An efficient Ru(II)-catalyzed C-H activation-based spiroannulation of benzoxazines with the easily available benzoquinone and N-sulfonyl quinone monoimine has been realized, providing a straightforward strategy to access NH-containing spiropyrans in moderate to good yields with good functional group compatibility. The procedure features atom- and step-economy, mild conditions, and excellent chemoselectivity. Moreover, a catalytically competent five-membered cycloruthenated complex has been isolated.
Lasiodiplodia is a widely distributed genus that is associated with a variety of diseases in many plant species, especially fruit trees. In this study, a disease survey of fruit trees growing in 12 orchards located in the Henan and Shandong provinces of China was conducted between 2020 and 2022. The symptoms observed included stem canker, branch dieback, and gummosis. Phylogenetic analyses of internal transcribed spacer, tub2, tef1, and rpb2 sequence data combined with morphological characteristics revealed that the 19 isolates collected during the survey belonged to five documented Lasiodiplodia species, namely, Lasiodiplodia citricola, L. chiangraiensis, L. huangyanensis, L. pseudotheobromae, and L. theobromae, and two previously undescribed species, L. xinyangensis and L. ziziphi. In addition, the survey identified three novel host-pathogen interactions: L. chiangraiensis on loquat, L. citricola on apple, and L. huangyanensis on grapevine. Furthermore, the detailed phylogenic analysis indicated that four previously described Lasiodiplodia species were genetically very closely related that they would be better classified as synonyms rather than distinct species, so L. paraphysoides and L. nanpingensis should be considered synonyms of L. citricola, L. fujianensis should be a synonym of L. iraniensis, and L. henanica should be a synonym of L. huangyanensis. Pathogenicity tests confirmed that representative isolates of the two novel species and three new host-pathogen interactions identified in the current study were pathogenic to their original hosts, thereby fulfilling Koch's postulates. Similarly, all of the isolates were found to be pathogenic on four alternative hosts, although a high degree of variation in virulence was observed.
Ascomycota , Malus , Mitosporic Fungi , Fruit , Phylogeny , China , Ascomycota/genetics
Plant endophytic fungi and rhizosphere soil fungi are often reported as biocontrol agents against plant pathogens or with plant growth promotion potential. Four treatments were performed in field and greenhouse experiments where cucumber plants were inoculated with Trichoderma harzianum and Fusarium oxysporum in 2022. The roots, stems and leaves of cucumber plants and their rhizosphere soil were collected twice individually from the field and greenhouse for isolation of cucumber endophytic and rhizosphere soil fungi. All fungal strains were identified through sequence similarity of the ITS1-5.8s-ITS2 rDNA region. The potato dextrose agar (PDA) media yielded the highest number of genera isolated from cucumber plants, rhizosphere soil and both compared to other media. There were no significant differences among the four media for the isolation of all cucumber endophytic fungi. However, in the roots, the number of endophytic fungi isolated by MRBA was significantly higher than that isolated on malt extract agar (MEA), while in the stems, the number of fungi isolated with PDA was significantly higher than that isolated with Martin's rose bengal agar medium (MRBA). PDA had significantly higher isolation efficiency for the rhizosphere soil fungi than MRBA. The 28 fungal genera had high isolation efficiency, and the endophytic Trichoderma strains were significantly more isolated by MEA than those of MRBA. It is suggested that PDA can be used as a basic medium, and different cultural media can be considered for specific fungal genera.
Ru(II)-catalyzed condition-controlled divergent coupling between 3-aryl-2H-benzoxazin-2-ones and benzoquinones has been realized under operationally simple conditions, affording a series of structurally stable spiropyrans and valuable arylquinones. The potential of this method is also demonstrated by scale-up synthesis and derivatization. Additionally, an unprecedented cycloruthenated complex has been identified as a key intermediate.
Vinylarenes represent an important class of core skeleton embedded in natural products, organic materials, and pharmaceutical molecules. Therefore, numerous efforts have been devoted to developing efficient methods for their preparation. Among them, transition-metal-catalyzed oxidative coupling of arenes and alkenes has proved to be a powerful method due to its high atom and step economy. Although a wide range of oxidative alkenylations of arenes have been developed, the alkenes employed in most cases are still limited to electron-deficient alkenes. Reported herein is a Rh(III)-catalyzed C-H cross-coupling of benzoxazinones and simple unactivated styrenes to furnish a variety of vinylarene scaffolds. This established protocol is characterized by wide functional group compatibility, high yields, and excellent regio- and chemo-selectivity. Mechanistic studies and gram-scale experiments on this high-value conversion are disclosed. Moreover, the potential utility of this method was highlighted by a series of further transformations.
Tup1, a conserved transcriptional repressor, plays a critical role in the growth and development of fungi. Here, we identified a BsTup1 gene from the plant pathogenic fungus Bipolaris sorokiniana. The expression of BsTup1 showed a more than three-fold increase during the conidial stage compared with mycelium stage. Deletion of BsTup1 led to decrease hyphal growth and defect in conidia formation. A significant difference was detected in osmotic, oxidative, or cell wall stress responses between the WT and ΔBsTup1 strains. Pathogenicity assays showed that virulence of the ΔBsTup1 mutant was dramatically decreased on wheat and barely leaves. Moreover, it was observed that hyphal tips of the mutants could not form appressorium-like structures on the inner epidermis of onion and barley coleoptile. Yeast two-hybrid assays indicated that BsTup1 could interact with the BsSsn6. RNAseq revealed significant transcriptional changes in the ΔBsTup1 mutant with 2369 genes down-regulated and 2962 genes up-regulated. In these genes, we found that a subset of genes involved in fungal growth, sporulation, cell wall integrity, osmotic stress, oxidation stress, and pathogenicity, which were misregulated in the ΔBsTup1 mutant. These data revealed that BsTup1 has multiple functions in fungal growth, development, stress response and pathogenesis in B. sorokiniana.
Bipolaris , Hordeum , Fungal Proteins/genetics , Fungal Proteins/metabolism , Hordeum/genetics , Hordeum/metabolism , Plant Diseases/microbiology , Spores, Fungal/genetics , Virulence/genetics
Exserohilum turcicum and E. rostratum, two closely related fungal species, are both economically important pathogens but have quite different target hosts (specific to plants and cross-kingdom infection, respectively). In the present study, complete circular mitochondrial genomes of the two Exserohilum species were sequenced and de novo assembled, which mainly comprised the same set of 13 core protein-coding genes (PCGs), two rRNAs, and a certain number of tRNAs and unidentified open reading frames (ORFs). Comparative analyses indicated that these two fungi had significant mitogenomic collinearity and consistent mitochondrial gene arrangement, yet with vastly different mitogenome sizes, 264,948 bp and 64,620 bp, respectively. By contrast with the 17 introns containing 17 intronic ORFs (one-to-one) in the E. rostratum mitogenome, E. turcicum involved far more introns (70) and intronic ORFs (126), which was considered as the main contributing factors of their mitogenome expansion/contraction. Within the generally intron-rich gene cox1, a total of 18 and 10 intron position classes (Pcls) were identified separately in the two mitogenomes. Moreover, 16.16% and 10.85% ratios of intra-mitogenomic repetitive regions were detected in E. turcicum and E. rostratum, respectively. Based on the combined mitochondrial gene dataset, we established a well-supported topology of phylogeny tree of 98 ascomycetes, implying that mitogenomes may act as an effective molecular marker for fungal phylogenetic reconstruction. Our results served as the first report on mitogenomes in the genus Exserohilum, and would have significant implications in understanding the origin, evolution and pathogenic mechanisms of this fungal lineage.
Corynespora cassiicola, the causal agent of an extensive range of plant diseases worldwide, is a momentous fungus with diverse lifestyles and rich in intraspecies variations. In the present study, a total of 56 mitochondrial genomes of C. cassiicola were assembled (except two available online) and analyzed, of which 16 mitogenomes were newly sequenced here. All these circular mitochondrial DNA (mtDNA) molecules, ranging from 39,223 bp to 45,786 bp in length, comprised the same set of 13 core protein-coding genes (PCGs), two rRNAs and 27 tRNAs arranged in identical order. Across the above conserved genes, nad3 had the largest genetic distance between different isolates and was possibly subjected to positive selection pressure. Comparative mitogenomic analysis indicated that seven group I (IB, IC1, and IC2) introns with a length range of 1013-1876 bp were differentially inserted in three core PCGs (cox1, nad1, and nad5), resulting in the varied mitogenome sizes among C. cassiicola isolates. In combination with dynamic distribution of the introns, a well-supported mitogenome-wide phylogeny of the 56 C. cassiicola isolates revealed eight phylogenetic groups, which only had weak correlations with host range and toxin class. Different groups of isolates exhibited obvious differences in length and GC content of some genes, while a degree of variance in codon usage and tRNA structure was also observed. This research served as the first report on mitogenomic comparisons within C. cassiicola, and could provide new insights into its intraspecific microevolution and genetic diversity.
Corynespora cassiicola is a ubiquitous pathogenic fungus that can infect a broad range of plant hosts. Corynespora leaf fall, caused by C. cassiicola, is one of the major diseases on rubber tree in China. This disease is having an increasing affect on natural rubber production worldwide. In this study, by combining the Nanopore and Illumina sequencing technologies, we present the chromosome-scale genome sequence of the rubber tree-sampled C. cassiicola strain XJ collected in the subtropical region of China. The assembly consists of 23 scaffolds (N50 = 4.62 Mb) with an estimated genome size of 44.42 Mb (only 166 non-ATCG bases) and 16,108 protein-coding genes. The genome will provide a valuable resource for further research on the pathogenesis and comparative genomics of C. cassiicola on rubber tree and other hosts.
Hevea , Nanopores , Ascomycota , High-Throughput Nucleotide Sequencing , Plant Diseases
A novel mycovirus, named "Corynespora cassiicola bipartite mycovirus 1" (CcBV1), was isolated from a phytopathogenic fungus, Corynespora cassiicola, the causal agent of rubber leaf fall disease. The nucleotide sequence of the complete genome of CcBV1, which consists of two double-stranded RNA (dsRNA) segments, was determined. The first dsRNA is 2,002 bp in length and contains a single open reading frame (ORF) encoding a putative RNA-dependent RNA polymerase (RdRp) (69 kDa), while the second is 1,738 bp in length and contains a single ORF encoding a hypothetical protein of unknown function, with an approximately molecular weight of 36 kDa. The amino acid sequences of the both deduced proteins are most similar (58.9% and 45.1% identity, respectively) to those of Cryphonectria parasitica bipartite mycovirus 1 (CpBV1). Phylogenetic analysis indicated that CcBV1 clusters together with CpBV1 and other unassigned dsRNA mycoviruses. To the best of our knowledge, this represents the first report of a mycovirus infecting C. cassiicola.
Ascomycota/virology , Fungal Viruses/genetics , Genome, Viral , Phylogeny , RNA, Double-Stranded/genetics , RNA, Viral/genetics , Ascomycota/pathogenicity , Base Sequence , China , Fungal Viruses/classification , Fungal Viruses/isolation & purification , Open Reading Frames , Plant Diseases/microbiology , Plants/microbiology , RNA-Dependent RNA Polymerase/genetics , Sequence Alignment , Viral Proteins/genetics , Whole Genome Sequencing
A number of species in Bipolaris are important plant pathogens. Due to a limited number of synapomorphic characters, it is difficult to perform species identification and to estimate phylogeny of Bipolaris based solely on morphology. In this study, we sequenced the complete mitochondrial genome of Bipolaris sorokiniana, and presented the detailed annotation of the genome. The B. sorokiniana mitochondrial genome is 137,775 bp long, and contains two ribosomal RNA genes, 12 core protein-coding genes, 38 tRNA genes. In addition, two ribosomal protein genes (rps3 gene and rps5 gene) and the fungal mitochondrial RNase P gene (rnpB) are identified. The large genome size is mostly determined by the presence of numerous intronic and intergenic regions. A total of 28 introns are inserted in eight core protein-coding genes. Together with the published mitochondrial genome sequences, we conducted a preliminary phylogenetic inference of Dothideomycetes under various datasets and substitution models. The monophyly of Capnodiales, Botryosphaeriales and Pleosporales are consistently supported in all analyses. The Venturiaceae forms an independent lineage, with a distant phylogenetic relationship to Pleosporales. At the family level, the Mycosphaerellaceae, Botryosphaeriaceae. Phaeosphaeriaceae, and Pleosporaceae are recognized in the majority of trees.
BACKGROUND: To clarify the impact of IL-1B gene polymorphisms (IL-1B-511C/T, IL-1B-31C/T, IL-1B+3954C/T) in Helicobacter pylori (H. pylori) infection by mean of a meta-analysis. METHODS: The relevant studies were retrieved from PubMed, Web of Science, EMBASE, Cochrane Library, Chinese National Knowledge Infrastructure (CNKI) and Wanfang databases until September 9, 2018. Odds ratio (OR) and its 95% confidence intervals (CIs) were used to assess the associations. Statistical analyses of this meta-analysis were conducted by using STATA 12 software. RESULTS: Totally, 45 articles including 9606 cases and 5654 controls were enrolled in this meta-analysis. Our results indicated that IL-1B-511C/T polymorphism was significantly related to an increased the risk of H. pylori infection under recessive model (ORâ¯=â¯1.13, 95% CI: 1.00-1.27, Pâ¯=â¯0.048). However, no significant associations were obtained between H. pylori infection and IL-1B-31C/T as well as IL-1B+3954C/T polymorphisms under all models. In addition, subgroup analyses were also performed by country, study design, and detection methods of H. pylori. CONCLUSIONS: This meta-analysis suggested that IL-1B-511C/T polymorphism was related to the risk of H. pylori infection. Further larger studies with high quality are needed to conform these findings.
Helicobacter Infections/genetics , Helicobacter pylori/physiology , Interleukin-1beta/genetics , Polymorphism, Single Nucleotide , Genetic Predisposition to Disease , Helicobacter Infections/metabolism , Helicobacter Infections/microbiology , Helicobacter pylori/genetics , Humans , Interleukin-1beta/metabolism
Changes in electroencephalogram (EEG) signals under repetitive magnetic stimulation at the acupoint of Guangming (GB37) were analyzed using nonlinear dynamics complexity. C0 complexity is a statistical indicators which can quantify time dynamics of EEG signals. The study compared the C0 complexity under magnetic stimulation at GB37 with those at a mock point, as well as the C0 complexity under visual stimulation before and after magnetic stimulation at GB37. The results showed that the C0 complexity values of EEG signals in the electrode F3,Cz,C4 and P3 were different depending on whether magnetic stimulation was at the GB37 or a mock point (P <; 0.05),and the C0 complexity of magnetic stimulation on GB37 was generally higher than that on mock point. Moreover, EEG signals from visual stimulation before magnetic stimulation at GB37 were significantly different from those after magnetic stimulation at GB37 in two electrodes which were C4 and P3(P <; 0.05). The conclusion of the study is that magnetic stimulation at GB37 has a significant impact on EEG signals. First, EEG complexity during magnetic stimulation at GB37 was significantly higher than that at the mock point in frontal area, central area and parietal area (electrodes F3, Cz, C4 and P3). Second, EEG signals in central area and parietal area (electrodes C4 and P3) resulting from visual stimulation differed depending on whether magnetic stimulation at GB37 was given. The study has important significance for the application of magnetic stimulation on acupoints.
Acupuncture Points , Electroencephalography , Magnetics , Photic Stimulation , Systems Analysis
Here, we administered repeated-pulse transcranial magnetic stimulation to healthy people at the left Guangming (GB37) and a mock point, and calculated the sample entropy of electroencephalo-gram signals using nonlinear dynamics. Additionally, we compared electroencephalogram sample entropy of signals in response to visual stimulation before, during, and after repeated-pulse tran-scranial magnetic stimulation at the Guangming. Results showed that electroencephalogram sample entropy at left (F3) and right (FP2) frontal electrodes were significantly different depending on where the magnetic stimulation was administered. Additionally, compared with the mock point, electroencephalogram sample entropy was higher after stimulating the Guangming point. When visual stimulation at Guangming was given before repeated-pulse transcranial magnetic stimula-tion, significant differences in sample entropy were found at five electrodes (C3, Cz, C4, P3, T8) in parietal cortex, the central gyrus, and the right temporal region compared with when it was given after repeated-pulse transcranial magnetic stimulation, indicating that repeated-pulse transcranial magnetic stimulation at Guangming can affect visual function. Analysis of electroencephalogram revealed that when visual stimulation preceded repeated pulse transcranial magnetic stimulation, sample entropy values were higher at the C3, C4, and P3 electrodes and lower at the Cz and T8 electrodes than visual stimulation followed preceded repeated pulse transcranial magnetic stimula-tion. The findings indicate that repeated-pulse transcranial magnetic stimulation at the Guangming evokes different patterns of electroencephalogram signals than repeated-pulse transcranial mag-netic stimulation at other nearby points on the body surface, and that repeated-pulse transcranial magnetic stimulation at the Guangming is associated with changes in the complexity of visually evoked electroencephalogram signals in parietal regions, central gyrus, and temporal regions.
Annexin A2 and Cdc42 were identified by 2-dimensional electrophoresis (2-DE) and MALDI-TOF-MS between esophageal squamous cell carcinomas (ESCC) and corresponding normal esophagus mucosa in our previous study. To assess clinico-pathological pattern and Annexin A2 and Cdc42 status with respect to cell differentiation and lymphnode metastasis in patients with ESCC. The expression of Annexin A2 and Cdc42 in 22 pairs of fresh ESCC and matched tissues were detected by qRT-PCR and western blot, respectively. And it was further confirmed by immunohistochemistry with 175 pairs of formalin-fixed, paraffin-embedded ESCC. Results showed that Annexin A2 expression was significantly down-regulated, and Cdc42 was up-regulated in ESCC compared to matched control on both mRNA and protein level (P < 0.05), which was in accordance with our previous results on proteomics data. Additionally, Annexin A2 and Cdc42 expression was significantly correlated with lymphoid node metastasis (P < 0.05) and pathological differentiation (P < 0.05). Taken together, we proposed that the aberrant expression of Annexin A2 and Cdc42 played a role in carcinogenesis, differentiation and metastasis of ESCC, which implied its potential target for clinical biomarkers in differentiation and lymph node metastasis.
Annexin A2/metabolism , Carcinoma, Squamous Cell/metabolism , Esophageal Neoplasms/metabolism , Lymph Nodes/pathology , Lymphatic Metastasis/pathology , cdc42 GTP-Binding Protein/metabolism , Blotting, Western , Carcinoma, Squamous Cell/pathology , DNA Primers/genetics , Electrophoresis, Gel, Two-Dimensional , Esophageal Neoplasms/pathology , Humans , Immunohistochemistry , Lymph Nodes/metabolism , Real-Time Polymerase Chain Reaction , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Acupuncture is a useful irritation therapy clinically. Acupuncture at acupoints is proved to have specific function on human body. Magnetic stimulation is a new kind of stimulating technique of non-invasive, painless and effective. The effect of magnetic stimulation on acupoint is a new subject in recent years. In this paper, the electroencephalogram (EEG) evoked potentials (EPs) of magnetic stimulation by stimulating the acupoint of Shenmen(HT7) are studied. The experiments are divided into four groups: quiet, acupoint stimulation, mock stimulation and mock point stimulation. The EEG EPs are collected and an obvious P150 component is obtained. The amplitudes of P150s are analyzed and compared. The P150s are localized in brain by dipole model and the coronal, sagittal and axial plans are painted and observed. The results show that acupoint stimulation on acupoint of Shenmen(HT7) can evoke stronger nerve activity of somatosensory than stimulation on common surface. The dipole source of acupoint stimulation and common surface stimulation are both focused on the cingulated gyrus which related to somatosensory.
Acupuncture Points , Electroencephalography/methods , Evoked Potentials , Magnetics , Humans , Principal Component Analysis
Two acupuncture manipulations are clinically used: manual manipulation and electrical acupuncture. There is little published on the EEG changes during magnetic stimulation on an acupuncture site. In this study, EEG data in response to magnetic stimulation on HeGu (LI 4) acupoint were measured to determine whether magnetic acupoint stimulation might modulate ongoing EEG or not. Eighteen healthy volunteers (13 male, 5 female) 20 to 35 years old were chosen in this experiment, with consent obtained before the study. The highest evoked potential was recorded in FCZ electrode, at about 140-170 ms (P150) after acupoint stimulation, but not mock point stimulation. Comparison of the somatosensory-evoked potentials in response to acupoint stimulation and mock point stimulation showed that P150 was specific to acupoint stimulation. With regard to the location of P150 in the human brain, we suggest that magnetic stimulation on HeGu acupoint would affect specific brain areas compared with the mock point. The difference in the anatomical structure of acupoint and non-acupoint may explain the specific acupoint-brain correlation, and P150 may be a characteristic activation in response to acupoint afferent.
Acupuncture/methods , Brain Mapping/methods , Cerebral Cortex/pathology , Electroencephalography/methods , Evoked Potentials, Somatosensory/physiology , Magnetics , Adult , Brain/pathology , Brain Mapping/instrumentation , Computer Simulation , Electrodes , Female , Hand/pathology , Humans , Male , Middle Aged , Software
