langmuir volume ranges from 19.5 to 28.4 cm3/g, with an average of 22.2 cm3/g. With increasing ash yield, the Langmuir volume decreased linearly, indicating that methane molecules were mainly adsorbed in the organic matter of coal. As methane is mainly adsorbed in the micropores of coal organic matter and there are few mesopores and nanoscale macropores in the organic pores in coal, there are not enough tunnels to transport the adsorbed methane molecules to the outside. Thus, it is difficult to extract coalbed methane from anthracite. This study reveals the impact of pore system limitations on the storage and extraction of coalbed methane in anthracite coal. The findings can be applied to the extraction of coalbed methane from anthracite coal seams worldwide.

Abstract Rice is a staple crop feeding more than 50% of the world’s population and, its sustainable production is crucial to the global food security. However, blast disease caused by the filamentous fungus Magnaporthe oryzae (anamorph: Pyricularia oryzae) threatens sustainable rice production as it can reduce grain yields over 30% in epidemic years. Therefore, deciphering the molecular mechanisms of the M. oryzae-rice interaction and the mechanism that how M. oryzae overcomes rice defense is crucial for developing new strategies to make green fungicides and disease-resistant rice varieties. This review provides a comprehensive overview of the latest advances in understanding the molecular mechanisms by which M. oryzae perceives host surface signals and subsequently develops specific infection structure called appressoria. In addition, the review delves into the strategies that the fungus utilizes to overcome host immunity, which in turn allows it to colonize rice tissues. Finally, we propose the perspectives for dissecting the pathogenesis of the fungus and controlling rice blast disease.

ABSTRACTAutophagy is a central biodegradation pathway critical in eliminating intracellular cargo to maintain cellular homeostasis and improve stress resistance. At the same time, the key component of the mitogen-activated protein kinase cascade regulating cell wall integrity signaling MoMkk1 has an essential role in the autophagy of the rice blast fungus Magnaporthe oryzae. Still, the mechanism of how MoMkk1 regulates autophagy is unclear. Interestingly, we found that MoMkk1 regulates the autophagy protein MoAtg9 through phosphorylation. MoAtg9 is a transmembrane protein subjected to phosphorylation by autophagy-related protein kinase MoAtg1. Here, we provide evidence demonstrating that MoMkk1-dependent MoAtg9 phosphorylation is required for phospholipid translocation during isolation membrane stages of autophagosome formation, an autophagic process essential for the development and pathogenicity of the fungus. In contrast, MoAtg1-dependent phosphorylation of MoAtg9 negatively regulates this process, also impacting growth and pathogenicity. Our studies are the first to demonstrate that MoAtg9 is subject to MoMkk1 regulation through protein phosphorylation and that MoMkk1 and MoAtg1 dichotomously regulate autophagy to underlie the growth and pathogenicity of M. oryzae.IMPORTANCEMagnaporthe oryzae utilizes multiple signaling pathways to promote colonization of host plants. MoMkk1, a cell wall integrity signaling kinase, plays an essential role in autophagy governed by a highly conserved autophagy kinase MoAtg1-mediated pathway. How MoMkk1 regulates autophagy in coordination with MoAtg1 remains elusive. Here, we provide evidence that MoMkk1 phosphorylates MoAtg9 to positively regulate phospholipid translocation during the isolation membrane or smaller membrane structures stage of autophagosome formation. This is in contrast to the negative regulation of MoAtg9 by MoAtg1 for the same process. Intriguingly, MoMkk1-mediated MoAtg9 phosphorylation enhances the fungal infection of rice, whereas MoAtg1-dependant MoAtg9 phosphorylation significantly attenuates it. Taken together, we revealed a novel mechanism of autophagy and virulence regulation by demonstrating the dichotomous functions of MoMkk1 and MoAtg1 in the regulation of fungal autophagy and pathogenicity.

Abstract Oceanic transport of heat by ubiquitous mesoscale eddies plays a critical role in regulating climate variability and redistributing excess heat absorbed by ocean under global warming. Eddies have long been simplified as axisymmetric vortices and their influence on heat transport remains unclear. Here, we combine satellite and drifter data and show that oceanic mesoscale eddies are asymmetric and directionally-dependent, and are controlled by their self-sustaining nature and their dynamical environment. Both the direction and amplitude of eddy-induced heat fluxes are significantly influenced by eddy’s asymmetry and directional dependence. When the eddy velocity field is decomposed into asymmetric and symmetric components, the eddy kinetic energy exhibits a nearly equal partition between these two components. The total eddy-induced meridional heat flux similarly doubles the heat flux induced by the symmetric components, highlighting the crucial contribution of eddy asymmetry on the magnitude of eddy-induced oceanic heat transport.

Mesoscale eddies contribute to the global oceanic energy budget and cascade; they induce substantial mass and heat transport, thus playing a key role in the global climate system. The three-dimensional (3D) structures of mesoscale eddies should be studied in order to quantify their impact. In this paper, mesoscale eddies are reviewed from the perspective of their horizontal and vertical structures, temporal evolution, and fine structures. The universal 3D structure of mesoscale eddies is revealed via observations, and findings show that their tendency to achieve lowest-energy or minimum-dissipation states shapes their lowest-order coherent structure. Recent efforts also push forward to the higher-order structure of eddies: The understanding of horizontal structures is gradually evolving from symmetric to asymmetric, and the vertical structure based on separable variable assumption and vertical mode decomposition should further take the eddy vertical alignment and tilt into account. The temporal evolution of mesoscale eddies’ 3D structure is in a dynamical balance influenced by multiple competitive factors, such as eddies’ self-sustaining ability, background gradient and deformation, and generation of submesoscale fine structures. The submesoscale processes associated with mesoscale eddies have been intensively studied in recent years, and they should be considered as a fine-scale part of the 3D structure of eddies rather than separate instability processes. This review hopes to provide the readers with an enlightening, yet not thorough, review of the development of mesoscale eddies’ structures and brings up potential topics and unresolved difficulties for further research and development.

Autophagy and Cell wall integrity (CWI) signaling are critical stress-responsive processes during fungal infection of host plants. In the rice blast fungus Magnaporthe oryzae, autophagy-related (ATG) proteins phosphorylate CWI kinases to regulate virulence; however, how autophagy interplays with CWI signaling to coordinate such regulation remains unknown. Here, we have identified the phosphorylation of ATG protein MoAtg4 as an important process in the coordination between autophagy and CWI in M. oryzae. The ATG kinase MoAtg1 phosphorylates MoAtg4 to inhibit the deconjugation and recycling of the key ATG protein MoAtg8. At the same time, MoMkk1, a core kinase of CWI, also phosphorylates MoAtg4 to attenuate the C-terminal cleavage of MoAtg8. Significantly, these two phosphorylation events maintain proper autophagy levels to coordinate the development and pathogenicity of the rice blast fungus.

The Hurenbuqi Sag, located on the Bayinbaolige uplift in the northern Erlian Basin, is a secondary depression rich in oil. In order to study the influence on hydrocarbon accumulation of the Early Cretaceous structures in the southern Hurenbuqi Sag, the geological, drilling and geophysical data has been analyzed carefully to reveal the Early Cretaceous extensional structural characteristics, differential structural deformation characteristics, tectonic evolution process, and the hydrocarbon accumulation process. The study revealed that, extensional tectonic system is the mainly composition of Early Cretaceous structure, it has controlled the three main tectonic subsidence stages, early initial tectonic subsidence, middle rapid tectonic subsidence and late slow tectonic subsidence, of the southern Hurenbuqi Sag. During the tectonic evolution, the Early Cretaceous deformation obviously controlled the development of the source rocks and hydrocarbon accumulation. This study is of great significance for analyzing the enrichment regulations of hydrocarbon and guiding the next oil and gas exploration.

ABSTRACTDuring the early stages of host-pathogen interaction, the production of reactive oxygen species (ROS) in host cells is a critical strategy to halt the spread of pathogens. Pathogens have to minimize the deleterious effects of host-generated ROS for successful invasion. However, the underlying mechanism remains largely elusive. Here, we report that a peroxisomal 3-ketoacyl-CoA thiolase of rice blast fungus Magnaporthe oryzae, MoKat2, functions in host ROS homeostasis likely through regulating fungal peroxisome morphology during infection. M. oryzae peroxisome elongates in response to ROS stress, and this is attributed to the function of peroxisome localized MoKat2. MoKat2 forms homodimer in vivo while the homodimer is dissociated into monomer in response to host-derived ROS. The MoKat2 monomer binds to peroxisome membranes through its amphipathic helices to control peroxisome elongation, resulting in host ROS depletion and subsequent M. oryzae invasion in rice cells. Our findings shed light on the interplay between pathogenic peroxisome dynamics and host ROS accumulation, expanding the crucial role of organelle-mediated infection in plant-microbe interactions.IMPORTANCEThe interplay between plant and pathogen is a dynamic process, with the host’s innate defense mechanisms serving a crucial role in preventing infection. In response to many plant pathogen infections, host cells generate the key regulatory molecule, reactive oxygen species (ROS), to limit the spread of the invading organism. In this study, we reveal the effects of fungal peroxisome dynamics on host ROS homeostasis, during the rice blast fungus Magnaporthe oryzae infection. The elongation of the peroxisome appears contingent upon ROS and links to the accumulation of ROS within the host and the infectious growth of the pathogen. Importantly, we identify a peroxisomal 3-ketoacyl-CoA thiolase, MoKat2, responsible for the elongation of the peroxisome during the infection. In response to host-derived ROS, the homodimer of MoKat2 undergoes dissociation to bind peroxisome membranes for peroxisome elongation. This process, in turn, inhibits the accumulation of host ROS, which is necessary for successful infection. Overall, our study is the first to highlight the intricate relationship between fungal organelle dynamics and ROS-mediated host immunity, extending the fundamental knowledge of pathogen-host interaction.

Context Guanxin V (GX), a traditional Chinese medicine formula, is safe and effective in the treatment of coronary artery disease. However, its protective effect on myocardial ischaemia reperfusion injury (MIRI) is unclear.Objective To investigate the cardioprotective effect of GX on MIRI and explore the potential mechanism.Materials and methods Sprague-Dawley male rats were divided into Sham, MIRI and MIRI + GX groups. GX (6 g/kg) was administered to rats via intragastric administration for seven days before ischaemia reperfusion (IR) surgery. The infarct size, histopathology, serum enzyme activities, ultrastructure of the cardiac mitochondria were assessed. H9c2 cells were pre-treated with GX (0.5 mg/mL), and then exposed to hypoxia/reoxygenation (HR). The cell viability and LDH levels were measured. Network pharmacology was conducted to predict the potential mechanism. The related targets of GX were predicted using the TCMSP database, DrugBank database, etc. Finally, pharmacological experiments were used to validate the predicted results.Results In vivo, GX significantly reduced the myocardial infarct size from 56.33% to 17.18%, decreased the levels of AST (239.32 vs. 369.18 U/L), CK-MB (1324.61 vs. 2066.47 U/L) and LDH (1245.26 vs. 1969.62 U/L), and reduced mitochondrial damage. In vitro, GX significantly increased H9c2 cell viability (IC50 = 3.913 mg/mL) and inhibited the release of LDH (207.35 vs. 314.33). In addition, GX could maintain iron homeostasis and reduce oxidative stress level by regulating iron metabolism-associated proteins.Conclusions GX can attenuate MIRI via regulating iron homeostasis, indicating that GX may act as a potential candidate for the treatment of MIRI.

The emergence of fungicide resistance severely threatens crop production by limiting the availability and application of established fungicides. Therefore, it is urgent to identify new fungicidal targets for controlling plant diseases. Here, we characterized the function of a conserved homoserine O-acetyltransferase (HOA) from the rice blast fungus Magnaporthe oryzae that could serve as the candidate antifungal target. Deletion of the MoMET2 and MoCYS2 genes encoding HOAs perturbed the biosynthesis of methionine and S-adenyl methionine, a methyl group donor for epigenetic modifications, and severely attenuated the development and virulence of M. oryzae. The ∆Momet2 mutant is significantly increased in 5-methylcytosine (5mC) modification that represses the expression of genes required for pathogenicity, including MoGLIK and MoCDH-CYT. We further showed that host-induced gene silencing (HIGS) targeting MoMET2 and MoCYS2 effectively controls rice blasts. Our studies revealed the importance of HOA in the development and virulence of M. oryzae, which suggests the potential feasibility of HOA as new targets for novel anti-rice blast measurements.

Precision actuation is a foundational technology in high-end equipment domains, where stroke, velocity, and accuracy are critical for processing and/or detection quality, precision in spacecraft flight trajectories, and accuracy in weapon system strikes. Piezoelectric actuators (PEAs), known for their nanometer-level precision, flexible stroke, resistance to electromagnetic interference, and scalable structure, have been widely adopted across various fields. Therefore, this study focuses on extreme scenarios involving ultra-high precision (micrometer and beyond), minuscule scales, and highly complex operational conditions. It provides a comprehensive overview of the types, working principles, advantages, and disadvantages of PEAs, along with their potential applications in piezo-actuated smart mechatronic systems (PSMSs). To address the demands of extreme scenarios in high-end equipment fields, we have identified five representative application areas: positioning and alignment, biomedical device configuration, advanced manufacturing and processing, vibration mitigation, micro robot system. Each area is further divided into specific subcategories, where we explore the underlying relationships, mechanisms, representative schemes, and characteristics. Finally, we discuss the challenges and future development trends related to PEAs and PSMSs. This work aims to showcase the latest advancements in the application of PEAs and provide valuable guidance for researchers in this field.

Oceanic mesoscale eddies are important dynamical processes in the Southern Ocean. Using high-resolution (∼0.1° for the ocean) Community Earth System Model (CESM-HR) simulations under a high-carbon emission scenario, we investigate the role of mesoscale eddies in regulating the response of the Subantarctic Front (SAF) to global warming. The CESM-HR simulates more realistic oceanic fronts and mesoscale eddies in the Southern Ocean than a coarse-resolution (∼1° for the ocean) CESM. Under global warming, the SAF is projected to intensify. The mean flow temperature advection intensifies the front, whereas the mesoscale-eddy-induced temperature advection and atmospheric dampening play primary (∼67%) and secondary (∼28%) roles in counteracting the effect of mean flow temperature advection. Our study suggests the importance of mesoscale eddies on inhibiting the SAF intensification under global warming and necessity of mesoscale-eddy-resolving simulations for faithful projection of future climate changes in the Southern Ocean.

Abstract The mitotic exit network (MEN) pathway is a vital kinase cascade regulating the timely and correct progress of cell division. In the rice blast fungus Magnaporthe oryzae, the MEN pathway, consisting of conserved protein kinases MoSep1 and MoMob1-MoDbf2, is important in the development and pathogenicity of the fungus. We found that deletion of MoSEP1 affects the phosphorylation of MoMob1, but not MoDbf2, in contrast to what was found in the buddy yeast Saccharomyces cerevisiae, and verified this finding by in vitro phosphorylation assay and mass spectrometry (MS) analysis. We also found that S43 residue is the critical phosphor-site of MoMob1 by MoSep1, and proved that MoSep1-dependent MoMob1 phosphorylation is essential for cell division during the development of M. oryzae. We further provided evidence demonstrating that MoSep1 phosphorylates MoMob1 to maintain the cell cycle during vegetative growth and infection. Taken together, our results revealed that the MEN pathway has both distinct and conservative functions in regulating the cell cycle during the development and pathogenesis of M. oryzae.

The rice blast fungus Magnaporthe oryzae forms specialized infectious structures called appressoria that breach host cells to initiate infection. Previous studies demonstrated that the regulator of G-protein signaling (RGS)-like protein MoRgs7 undergoes endocytosis upon fungal sensing of hydrophobic environmental cues to activate cAMP signaling required for appressorium formation. However, the mechanism by which MoRgs7 internalizes and its fate remains undetermined. We here show that MoSep1, a conserved protein kinase of Mitotic Exit Network (MEN), phosphorylates MoRgs7 to regulate its function. MoRgs7 phosphorylation determines its interaction with MoCrn1, a coronin-like actin-binding protein homolog that also modulates the internalization of MoRgs7. Importantly, the endocytic transport of MoRgs7 is critical for its GTPase-activating protein (GAP) function important in cAMP signaling. Together, our findings revealed a novel mechanism by which M. oryzae activates MoRgs7-mediated hydrophobic cue-sensing signal transduction involving protein phosphorylation and endocytic transport to govern appressorium formation and fungal pathogenicity.

Magnaporthe oryzae causes rice blasts posing serious threats to food security worldwide. During infection, M. oryzae utilizes several transmembrane receptor proteins that sense cell surface cues to induce highly specialized infectious structures called appressoria. However, little is known about the mechanisms of intracellular receptor tracking and their function. Here, we described that disrupting the coat protein complex II (COPII) cargo protein MoErv14 severely affects appressorium formation and pathogenicity as the ΔMoerv14 mutant is defective not only in cAMP production but also in the phosphorylation of the mitogen-activated protein kinase (MAPK) MoPmk1. Studies also showed that either externally supplementing cAMP or maintaining MoPmk1 phosphorylation suppresses the observed defects in the ΔMoerv14 strain. Importantly, MoErv14 is found to regulate the transport of MoPth11, a membrane receptor functioning upstream of G-protein/cAMP signaling, and MoWish and MoSho1 function upstream of the Pmk1-MAPK pathway. In summary, our studies elucidate the mechanism by which the COPII protein MoErv14 plays an important function in regulating the transport of receptors involved in the appressorium formation and virulence of the blast fungus.

According to recent studies, the large-scale effect of bottom topography on the ocean overturning circulation can be considered as a result of bottom enhancement of turbulent mixing in the abyssal ocean. Here we show, using laboratory experiments of rotating horizontal convection, that even without spatial variation of mixing intensity, oceanic meridional ridges can strongly impact both the strength and the pattern of the overturning in some fundamental ways. For example, as suggested by experimental results, the existence of the mid-Atlantic ridge can lead to the formation of another deep jet, like the deep western boundary current (DWBC), along the ridge’s eastern edge as a pathway for southward export of newly formed deep water. In response to this interior (mid-basin) jet and the associated isopycnal displacement, adiabatic flow structures may occur in upper and lower layers, including two opposing jets located respectively above and below the interior DWBC. Though unable to contribute to the overturning, they can probably affect transport along isopycnals. In the latitudinal band of the Antarctic Circumpolar Current without side boundary but with multiple ridges lying over the bottom, multiple interior DWBCs may develop preferentially along higher ridges, carrying the Antarctic Bottom Water into the northern latitudes. Moreover, the overturning cell or the strong jets can migrate vertically with the grow or decay of the ridge. Therefore, presumably strong variations of both ocean circulation and stratification may have occurred more than once during the past millions of years, as consequences of plate tectonic evolution, and have caused substantial changes of earth climate.

Abstract Transcription factors (TFs) play critical roles in the control of development and pathogenicity of phytopathogens by directly or indirectly regulating the expression of downstream genes. Here, we identified and characterized a zinc finger TF UvMsn2 in Ustilaginoidea virens, a homolog of MoMsn2 from the rice blast fungus. Heterogenous complementation assay revealed that UvMsn2 fully restored the defects of the ∆Momsn2 mutant in vegetative growth, conidiation and pathogenicity. Deletion of UvMsn2 in U. virens led to a reduction of the pathogen in vegetative growth, aerial hyphae and conidiation. Additionally, the ∆Uvmsn2 mutant displayed defects in conidial morphology and germination, as well as mitochondrial morphology. Pathogenicity and toxicity assays revealed that the ∆Uvmsn2 mutant was non-pathogenic and less inhibitory to rice seed germination. The ∆Uvmsn2 mutant showed different sensitivity to various stresses. Further microscopic examination found that UvMsn2 was localized in both cytosol and nucleus, and translocated from cytosol to nucleus under the treatment of NaCl. Our results demonstrate that UvMsn2 is a critical TF that regulates the vegetative growth, conidiogenesis, stress response, mitochondrial morphology and virulence in the rice false smut fungus.

Returning straw to the soil is an effective way to improve the soil quality. As genetically modified (GM) crops experience expanded growing scales, returning straw to the soil could also be necessary. However, the impact of GM crop straws on soil safety remains unclear. The environment (including soil types, humidity and temperature) can result in a significant difference in the diversity of soil bacterial communities. Here, we compared the impacts of the straw from Bt maize IE09S034 (IE) and near-isogenic non-Bt maize Zong31 (CK) on soil bacterial community and microbial metabolic activity in three different environments. Sampling was carried out following 6-10 months of decomposition (May, June, July, and August) in three localities in Chinese cities (Changchun, Jinan, and Beijing). Our results showed that Bt maize residues posed no direct impact on soil bacterial communities in contrast to the environment and decomposed time. The microbial functional diversity and metabolic activity showed no significant difference between IE and CK. The results could be a reference for further assessing the effect of Bt maize residues on the soil that promotes the commercialisation of Bt maize IE09S034.

Ustilaginoidea virens is a fungal pathogen that infects rice via the panicles. Here, the authors show that U. virens SGP1, a conserved Ser-Thr-rich glycosyl-phosphatidyl-inositol-anchored protein, elicits immune responses in rice leaves while contributing to virulence in panicles.

Summary: Endoplasmic reticulum (ER) homeostasis is essential for plants to manage responses under environmental stress. Plant immune activation requires the ER, but how ER homeostasis is associated with plant immune activation is largely unexplored. Here we find that transcription of an HVA22 family gene, OsHLP1 (HVA22-like protein 1), is induced by Magnaporthe oryzae infection. Overexpression of OsHLP1 significantly enhances blast disease resistance but impairs ER morphology in rice (Oryza sativa), resulting in enhanced sensitivity to ER stress. OsHLP1 interacts with the NAC (NAM, ATAF, and CUC) transcription factor OsNTL6 at the ER. OsNTL6 localizes to the ER and is relocated to the nucleus after cleavage of the transmembrane domain. OsHLP1 suppresses OsNTL6 protein accumulation, whereas OsNTL6 counteracts OsHLP1 by alleviating sensitivity to ER stress and decreasing disease resistance in OsHLP1 overexpression plants. These findings unravel a mechanism whereby OsHLP1 promotes disease resistance by compromising ER homeostasis when plants are infected by pathogens.

Based on long-term dynamic tracing of dissolved inorganic carbon (DIC) and stable carbon isotope (δ13CDIC) in produced water from 20 coalbed methane (CBM) wells in western Guizhou, the spatial-temporal dynamic variations of δ13CDIC of the GP well group produced in multi-layer commingled manner were analyzed, and the relationship between the value of δ13CDIC and CBM productivity was examined. The produced water samples of typical wells in the GP well group were amplified and sequenced using 16S rDNA, and a geological response model of δ13CDIC in produced water from CBM wells with multi-coal seams was put forward. The research shows that: δ13CDIC in produced water from medium-rank coal seams commonly show positive anomalies, the produced water contains more than 15 species of methanogens, and Methanobacterium is the dominant genus. The dominant methanogens sequence numbers in the produced water are positively correlated with δ13CDIC, and the positive anomaly of δ13CDIC is caused by reduction of methanogens, and especially hydrogenotrophic methanogens. Vertical segmentation of sedimentary facies and lithology in stratum with multi-coal seams will result in permeability and water cut segmentation, which will lead to the segmentation of δ13CDIC and archaea community in produced water, so in the strata with better permeability and high water cut, the δ13CDIC of the produced water is abnormally enriched, and the dominant archaea is mainly Methanobacterium. In the strata with weak permeability and low water cut, the δ13CDIC of the produced water is small, and the microbial action is weak. The shallow layer close to the coal seam outcrop is likely to be affected by meteoric precipitation, so the δ13CDIC of the produced water is smaller. The geological response model of δ13CDIC in produced water from multi-coal seams CBM wells in the medium-rank coal reveals the geological mechanism and microbial action mechanism of the δ13CDIC difference in the produced water from the multi-coal seams CBM wells. It also provides effective geochemical evidence for the superimposed fluid system controlled by sedimentary facies, and can also be used for the contribution analysis of the produced gas and water by the multi-layer CBM wells.

Genetically modified (GM) crops have brought huge economic benefits to mankind, however, at the same time, their safety issues are drawing growing attention. This investigation was conducted to assess whether the long-term cultivation of GM glyphosate resistant maize CC-2 effects bacterial communities in the rhizosphere soil. A 2-year follow-up trial was conducted, and soils were sampled at various plant developmental stages. The bacterial community structure of the rhizosphere soil was analysed by the high-throughput sequencing and compared with the near-isogenic non-GM maize Zheng 58. We showed here that long-term cultivation of CC-2 has no significant effect on the structure and diversity of bacterial communities, while different growth stages had significant effect. These results provided a reliable theoretical basis for the future cultivation and increased commercialisation of CC-2.

Ocean mesoscale eddies contribute significantly to water transport on a global scale, constituting the ubiquitous, irregular, discrete, nonlinear components. In this manuscript, we propose to explore whether and how the topographic effect of one meridional ridge, could exert considerable influences on the evolution and propagation of mesoscale eddies through their life cycle, directly from the perspectives of real observation statistics. We systematically investigate the known variability of mesoscale eddy trajectories, derived by multimission satellite altimetry from 1993 to 2018, of a life cycle more than 6 months, over the Izu-Ogasawara Ridge, and quantitatively examined the eddy-ridge interaction by observation statistics and wavelet coherence map, with respect to the intrinsic attributes, namely, the amplitude, the rotation speed, the radius. Due to the spatial-temporal diversity, a series of correlative steps have been particularly designed along time-frequency domain to trace back mesoscale eddy trajectories in a variety of origins, location, lifespan, polarity, either completely or partially passing over the ridge, and to facilitate the standardization in statistics across three phases of their life cycle, i.e., before, during and after the interaction with the ridge. It has been revealed in our experiment that three intrinsic attributes of mesoscale eddies within 25 years, all demonstrated noticeable correlation with the variation of topographic relief over the ridge. We observed that most of the cyclonic eddies obviously tended to begin to decay or even demise, while on the contrary, some of the anticyclonic eddies preferred to intensify slightly, or making no significant difference when encountering the upslope until climbing across the top, basically consistent with the expectation of potential vorticity (PV) conservation. The drifting velocity agreed with the tendency that the direction would be more probably modified toward equatorward or poleward by forcing to meridional component, with zonal component reduced at the beginning. The mesoscale eddies with the passage over the ridge exhibited the relatively high average horizontal scales, amplitude, rotation speed on the whole, compared to those with only partially passage. The developed scheme could integrate more evidences on how mesoscale eddies response to the topographic effects during their time-varying evolution and propagation process, and help provide opportunities to potentially identify and predict the underlying dynamic patterns and mechanisms that mesoscale eddies engage in ocean dynamics when proceeding toward meridional ridges on a global scale, with the promise of the end-to-end data-driven solution, such as deep learning architecture involved in the future.

Fatty acid metabolism is important for the maintenance of fatty acid homeostasis. Free fatty acids, which are toxic in excess, are activated by esterification with coenzyme A (CoA) and then subjected to β-oxidization. Fatty acid β-oxidation-related genes play critical roles in the development and virulence of several phytopathogens. In this study, we identified and characterized a peroxisomal-CoA synthetase in the rice blast fungus Magnaporthe oryzae, MoPCS60, which is a homolog of PCS60 in budding yeast. MoPCS60 was highly expressed during the conidial and early infectious stages and was induced under oleate treatment. Targeted deletion of MoPCS60 resulted in a significant reduction in growth rate when oleate and olive oil were used as the sole carbon sources. Compared with the wild-type strain Guy11, the ΔMopcs60 mutant exhibited fewer peroxisomes, more lipid droplets, and decreased pathogenicity. The distribution of MoPcs60 varied among developmental stages and was mainly localized to peroxisomes in the hyphae, conidia, and appressoria when treated with oleate. Our results suggest that MoPcs60 is a key peroxisomal-CoA synthetase involved in fatty acid β-oxidation and pathogenicity in rice blast fungi.

Ocean mesoscale eddies are ubiquitous in world ocean and account for 90% oceanic kinetic energy, which dominate the upper ocean flow field. Accurately predicting the variation of ocean mesoscale eddies is the key to understand the oceanic flow field and circulation system. In this article, we propose to make an initial attempt to explore spatio-temporal predictability of mesoscale eddies, employing deep learning architecture, which primarily establishes Memory In Memory (MIM) for sea level anomaly (SLA) prediction, combined with the existing mesoscale eddy detection. Oriented to the western Pacific ocean (125°−137.5°E and 15°−27.5°N), we quantitatively investigate the historic daily SLA variability at a 0.25° spatial resolution from 2000 to 2018, derived by satellite altimetry. We develop the enhanced MIM prediction strategies, equipped with Gated Recurrent Unit (GRU) and spatial attention module, in a scheduled sampling manner, which overcomes the gradient vanishing and complements to strengthen spatio-temporal features for long-term dependencies. At the early stage, the real value SLA input guides the model training process for initialization, while the scheduled sampling intentionally feeds the newly predicted value, to resolve the distribution inconsistency of inference. It has been demonstrated in our experiment results that our proposed prediction scheme outperformed the state-of-art approaches for SLA time series, with MAPE, RMSE of the 14-day prediction duration, respectively, 5.1%, 0.023 m on average, even up to 4.6%, 0.018 m for the effective sub-regions, compared to 19.8%, 0.086 m in ConvLSTM and 8.3%, 0.040 m in original MIM, which greatly facilitated the mesoscale eddy prediction. This proposed scheme will be beneficial to understand of the underlying dynamical mechanism behind the predictability of mesoscale eddies in the future, and help the deployment of ARGO, glider, AUV and other observational platforms.

Based on the productivity equation of coalbed methane (CBM) well, considering the impact of coal reservoir reformability on gas well productivity, the main production layer optimization index in the “three-step method” of optimal combination of production layers is corrected, and then the CBM production layer potential index is introduced to evaluate favorable areas for commingled multi-coal seam production. Through analysis of the key parameters of coal reservoirs affecting the CBM productivity index, a development unit division method for areas with multi-coal seams is established, and a quantitative grading index system is proposed. On this basis, the evaluation process of CBM development favorable area is developed: the mature 3-D modeling technology is used to characterize the reservoir physical properties of multi-coal seams in full-scale; the production layer potential index of each grid is calculated, and the production layer potential index contour under single-layer or commingled multi-layer production are plotted; according to the distribution of the contour of production layer potential index, the quantitative index of CBM development unit is adopted to outline the grade I, II, III coal reservoir distribution areas, and thus to pick out the favorable development areas. The practical application in the Yuwang block of Laochang in Yunnan proved that the favorable area evaluation process proposed can effectively overcome the defects of selecting favorable development areas only relying on evaluation results of a major coal seam pay, and enhance the accuracy of the evaluation results, meeting the requirements of selecting favorable areas for multi-coal seam commingled CBM production. Key words: coalbed methane, reservoir physical property, joint mining, production layer potential index, development unit division, quantitative index, favorable area evaluation

The ocean’s swirling eddies are known to influence primary productivity, yet understanding of these processes has been hampered by a lack of resolution. Here the authors combine in-situ drifter, altimeter-based feature tracking, and satellite chlorophyll data to quantify how phytoplankton respond to surface mesoscale phenomena across the globe.

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.

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.

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 Magnaporthe oryzae, 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 M. oryzae utilizes a phosphor-regulatory circuitry to face plant immunity during infection.

Membrane vesicles are considered virulence cargoes as they carry capsular and melanin components whose secretory transport is critical for the virulence of the human fungal pathogen Cryptococcus species. However, other components of the vesicles and their function in the growth and virulence of the fungus remain unclear. We have previously found that the cryptococcal intersectin protein Cin1 governs a unique Cin1-Wsp1-Cdc42 endocytic pathway required for intracellular transport and virulence. Using RNA sequencing, we compared the profiles of extracellular RNA (exRNA), including microRNA (miRNA), small interference RNA (siRNA), long noncoding RNA (lncRNA), and messenger RNA (mRNA) between the wild-type (WT), and derived Δcin1 mutant strains of Cryptococcus deneoformans. Seven hundred twelve miRNAs and 88 siRNAs were identified from WT, whereas 799 miRNAs and 66 siRNAs were found in Δcin1. Also, 572 lncRNAs and 7,721 mRNAs were identified from WT and 584 lncRNAs and 7,703 mRNAs from Δcin1. Differential expression analysis revealed that the disruption of CIN1 results in many important cellular changes, including those in exRNA expression, transport, and function. First, for miRNA target genes, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis revealed that cellular processes, components, and macromolecular functions are the most affected pathways. A higher number of genes were involved in the intracellular transport of endocytosis. Second, the results of GO term and KEGG analysis of differentially expressed lncRNA target genes and mRNA genes were consistent with those of miRNA targets. In particular, protein export is the topmost affected pathway among lncRNA target genes and one of the affected pathways among mRNA genes. The result of quantitative real-time reverse transcription PCR (qRT-PCR) from 12 mRNAs tested is largely agreeable with that of RNA-Seq. Taken together, our studies provide a comprehensive reference that Cryptococcus secretes abundant RNAs and that Cin1 plays a critical role in regulating their secretion. Given the growing clinical importance of exRNAs, our studies illuminate the significance of exploring this cutting-edge technology in studies of cryptococcal pathogenesis for the discovery of novel therapeutic strategies.

ABSTRACT The appressoria that are generated by the rice blast fungus Magnaporthe oryzae in response to surface cues are important for successful colonization. Previous work showed that regulators of G-protein signaling (RGS) and RGS-like proteins play critical roles in appressorium formation. However, the mechanisms by which these proteins orchestrate surface recognition for appressorium induction remain unclear. Here, we performed comparative transcriptomic studies of ΔMorgs mutant and wild-type strains and found that M. oryzae Aa91 (MoAa91), a homolog of the auxiliary activity family 9 protein (Aa9), was required for surface recognition of M. oryzae. We found that MoAA91 was regulated by the MoMsn2 transcription factor and that its disruption resulted in defects in both appressorium formation on the artificial inductive surface and full virulence of the pathogen. We further showed that MoAa91 was secreted into the apoplast space and was capable of competing with the immune receptor chitin elicitor-binding protein precursor (CEBiP) for chitin binding, thereby suppressing chitin-induced plant immune responses. In summary, we have found that MoAa91 is a novel signaling molecule regulated by RGS and RGS-like proteins and that MoAa91 not only governs appressorium development and virulence but also functions as an effector to suppress host immunity. IMPORTANCE The rice blast fungus Magnaporthe oryzae generates infection structure appressoria in response to surface cues largely due to functions of signaling molecules, including G-proteins, regulators of G-protein signaling (RGS), mitogen-activated protein (MAP) kinase pathways, cAMP signaling, and TOR signaling pathways. M. oryzae encodes eight RGS and RGS-like proteins (MoRgs1 to MoRgs8), and MoRgs1, MoRgs3, MoRgs4, and MoRgs7 were found to be particularly important in appressorium development. To explore the mechanisms by which these proteins regulate appressorium development, we have performed a comparative in planta transcriptomic study and identified an auxiliary activity family 9 protein (Aa9) homolog that we named MoAa91. We showed that MoAa91 was secreted from appressoria and that the recombinant MoAa91 could compete with a chitin elicitor-binding protein precursor (CEBiP) for chitin binding, thereby suppressing chitin-induced plant immunity. By identifying MoAa91 as a novel signaling molecule functioning in appressorium development and an effector in suppressing host immunity, our studies revealed a novel mechanism by which RGS and RGS-like proteins regulate pathogen-host interactions.

Rhein is one of active anthraquinone components in traditional Chinese herbal medicine Rheum palmatum L., possessing anti-inflammatory, antioxidant, antitumor, antiviral, and hepatoprotective activities. Human respiratory syncytial virus (RSV), a common virus, is able to result in pneumonia and bronchitis, which usually can be seen in infants. However, so far the effects of Rhein on RSV-induced pneumonia are still unknown. As the NLRP3 inflammasome is activated excessively, it is able to lead to inflammatory response and tissue injury in most viral infection process (including RSV infection) of respiratory tract. Therefore, we designed experiments to reveal whether Rhein can treat RSV-induced pneumonia by inhibiting NLRP3 inflammasome activation. In present research, we established the pneumonia model of BALB/C mice caused by RSV. First of all, the pathology of lung tissue and the weight of mice were evaluated, and the corresponding lung index was calculated. Additionally, the expression of pro-inflammatory mediators in serum and lung tissues, and related proteins (NLRP3, ASC and Caspase-1) of NLRP3 inflammasome and NF-κB pathway were detected by Enzyme-linked immunosorbent assay (ELISA), Real-time PCR (RT-PCR), Immunohistochemistry (IHC), and Western blot (WB), respectively. The determination of lung index and lung tissue pathological evaluation revealed that Rhein was able to alleviate lung infection and injury caused by RSV. The results of ELISA showed that Rhein was able to reduce the release of pro-inflammatory cytokines in the serum and lung tissues of RSV-induced BALB/c mice, including IL-1β, IL-6, TNF-α, IL-18, and IL-33. Additionally, it was revealed that Rhein inhibited the immune inflammatory response of RSV-infected mice, which was likely to be associated with the inhibition the NLRP3 inflammasome activation via NF-κB pathway. To sum up, our results indicated that Rhein may inhibit RSV-induced pulmonary inflammatory response effectively; meanwhile, it is emphasized that Rhein therapy is likely to be a promising treatment on the RSV-infected lung inflammation and avoidance of lung tissue damage.

The release of genetically modified (GM) crops has potential to alter the bacterial population within rhizosphere. Here, the potential effect of GM maize cv. IE09S034 containing the Cry1Ie toxin gene from Bacillus thuringiensis (Bt) was investigated under the field conditions. The community composition and the relative abundance of the bacteria in rhizosphere soil were estimated by analysing 16S rRNA PCR amplicons. Our results indicated that Bt maize IE09S034 has no significant effects on the rhizosphere bacterial community. Instead, it was found that factors such as plant growth stage and year have a stronger effect on the bacterial population dynamics. Our findings therefore provide reliable evidence supporting the potential commercial cultivation of the cv. IE09S034.

Based on the productivity equation of coalbed methane (CBM) wells, three indexes, main production layer optimization index, main production layer expansion index and capacity contribution index are proposed, with which the three – step optimization method of production-layer combination is established. In selecting main production layer, the coal seam thickness, CBM content, coal seam permeability, coal seam reservoir pressure and coal structure are considered comprehensively to evaluate the potential of the production layer. In selecting expansion of the main production layer combination, on the premise of ensuring full and slow desorption of the main production layer and non-exposure of the main production layer out of liquid surface, the degree of mutual interference between the main and non-main production layers is comprehensively evaluated by coupling the critical desorption pressure, layer spacing and reservoir pressure gradient difference. In optimizing production layer combination, the main concern is the economic efficiency of the combined layers. Only when the contribution coefficient of the main production layer is greater than 30% and the contribution index of the other production layers is more than 10%, the economic benefit of a CBM well after being put into production can be ensured. Based on the comparative analysis of the development effect of the development test wells in Songhe of Guizhou province, it is proved that the “three-step method” for the optimization of production-layer combination is scientific and practical, and can be used to design the multi-layer commingling scheme of coalbed methane. Key words: CBM, multi-layer commingled production, selection of main production layer, combination optimization, production contribution

Transgenic plants may induce shifts in the microbial community composition that in turn alter microbially-mediated nutrient cycling in soil. Studies of how specific microbial groups respond to genetically modified (GM) planting help predict potential impacts upon processes performed by these groups. This study investigated the effect of transgenic high-methionine soybean cv. ZD91 on nitrogen-fixing and ammonia-oxidizing bacterial populations. A difference in nitrogen-fixing or ammonia-oxidizing bacteria community composition was not found, suggesting that cv. ZD91 does not alter the bacterial populations in rhizosphere soil. This study increases our understanding of the potential effect of transgenic soybean on microbial functional groups within soil by suggesting that nitrogen-transforming bacteria may be useful for future investigations on the GM crops impact in the soil ecosystem.

Various effectors of plant pathogens modulate host cell biology. Here, Huang et al. show PsAvr3c, an avirulence effector from oomycete plant pathogen Phytophthora sojae, can reprogram host pre-mRNA splicing for immune modulation.

Vesicle trafficking is an important event in eukaryotic organisms. Many proteins and lipids transported between different organelles or compartments are essential for survival. These processes are mediated by soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins, Rab-GTPases, and multisubunit tethering complexes such as class C core vacuole or endosome tethering and homotypic fusion or vacuole protein sorting (HOPS). Our previous study has demonstrated that FgVam7, which encodes a SNARE protein involving in vesicle trafficking, plays crucial roles in growth, asexual or sexual development, deoxynivalenol production, and pathogenicity in Fusarium graminearum. Here, the affinity purification approach was used to identify FgVam7-interacting proteins to explore its regulatory mechanisms during vesicle trafficking. The orthologs of yeast Vps39, a HOPS tethering complex subunit, and Sso1, a SNARE protein localized to the vacuole or endosome, were identified and selected for further characterization. In yeast two-hybrid and glutathione-S-transferase pull-down assays, FgVam7, FgVps39, and FgSso1 interacted with each other as a complex. The ∆Fgvps39 mutant generated by targeted deletion was significantly reduced in vegetative growth and asexual development. It failed to produce sexual spores and was defective in plant infection and deoxynivalenol production. Further cellular localization and cytological examinations suggested that FgVps39 is involved in vesicle trafficking from early or late endosomes to vacuoles in F. graminearum. Additionally, the ∆Fgvps39 mutant was defective in vacuole morphology and autophagy, and it was delayed in endocytosis. Our results demonstrate that FgVam7 interacts with FgVps39 and FgSso1 to form a unique complex, which is involved in vesicle trafficking and modulating the proper development of infection-related morphogenesis in F. graminearum.

[This corrects the article DOI: 10.1371/journal.ppat.1001302.].

[This corrects the article DOI: 10.1371/journal.ppat.1002450.].

ABSTRACT ADP ribosylation factor (Arf) small GTPase family members are involved in vesicle trafficking and organelle maintenance in organisms ranging from Saccharomyces cerevisiae to humans. A previous study identified Magnaporthe oryzae Arf6 (MoArf6) as one of the Arf proteins that regulates growth and conidiation in the rice blast fungus M. oryzae, but the remaining family proteins remain unknown. Here, we identified six additional Arf proteins, including MoArf1, MoArl1, MoArl3, MoArl8, MoCin4, and MoSar1, as well as their sole adaptor protein, MoGga1, and determined their shared and specific functions. We showed that the majority of these proteins exhibit positive regulatory functions, most notably, in growth. Importantly, MoArl1, MoCin4, and MoGga1 are involved in pathogenicity through the regulation of host penetration and invasive hyphal growth. MoArl1 and MoCin4 also regulate normal vesicle trafficking, and MoCin4 further controls the formation of the biotrophic interfacial complex (BIC). Moreover, we showed that Golgi-cytoplasm cycling of MoArl1 is required for its function. Finally, we demonstrated that interactions between MoArf1 and MoArl1 with MoGga1 are important for Golgi localization and pathogenicity. Collectively, our findings revealed the shared and specific functions of Arf family members in M. oryzae and shed light on how these proteins function through conserved mechanisms to govern growth, transport, and virulence of the blast fungus. IMPORTANCE Magnaporthe oryzae is the causal agent of rice blast, representing the most devastating diseases of rice worldwide, which results in losses of amounts of rice that could feed more than 60 million people each year. Arf (ADP ribosylation factor) small GTPase family proteins are involved in vesicle trafficking and organelle maintenance in eukaryotic cells. To investigate the function of Arf family proteins in M. oryzae, we systematically characterized all seven Arf proteins and found that they have shared and specific functions in governing the growth, development, and pathogenicity of the blast fungus. We have also identified the pathogenicity-related protein MoGga1 as the common adaptor of MoArf1 and MoArl1. Our findings are important because they provide the first comprehensive characterization of the Arf GTPase family proteins and their adaptor protein MoGga1 functioning in a plant-pathogenic fungus, which could help to reveal new fungicide targets to control this devastating disease.

Endocytosis is an intracellular trafficking pathway that occurs in nutrient uptake, signal transduction and reconstruction of cell polarity and is conserved in eukaryotic cells. In fungi, endocytosis plays crucial roles in the physiology of hyphal growth and pathogenicity. vidence for endocytosis in filamentous fungi is detected by the membrane-selective dyes FM4-64. Cells of a range of filamentous fungal species readily take up these dyes. However, the method for endocytosis detection has not been well established in Magnaporthe oryzae. Here, we provide a protocol for tracking endocytosis in Magnaporthe oryzae.

Regulator of G-protein signaling (RGS) proteins primarily function as GTPase-accelerating proteins (GAPs) to promote GTP hydrolysis of Gα subunits, thereby regulating G-protein mediated signal transduction. RGS proteins could also contain additional domains such as GoLoco to inhibit GDP dissociation. The rice blast fungus Magnaporthe oryzae encodes eight RGS and RGS-like proteins (MoRgs1 to MoRgs8) that have shared and distinct functions in growth, appressorium formation and pathogenicity. Interestingly, MoRgs7 and MoRgs8 contain a C-terminal seven-transmembrane domain (7-TM) motif typical of G-protein coupled receptor (GPCR) proteins, in addition to the conserved RGS domain. We found that MoRgs7, but not MoRgs8, couples with Gα MoMagA to undergo endocytic transport from the plasma membrane to the endosome upon sensing of surface hydrophobicity. We also found that MoRgs7 can interact with hydrophobic surfaces via a hydrophobic interaction, leading to the perception of environmental hydrophobiccues. Moreover, we found that MoRgs7-MoMagA endocytosis is regulated by actin patch-associated protein MoCrn1, linking it to cAMP signaling. Our studies provided evidence suggesting that MoRgs7 could also function in a GPCR-like manner to sense environmental signals and it, together with additional proteins of diverse functions, promotes cAMP signaling required for developmental processes underlying appressorium function and pathogenicity.

On the basis of its downy mildew–like morphology, the litchi downy blight pathogen was previously named Peronophythora litchii. Recently, however, it was proposed to transfer this pathogen to Phytophthora clade 4. To better characterize this unusual oomycete species and important fruit pathogen, we obtained the genome sequence of Phytophthora litchii and compared it to those from other oomycete species. P. litchii has a small genome with tightly spaced genes. On the basis of a multilocus phylogenetic analysis, the placement of P. litchii in the genus Phytophthora is strongly supported. Effector proteins predicted included 245 RxLR, 30 necrosis-and-ethylene-inducing protein-like, and 14 crinkler proteins. The typical motifs, phylogenies, and activities of these effectors were typical for a Phytophthora species. However, like the genome features of the analyzed downy mildews, P. litchii exhibited a streamlined genome with a relatively small number of genes in both core and species-specific protein families. The low GC content and slight codon preferences of P. litchii sequences were similar to those of the analyzed downy mildews and a subset of Phytophthora species. Taken together, these observations suggest that P. litchii is a Phytophthora pathogen that is in the process of acquiring downy mildew–like genomic and morphological features. Thus P. litchii may provide a novel model for investigating morphological development and genomic adaptation in oomycete pathogens.