41 results on '"Chenlei Hua"'
Search Results
2. The function of MoGlk1 in integration of glucose and ammonium utilization in Magnaporthe oryzae.
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Lisha Zhang, Ruili Lv, Xianying Dou, Zhongqiang Qi, Chenlei Hua, Haifeng Zhang, Zhengyi Wang, Xiaobo Zheng, and Zhengguang Zhang
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Medicine ,Science - Abstract
Hexokinases are conserved proteins functioning in glucose sensing and signaling. The rice blast fungus Magnaporthe oryzae contains several hexokinases, including MoHxk1 (hexokinase) and MoGlk1 (glucokinase) encoded respectively by MoHXK1 and MoGLK1 genes. The heterologous expression of MoGlk1 and MoHxk1 in Saccharomyces cerevisiae confirmed their conserved functions. Disruption of MoHXK1 resulted in growth reduction in medium containing fructose as the sole carbon source, whereas disruption of MoGLK1 did not cause the similar defect. However, the ΔMoglk1 mutant displayed decreased proton extrusion and a lower biomass in the presence of ammonium, suggesting a decline in the utilization of ammonium. Additionally, the MoGLK1 allele lacking catalytic activity restored growth to the ΔMoglk1 mutant. Moreover, the expression of MoPMA1 encoding a plasma membrane H(+)-ATPase decreased in the ΔMoglk1 mutant that can be suppressed by glucose and G-6-P. Thus, MoGlk1, but not MoHxk1, regulates ammonium utilization through a mechanism that is independent from its catalytic activity.
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- 2011
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3. Functional diversification of a wild potato immune receptor at its center of origin.
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Ascurra, Yerisf C. Torres, Lisha Zhang, Toghani, AmirAli, Chenlei Hua, Rangegowda, Nandeesh Jalahalli, Posbeyikian, Andres, Hsuan Pai, Xiao Lin, Wolters, Pieter J., Wouters, Doret, de Blok, Reinhoud, Steigenga, Niels, Paillart, Maxence J. M., Visser, Richard G. F., Kamoun, Sophien, Nürnberger, Thorsten, and Vleeshouwers, Vivianne G. A. A.
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- 2023
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4. The Arabidopsis TIR-NBS-LRR protein CSA1 guards BAK1-BIR3 homeostasis and mediates convergence of pattern- and effector-induced immune responses
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Sarina Schulze, Liping Yu, Chenlei Hua, Lisha Zhang, Dagmar Kolb, Hannah Weber, Alexandra Ehinger, Svenja C. Saile, Mark Stahl, Mirita Franz-Wachtel, Lei Li, Farid El Kasmi, Thorsten Nürnberger, Volkan Cevik, and Birgit Kemmerling
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Virology ,Parasitology ,Microbiology - Abstract
Arabidopsis BAK1/SERK3, a co-receptor of leucine-rich repeat pattern recognition receptors (PRRs), mediates pattern-triggered immunity (PTI). Genetic inactivation of BAK1 or BAK1-interacting receptor-like kinases (BIRs) causes cell death, but the direct mechanisms leading to such deregulation remains unclear. Here, we found that the TIR-NBS-LRR protein CONSTITUTIVE SHADE AVOIDANCE 1 (CSA1) physically interacts with BIR3, but not with BAK1. CSA1 mediates cell death in bak1-4 and bak1-4 bir3-2 mutants via components of effector-triggered immunity-(ETI) pathways. Effector HopB1-mediated perturbation of BAK1 also results in CSA1-dependent cell death. Likewise, microbial pattern pg23-induced cell death, but not PTI responses, requires CSA1. Thus, we show that CSA1 guards BIR3 BAK1 homeostasis and integrates pattern- and effector-mediated cell death pathways downstream of BAK1. De-repression of CSA1 in the absence of intact BAK1 and BIR3 triggers ETI cell death. This suggests that PTI and ETI pathways are activated downstream of BAK1 for efficient plant immunity.
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- 2022
5. The EDS1–PAD4–ADR1 node mediates Arabidopsis pattern-triggered immunity
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Anna Joe, Shaofei Rao, Svenja C. Saile, Bart P. H. J. Thomma, Rory N Pruitt, Farid El Kasmi, Klaus Harter, Jianmin Zhou, Jane E. Parker, Katja Fröhlich, Sara C. Stolze, Lisha Zhang, Darya Karelina, Chenlei Hua, Federica Locci, Jeffery L. Dangl, Wei Lin Wan, Hirofumi Nakagami, Claudia Oecking, Matthieu H. A. J. Joosten, Andrea A. Gust, Anne Harzen, Friederike Wanke, Thorsten Nürnberger, Meijuan Hu, and Detlef Weigel
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Arabidopsis ,Plant Immunity ,Receptors, Cell Surface ,chemical and pharmacologic phenomena ,Protein Serine-Threonine Kinases ,Immune system ,Protein Domains ,Life Science ,Receptor ,Gene ,Multidisciplinary ,biology ,Arabidopsis Proteins ,Effector ,Kinase ,fungi ,biology.organism_classification ,Cell biology ,Laboratorium voor Phytopathologie ,DNA-Binding Proteins ,Crosstalk (biology) ,Laboratory of Phytopathology ,Protein Multimerization ,EPS ,Carboxylic Ester Hydrolases ,Protein Kinases - Abstract
Plants deploy cell-surface and intracellular leucine rich-repeat domain (LRR) immune receptors to detect pathogens1. LRR receptor kinases and LRR receptor proteins at the plasma membrane recognize microorganism-derived molecules to elicit pattern-triggered immunity (PTI), whereas nucleotide-binding LRR proteins detect microbial effectors inside cells to confer effector-triggered immunity (ETI). Although PTI and ETI are initiated in different host cell compartments, they rely on the transcriptional activation of similar sets of genes2, suggesting pathway convergence upstream of nuclear events. Here we report that PTI triggered by the Arabidopsis LRR receptor protein RLP23 requires signalling-competent dimers of the lipase-like proteins EDS1 and PAD4, and of ADR1 family helper nucleotide-binding LRRs, which are all components of ETI. The cell-surface LRR receptor kinase SOBIR1 links RLP23 with EDS1, PAD4 and ADR1 proteins, suggesting the formation of supramolecular complexes containing PTI receptors and transducers at the inner side of the plasma membrane. We detected similar evolutionary patterns in LRR receptor protein and nucleotide-binding LRR genes across Arabidopsis accessions; overall higher levels of variation in LRR receptor proteins than in LRR receptor kinases are consistent with distinct roles of these two receptor families in plant immunity. We propose that the EDS1–PAD4–ADR1 node is a convergence point for defence signalling cascades, activated by both surface-resident and intracellular LRR receptors, in conferring pathogen immunity. The authors provide mechanistic insights into the crosstalk between signalling components of pattern-triggered immunity and effector-triggered immunity and their molecular linkers.
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- 2021
6. Distinct immune sensor systems for fungal endopolygalacturonases in closely related Brassicaceae
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Lei Wang, Isabell Albert, Markus Albert, Si Qin, Chenlei Hua, Jan A. L. van Kan, Lisha Zhang, Thorsten Nürnberger, and Rory N Pruitt
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Genetics ,biology ,Complex formation ,fungi ,Pattern recognition receptor ,Plant Immunity ,food and beverages ,Brassicaceae ,Plant Science ,biology.organism_classification ,Arabidopsis arenosa ,Laboratorium voor Phytopathologie ,Immune system ,Brassica rapa ,Laboratory of Phytopathology ,Arabidopsis thaliana ,Life Science ,EPS - Abstract
Plant pattern recognition receptors (PRRs) facilitate recognition of microbial patterns and mediate activation of plant immunity. Arabidopsis thaliana RLP42 senses fungal endopolygalacturonases (PGs) and triggers plant defence through complex formation with SOBIR1 and SERK co-receptors. Here, we show that a conserved 9-amino-acid fragment pg9(At) within PGs is sufficient to activate RLP42-dependent plant immunity. Structure-function analysis reveals essential roles of amino acid residues within the RLP42 leucine-rich repeat and island domains for ligand binding and PRR complex assembly. Sensitivity to pg9(At), which is restricted to A. thaliana and exhibits scattered accession specificity, is unusual for known PRRs. Arabidopsis arenosa and Brassica rapa, two Brassicaceae species closely related to A. thaliana, respectively perceive immunogenic PG fragments pg20(Aa) and pg36(Bra), which are structurally distinct from pg9(At). Our study provides evidence for rapid evolution of polymorphic PG sensors with distinct pattern specificities within a single plant family.
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- 2021
7. Characteristics of demersal fish community structure during summer hypoxia in the Pearl River Estuary, China
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Han Lai, Sheng Bi, Huadong Yi, Haiyang Li, Xuchong Wei, Gongpei Wang, Dingli Guo, Xuange Liu, Jiahui Chen, Qiuxian Chen, Zhilun Zhang, Shuang Liu, Chenlei Huang, Li Lin, and Guifeng Li
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dissolved oxygen ,environmental factors ,fish community ,hypoxia ,Pearl River Estuary ,Ecology ,QH540-549.5 - Abstract
Abstract In recent decades, hypoxic areas have rapidly expanded worldwide in estuaries and coastal zones. The Pearl River Estuary (PRE), one of China's largest estuaries, experiences frequent seasonal hypoxia due to intense human activities and eutrophication. However, the ecological effects of hypoxia in the PRE, particularly on fish communities, remain unclear. To explore these effects, we collected fish community and environmental data in July 2021 during the summer hypoxia development period. The results revealed that bottom‐layer dissolved oxygen (DO) in the PRE ranged from 0.08 to 5.71 mg/L, with extensive hypoxic zones (DO ≤ 2 mg/L) observed. Hypoxia has varied effects on fish community composition, distribution, species, and functional diversity in the PRE. A total of 104 fish species were collected in this study, with approximately 30 species (28.6%) exclusively found in hypoxic areas. Species responses to hypoxia varied: species such as Sardinella zunasi, Coilia mystus, and Nuchequula nuchalis were sensitive, while Decapterus maruadsi, Siganus fuscescens, and Lagocephalus spadiceus showed higher tolerance. Within the hypoxia area, dissolved oxygen was the main limiting factor for fish community diversity. Functional diversity (FDiv) decreased with higher dissolved oxygen levels, indicating a potential shift in the functional traits and ecological roles of fish species in response to changing oxygen conditions. Further analysis demonstrated that dissolved oxygen had a significantly stronger effect on fish community structure at hypoxic sites than in the whole PRE. Moreover, other environmental variables also had significant effects on the fish community structure and interacted with dissolved oxygen in the hypoxia area. These findings suggest that maintaining sufficient dissolved oxygen levels is essential for sustaining fish communities and ecosystem health in the PRE. This study provides novel insights into the effects of hypoxia on fish communities in estuarine ecosystems and has significant implications for the ecological health and management of the PRE.
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- 2024
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8. Surface Sensor Systems in Plant Immunity
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Rory N Pruitt, Thorsten Nürnberger, Chenlei Hua, Lisha Zhang, and Isabell Albert
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Physiology ,Cell Membrane ,Cell ,Plant Immunity ,Plant Science ,Biology ,Models, Biological ,Cell biology ,medicine.anatomical_structure ,Immune system ,Post translational ,Receptors, Pattern Recognition ,Protein processing ,Genetics ,medicine ,Signal transduction ,Receptor ,Protein Processing, Post-Translational ,human activities ,Updates - Focus Issue ,Intracellular ,Signal Transduction - Abstract
Protein complexes at the cell surface facilitate the detection of danger signals from diverse pathogens and initiate a series of complex intracellular signaling events that result in various immune responses.
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- 2019
9. Theoretical and numerical research on the dynamic launch response of carbon fiber composite cartridges
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Ruijie Zhang, Hui Xu, Chenlei Huang, Kun Liu, and Zhilin Wu
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Composite material cartridges ,Dynamic response ,Thick-walled cylinder theory ,Finite element simulation ,Toughing-chamber behavior ,Military Science - Abstract
Understanding the dynamic response of composite material cartridges during the firing process is of great significance for improving their reliability and safety. A theoretical model describing the dynamic response of composite material cartridges is established based on the thick-walled cylinder theory and rate-dependent constitutive model of composite materials. The correctness of the theoretical model is validated through finite element simulations of cartridge deformation. The influence of chamber pressure and cartridge wall thickness on the cartridge’s deformation process and stress distribution is analyzed. The results indicate that the primary deformation of composite material cartridges inside the chamber is elastic deformation. Compared to metal cartridges, composite material cartridges require higher pressure for touching-chamber and are more prone to developing gaps after unloading to ensure smooth extraction. During the deformation process, the touching-chamber behavior of the cartridge can improve the stress distribution. Under the same chamber pressure, the touching-chamber behavior can reduce the circumferential stress by approximately 30%. The inner wall surface of the cartridge is a critical area that requires attention. The touching-chamber behavior can be facilitated by appropriately reducing the cartridge wall thickness while ensuring overall strength. This study can provide guidance for the optimization design of composite material cartridges.
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- 2024
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10. Phytophthora infestans small phospholipase D-like proteins elicit plant cell death and promote virulence
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C. Schoina, Harold J. G. Meijer, Chenlei Hua, Francine Govers, Shutong Wang, and Klaas Bouwmeester
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0106 biological sciences ,0301 basic medicine ,Signal peptide ,Oomycete ,Phospholipase D ,Soil Science ,Virulence ,Plant Science ,Phospholipase ,Biology ,biology.organism_classification ,01 natural sciences ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,Haustorium ,Phytophthora infestans ,lipids (amino acids, peptides, and proteins) ,Agronomy and Crop Science ,Molecular Biology ,Pathogen ,010606 plant biology & botany - Abstract
The successful invasion of host tissue by (hemi-)biotrophic plant pathogens is dependent on modifications of the host plasma membrane to facilitate the two-way transfer of proteins and other compounds. Haustorium formation and the establishment of extrahaustorial membranes are probably dependent on a variety of enzymes that modify membranes in a coordinated fashion. Phospholipases, enzymes that hydrolyse phospholipids, have been implicated as virulence factors in several pathogens. The oomycete Phytophthora infestans is a hemibiotrophic pathogen that causes potato late blight. It possesses different classes of phospholipase D (PLD) proteins, including small PLD-like proteins with and without signal peptide (sPLD-likes and PLD-likes, respectively). Here, we studied the role of sPLD-like-1, sPLD-like-12 and PLD-like-1 in the infection process. They are expressed in expanding lesions on potato leaves and during in vitro growth, with the highest transcript levels in germinating cysts. When expressed in planta in the presence of the silencing suppressor P19, all three elicited a local cell death response that was visible at the microscopic level as autofluorescence and strongly boosted in the presence of calcium. Moreover, inoculation of leaves expressing the small PLD-like genes resulted in increased lesion growth and greater numbers of sporangia, but this was abolished when mutated PLD-like genes were expressed with non-functional PLD catalytic motifs. These results show that the three small PLD-likes are catalytically active and suggest that their enzymatic activity is required for the promotion of virulence, possibly by executing membrane modifications to support the growth of P. infestans in the host.
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- 2018
11. Genotyping-by-sequencing-based identification of Arabidopsis pattern recognition receptor RLP32 recognizing proteobacterial translation initiation factor IF1
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Li, Fan, Katja, Fröhlich, Eric, Melzer, Rory N, Pruitt, Isabell, Albert, Lisha, Zhang, Anna, Joe, Chenlei, Hua, Yanyue, Song, Markus, Albert, Sang-Tae, Kim, Detlef, Weigel, Cyril, Zipfel, Eunyoung, Chae, Andrea A, Gust, and Thorsten, Nürnberger
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Genotype ,Arabidopsis Proteins ,Receptors, Pattern Recognition ,Proteobacteria ,Arabidopsis ,Pseudomonas syringae ,Plant Immunity ,Prokaryotic Initiation Factors ,Plant Diseases - Abstract
Activation of plant pattern-triggered immunity (PTI) relies on the recognition of microbe-derived structures, termed patterns, through plant-encoded surface-resident pattern recognition receptors (PRRs). We show that proteobacterial translation initiation factor 1 (IF1) triggers PTI in Arabidopsis thaliana and related Brassicaceae species. Unlike for most other immunogenic patterns, IF1 elicitor activity cannot be assigned to a small peptide epitope, suggesting that tertiary fold features are required for IF1 receptor activation. We have deployed natural variation in IF1 sensitivity to identify Arabidopsis leucine-rich repeat (LRR) receptor-like protein 32 (RLP32) as IF1 receptor using a restriction site-associated DNA sequencing approach. RLP32 confers IF1 sensitivity to rlp32 mutants, IF1-insensitive Arabidopsis accessions and IF1-insensitive Nicotiana benthamiana, binds IF1 specifically and forms complexes with LRR receptor kinases SOBIR1 and BAK1 to mediate signaling. Similar to other PRRs, RLP32 confers resistance to Pseudomonas syringae, highlighting an unexpectedly complex array of bacterial pattern sensors within a single plant species.
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- 2021
12. Distinct immune sensor systems for fungal endopolygalacturonases in closely related Brassicaceae
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Lisha, Zhang, Chenlei, Hua, Rory N, Pruitt, Si, Qin, Lei, Wang, Isabell, Albert, Markus, Albert, Jan A L, van Kan, and Thorsten, Nürnberger
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Polygalacturonase ,Genotype ,Gene Expression Regulation, Plant ,Tobacco ,Arabidopsis ,Genetic Variation ,Plant Immunity ,Brassica ,Genes, Plant ,Plants, Genetically Modified ,Plant Diseases - Abstract
Plant pattern recognition receptors (PRRs) facilitate recognition of microbial patterns and mediate activation of plant immunity. Arabidopsis thaliana RLP42 senses fungal endopolygalacturonases (PGs) and triggers plant defence through complex formation with SOBIR1 and SERK co-receptors. Here, we show that a conserved 9-amino-acid fragment pg9(At) within PGs is sufficient to activate RLP42-dependent plant immunity. Structure-function analysis reveals essential roles of amino acid residues within the RLP42 leucine-rich repeat and island domains for ligand binding and PRR complex assembly. Sensitivity to pg9(At), which is restricted to A. thaliana and exhibits scattered accession specificity, is unusual for known PRRs. Arabidopsis arenosa and Brassica rapa, two Brassicaceae species closely related to A. thaliana, respectively perceive immunogenic PG fragments pg20(Aa) and pg36(Bra), which are structurally distinct from pg9(At). Our study provides evidence for rapid evolution of polymorphic PG sensors with distinct pattern specificities within a single plant family.
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- 2020
13. Phytophthora infestans RXLR effector SFI5 requires association with calmodulin for PTI/MTI suppressing activity
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Frédéric Brunner, Petra C. Boevink, Chenlei Hua, Paul R. J. Birch, Nadine Wagener, Hazel McLellan, and Xiangzi Zheng
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RXLR effector ,0301 basic medicine ,calmodulin ,animal structures ,Calmodulin ,Phytophthora infestans ,MAMP‐triggered immunity (MTI) ,Physiology ,In silico ,Amino Acid Motifs ,Plant Science ,SFI5 ,03 medical and health sciences ,Immune system ,Solanum lycopersicum ,Downregulation and upregulation ,Plant Immunity ,Secretion ,Amino Acid Sequence ,MAMP ,calcium ,Innate immune system ,Full Paper ,biology ,tomato protoplast ,Chemistry ,Effector ,Research ,Cell Membrane ,Pathogen-Associated Molecular Pattern Molecules ,Proteins ,Full Papers ,Cell biology ,virulence ,030104 developmental biology ,biology.protein ,Peptides ,Protein Binding - Abstract
Summary Pathogens secrete effector proteins to interfere with plant innate immunity, in which Ca2+/calmodulin (CaM) signalling plays key roles. Thus far, few effectors have been identified that directly interact with CaM for defence suppression. Here, we report that SFI5, an RXLR effector from Phytophthora infestans, suppresses microbe‐associated molecular pattern (MAMP)‐triggered immunity (MTI) by interacting with host CaMs.We predicted the CaM‐binding site in SFI5 using in silico analysis. The interaction between SFI5 and CaM was tested by both in vitro and in vivo assays. MTI suppression by SFI5 and truncated variants were performed in a tomato protoplast system.We found that both the predicted CaM‐binding site and the full‐length SFI5 protein interact with CaM in the presence of Ca2+. MTI responses, such as FRK1 upregulation, reactive oxygen species accumulation, and mitogen‐activated protein kinase activation were suppressed by truncated SFI5 proteins containing the C‐terminal CaM‐binding site but not by those without it. The plasma membrane localization of SFI5 and its ability to enhance infection were also perturbed by loss of the CaM‐binding site.We conclude that CaM‐binding is required for localization and activity of SFI5. We propose that SFI5 suppresses plant immunity by interfering with immune signalling components after activation by CaMs.
- Published
- 2018
14. Expression of pathogenesis-related genes in cotton roots in response to Verticillium dahliae PAMP molecules
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Chenlei Hua, Lisha Zhang, Xuan Du, Jianhua Zhao, Sheng Wang, Hui-Shan Guo, and Feng Gao
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0301 basic medicine ,Verticillium ,Genes, Plant ,Gossypium ,Plant Roots ,Genome ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,Gene Expression Regulation, Plant ,Botany ,Gene family ,Verticillium dahliae ,Gene ,General Environmental Science ,Genetics ,biology ,Gene Expression Profiling ,Pathogen-Associated Molecular Pattern Molecules ,food and beverages ,biology.organism_classification ,Gene expression profiling ,030104 developmental biology ,Host-Pathogen Interactions ,Verticillium wilt ,Peptides ,General Agricultural and Biological Sciences - Abstract
Verticillium wilt disease becomes a major threat to many economically important crops. It is unclear whether and how plant immunity takes place during cotton-Verticillium interaction due to the lack of marker genes. Taking advantage of cotton (Gossypium hirsutum) genome, we discovered pathogenesis-related (PR) gene families, which have been widely used as markers of immune responses in plants. To profile the expression of G. hirsutum PR genes in the process of plant immunity, we treated cotton roots with two immunogenic peptides, flg22 and nlp20 known as pathogen-associated molecular patterns, as well as three Verticillium dahliae-derived peptides, nlp20Vd2, nlp23Vd3, and nlp23Vd4 which are highly identical to nlp20. Quantitative real-time PCR results revealed that 14 G. hirsutum PR gene (GhPR) families were induced or suppressed independently in response to flg22, nlp20, nlp20Vd2, nlp23Vd3, and nlp23Vd4. Most GhPR genes are expressed highest at 3 h post incubation of immunogenic peptides. Compared to flg22 and nlp20, nlp20Vd2 is more effective to trigger up-regulated expression of GhPR genes. Notably, both nlp23Vd3 and nlp23Vd4 are able to induce GhPR gene up-regulation, although they do not induce necrosis on cotton leaves. Thus, our results provide marker genes and new immunogenic peptides for further investigation of cotton-V. dahliae interaction.
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- 2017
15. VdPKS1 is required for melanin formation and virulence in a cotton wilt pathogen Verticillium dahliae
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Zhirong Chen, Jia-Feng Huang, Ting Zhang, Feng Gao, Chenlei Hua, Pei Meng, Bosen Zhang, Sheng Wang, and Yejuan Du
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0106 biological sciences ,0301 basic medicine ,Mutant ,Conidiation ,Virulence ,Fungus ,Verticillium ,Biology ,Genes, Plant ,01 natural sciences ,General Biochemistry, Genetics and Molecular Biology ,Microbiology ,Fungal Proteins ,Melanin ,03 medical and health sciences ,Verticillium dahliae ,Pathogen ,General Environmental Science ,Wilt disease ,Melanins ,Gossypium ,Gene Expression Profiling ,biology.organism_classification ,030104 developmental biology ,Host-Pathogen Interactions ,General Agricultural and Biological Sciences ,Polyketide Synthases ,010606 plant biology & botany - Abstract
Verticillium dahliae is a soil-borne phytopathogenic fungus that causes vascular wilt disease in a broad range of hosts. This pathogen survives for many years in soil in the form of melanized microsclerotia. To investigate the melanin synthesis in V. dahliae, we identified a polyketide synthase gene in V. dahliae, namely VdPKS1. PKS1 is known to involve in the dihydroxynaphthalene melanin synthesis pathway in many fungi. We found that VdPKS1 was required for melanin formation but not for microsclerotial production in V. dahliae. The VdPKS1 gene-disruption mutant (vdpks1) formed melanin-deficient albino microsclerotia, which did not affect the fungal colonization in host tissues but significantly reduced the disease severity. Gene transcription analysis in the wild-type and the vdpks1 strains suggested that VdPKS1 gene-disruption influenced the expression of a series of genes involved in ethylene biosynthesis, microsclerotial formation and pathogenesis. Our results suggest that the VdPKS1-mediated melanin synthesis is important for virulence and developmental traits of V. dahliae.
- Published
- 2017
16. The Verticillium ‐specific protein VdSCP7 localizes to the plant nucleus and modulates immunity to fungal infections
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Hui-Shan Guo, Xuan Du, Xiao-Wei Ma, Hao Ni, Chenlei Hua, Thorsten Nürnberger, Sheng Wang, and Lisha Zhang
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0106 biological sciences ,0301 basic medicine ,Hypersensitive response ,Physiology ,Plant Immunity ,Plant Science ,Verticillium ,Biology ,01 natural sciences ,Green fluorescent protein ,Fungal Proteins ,03 medical and health sciences ,Gene Expression Regulation, Fungal ,Verticillium dahliae ,Nuclear export signal ,Gene knockout ,Plant Diseases ,Cell Nucleus ,Effector ,fungi ,food and beverages ,biology.organism_classification ,Virology ,Cell biology ,030104 developmental biology ,Reactive Oxygen Species ,Nuclear localization sequence ,010606 plant biology & botany - Abstract
Summary Fungal pathogens secrete effector proteins to suppress plant basal defense for successful colonization. Resistant plants, however, can recognize effectors by cognate R proteins to induce effector-triggered immunity (ETI). By analyzing secretomes of the vascular fungal pathogen Verticillium dahliae, we identified a novel secreted protein VdSCP7 that targets the plant nucleus. The green fluorescent protein (GFP)-tagged VdSCP7 gene with either a mutated nuclear localization signal motif or with additional nuclear export signal was transiently expressed in Nicotiana benthamiana, and investigated for induction of plant immunity. The role of VdSCP7 in V. dahliae pathogenicity was characterized by gene knockout and complementation, and GFP labeling. Expression of the VdSCP7 gene in N. benthamiana activated both salicylic acid and jasmonate signaling, and altered the plant's susceptibility to the pathogens Botrytis cinerea and Phytophthora capsici. The immune response activated by VdSCP7 was highly dependent on its initial extracellular secretion and subsequent nuclear localization in plants. Knockout of the VdSCP7 gene significantly enhanced V. dahliae aggressiveness on cotton. GFP-labeled VdSCP7 is secreted by V. dahliae and accumulates in the plant nucleus. We conclude that VdSCP7 is a novel effector protein that targets the host nucleus to modulate plant immunity, and suggest that plants can recognize VdSCP7 to activate ETI during fungal infection.
- Published
- 2017
17. An Improved Single-Step Cloning Strategy Simplifies the Agrobacterium tumefaciens-Mediated Transformation (ATMT)-Based Gene-Disruption Method for Verticillium dahliae
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Chenlei Hua, Sheng Wang, Haiying Xing, Hui-Shan Guo, and Jie Zhang
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0301 basic medicine ,Cloning ,biology ,Agrobacterium ,Plant Science ,Agrobacterium tumefaciens ,Computational biology ,Verticillium ,biology.organism_classification ,Microbiology ,03 medical and health sciences ,Transformation (genetics) ,Transformation, Genetic ,030104 developmental biology ,Vector (molecular biology) ,Verticillium dahliae ,Cloning, Molecular ,Agronomy and Crop Science ,Gene ,Gene Deletion - Abstract
The soilborne fungal pathogen Verticillium dahliae infects a broad range of plant species to cause severe diseases. The availability of Verticillium genome sequences has provided opportunities for large-scale investigations of individual gene function in Verticillium strains using Agrobacterium tumefaciens-mediated transformation (ATMT)-based gene-disruption strategies. Traditional ATMT vectors require multiple cloning steps and elaborate characterization procedures to achieve successful gene replacement; thus, these vectors are not suitable for high-throughput ATMT-based gene deletion. Several advancements have been made that either involve simplification of the steps required for gene-deletion vector construction or increase the efficiency of the technique for rapid recombinant characterization. However, an ATMT binary vector that is both simple and efficient is still lacking. Here, we generated a USER-ATMT dual-selection (DS) binary vector, which combines both the advantages of the USER single-step cloning technique and the efficiency of the herpes simplex virus thymidine kinase negative-selection marker. Highly efficient deletion of three different genes in V. dahliae using the USER-ATMT-DS vector enabled verification that this newly-generated vector not only facilitates the cloning process but also simplifies the subsequent identification of fungal homologous recombinants. The results suggest that the USER-ATMT-DS vector is applicable for efficient gene deletion and suitable for large-scale gene deletion in V. dahliae.
- Published
- 2016
18. Intestinal microbiota of the four omnivorous fishes revealed by 16S rRNA metabarcoding from the habitats of oyster reefs
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Sheng Bi, Huadong Yi, Han Lai, Haiyang Li, Xuange Liu, Qiuxian Chen, Jiahui Chen, Zhilun Zhang, Xuchong Wei, Chenlei Huang, Li Lin, Guorong Xin, and Guifeng Li
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Microbial ecology ,Oyster reefs habitats ,Intestinal microbiota ,Pearl River Estuary ,16S rRNA ,Ecology ,QH540-549.5 - Abstract
The habitats of oyster reefs had played various ecological services functions, as an important means of marine ecological restoration and fishery resources conservation, which had been widely used worldwide. And the characteristics of microbial communities would be inevitably affected by habitat of oyster reefs. The aim of this study was to reveal the relationship between oyster reefs and the intestinal microbiota of fish (Takifugu ocellatus, Acanthopagrus latus, Clupanodon thrissa, and Mugil cephalus). Here, based on 16S rRNA sequencing, this research analyzed the microbial communities of fishes gut in the oyster reefs from the most crucial estuary economic zone in Southern China. It was significantly lower that the bacterial alpha-diversity in the fish intestines than in the sediments of oyster reefs. The non-metric multidimensional scaling (NMDS) and analysis of similarities (ANOSIM) results showed that intestines microbiota of T. ocellatus and A. latus were more homologous to sediments microbiota, but intestines microbiota of C. thrissa and M. cephalus were more similar to the water microbiota. There were obvious differences in the contribution of oyster reefs habitat to intestinal microorganisms of four fishes. Source tracking analysis as well reached the similar conclusion. For instance, contribution of water to T. ocellatus and A. latus was lesser than sediments, which was related to their benthic habits. Based on the differences in microbial community compositions, the functional prediction analysis had demonstrated that special functional pathways and independent core indicators had been formed separately from oyster reefs and fish intestines. Overall, these findings highlighted the characteristics of microbial diversity and functions from the omnivorous fish living in oyster reefs habitats. This research proposed new insights into the function of oyster reefs habitats, which have enriched the ecological service value of oyster reefs from microbial ecology and provided a reference for the application of oyster reefs habitats in other estuaries.
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- 2023
- Full Text
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19. Phytophthora sojae and soybean isoflavones, a model to study zoospore chemotaxis
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Chenlei Hua, Xinyu Yang, and Yuanchao Wang
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Genetics ,biology ,Zoospore ,food and beverages ,Chemotaxis ,Plant Science ,Isoflavones ,biology.organism_classification ,Microbiology ,chemistry.chemical_compound ,chemistry ,Molecular mechanism ,Gene silencing ,Phytophthora sojae ,Signal transduction ,Functional genomics - Abstract
The ability of sensing the signals from hosts, aggregating, and searching for proper sites of invasion is crucial for many soil-borne plant pathogens. One of these is Phytophthora sojae that specifically infects soybean and causes severe yield loss. In the field P. sojae zoospores attracted to soybean exudates isoflavones is an essential step for early infection. Recent studies based on genome mining and gene silencing technologies have shown that this special zoospore behavior relies on G-protein mediated signaling pathways. This review is to summarize researches on P. sojae functional genomics related to G-protein signaling and briefly describe the molecular mechanism of P. sojae zoospore chemotaxis to root exudates, especially isoflavones.
- Published
- 2015
20. PsHint1, associated with the G-protein α subunit PsGPA1, is required for the chemotaxis and pathogenicity ofPhytophthora sojae
- Author
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Pingping Nie, Yuanchao Wang, Min Qiu, Xin Zhang, Chenlei Hua, Wenwu Ye, Yujuan Hao, and Chunhua Zhai
- Subjects
0301 basic medicine ,Oomycete ,biology ,GTP' ,G protein ,food and beverages ,Soil Science ,Germ tube ,Chemotaxis ,Plant Science ,Guanosine triphosphate ,biology.organism_classification ,03 medical and health sciences ,chemistry.chemical_compound ,030104 developmental biology ,chemistry ,Biochemistry ,Guanosine diphosphate ,Phytophthora sojae ,Agronomy and Crop Science ,Molecular Biology - Abstract
Summary Zoospore chemotaxis to soybean isoflavones is essential in the early stages of infection by the oomycete pathogen Phytophthora sojae. Previously, we have identified a G-protein α subunit encoded by PsGPA1 which regulates the chemotaxis and pathogenicity of P. sojae. In the present study, we used affinity purification to identify PsGPA1-interacting proteins, including PsHint1, a histidine triad (HIT) domain-containing protein orthologous to human HIT nucleotide-binding protein 1 (HINT1). PsHint1 interacted with both the guanosine triphosphate (GTP)- and guanosine diphosphate (GDP)-bound forms of PsGPA1. An analysis of the gene-silenced transformants revealed that PsHint1 was involved in the chemotropic response of zoospores to the isoflavone daidzein. During interaction with a susceptible soybean cultivar, PsHint1-silenced transformants displayed significantly reduced infectious hyphal extension and caused a strong cell death in plants. In addition, the transformants displayed defective cyst germination, forming abnormal germ tubes that were highly branched and exhibited apical swelling. These results suggest that PsHint1 not only regulates chemotaxis by interacting with PsGPA1, but also participates in a Gα-independent pathway involved in the pathogenicity of P. sojae.
- Published
- 2015
21. Effect of Flumorph on F-Actin Dynamics in the Potato Late Blight Pathogen Phytophthora infestans
- Author
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Francine Govers, K. Kots, Chenlei Hua, Tijs Ketelaar, and Harold J. G. Meijer
- Subjects
Hyphal growth ,Hypha ,Phytophthora infestans ,Morpholines ,domain ,Green Fluorescent Proteins ,Hyphae ,Hyphal tip ,macromolecular substances ,Plant Science ,in-vitro ,Microbiology ,resistance ,fungicide ,Cytoskeleton ,Plant Diseases ,Solanum tuberosum ,Oomycete ,biology ,EPS-2 ,fungi ,cytoskeleton ,Laboratorium voor Celbiologie ,organization ,oomycete aphanomyces-euteiches ,Bridged Bicyclo Compounds, Heterocyclic ,biology.organism_classification ,Actin cytoskeleton ,plant pathogen ,proteins ,Actins ,Laboratorium voor Phytopathologie ,Laboratory of Cell Biology ,Laboratory of Phytopathology ,identification ,Thiazolidines ,Phytophthora ,Agronomy and Crop Science - Abstract
Oomycetes are fungal-like pathogens that cause notorious diseases. Protecting crops against oomycetes requires regular spraying with chemicals, many with an unknown mode of action. In the 1990s, flumorph was identified as a novel crop protection agent. It was shown to inhibit the growth of oomycete pathogens including Phytophthora spp., presumably by targeting actin. We recently generated transgenic Phytophthora infestans strains that express Lifeact-enhanced green fluorescent protein (eGFP), which enabled us to monitor the actin cytoskeleton during hyphal growth. For analyzing effects of oomicides on the actin cytoskeleton in vivo, the P. infestans Lifeact-eGFP strain is an excellent tool. Here, we confirm that flumorph is an oomicide with growth inhibitory activity. Microscopic analyses showed that low flumorph concentrations provoked hyphal tip swellings accompanied by accumulation of actin plaques in the apex, a feature reminiscent of tips of nongrowing hyphae. At higher concentrations, swelling was more pronounced and accompanied by an increase in hyphal bursting events. However, in hyphae that remained intact, actin filaments were indistinguishable from those in nontreated, nongrowing hyphae. In contrast, in hyphae treated with the actin depolymerizing drug latrunculin B, no hyphal bursting was observed but the actin filaments were completely disrupted. This difference demonstrates that actin is not the primary target of flumorph.
- Published
- 2015
22. The heat shock transcription factor PsHSF1 ofPhytophthora sojaeis required for oxidative stress tolerance and detoxifying the plant oxidative burst
- Author
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Kai Tao, Yuting Sheng, Yuanchao Wang, Francine Govers, Xiaoyun Liu, Harold J. G. Meijer, Wenwu Ye, Chenlei Hua, Xinyu Yang, and Yonglin Wang
- Subjects
Laccase ,chemistry.chemical_classification ,Reactive oxygen species ,biology ,fungi ,food and beverages ,medicine.disease_cause ,biology.organism_classification ,Microbiology ,Respiratory burst ,Heat shock factor ,chemistry ,medicine ,Phytophthora sojae ,Pathogen ,Transcription factor ,Ecology, Evolution, Behavior and Systematics ,Oxidative stress - Abstract
In the interaction between plant and microbial pathogens, reactive oxygen species (ROS) rapidly accumulate upon pathogen recognition at the infection site and play a central role in plant defence. However, the mechanisms that plant pathogens use to counteract ROS are still poorly understood especially in oomycetes, filamentous organisms that evolved independently from fungi. ROS detoxification depends on transcription factors (TFs) that are highly conserved in fungi but much less conserved in oomycetes. In this study, we identified the TF PsHSF1 that acts as a modulator of the oxidative stress response in the soybean stem and root rot pathogen Phytophthora sojae. We found that PsHSF1 is critical for pathogenicity in P.?sojae by detoxifying the plant oxidative burst. ROS produced in plant defence can be detoxified by extracellular peroxidases and laccases which might be regulated by PsHSF1. Our study extends the understanding of ROS detoxification mechanism mediated by a heat shock TF in oomycetes.
- Published
- 2014
23. Trans-Kingdom RNA Silencing in Plant-Fungal Pathogen Interactions
- Author
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Hui-Shan Guo, Chenlei Hua, and Jianhua Zhao
- Subjects
0301 basic medicine ,Genetics ,Small RNA ,fungi ,food and beverages ,RNA ,Plant Immunity ,Plant Science ,Biology ,Verticillium ,biology.organism_classification ,03 medical and health sciences ,RNA silencing ,MicroRNAs ,030104 developmental biology ,RNA interference ,microRNA ,Gene silencing ,RNA Interference ,Verticillium dahliae ,Botrytis ,Gene Silencing ,RNA, Small Interfering ,Molecular Biology ,Plant Diseases - Abstract
Fungal pathogens represent a major group of plant invaders that are the causative agents of many notorious plant diseases. Large quantities of RNAs, especially small RNAs involved in gene silencing, have been found to transmit bidirectionally between fungal pathogens and their hosts. Although host-induced gene silencing (HIGS) technology has been developed and applied to protect crops from fungal infections, the mechanisms of RNA transmission, especially small RNAs regulating trans-kingdom RNA silencing in plant immunity, are largely unknown. In this review, we summarize and discuss recent important findings regarding trans-kingdom sRNAs and RNA silencing in plant-fungal pathogen interactions compared with the well-known RNAi mechanisms in plants and fungi. We focus on the interactions between plant and fungal pathogens with broad hosts, represented by the vascular pathogen Verticillium dahliae and non-vascular pathogen Botrytis cinerea, and discuss the known instances of natural RNAi transmission between fungal pathogens and host plants. Given that HIGS has been developed and recently applied in controlling Verticillium wilt diseases, we propose an ideal research system exploiting plant vasculature-Verticillium interaction to further study trans-kingdom RNA silencing.
- Published
- 2017
24. Surface Sensor Systems in Plant Immunity.
- Author
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Albert, Isabell, Chenlei Hua, Nürnberger, Thorsten, Pruitt, Rory N., and Lisha Zhang
- Published
- 2020
- Full Text
- View/download PDF
25. Chemotaxis and oospore formation inPhytophthora sojaeare controlled by G-protein-coupled receptors with a phosphatidylinositol phosphate kinase domain
- Author
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Xiaobo Zheng, M. Jing, Wei Zhao, D. Li, Francine Govers, Chenlei Hua, Xinyu Yang, Yuanchao Wang, and Harold J. G. Meijer
- Subjects
biology ,Kinase ,Zoospore ,Chemotaxis ,biology.organism_classification ,Microbiology ,chemistry.chemical_compound ,chemistry ,Biochemistry ,Oospore ,Phytophthora sojae ,Phosphatidylinositol ,Oospore formation ,Molecular Biology ,G protein-coupled receptor - Abstract
G-protein-coupled receptors (GPCRs) are key cellular components that mediate extracellular signals into intracellular responses. Genome mining revealed that Phytophthora spp. have over 60 GPCR genes among which a prominent class of 12 encoding novel proteins with an N-terminal GPCR domain fused to a C-terminal phosphatidylinositol phosphate kinase (PIPK) domain. This study focuses on two GPCR-PIPKs (GKs) in Phytophthora sojae. PsGK4 and PsGK5 are differentially expressed during the life cycle with the highest expression in cysts and during cyst germination, and at late infection stages. In P.?sojae transformants that constitutively express RFP-tagged PsGK4 and PsGK5, the fusion proteins in hyphae reside in small, rapidly moving vesicular-like structures. Functional analysis using gene silencing showed that PsGK4-silenced transformants displayed higher levels of encystment and a reduced cyst germination rate when compared with the recipient strain. Moreover, GK4 deficiency (or reduction) resulted in severe defects in zoospore chemotaxis towards isoflavones and soybean roots. In contrast, PsGK5-silenced transformants exhibited no obvious defects in asexual development but oospore production was severely impaired. Both, PsGK4- and PsGK5-silenced transformants showed reduced pathogenicity. These results point to involvement of GKs in zoospore behaviour, chemotaxis and oospore development, and suggest that PsGK4 and PsGK5 each head independent signalling pathways.
- Published
- 2013
26. GK4, a G-protein-coupled receptor with a phosphatidylinositol phosphate kinase domain inPhytophthora infestans, is involved in sporangia development and virulence
- Author
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Wei Zhao, Yuanchao Wang, Francine Govers, Chenlei Hua, Jeroen de Keijzer, and Harold J. G. Meijer
- Subjects
biology ,Kinase ,fungi ,biology.organism_classification ,Microbiology ,chemistry.chemical_compound ,Biochemistry ,chemistry ,Protein kinase domain ,Phytophthora infestans ,Spore germination ,Phytophthora sojae ,Phosphatidylinositol ,Molecular Biology ,G protein-coupled receptor ,G alpha subunit - Abstract
For dispersal and host infection plant pathogens largely depend on asexual spores. Pathogenesis and sporulation are complex processes that are governed by cellular signalling networks including G-protein and phospholipid signalling. Oomycetes possess a family of novel proteins called GPCR-PIPKs (GKs) that are composed of a seven-transmembrane spanning (7-TM) domain fused to a phosphatidylinositol phosphate kinase (PIPK) domain. Based on this domain structure GKs are anticipated to link G-protein and phospholipid signal pathways; however, their functions are currently unknown. Expression analyses of the 12 GK genes in Phytophthora infestans and their orthologues in Phytophthora sojae, revealed differential expression during asexual development. PiGK1 and PiGK4 were fused to monomeric red fluorescent protein (mRFP) and ectopically expressed in P.?infestans. In growing hyphae different subcellular distribution patterns were observed indicating that these two GKs act independently during development. We focused on the functional analyses of PiGK4. Its localization suggested involvement in cell differentiation and elongation and its 7-TM domain showed a canonical GPCR membrane topology. Silencing of GK4 and overexpression of full-length and truncated constructs in P.?infestans revealed that PiGK4 is not only involved in spore germination and hyphal elongation but also in sporangia cleavage and infection.
- Published
- 2013
27. Cotton plants export microRNAs to inhibit virulence gene expression in a fungal pathogen
- Author
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Hui-Shan Guo, Tao Zhang, Yun Jin, Sheng Wang, Yuan-Yuan Fang, Zhong-Qi Chen, Yunlong Zhao, Shou-Wei Ding, Jianhua Zhao, and Chenlei Hua
- Subjects
0301 basic medicine ,Crop and Pasture Production ,Hyphae ,Virulence ,Plant Biology ,Plant Science ,Plant disease resistance ,Biology ,Verticillium ,Microbiology ,03 medical and health sciences ,RNA interference ,Gene Expression Regulation, Fungal ,Genetics ,Gene silencing ,2.1 Biological and endogenous factors ,2.2 Factors relating to the physical environment ,Verticillium dahliae ,Gene Silencing ,Aetiology ,Gene ,Plant Diseases ,Disease Resistance ,Regulation of gene expression ,Gossypium ,fungi ,food and beverages ,biology.organism_classification ,RNA silencing ,MicroRNAs ,030104 developmental biology ,Fungal ,Infectious Diseases ,Gene Expression Regulation ,Zero Hunger ,Infection ,Biotechnology - Abstract
Plant pathogenic fungi represent the largest group of disease-causing agents on crop plants, and are a constant and major threat to agriculture worldwide. Recent studies have shown that engineered production of RNA interference (RNAi)-inducing dsRNA in host plants can trigger specific fungal gene silencing and confer resistance to fungal pathogens1-7. Although these findings illustrate efficient uptake of host RNAi triggers by pathogenic fungi, it is unknown whether or not such an uptake mechanism has been evolved for a natural biological function in fungus-host interactions. Here, we show that in response to infection with Verticillium dahliae (a vascular fungal pathogen responsible for devastating wilt diseases in many crops) cotton plants increase production of microRNA 166 (miR166) and miR159 and export both to the fungal hyphae for specific silencing. We found that two V. dahliae genes encoding a Ca2+-dependent cysteine protease (Clp-1) and an isotrichodermin C-15 hydroxylase (HiC-15), and targeted by miR166 and miR159, respectively, are both essential for fungal virulence. Notably, V. dahliae strains expressing either Clp-1 or HiC-15 rendered resistant to the respective miRNA exhibited drastically enhanced virulence in cotton plants. Together, our findings identify a novel defence strategy of host plants by exporting specific miRNAs to induce cross-kingdom gene silencing in pathogenic fungi and confer disease resistance.
- Published
- 2016
28. The PsCZF1 gene encoding a C2H2 zinc finger protein is required for growth, development and pathogenesis in Phytophthora sojae
- Author
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Chenlei Hua, Yuanchao Wang, Daolong Dou, Binyan Cheng, Yonglin Wang, Xiao-Ren Chen, Aining Li, Yuting Sheng, Xiaobo Zheng, and Xiaoli Wang
- Subjects
Phytophthora ,Hyphal growth ,Molecular Sequence Data ,Mutant ,Biology ,Microbiology ,Evolution, Molecular ,Phytophthora sojae ,Amino Acid Sequence ,Phylogeny ,Plant Diseases ,Regulation of gene expression ,Genetics ,Zinc finger ,Oomycete ,Reporter gene ,Virulence ,C2H2 Zinc Finger ,Algal Proteins ,fungi ,Zinc Fingers ,biology.organism_classification ,Cell biology ,Infectious Diseases ,Gene Expression Regulation ,Soybeans ,Sequence Alignment - Abstract
The C(2)H(2) zinc finger proteins form one of the largest families of transcriptional regulators in eukaryotes. We identified a Phytophthora sojae C(2)H(2) zinc finger (PsCZF1), that is highly conserved in sequenced oomycete pathogens. In transformants of P. sojae containing the PsCZF1 promoter fused to the beta-glucuronidase (GUS) reporter gene, GUS activity was highly induced in the P. sojae oospore stage and upregulated after infection. To elucidate the function of PsCZF1, its expression was silenced by introducing anti-sense constructs into P sojae. PsCZF1-silenced transformants did not exhibit altered cell size or morphology of sporangia and hyphae; however, hyphal growth rate was reduced by around 50% in the mutants. PsCZF1-deficient mutants were also impaired in production of oospores, swimming zoospores and germinating cysts, indicating that the gene is involved in various stages of the life cycle. Furthermore, we found that PsCZF1-deficient mutants lost virulence on host soybean cultivars. Our results suggest that this oomycete-specific C(2)H(2)-type zinc finger protein plays an important role in growth, development, and pathogenesis; therefore, PsCZF1 might be an attractive oomycete-specific target for chemical fungicide screening.
- Published
- 2009
29. A Phytophthora sojae G protein alpha subunit is involved in chemotaxis to soybean isoflavones
- Author
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Zhengguang Zhang, Francine Govers, Xiaobo Zheng, Yonglin Wang, Yuanchao Wang, Chenlei Hua, and Daolong Dou
- Subjects
Hyphal growth ,Phytophthora ,Spores ,signal-transduction ,Zoospore ,Protein subunit ,Mutant ,Molecular Sequence Data ,cyst germination ,Gene Dosage ,Gene Expression ,hyphal growth ,Microbiology ,oomycete ,Phytophthora sojae ,Calcium Signaling ,Molecular Biology ,Gene ,transcription factor ,Plant Diseases ,zoospore encystment ,biology ,Virulence ,EPS-2 ,Chemotaxis ,Algal Proteins ,fungi ,food and beverages ,General Medicine ,Articles ,biology.organism_classification ,Isoflavones ,GTP-Binding Protein alpha Subunits ,Laboratorium voor Phytopathologie ,Biochemistry ,Phytophthora infestans ,Laboratory of Phytopathology ,beta-subunit ,saccharomyces-cerevisiae ,Soybeans ,botrytis-cinerea ,infestans - Abstract
For the soybean pathogen Phytophthora sojae , chemotaxis of zoospores to isoflavones is believed to be critical for recognition of the host and for initiating infection. However, the molecular mechanisms underlying this chemotaxis are largely unknown. To investigate the role of G-protein and calcium signaling in chemotaxis, we analyzed the expression of several genes known to be involved in these pathways and selected one that was specifically expressed in sporangia and zoospores but not in mycelium. This gene, named PsGPA1 , is a single-copy gene in P. sojae and encodes a G-protein α subunit that shares 96% identity in amino acid sequence with that of Phytophthora infestans . To elucidate the function, expression of PsGPA1 was silenced by introducing antisense constructs into P. sojae. PsGPA1 silencing did not disturb hyphal growth or sporulation but severely affected zoospore behavior, including chemotaxis to the soybean isoflavone daidzein. Zoospore encystment and cyst germination were also altered, resulting in the inability of the PsGPA1 -silenced mutants to infect soybean. In addition, the expressions of a calmodulin gene, PsCAM1 , and two calcium- and calmodulin-dependent protein kinase genes, PsCMK3 and PsCMK4 , were increased in the mutant zoospores, suggesting that PsGPA1 negatively regulates the calcium signaling pathways that are likely involved in zoospore chemotaxis.
- Published
- 2008
30. The dual edge of RNA silencing suppressors in the virus-host interactions
- Author
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Hui-Shan Guo, Jianhua Zhao, Chenlei Hua, and Yuan-Yuan Fang
- Subjects
0106 biological sciences ,0301 basic medicine ,Cytoplasm ,viruses ,Biology ,Virus Replication ,01 natural sciences ,Virus ,law.invention ,Plant Viruses ,03 medical and health sciences ,Viral Proteins ,RNA interference ,law ,Virology ,Plant virus ,Gene silencing ,RNA Viruses ,Symbiosis ,RNA ,Plants ,RNA silencing ,030104 developmental biology ,Viral replication ,Host-Pathogen Interactions ,Suppressor ,RNA, Viral ,RNA Interference ,010606 plant biology & botany - Abstract
RNA silencing (or RNA interference, RNAi) plays a key role in the plant antiviral defense. To facilitate infection, viruses encode suppressors of RNA silencing (VSRs) to counteract antiviral defense. In the co-evolutionary arms race between hosts and viruses, extreme viral accumulation does not benefit either hosts or viruses. During viral infection, antiviral silencing and VSRs have dual effects to maintain the balance between plant development and virus accumulation. Here, we summarize and discuss the multiple functions of the antiviral RNAi defense and VSRs, revealing the central hub regulators of VSRs in dynamically integrated connections between hosts and viruses.
- Published
- 2015
31. The heat shock transcription factor PsHSF1 of Phytophthora sojae is required for oxidative stress tolerance and detoxifying the plant oxidative burst
- Author
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Yuting, Sheng, Yonglin, Wang, Harold J G, Meijer, Xinyu, Yang, Chenlei, Hua, Wenwu, Ye, Kai, Tao, Xiaoyun, Liu, Francine, Govers, and Yuanchao, Wang
- Subjects
DNA-Binding Proteins ,Phytophthora ,Oxidative Stress ,Heat Shock Transcription Factors ,Host-Pathogen Interactions ,Molecular Sequence Data ,Soybeans ,Reactive Oxygen Species ,Plant Roots ,Plant Diseases ,Transcription Factors - Abstract
In the interaction between plant and microbial pathogens, reactive oxygen species (ROS) rapidly accumulate upon pathogen recognition at the infection site and play a central role in plant defence. However, the mechanisms that plant pathogens use to counteract ROS are still poorly understood especially in oomycetes, filamentous organisms that evolved independently from fungi. ROS detoxification depends on transcription factors (TFs) that are highly conserved in fungi but much less conserved in oomycetes. In this study, we identified the TF PsHSF1 that acts as a modulator of the oxidative stress response in the soybean stem and root rot pathogen Phytophthora sojae. We found that PsHSF1 is critical for pathogenicity in P. sojae by detoxifying the plant oxidative burst. ROS produced in plant defence can be detoxified by extracellular peroxidases and laccases which might be regulated by PsHSF1. Our study extends the understanding of ROS detoxification mechanism mediated by a heat shock TF in oomycetes.
- Published
- 2014
32. Genome-wide analysis of pectate-induced gene expression in Botrytis cinerea: identification and functional analysis of putative d-galacturonate transporters
- Author
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Joost H. M. Stassen, Sayantani Chatterjee, Jan A. L. van Kan, Maxim Cornelissen, Chenlei Hua, and Lisha Zhang
- Subjects
Mutant ,Nicotiana benthamiana ,Gene Expression ,confocal microscopy ,Real-Time Polymerase Chain Reaction ,Microbiology ,Cell wall ,chemistry.chemical_compound ,Gene Knockout Techniques ,Solanum lycopersicum ,Gene expression ,Tobacco ,Genetics ,endopolygalacturonase genes ,Gene ,hexose transporters ,Botrytis cinerea ,wall degrading enzymes ,biology ,Virulence ,EPS-2 ,Gene Expression Profiling ,Hexuronic Acids ,filamentous fungi ,fungi ,Cell Membrane ,Cytoplasmic Vesicles ,food and beverages ,Membrane Transport Proteins ,biology.organism_classification ,Fusion protein ,Laboratorium voor Phytopathologie ,Culture Media ,virulence ,chemistry ,Biochemistry ,rna-seq ,Laboratory of Phytopathology ,colletotrichum fungi ,saccharomyces-cerevisiae ,Pectins ,acid ,Botrytis ,Genome, Fungal ,D-Galacturonic acid - Abstract
The fungal plant pathogen Botrytis cinerea produces a spectrum of cell wall degrading enzymes for the decomposition of host cell wall polysaccharides and the consumption of the monosaccharides that are released. Especially pectin is an abundant cell wall component, and the decomposition of pectin by B. cinerea has been extensively studied. An effective concerted action of the appropriate pectin depolymerising enzymes, monosaccharide transporters and catabolic enzymes is important for complete d-galacturonic acid utilization by B. cinerea. In this study, we performed RNA sequencing to compare genome-wide transcriptional profiles between B. cinerea cultures grown in media containing pectate or glucose as sole carbon source. Transcript levels of 32 genes that are induced by pectate were further examined in cultures grown on six different monosaccharides, by means of quantitative RT-PCR, leading to the identification of 8 genes that are exclusively induced by d-galacturonic acid. Among these, the hexose transporter encoding genes Bchxt15 and Bchxt19 were functionally characterised. The subcellular location was studied of BcHXT15-GFP and BcHXT19-GFP fusion proteins expressed under control of their native promoter, in a B. cinerea wild-type strain. Both genes are expressed during growth on d-galacturonic acid and the fusion proteins are localized in plasma membranes and intracellular vesicles. Target gene knockout analysis revealed that BcHXT15 contributes to d-galacturonic acid uptake at pH 5~5.6. The virulence of all B. cinerea hexose transporter mutants tested was unaltered on tomato and Nicotiana benthamiana leaves.
- Published
- 2013
33. Actin dynamics in Phytophthora infestans; rapidly reorganizing cables and immobile, long-lived plaques
- Author
-
Harold J G, Meijer, Chenlei, Hua, Kiki, Kots, Tijs, Ketelaar, and Francine, Govers
- Subjects
Microscopy, Fluorescence ,Staining and Labeling ,Phytophthora infestans ,Hyphae ,Protein Multimerization ,Actins - Abstract
The actin cytoskeleton is a dynamic but well-organized intracellular framework that is essential for proper functioning of eukaryotic cells. Here, we use the actin binding peptide Lifeact to investigate the in vivo actin cytoskeleton dynamics in the oomycete plant pathogen Phytophthora infestans. Lifeact-eGFP labelled thick and thin actin bundles and actin filament plaques allowing visualization of actin dynamics. All actin structures in the hyphae were cortically localized. In growing hyphae actin filament cables were axially oriented in the sub-apical region whereas in the extreme apex in growing hyphae, waves of fine F-actin polymerization were observed. Upon growth termination, actin filament plaques appeared in the hyphal tip. The distance between a hyphal tip and the first actin filament plaque correlated strongly with hyphal growth velocity. The actin filament plaques were nearly immobile with average lifetimes exceeding 1 h, relatively long when compared to the lifetime of actin patches known in other eukaryotes. Plaque assembly required ∼30 s while disassembly was accomplished in ∼10 s. Remarkably, plaque disassembly was not accompanied with internalization and the formation of endocytic vesicles. These findings suggest that the functions of actin plaques in oomycetes differ from those of actin patches present in other organisms.
- Published
- 2013
34. GK4, a G-protein-coupled receptor with a phosphatidylinositol phosphate kinase domain in Phytophthora infestans, is involved in sporangia development and virulence
- Author
-
Chenlei, Hua, Harold J G, Meijer, Jeroen, de Keijzer, Wei, Zhao, Yuanchao, Wang, and Francine, Govers
- Subjects
Plant Leaves ,Spores ,Luminescent Proteins ,Phosphatidylinositol Phosphates ,Phytophthora infestans ,Sporangia ,Phosphotransferases ,Tobacco ,Hyphae ,Gene Expression Regulation, Developmental ,Plant Diseases ,Receptors, G-Protein-Coupled ,Solanum tuberosum - Abstract
For dispersal and host infection plant pathogens largely depend on asexual spores. Pathogenesis and sporulation are complex processes that are governed by cellular signalling networks including G-protein and phospholipid signalling. Oomycetes possess a family of novel proteins called GPCR-PIPKs (GKs) that are composed of a seven-transmembrane spanning (7-TM) domain fused to a phosphatidylinositol phosphate kinase (PIPK) domain. Based on this domain structure GKs are anticipated to link G-protein and phospholipid signal pathways; however, their functions are currently unknown. Expression analyses of the 12 GK genes in Phytophthora infestans and their orthologues in Phytophthora sojae, revealed differential expression during asexual development. PiGK1 and PiGK4 were fused to monomeric red fluorescent protein (mRFP) and ectopically expressed in P. infestans. In growing hyphae different subcellular distribution patterns were observed indicating that these two GKs act independently during development. We focused on the functional analyses of PiGK4. Its localization suggested involvement in cell differentiation and elongation and its 7-TM domain showed a canonical GPCR membrane topology. Silencing of GK4 and overexpression of full-length and truncated constructs in P. infestans revealed that PiGK4 is not only involved in spore germination and hyphal elongation but also in sporangia cleavage and infection.
- Published
- 2013
35. The function of MoGlk1 in integration of glucose and ammonium utilization in Magnaporthe oryzae
- Author
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Xianying Dou, Zhengyi Wang, Haifeng Zhang, Lisha Zhang, Zhongqiang Qi, Zhengguang Zhang, Xiaobo Zheng, Ruili Lv, and Chenlei Hua
- Subjects
Magnaporthe ,Fungal Physiology ,Mutant ,Saccharomyces cerevisiae ,Genes, Fungal ,lcsh:Medicine ,Mycology ,Microbiology ,Fungal Proteins ,chemistry.chemical_compound ,Gene Expression Regulation, Fungal ,Microbial Physiology ,Genetics ,Ammonium ,lcsh:Science ,Biology ,Hexokinase ,Multidisciplinary ,biology ,Glucokinase ,lcsh:R ,Genetic Complementation Test ,Fructose ,Sequence Analysis, DNA ,biology.organism_classification ,Culture Media ,Quaternary Ammonium Compounds ,Glucose ,Phenotype ,chemistry ,Biochemistry ,Mutation ,Biocatalysis ,lcsh:Q ,Heterologous expression ,Gene Function ,Acids ,Research Article - Abstract
Hexokinases are conserved proteins functioning in glucose sensing and signaling. The rice blast fungus Magnaporthe oryzae contains several hexokinases, including MoHxk1 (hexokinase) and MoGlk1 (glucokinase) encoded respectively by MoHXK1 and MoGLK1 genes. The heterologous expression of MoGlk1 and MoHxk1 in Saccharomyces cerevisiae confirmed their conserved functions. Disruption of MoHXK1 resulted in growth reduction in medium containing fructose as the sole carbon source, whereas disruption of MoGLK1 did not cause the similar defect. However, the ΔMoglk1 mutant displayed decreased proton extrusion and a lower biomass in the presence of ammonium, suggesting a decline in the utilization of ammonium. Additionally, the MoGLK1 allele lacking catalytic activity restored growth to the ΔMoglk1 mutant. Moreover, the expression of MoPMA1 encoding a plasma membrane H(+)-ATPase decreased in the ΔMoglk1 mutant that can be suppressed by glucose and G-6-P. Thus, MoGlk1, but not MoHxk1, regulates ammonium utilization through a mechanism that is independent from its catalytic activity.
- Published
- 2011
36. Genome-wide identification of Phytophthora sojae SNARE genes and functional characterization of the conserved SNARE PsYKT6
- Author
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Harold J. G. Meijer, Yuanchao Wang, Xianying Dou, Suomeng Dong, Wei Zhao, Francine Govers, Wenwu Ye, and Chenlei Hua
- Subjects
Phytophthora ,fusion ,Molecular Sequence Data ,systematic analysis ,Microbiology ,Genome ,localization ,Fungal Proteins ,eukaryotes ,Genetics ,Gene silencing ,root-rot ,Phytophthora sojae ,Amino Acid Sequence ,Gene Silencing ,Gene ,Peptide sequence ,Plant Diseases ,Whole genome sequencing ,Oomycete ,mechanisms ,Sequence Homology, Amino Acid ,biology ,EPS-2 ,food and beverages ,biology.organism_classification ,Phenotype ,proteins ,Laboratorium voor Phytopathologie ,pathogen phytophthora ,Laboratory of Phytopathology ,Soybeans ,Genome, Fungal ,SNARE Proteins ,complex ,infestans - Abstract
Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) are central components of the machinery mediating membrane fusion and key factors for vesicular trafficking in all eukaryotic cells. Taking advantage of the available whole genome sequence of the oomycete plant pathogen Phytophthora sojae, 35 genes encoding putative SNARE proteins were identified in the genome of this organism. PsYKT6, one of the most conserved SNARE proteins, was functionally characterized by homology-dependent gene silencing. The phenotype analysis showed that PsYKT6 is important for proper asexual development, sexual reproduction, and pathogenesis on host soybean cultivars.
- Published
- 2011
37. G protein alpha subunit may help zoospore to find the infection site and influence the expression of RGS protein
- Author
-
Xiaobo Zheng, Yuanchao Wang, and Chenlei Hua
- Subjects
Signal Pathways ,biology ,G protein ,Zoospore ,Mutant ,fungi ,chemistry.chemical_element ,Chemotaxis ,Calcium ,biology.organism_classification ,Article Addendum ,G-Protein alpha Subunit ,Biochemistry ,chemistry ,sense organs ,Phytophthora ,General Agricultural and Biological Sciences - Abstract
Sensing chemical signal secreted from host root and find the best site for penetration are crucial for initiating infection of Phytophthora zoospore. G protein alpha subunit of P. sojae participates in not only the chemotaxis to soybean isoflavone, but also finding penetrating site. Furthermore, although calcium signal pathways are influenced by Galpha, other signal pathways also influenced by G protein remain to be discovered. In this addendum, we describe an RGS protein, PsRGS6, is expressed downregulated in zoospores of Galpha silenced mutant. This result indicates that the expression of Galpha and RGS protein may be influenced by each other. Some differences between Galpha mutants of P. infestans and P. sojae may be due to the different developmental procedures.
- Published
- 2008
38. Hemodynamic and Morphological Parameters of Ruptured Mirror Posterior Communicating Artery Aneurysms
- Author
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Jinlong Yuan, Chenlei Huang, Zhenbao Li, Xiaochun Jiang, Xintong Zhao, Degang Wu, Nianshen Lai, Jiaqiang Liu, Bingbing Zhang, Feiyun Qin, Dayong Xia, and Xinggen Fang
- Subjects
computational fluid dynamic ,hemodynamics ,morphological ,mirror aneurysms ,posterior communicating artery ,Neurology. Diseases of the nervous system ,RC346-429 - Abstract
Objective: Morphological and hemodynamic parameters might predict rupture of intracranial aneurysms (IAs). A practical model for the study is patients with ruptured mirror IAs in which one is ruptured and the other is unruptured. Although there have been analyses of the morphology and hemodynamics of ruptured mirror posterior communicating artery aneurysms (PComAAs), the sample sizes in these studies were small and only considered hemodynamics or morphological characters. Therefore, this study aimed to investigate the morphological and hemodynamic parameters associated with ruptured mirror PComAAs.Methods: We considered 72 patients with ruptured mirror PComAAs using computational fluid dynamics (CFDs). Ruptured mirror PComAAs were divided into ruptured and unruptured groups. Fourteen morphological and eight hemodynamic parameters were calculated and compared. Significant parameters were analyzed by the multivariate logistic regression to identify independent risk factors. Receiver operating characteristic (ROC) analysis was performed, and the area under the ROC curve (AUC) was calculated for all independent risk factors to determine the predictability and identify the optimal threshold.Results: Four hemodynamic and three morphological parameters were significantly different between ruptured and unruptured groups: normalized wall shear stress (NWSS), mean WSS, low wall shear WSS area (LSA%), size, aspect ratio (AR), size ratio (SR), and inflow angle (IA). Multivariate logistic regression analysis showed that AR, SR, NWSS, mean WSS, and LSA% were all independent factors significantly associated with PComAAs rupture. The ROC analysis for independent risk factors indicated that AR (0.751), NWSS (0.755), mean WSS (0.69), and LSA (0.778) had merely acceptable AUC values. Only SR (0.803) had a high acceptable AUC value. The threshold value of SR was 1.96.Conclusions: SR (>1.96) was the most significant parameter associated with IA rupture, whereas AR, NWSS, mean WSS, and LSA independently characterized the status of IA rupture.
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- 2021
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39. Effect of Flumorph on F-Actin Dynamics in the Potato Late Blight Pathogen Phytophthora infestans.
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Chenlei Hua, Kots, Kiki, Ketelaar, Tijs, Govers, Francine, and Meijer, Harold J. G.
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FLUMORPH , *MORPHOLINE , *F-actin , *ACTIN , *PHYTOPHTHORA infestans - Abstract
Oomycetes are fungal-like pathogens that cause notorious diseases. Protecting crops against oomycetes requires regular spraying with chemicals, many with an unknown mode of action. In the 1990s, flumorph was identified as a novel crop protection agent. It was shown to inhibit the growth of oomycete pathogens including Phytophthora spp., presumably by targeting actin. We recently generated transgenic Phytophthora infestans strains that express Lifeact-enhanced green fluorescent protein (eGFP), which enabled us to monitor the actin cytoskeleton during hyphal growth. For analyzing effects of oomicides on the actin cytoskeleton in vivo, the P. infestans Lifeact-eGFP strain is an excellent tool. Here, we confirm that flumorph is an oomicide with growth inhibitory activity. Microscopic analyses showed that low flumorph concentrations provoked hyphal tip swellings accompanied by accumulation of actin plaques in the apex, a feature reminiscent of tips of nongrowing hyphae. At higher concentrations, swelling was more pronounced and accompanied by an increase in hyphal bursting events. However, in hyphae that remained intact, actin filaments were indistinguishable from those in nontreated, nongrowing hyphae. In contrast, in hyphae treated with the actin depolymerizing drug latrunculin B, no hyphal bursting was observed but the actin filaments were completely disrupted. This difference demonstrates that actin is not the primary target of flumorph. [ABSTRACT FROM AUTHOR]
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- 2015
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40. Hemodynamic and Morphological Differences Between Unruptured Carotid-Posterior Communicating Artery Bifurcation Aneurysms and Infundibular Dilations of the Posterior Communicating Artery
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Jinlong Yuan, Zhenbao Li, Xiaochun Jiang, Niansheng Lai, Xuanzhi Wang, Xintong Zhao, Degang Wu, Jiaqiang Liu, Dayong Xia, Chenlei Huang, and Xinggen Fang
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computational fluid dynamic ,hemodynamics ,morphological ,communicating artery aneurysms ,infundibular dilations ,Neurology. Diseases of the nervous system ,RC346-429 - Abstract
Objective: Posterior communicating artery bifurcation aneurysms (PcomA-BAs) and infundibular dilations (PcomA-IDs) are found at the junction between the internal carotid artery (ICA) and the posterior communicating artery (PcomA). Several studies found that PcomA-IDs potentially progress to aneurysms and can even rupture. In our clinical practice, digital subtraction angiography (DSA) helps differentiate PcomA-IDs from unruptured PcomA-BAs. However, when PcomA-IDs are >3 mm in diameter or PcomA are absent on DSA, it is challenging to use DSA to differentiate PcomA-IDs from unruptured PcomA-BAs. Hemodynamic and morphological factors are thought to play important roles in the pathogenesis, progression, and rupture of cerebral aneurysms. We compared hemodynamic and morphological differences in unruptured PcomA-BAs and PcomA-IDs to better manage PcomA-IDs.Methods: We included 83 PcomA-IDs and 115 unruptured PcomA-BAs dignosed and measured using DSA from January 2015 to January 2019. Computational fluid dynamics was performed on these patient-specific models reconstructed using axial slices in DICOM format. Clinical, hemodynamic, and morphological factors were compared between the PcomA-IDs and PcomA-BAs. Significant parameters were analyzed using binary logistic regression analysis to identify independent risk factors. Receiver operating characteristic (ROC) analysis was performed on the independent risk factors to acquire cutoff values.Results: One hemodynamic and four morphyological parameters were significantly different between PcomA-IDs and PcomA-BAs: normalized wall shear stress (NWSS), size, the angle between the ophthalmic segment of the ICA and the PcomA (APcomA), the angle between the ophthalmic and the communicating segment of the ICA (AICA) and the diameter of the PcomA (DPcomA). Binary logistic regression analysis showed that small size and DPcomA as well as APcomA were all independent significant factors characterizing the status of PcomA-IDs and the ROC analysis for independent risk factors indicated the cutoff values of size, APcomA, and DPcomA were 3.45 mm, 66.27°, and 1.24 mm, respectively.Conclusions: Size, DpcomA, and ApcomA could independently characterize the status of PcomA-IDs. These might help us better differentiate them from real aneurysms and guide its management.
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- 2020
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41. An Improved Single-Step Cloning Strategy Simplifies the Agrobacterium tumefaciens-Mediated Transformation (ATMT)-Based Gene-Disruption Method for Verticillium dahliae.
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Sheng Wang, Haiying Xing, Chenlei Hua, Hui-Shan Guo, and Jie Zhang
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VERTICILLIUM dahliae , *PLANT disease research , *AGROBACTERIUM tumefaciens , *PLANT genetics , *GENETIC research , *PLANT species - Abstract
The soilbome fungal pathogen Verticillium dahliae infects a broad range of plant species to cause severe diseases. The availability of Verticillium genome sequences has provided opportunities for large-scale investigations of individual gene function in Verticillium strains using Agrobacterium tumefaciens-mediated transformation (ATMT)- based gene-disruption strategies. Traditional ATMT vectors require multiple cloning steps and elaborate characterization procedures to achieve successful gene replacement; thus, these vectors are not suitable for high-throughput ATMT-based gene deletion. Several advancements have been made that either involve simplification of the steps required for gene-deletion vector construction or increase the efficiency of the technique for rapid recombinant characterization. However, an ATMT binary vector that is both simple and efficient is still lacking. Here, we generated a USER-ATMT dual-selection (DS) binary vector, which combines both the advantages of the USER single-step cloning technique and the efficiency of the herpes simplex virus thymidine kinase negative-selection marker. Highly efficient deletion of three different genes in V. dahliae using the USER-ATMT-DS vector enabled verification that this newly-generated vector not only facilitates the cloning process but also simplifies the subsequent identification of fungal homologous recombinants. The results suggest that the USER-ATMT-DS vector is applicable for efficient gene deletion and suitable for large-scale gene deletion in V. dahliae. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
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