In situ structural analysis reveals membrane shape transitions during autophagosome formation.

Autophagosomes are unique organelles that form de novo as double-membrane vesicles engulfing cytosolic material for destruction. Their biogenesis involves membrane transformations of distinctly shaped intermediates whose ultrastructure is poorly understood. Here, we combine cell biology, correlative cryo-electron tomography (cryo-ET), and extensive data analysis to reveal the step-by-step structural progression of autophagosome biogenesis at high resolution directly within yeast cells. The analysis uncovers an unexpectedly thin intermembrane distance that is dilated at the phagophore rim. Mapping of individual autophagic structures onto a timeline based on geometric features reveals a dynamical change of membrane shape and curvature in growing phagophores. Moreover, our tomograms show the organelle interactome of growing autophagosomes, highlighting a polar organization of contact sites between the phagophore and organelles, such as the vacuole and the endoplasmic reticulum (ER). Collectively, these findings have important implications for the contribution of different membrane sources during autophagy and for the forces shaping and driving phagophores toward closure without a templating cargo.

Declining coupling between vegetation and drought over the past three decades.

Droughts have been implicated as the main driver behind recent vegetation die-off and are projected to drive greater mortality under future climate change. Understanding the coupling relationship between vegetation and drought has been of great global interest. Currently, the coupling relationship between vegetation and drought is mainly evaluated by correlation coefficients or regression slopes. However, the optimal drought timescale of vegetation response to drought, as a key indicator reflecting vegetation sensitivity to drought, has largely been ignored. Here, we apply the optimal drought timescale identification method to examine the change in coupling between vegetation and drought over the past three decades (1982-2015) with long-term satellite-derived Normalized Difference Vegetation Index and Standardized Precipitation-Evapotranspiration Index data. We find substantial increasing response of vegetation to drought timescales globally, and the correlation coefficient between vegetation and drought under optimal drought timescale overall declines between 1982 and 2015. This decrease in vegetation-drought coupling is mainly observed in regions with water deficit, although its initial correlation is relatively high. However, vegetation in water-surplus regions, with low coupling in earlier stages, is prone to show an increasing trend. The observed changes may be driven by the increasing trend of atmospheric CO2 . Our findings highlight more pressing drought risk in water-surplus regions than in water-deficit regions, which advances our understanding of the long-term vegetation-drought relationship and provides essential insights for mapping future vegetation sensitivity to drought under changing climate conditions.

Bone Marrow Mesenchymal Stem Cells-Derived Exosomes Promote Survival of Random Flaps in Rats through NRF2-Mediated Antioxidative Stress.

BACKGROUND: Random flaps are the most used defect repair method for head and neck tumors and trauma plastic surgery. The distal part of the flap often undergoes oxidative stress (OS), ultimately leading to flap necrosis. Stem cells exosomes exhibit potential effects related to anti-inflammatory, regenerative, and antioxidant properties. Nuclear factor erythroid-2-related factor 2 (Nrf2) is an important factor in regulating oxidative balance. Exosomes have been reported to monitor its transcription to alleviate OS. This study examined the impacts and underlying mechanisms of antioxidant actions of exosomes derived from bone marrow mesenchymal stem cells (BMSCs-Exo) on random flaps. METHODS: BMSCs-Exo was injected into the tail veins of rats on days 0, 1, and 2 after surgery of random flaps. The rats were euthanized on day 3 to calculate the survival rate. Immunohistochemical staining, western blotting, dihydroethidium probe, superoxide dismutase, and malondialdehyde assay kits were used to detect OS level. Human umbilical vein endothelial cells were co-cultured with BMSCs-Exo and ML385 (an inhibitor of Nrf2) in vitro. RESULTS: BMSCs-Exo may significantly improve the survival rate of the random flaps by reducing apoptosis, inflammation, and OS while increasing angiogenesis. Besides, BMSCs-Exo can also increase mitochondrial membrane potential and reduce reactive oxygen species levels in vitro. These therapeutic effects might stem from the activation of the Keap1/Nrf2 signaling pathway. CONCLUSION: BMSCs-Exo improved the tissue antioxidant capacity by regulating the keap1/Nrf2 signaling pathway. BMSCs-Exo may be a new strategy to solve the problem of random flap necrosis.

OCT4 regulates WNT/ß-catenin signaling and prevents mesoendoderm differentiation by repressing EOMES in porcine pluripotent stem cells.

The regulatory network between signaling pathways and transcription factors (TFs) is crucial for the maintenance of pluripotent stem cells. However, little is known about how the key TF OCT4 coordinates signaling pathways to regulate self-renewal and lineage differentiation of porcine pluripotent stem cells (pPSCs). Here, we explored the function of OCT4 in pPSCs by transcriptome and chromatin accessibility analysis. The TFs motif enrichment analysis revealed that, following OCT4 knockdown, the regions of increased chromatin accessibility were enriched with EOMES, GATA6, and FOXA1, indicating that pPSCs differentiated toward the mesoendoderm (ME) lineage. Besides, pPSCs rapidly differentiated into ME when the WNT/ß-catenin inhibitor XAV939 was removed. However, the ME differentiation of pPSCs caused by OCT4 knockdown did not rely on the activation of WNT/ß-catenin signaling because the target gene of WNT/ß-catenin signaling, AXIN2 was not upregulated after OCT4 knockdown, despite significant upregulation of WLS and some WNT ligands. Importantly, OCT4 is directly bound to the promoter and enhancers of EOMES and repressed its transcription. Overexpression of EOMES was sufficient to induce ME differentiation in the presence of XAV939. These results demonstrate that OCT4 can regulate WNT/ß-catenin signaling and prevent ME differentiation of pPSCs by repressing EOMES transcription.

Metformin attenuates osteoarthritis by targeting chondrocytes, synovial macrophages and adipocytes.

OBJECTIVE: To investigate the therapeutic effect and mechanism of metformin on knee OA in normal diet (ND) mice or high-fat diet (HFD)-induced obese mice. METHODS: Destabilization of the medial meniscus surgery was performed in ND mice or HFD mice, and metformin was administrated in drinking water or not. The changes of OA joint structure, infiltration and polarization of synovial macrophages and circulating and local levels of leptin and adiponectin were evaluated. In vitro, the effects of metformin on chondrocytes and macrophages, and of conditioned mediums derived from mouse abdominal fat on murine chondrogenic cell line ATDC5 and murine macrophage cell line RAW264.7, were detected. RESULTS: Metformin showed protective effects on OA, characterized by reductions on OARSI score [2.00, 95% CI (1.15, 2.86) for ND mice and 3.17, 95% CI (2.37, 3.96) for HFD mice] and synovitis score [1.17, 95% CI (0.27, 2.06) for ND mice and 2.50, 95% CI (1.49, 3.51) for HFD mice] after 10 weeks of treatment, and the effects were more significant in HFD mice than in ND mice. Mechanistically, in addition to decreasing apoptosis and matrix-degrading enzymes expression in chondrocytes as well as infiltration and pro-inflammatory differentiation of synovial macrophages, metformin reduced leptin secretion by adipose tissue in HFD mice. CONCLUSIONS: Metformin protects against knee OA which could be through reducing apoptosis and catabolism of chondrocytes, and suppressing infiltration and pro-inflammatory polarization of synovial macrophages. For obese mice, metformin has a greater protective effect in knee OA additionally through reducing leptin secretion from adipose tissue.

Silencing of Long Noncoding RNA TUG1 Ameliorates Atherosclerosis-Induced Myocardial Injury by Upregulating microRNA-30b-3p and Downregulating Brd4.

ABSTRACT: Long noncoding RNAs and microRNAs (miRNAs) are emerging biomarkers involved in human diseases, and we focused on the roles of long noncoding RNA taurine upregulated gene 1 (TUG1) and miR-30b-3p in the related mechanisms of atherosclerosis-induced myocardial injury. ApoE-deficient mice were fed with high-fat diet to establish atherosclerotic models and then were subjected to either TUG1 downregulation or miR-30b-3p upregulation treatment. The serum myocardial enzymes, inflammatory biomarkers, pathological changes, intramyocardial macrophage infiltration, and apoptosis of cardiomyocytes in atherosclerotic mice were determined. The expression of TUG1, miR-30b-3p, and bromodomain protein 4 (Brd4) in atherosclerotic models was evaluated. Moreover, the correlations of TUG1, miR-30b-3p, and Brd4 were verified. TUG1 and Brd4 were increased while miR-30b-3p was decreased in atherosclerotic mice. The silenced TUG1 or elevated miR-30b-3p attenuated atherosclerosis-induced myocardial injury mainly by reducing serum myocardial enzyme content and inflammatory response, improving pathological changes, and preventing macrophage infiltration and cardiomyocyte apoptosis in atherosclerotic mice. Mechanistically, TUG1 could competitively bind with miR-30b-3p to prevent the degradation of its target gene Brd4. This study reveals that the silencing of TUG1 ameliorates atherosclerosis-induced myocardial injury by upregulating miR-30b-3p and downregulating Brd4, which may provide novel targets for atherosclerosis treatment.

The significance of elective neck dissection for patients with T2N0M0 OSCC at different ages.

BACKGROUND: The aim of this study was to determine the significance of elective neck dissection (END) for patients of different ages with T2N0M0 oral squamous cell carcinoma (OSCC) and sought to analyze the reasons behind it and its value for clinical guidance. METHODS: This study enrolled 391 patients with T2N0M0 OSCC who were surgically treated in our hospital and were divided into young-, moderate-, and advanced-age groups according to our previous study. The Chi-square test and Kaplan-Meier analysis were performed for statistical analysis. RESULTS: Compared with moderate- and advanced-age patients, young patients with T2N0M0 OSCC had higher lymph node metastasis rates and lymph node ratios. Therefore, END significantly improved the recurrence (p = 0.001) and survival (p = 0.001) for young patients, but not for moderate-age patients. Advanced-age patients even benefit from watchful waiting. END significantly improved recurrence and survival in young patients with smoking or alcohol consumption habits. CONCLUSIONS: END improved the prognosis of young patients, and it was related to their higher metastasis rate. However, advanced-age patients benefited from the wait-and-see policy. END is essential for the young patients with smoking or drinking habit, it is also highly recommended for nonsmokers and nondrinkers.

Chemotherapy for primary cancer can reduce the risk of head and neck second primary malignancy: a propensity-matched analysis.

OBJECTIVES: To investigate the effect of chemotherapy versus no chemotherapy on the risk of second primary head and neck malignancies (SPHNMs) in patients with locally advanced oral squamous cell carcinoma (OSCC) and to assess the survival outcomes of patients with SPHNM. MATERIALS AND METHODS: A total of 937 OSCC patients were divided into chemotherapy and nonchemotherapy groups by propensity score matching (PSM). In the presence of the competing event of non-SPHNM death, the fine and gray modified Cox proportional hazard model was fitted to detect the impact of various factors, including the history of chemotherapy, on SPHNM risk. The Kaplan-Meier method was used to assess the survival outcomes of patients. RESULTS: After PSM, the 10-year cumulative probability of SPHNM was 10.7% for patients who received chemotherapy and 22.1% for patients who did not. The fine and gray regression model showed that prior chemotherapy was associated with a 51% reduced risk of SPHNM (adjusted subdistribution hazard ratio (sHR): 0.49, 95% confidence interval (CI): 0.29-0.84, P = 0.1). The disease-free survival (DFS) rates did not differ significantly between the SPHNM and non-SPHNM groups. And there were no significant differences in DFS rates between the patients with and those without prior chemotherapy in the SPHNM group. CONCLUSIONS: Chemotherapy for locally advanced primary OSCC is associated with a decreased incidence of subsequent SPHNM. However, chemotherapy for the primary cancer does not improve DFS in patients with SPHNM. CLINICAL RELEVANCE: Chemotherapy plays a positive role in preventing SPHNMs for patients with oral squamous cell carcinoma. CLINICAL TRIAL REGISTRATION: Before January 2015, the data were retrieved retrospectively, while after January 2015, the data were collected prospectively in a POROMS database (ClinicalTrials.gov ID: NCT02395367).

A threshold effect of COVID-19 risk on oil price returns.

Using U.S. data, we investigate how the COVID-19 pandemic influences oil price returns in an asset pricing framework. Unlike earlier studies, we consider a threshold model to allow for the possibility that COVID-19 risk may not play a role until it reaches a certain level. Based on WTI crude oil spot price data from January 2020 to December 2021, our findings show that oil returns significantly decline with the daily number of COVID-19 deaths but only if the daily death toll exceeds approximately 2100. In addition, a more severe COVID-19 pandemic can substantially increase the exposure of oil returns to various systematic risk factors, which has not been documented in previous literature.

Prognostic factors and related complications/sequalae of squamous cell carcinoma located in the gingivobuccal complex.

BACKGROUND: Gingivobuccal complex (GBC) was a relatively new concept of oral subsite that was comprises of the upper and/or lower gingiva, gingival buccal sulcus, and adjacent buccal mucosa. Squamous cell carcinoma (SCC) of the GBC had a poor prognosis, with few studies analyzing this particular entity. The objective of this study was to analyze the risk factors affecting the prognosis and complications/sequalae of gingivobuccal complex cancer. METHODS: Between December 2014 and August 2019, a total of 122 patients diagnosed with primary gingivobuccal complex cancer in Beijing Stomatological Hospital, Capital Medical University were enrolled in the study. Through outpatient reviewed and telephone followed-up for 2-5 years postoperatively, postoperative relapse and complications/sequalae were assessed. The primary outcome parameter was 2-year disease-free survival. RESULTS: The most common central site of the tumor was the buccal mucosa (45.1%), followed by the lower gingiva (36.9%). The most diseases were pT4a (45.1%) and there was lymph node invasion (pN+) in 41.8% of patients. Moderate differentiated squamous carcinoma (77.9%) accounted for the vast majority of the histopathological differentiation. A total of 62.3% of tumors invaded the bone, while, 5.7% invaded the skin layer. Survival analysis found that 44.3% of patients experienced relapse within two years postoperatively and the mortality rate after relapse was 75.9%. Almost 60.0% of the tumors involving the maxilla and/or mandible developed relapse. Cox proportional hazards model found that pN stage (p= 0.002) and bone invasion (p= 0.007) were significant independent predictors of 2-year disease-free survival. Importantly, 63.1% of patients had postoperative (and postradiotherapy) complications/sequalae. It was noteworthy that 18 of 43 patients (41.9%) who implanted with titanium plates had hardware-related complications/sequalae, and the most of them were titanium plate exposure (61.1%). CONCLUSIONS: Squamous cell carcinoma of the gingivobuccal complex cancer, as a new subsite worthy of attention in oral cancer, has a high complication/sequalae rate, high relapse rate and poor prognosis. TRIAL REGISTRATION: Prospective, Observational, Real-world Oral Malignant Tumors Study ( clinicaltrials.gov identifier: NCT02395367). The approval of the Institutional Review Board of the Beijing Stomatological Hospital of Capital Medical University (Approval number: CMUSH-IRB-KJPJ-2015-08).

Non-linear effects of natural and anthropogenic drivers on ecosystem services: Integrating thresholds into conservation planning.

Ecosystem services (ESs) have been widely used for ecological protection and land spatial planning. Natural and anthropogenic drivers exhibit a strong dynamic coupling relationship with ESs. However, current ESs-related research focused on mapping the ESs spatially or investing the trade-offs and synergies relationship between ES, ignoring the nonlinear response of ESs to natural and anthropogenic drivers. Here we aimed to investigate the nonlinear effect of 14 potential drivers (8 natural and 6 anthropogenic) on the total value of six typical ESs (ESV). Taking Beijing-Tianjin-Hebei urban agglomeration (BTH) in China as an example, we established 14 constrain lines and identified critical thresholds through the restricted cubic splines (RCS) regression. We found strong non-linear impacts of natural and anthropogenic drivers on ESV and critical thresholds existed among all the 14 constrain lines. The RCS plots showed that the overall ESV was kept at a high level before or after certain thresholds (e.g., altitude >687 m, slope >13.4°, NDVI >0.7, distance from water <31.2 km, etc.). We categorized these threshold combinations and found the potentially high ES delivery areas were mainly distributed in the Yanshan Mountian, accounting for approximately 5% of the total BTH region. These critical thresholds offer a new method to delineate conservation and restoration priority areas.

Osteoarthritic infrapatellar fat pad aggravates cartilage degradation via activation of p38MAPK and ERK1/2 pathways.

OBJECTIVE: This study aimed to investigate the biochemical effects of osteoarthritic infrapatellar fat pad (IPFP) on cartilage and the underlying mechanisms. METHODS: Human IPFP and articular cartilage were collected from end-stage osteoarthritis (OA) patients during total knee arthroplasty. IPFP-derived fat-conditioned medium (FCM) was used to stimulate human primary chondrocytes and cartilage explants. Functional effect of osteoarthritic IPFP was explored in human primary chondrocytes and articular cartilage in vitro and ex vivo. Activation of relative pathways and its effects on chondrocytes were assessed through immunoblotting and inhibition experiments, respectively. Neutralization test was performed to identify the main factors and their associated pathways responsible for the effects of IPFP. RESULTS: Osteoarthritic IPFP-derived FCM significantly induced extracellular matrix (ECM) degradation in both human primary chondrocytes and cartilage explants. Several pathways, such as NF-κB, mTORC1, p38MAPK, JNK, and ERK1/2 signaling, were significantly activated in human chondrocytes with osteoarthritic IPFP-derived FCM stimulation. Interestingly, inhibition of p38MAPK and ERK1/2 signaling pathway could alleviate the detrimental effects of FCM on chondrocytes, while inhibition of other signaling pathways had no similar results. In addition, IL-1ß and TNF-α instead of IL-6 in osteoarthritic IPFP-derived FCM played key roles in cartilage degradation via activating p38MAPK rather than ERK1/2 signaling pathway. CONCLUSION: Osteoarthritic IPFP induces the degradation and inflammation of cartilage via activation of p38MAPK and ERK1/2 pathways, in which IL-1ß and TNF-α act as the key factors. Our study suggests that modulating the effects of IPFP on cartilage may be a promising strategy for knee OA intervention.

A review on two-dimensional materials for chemiresistive- and FET-type gas sensors.

Two-dimensional (2D) materials have shown great potential for gas sensing applications due to their large specific surface areas and strong surface activities. In addition to the commonly reported chemiresistive-type gas sensors, field-effect transistor (FET)-type gas sensors have attracted increased attention due to their miniaturized size, low power consumption, and good compatibility with CMOS technology. In this review, we aim to discuss the recent developments in chemiresistive- and FET-type gas sensors based on 2D materials, including graphene, transition metal dichalcogenides, MXenes, black phosphorene, and other layered materials. Firstly, the device structure and the corresponding fabrication process of the two types of sensors are given, and then the advantages and disadvantages are also discussed. Secondly, the effects of intrinsic and extrinsic factors on the sensing performance of 2D material-based chemiresistive and FET-type gas sensors are also detailed. Subsequently, the current gas-sensing applications of 2D material-based chemiresistive- and FET-type gas sensors are systematically presented. Finally, the future prospects of 2D materials in chemiresistive- and FET-type gas sensing applications as well as the current existing problems are pointed out, which could be helpful for the development of 2D material-based gas sensors with better sensing performance to meet the requirements for practical application.

Tetrandrine, a Potent Antifungal Agent, Inhibits Mycelial Growth and Virulence of Botrytis cinerea.

Tetrandrine (TET) is a potent calcium channel blocker used to treat hypertension and inflammation. Currently, TET is predominantly used to treat a variety of human diseases, and there is little information regarding the use of TET against plant pathogens. In this study, we explored the antifungal activity of TET on a plant pathogen, Botrytis cinerea. We show that administration of low concentrations of TET effectively inhibited hyphal growth of fungus grown on potato dextrose agarose and decreased the virulence of B. cinerea in tomato plants. Real-time PCR revealed that the expression of drug efflux pump-related genes (alcohol dehydrogenase 1, multidrug/pheromone exporter, pleiotropic drug resistance protein 1, and synaptic vesicle transporter) were downregulated in the presence of TET. Finally, we show that TET acts synergistically with iprodione, resulting in increased inhibition of B. cinerea both in vitro and in vivo. These results indicate that TET might act as an effective antifungal agent in reducing gray mold disease.

Identification and Characterization of Nothophoma quercina Causing Bud Blight on Photinia × fraseri in China.

Photinia (Photinia × fraseri Dress) is a well-known green plant that has high ornamental value and is widely distributed around the world. An outbreak of typical bud blight disease was observed between May and August in photinia in 2017 in Qingdao, China. The causal agent for this blight was subsequently isolated from symptomatic samples and identified as Nothophoma quercina based on morphological characterization and molecular analyses (ITS, LSU, RPB2, and TUB2). Results of pathogenicity tests on isolated fungi also supported the conclusion that N. quercina is the pathogen responsible for this condition. To our knowledge, this is the first report of bud blight on P. fraseri caused by N. quercina in China.

Comparative transcriptome analysis reveals multiple functions for Mhy1p in lipid biosynthesis in the oleaginous yeast Yarrowia lipolytica.

Yarrowia lipolytica is considered as a promising microbial cell factory for bio-oil production due to its ability to accumulate a large amount of lipid. However, the regulation of lipid metabolism in this oleaginous yeast is elusive. In this study, the MHY1 gene was disrupted, and 43.1% (w/w) intracellular oil based on cell dry weight was obtained from the disruptant M-MHY1, while only 30.2% (w/w) lipid based on cell dry weight was obtained from the reference strain. RNA-seq was then performed to analyze transcriptional changes during lipid biosynthesis after MHY1 gene inactivation. The expression of 1597 genes, accounting for 24.7% of annotated Y. lipolytica genes, changed significantly in the disruptant M-MHY1 during lipid biosynthesis. Differential gene expression analysis indicated that Mhy1p performs multiple functions and participates in a wide variety of biological processes, including lipid, amino acid and nitrogen metabolism. Notably, data analysis revealed increased carbon flux through lipid biosynthesis following MHY1 gene inactivation, accompanied by decreased carbon flux through amino acid biosynthesis. Moreover, Mhy1p regulates the cell cycle, and the cell cycle rate was enhanced in the disruptant M-MHY1. These results suggest that Mhy1p plays critical regulatory roles in diverse aspects of various biological processes, especially in lipid biosynthesis, amino acid and nitrogen metabolism and cell cycle. Our dataset appears to elucidate the crucial role of Mhy1p in lipid biosynthesis and serves as a resource for exploring physiological dimorphic growth in Y. lipolytica.

Vulnerability of the global terrestrial ecosystems to climate change.

Climate change has far-reaching impacts on ecosystems. Recent attempts to quantify such impacts focus on measuring exposure to climate change but largely ignore ecosystem resistance and resilience, which may also affect the vulnerability outcomes. In this study, the relative vulnerability of global terrestrial ecosystems to short-term climate variability was assessed by simultaneously integrating exposure, sensitivity, and resilience at a high spatial resolution (0.05°). The results show that vulnerable areas are currently distributed primarily in plains. Responses to climate change vary among ecosystems and deserts and xeric shrublands are the most vulnerable biomes. Global vulnerability patterns are determined largely by exposure, while ecosystem sensitivity and resilience may exacerbate or alleviate external climate pressures at local scales; there is a highly significant negative correlation between exposure and sensitivity. Globally, 61.31% of the terrestrial vegetated area is capable of mitigating climate change impacts and those areas are concentrated in polar regions, boreal forests, tropical rainforests, and intact forests. Under current sensitivity and resilience conditions, vulnerable areas are projected to develop in high Northern Hemisphere latitudes in the future. The results suggest that integrating all three aspects of vulnerability (exposure, sensitivity, and resilience) may offer more comprehensive and spatially explicit adaptation strategies to reduce the impacts of climate change on terrestrial ecosystems.

Sequencing of the Litchi Downy Blight Pathogen Reveals It Is a Phytophthora Species With Downy Mildew-Like Characteristics.

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.

PsMPK1, an SLT2-type mitogen-activated protein kinase, is required for hyphal growth, zoosporogenesis, cell wall integrity, and pathogenicity in Phytophthora sojae.

Mitogen-activated protein kinases (MAPKs) play important roles in the regulation of vegetative and pathogenic growth in plant pathogens. Here, we identified an SLT2-type MAP kinase in Phytophthora sojae, PsMPK1, which was transcriptionally induced in sporulating hyphae and the early stages of infection. Silencing of PsMPK1 caused defects in growth and zoosporogenesis, and increased hyphal swellings after the induction of sporangia formation, along with increasing hypersensitivity to cell wall-degrading enzymes. Transmission electron microscopy showed that the cell wall of PsMPK1-silenced mutants was also deleteriously affected. A dark outermost layer in the cell walls disappeared in the mutants, and an additional layer of the mutant cell wall that was deposited abnormally inside an inner bright layer appeared nonhomogeneous and rough compared to the wild type. Pathogenicity assays showed that PsMPK1-silenced transformants lost their pathogenicity on susceptible soybean host plants and triggered stronger cell death. Overall, PsMPK1 is involved in growth, differentiation, cell wall integrity, and pathogenicity in P. sojae.

Molecular characteristics of the complete genome of a J-subgroup avian leukosis virus strain isolated from Eurasian teal in China.

The J-subgroup avian leukosis virus (ALV-J) strain WB11098J was isolated from a wild Eurasian teal, and its proviral genomic sequences were determined. The complete proviral sequence of WB11098J was 7868 nt long. WB11098J was 95.3.9 % identical to the prototype strain HPRS-103, 94.2 % identical to the American strain ADOL-7501, 94.5-94.7 % identical to Chinese broiler isolates, 94.8-97.5 % identical to layer chicken isolates, and 94.4-95.0 % identical to Chinese local chicken isolates at the nucleotide level. Phylogenetic analysis showed that the WB11098J isolate shared the greatest homology with the layer strain SD09DP03 and was included in the same cluster. Interestingly, two 19-bp insertions in the U3 regions of the 5'LTR and 5'UTR that were most likely derived from other retroviruses were found in the WB11098J isolate. These insertions separately introduced one E2BP-binding site in the U3 region of the 5'LTR and a RNA polymerase II transcription factor IIB and core promoter motif of ten elements in the 5'UTR. A 5-bp deletion was identified in the U3 region of the 5'LTR. No nucleotides were deleted in the rTM or DR-1 regions in the 3'UTR. A 1-bp deletion was detected in the E element and introduced a specific and distinct binding site for c-Ets-1. Our study is the first to report the molecular characteristics of the complete genome of an ALV-J that was isolated from a wild bird and will provide necessary information for further understanding of the evolution of ALV-J.