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1.
Proc Natl Acad Sci U S A ; 120(39): e2300587120, 2023 09 26.
Article in English | MEDLINE | ID: mdl-37725647

ABSTRACT

Oomycetes were recently discovered as natural pathogens of Caenorhabditis elegans, and pathogen recognition alone was shown to be sufficient to activate a protective transcriptional program characterized by the expression of multiple chitinase-like (chil) genes. However, the molecular mechanisms underlying oomycete recognition in animals remain fully unknown. We performed here a forward genetic screen to uncover regulators of chil gene induction and found several independent loss-of-function alleles of old-1 and flor-1, which encode receptor tyrosine kinases belonging to the C. elegans-specific KIN-16 family. We report that OLD-1 and FLOR-1 are both necessary for mounting the immune response and act in the epidermis. FLOR-1 is a pseudokinase that acts downstream of the active kinase OLD-1 and regulates OLD-1 levels at the plasma membrane. Interestingly, the old-1 locus is adjacent to the chil genes in the C. elegans genome, thereby revealing a genetic cluster important for oomycete resistance. Furthermore, we demonstrate that old-1 expression at the anterior side of the epidermis is regulated by the VAB-3/PAX6 transcription factor, well known for its role in visual system development in other animals. Taken together, our study reveals both conserved and species-specific factors shaping the activation and spatial characteristics of the immune response to oomycete recognition.


Subject(s)
Caenorhabditis elegans , Chitinases , Animals , Caenorhabditis elegans/genetics , Receptor Protein-Tyrosine Kinases , Cell Membrane , Alleles
2.
Proc Natl Acad Sci U S A ; 119(35): e2114064119, 2022 08 30.
Article in English | MEDLINE | ID: mdl-35994659

ABSTRACT

Plants are resistant to most microbial species due to nonhost resistance (NHR), providing broad-spectrum and durable immunity. However, the molecular components contributing to NHR are poorly characterised. We address the question of whether failure of pathogen effectors to manipulate nonhost plants plays a critical role in NHR. RxLR (Arg-any amino acid-Leu-Arg) effectors from two oomycete pathogens, Phytophthora infestans and Hyaloperonospora arabidopsidis, enhanced pathogen infection when expressed in host plants (Nicotiana benthamiana and Arabidopsis, respectively) but the same effectors performed poorly in distantly related nonhost pathosystems. Putative target proteins in the host plant potato were identified for 64 P. infestans RxLR effectors using yeast 2-hybrid (Y2H) screens. Candidate orthologues of these target proteins in the distantly related non-host plant Arabidopsis were identified and screened using matrix Y2H for interaction with RxLR effectors from both P. infestans and H. arabidopsidis. Few P. infestans effector-target protein interactions were conserved from potato to candidate Arabidopsis target orthologues (cAtOrths). However, there was an enrichment of H. arabidopsidis RxLR effectors interacting with cAtOrths. We expressed the cAtOrth AtPUB33, which unlike its potato orthologue did not interact with P. infestans effector PiSFI3, in potato and Nicotiana benthamiana. Expression of AtPUB33 significantly reduced P. infestans colonization in both host plants. Our results provide evidence that failure of pathogen effectors to interact with and/or correctly manipulate target proteins in distantly related non-host plants contributes to NHR. Moreover, exploiting this breakdown in effector-nonhost target interaction, transferring effector target orthologues from non-host to host plants is a strategy to reduce disease.


Subject(s)
Arabidopsis , Disease Resistance , Host Specificity , Nicotiana , Plant Diseases , Plant Proteins , Arabidopsis/metabolism , Arabidopsis/parasitology , Oomycetes/metabolism , Phytophthora infestans/metabolism , Plant Diseases/parasitology , Plant Diseases/prevention & control , Plant Proteins/metabolism , Solanum tuberosum/parasitology , Nicotiana/metabolism , Nicotiana/parasitology , Two-Hybrid System Techniques
3.
BMC Biol ; 22(1): 100, 2024 Apr 29.
Article in English | MEDLINE | ID: mdl-38679707

ABSTRACT

BACKGROUND: Plant pathogens secrete effector proteins into host cells to suppress immune responses and manipulate fundamental cellular processes. One of these processes is autophagy, an essential recycling mechanism in eukaryotic cells that coordinates the turnover of cellular components and contributes to the decision on cell death or survival. RESULTS: We report the characterization of AVH195, an effector from the broad-spectrum oomycete plant pathogen, Phytophthora parasitica. We show that P. parasitica expresses AVH195 during the biotrophic phase of plant infection, i.e., the initial phase in which host cells are maintained alive. In tobacco, the effector prevents the initiation of cell death, which is caused by two pathogen-derived effectors and the proapoptotic BAX protein. AVH195 associates with the plant vacuolar membrane system and interacts with Autophagy-related protein 8 (ATG8) isoforms/paralogs. When expressed in cells from the green alga, Chlamydomonas reinhardtii, the effector delays vacuolar fusion and cargo turnover upon stimulation of autophagy, but does not affect algal viability. In Arabidopsis thaliana, AVH195 delays the turnover of ATG8 from endomembranes and promotes plant susceptibility to P. parasitica and the obligate biotrophic oomycete pathogen Hyaloperonospora arabidopsidis. CONCLUSIONS: Taken together, our observations suggest that AVH195 targets ATG8 to attenuate autophagy and prevent associated host cell death, thereby favoring biotrophy during the early stages of the infection process.


Subject(s)
Autophagy , Nicotiana , Phytophthora , Plant Diseases , Phytophthora/physiology , Plant Diseases/microbiology , Plant Diseases/parasitology , Nicotiana/microbiology , Autophagy-Related Protein 8 Family/metabolism , Autophagy-Related Protein 8 Family/genetics , Host-Pathogen Interactions
4.
Mol Plant Microbe Interact ; 37(1): 62-71, 2024 Jan.
Article in English | MEDLINE | ID: mdl-37889205

ABSTRACT

Microtubule-associated protein 65-1 (MAP65-1) protein plays an essential role in plant cellular dynamics through impacting stabilization of the cytoskeleton by serving as a crosslinker of microtubules. The role of MAP65-1 in plants has been associated with phenotypic outcomes in response to various environmental stresses. The Arabidopsis MAP65-1 (AtMAP65-1) is a known virulence target of plant bacterial pathogens and is thus a component of plant immunity. Soybean events were generated that carry transgenic alleles for both AtMAP65-1 and GmMAP65-1, the soybean AtMAP65-1 homolog, under control of cauliflower mosaic virus 35S promoter. Both AtMAP65-1 and GmMAP65-1 transgenic soybeans are more resistant to challenges by the soybean bacterial pathogen Pseudomonas syringae pv. glycinea and the oomycete pathogen Phytophthora sojae, but not the soybean cyst nematode, Heterodera glycines. Soybean plants expressing AtMAP65-1 and GmMAP65-1 also display a tolerance to the herbicide oryzalin, which has a mode of action to destabilize microtubules. In addition, GmMAP65-1-expressing soybean plants show reduced cytosol ion leakage under freezing conditions, hinting that ectopic expression of GmMAP65-1 may enhance cold tolerance in soybean. Taken together, overexpression of AtMAP65-1 and GmMAP65-1 confers tolerance of soybean plants to various biotic and abiotic stresses. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Microtubule-Associated Proteins/genetics , Microtubule-Associated Proteins/metabolism , Glycine max/genetics , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Microtubules/metabolism , Plant Diseases/microbiology , Gene Expression Regulation, Plant
5.
Mol Plant Microbe Interact ; 37(3): 239-249, 2024 Mar.
Article in English | MEDLINE | ID: mdl-37921637

ABSTRACT

Plant pathogens manipulate the cellular environment of the host to facilitate infection and colonization that often lead to plant diseases. To accomplish this, many specialized pathogens secrete virulence proteins called effectors into the host cell, which subvert processes such as immune signaling, gene transcription, and host metabolism. Phytophthora infestans, the causative agent of potato late blight, employs an expanded repertoire of RxLR effectors with WY domains to manipulate the host through direct interaction with protein targets. However, our understanding of the molecular mechanisms underlying the interactions between WY effectors and their host targets remains limited. In this study, we performed a structural and biophysical characterization of the P. infestans WY effector Pi04314 in complex with the potato Protein Phosphatase 1-c (PP1c). We elucidate how Pi04314 uses a WY domain and a specialized C-terminal loop carrying a KVxF motif that interact with conserved surfaces on PP1c, known to be used by host regulatory proteins for guiding function. Through biophysical and in planta analyses, we demonstrate that Pi04314 WY or KVxF mutants lose their ability to bind PP1c. The loss of PP1c binding correlates with changes in PP1c nucleolar localization and a decrease in lesion size in plant infection assays. This study provides insights into the manipulation of plant hosts by pathogens, revealing how effectors exploit key regulatory interfaces in host proteins to modify their function and facilitate disease. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.


Subject(s)
Phytophthora infestans , Phytophthora infestans/genetics , Phosphoric Monoester Hydrolases/metabolism , Plants/metabolism , Transcription Factors/metabolism , Protein Binding , Plant Diseases
6.
Mol Plant Microbe Interact ; 37(9): 653-661, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38949402

ABSTRACT

Effector secretion by different routes mediates the molecular interplay between host plant and pathogen, but mechanistic details in eukaryotes are sparse. This may limit the discovery of new effectors that could be utilized for improving host plant disease resistance. In fungi and oomycetes, apoplastic effectors are secreted via the conventional endoplasmic reticulum (ER)-Golgi pathway, while cytoplasmic effectors are packaged into vesicles that bypass Golgi in an unconventional protein secretion (UPS) pathway. In Magnaporthe oryzae, the Golgi bypass UPS pathway incorporates components of the exocyst complex and a t-SNARE, presumably to fuse Golgi bypass vesicles to the fungal plasma membrane. Upstream, cytoplasmic effector mRNA translation in M. oryzae requires the efficient decoding of AA-ending codons. This involves the modification of wobble uridines in the anticodon loop of cognate tRNAs and fine-tunes cytoplasmic effector translation and secretion rates to maintain biotrophic interfacial complex integrity and permit host infection. Thus, plant-fungal interface integrity is intimately tied to effector codon usage, which is a surprising constraint on pathogenicity. Here, we discuss these findings within the context of fungal and oomycete effector discovery, delivery, and function in host cells. We show how cracking the codon code for unconventional cytoplasmic effector secretion in M. oryzae has revealed AA-ending codon usage bias in cytoplasmic effector mRNAs across kingdoms, including within the RxLR-dEER motif-encoding sequence of a bona fide Phytophthora infestans cytoplasmic effector, suggesting its subjection to translational speed control. By focusing on recent developments in understanding unconventional effector secretion, we draw attention to this important but understudied area of host-pathogen interactions. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


Subject(s)
Oomycetes , Plant Diseases , Plant Diseases/microbiology , Oomycetes/pathogenicity , Oomycetes/genetics , Fungal Proteins/metabolism , Fungal Proteins/genetics , Disease Resistance/genetics , Fungi/physiology , Fungi/pathogenicity , Host-Pathogen Interactions , Plants/microbiology
7.
BMC Genomics ; 25(1): 710, 2024 Jul 23.
Article in English | MEDLINE | ID: mdl-39044130

ABSTRACT

BACKGROUND: Identifying the DNA-binding specificities of transcription factors (TF) is central to understanding gene networks that regulate growth and development. Such knowledge is lacking in oomycetes, a microbial eukaryotic lineage within the stramenopile group. Oomycetes include many important plant and animal pathogens such as the potato and tomato blight agent Phytophthora infestans, which is a tractable model for studying life-stage differentiation within the group. RESULTS: Mining of the P. infestans genome identified 197 genes encoding proteins belonging to 22 TF families. Their chromosomal distribution was consistent with family expansions through unequal crossing-over, which were likely ancient since each family had similar sizes in most oomycetes. Most TFs exhibited dynamic changes in RNA levels through the P. infestans life cycle. The DNA-binding preferences of 123 proteins were assayed using protein-binding oligonucleotide microarrays, which succeeded with 73 proteins from 14 families. Binding sites predicted for representatives of the families were validated by electrophoretic mobility shift or chromatin immunoprecipitation assays. Consistent with the substantial evolutionary distance of oomycetes from traditional model organisms, only a subset of the DNA-binding preferences resembled those of human or plant orthologs. Phylogenetic analyses of the TF families within P. infestans often discriminated clades with canonical and novel DNA targets. Paralogs with similar binding preferences frequently had distinct patterns of expression suggestive of functional divergence. TFs were predicted to either drive life stage-specific expression or serve as general activators based on the representation of their binding sites within total or developmentally-regulated promoters. This projection was confirmed for one TF using synthetic and mutated promoters fused to reporter genes in vivo. CONCLUSIONS: We established a large dataset of binding specificities for P. infestans TFs, representing the first in the stramenopile group. This resource provides a basis for understanding transcriptional regulation by linking TFs with their targets, which should help delineate the molecular components of processes such as sporulation and host infection. Our work also yielded insight into TF evolution during the eukaryotic radiation, revealing both functional conservation as well as diversification across kingdoms.


Subject(s)
Evolution, Molecular , Phylogeny , Phytophthora infestans , Transcription Factors , Phytophthora infestans/genetics , Phytophthora infestans/metabolism , Transcription Factors/metabolism , Transcription Factors/genetics , Binding Sites , Protein Binding
8.
Appl Environ Microbiol ; 90(5): e0222223, 2024 05 21.
Article in English | MEDLINE | ID: mdl-38624199

ABSTRACT

Fungal phytopathogens cause significant reductions in agricultural yields annually, and overusing chemical fungicides for their control leads to environmental pollution and the emergence of resistant pathogens. Exploring natural isolates with strong antagonistic effects against pathogens can improve our understanding of their ecology and develop new treatments for the future. We isolated and characterized a novel bacterial strain associated with the species Burkholderia cenocepacia, termed APO9, which strongly inhibits Zymoseptoria tritici, a commercially important pathogenic fungus causing Septoria tritici blotch in wheat. Additionally, this strain exhibits inhibitory activity against four other phytopathogens. We found that physical contact plays a crucial role for APO9's antagonistic capacity. Genome sequencing of APO9 and biosynthetic gene cluster (BGC) analysis identified nine classes of BGCs and three types of secretion systems (types II, III, and IV), which may be involved in the inhibition of Z. tritici and other pathogens. To identify genes driving APO9's inhibitory activity, we screened a library containing 1,602 transposon mutants and identified five genes whose inactivation reduced inhibition efficiency. One such gene encodes for a diaminopimelate decarboxylase located in a terpenoid biosynthesis gene cluster. Phylogenetic analysis revealed that while some of these genes are also found across the Burkholderia genus, as well as in other Betaproteobacteria, the combination of these genes is unique to the Burkholderia cepacia complex. These findings suggest that the inhibitory capacity of APO9 is complex and not limited to a single mechanism, and may play a role in the interaction between various Burkholderia species and various phytopathogens within diverse plant ecosystems. IMPORTANCE: The detrimental effects of fungal pathogens on crop yields are substantial. The overuse of chemical fungicides contributes not only to environmental pollution but also to the emergence of resistant pathogens. Investigating natural isolates with strong antagonistic effects against pathogens can improve our understanding of their ecology and develop new treatments for the future. We discovered and examined a unique bacterial strain that demonstrates significant inhibitory activity against several phytopathogens. Our research demonstrates that this strain has a wide spectrum of inhibitory actions against plant pathogens, functioning through a complex mechanism. This plays a vital role in the interactions between plant microbiota and phytopathogens.


Subject(s)
Ascomycota , Burkholderia cenocepacia , Plant Diseases , Ascomycota/genetics , Burkholderia cenocepacia/genetics , Burkholderia cenocepacia/drug effects , Plant Diseases/microbiology , Triticum/microbiology , Antibiosis , Multigene Family
9.
New Phytol ; 242(6): 2682-2701, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38622771

ABSTRACT

Plant cell death is regulated in plant-pathogen interactions. While some aspartic proteases (APs) participate in regulating programmed cell death or defense responses, the defense functions of most APs remain largely unknown. Here, we report on a virulence factor, PlPeL8, which is a pectate lyase found in the hemibiotrophic pathogen Peronophythora litchii. Through in vivo and in vitro assays, we confirmed the interaction between PlPeL8 and LcAP1 from litchi, and identified LcAP1 as a positive regulator of plant immunity. PlPeL8 induced cell death associated with NbSOBIR1 and NbMEK2. The 11 conserved residues of PlPeL8 were essential for inducing cell death and enhancing plant susceptibility. Twenty-three LcAPs suppressed cell death induced by PlPeL8 in Nicotiana benthamiana depending on their interaction with PlPeL8. The N-terminus of LcAP1 was required for inhibiting PlPeL8-triggered cell death and susceptibility. Furthermore, PlPeL8 led to higher susceptibility in NbAPs-silenced N. benthamiana than the GUS-control. Our results indicate the crucial roles of LcAP1 and its homologs in enhancing plant resistance via suppression of cell death triggered by PlPeL8, and LcAP1 represents a promising target for engineering disease resistance. Our study provides new insights into the role of plant cell death in the arms race between plants and hemibiotrophic pathogens.


Subject(s)
Ascomycota , Aspartic Acid Proteases , Disease Resistance , Litchi , Plant Proteins , Polysaccharide-Lyases , Amino Acid Sequence , Ascomycota/genetics , Ascomycota/pathogenicity , Aspartic Acid Proteases/metabolism , Aspartic Acid Proteases/genetics , Cell Death , Disease Resistance/genetics , Gene Expression Regulation, Plant , Litchi/genetics , Nicotiana/genetics , Plant Diseases/microbiology , Plant Diseases/immunology , Plant Immunity/genetics , Plant Proteins/metabolism , Plant Proteins/genetics , Polysaccharide-Lyases/metabolism , Polysaccharide-Lyases/genetics , Protein Binding
10.
New Phytol ; 243(4): 1490-1505, 2024 Aug.
Article in English | MEDLINE | ID: mdl-39021210

ABSTRACT

Grapevine downy mildew, caused by the oomycete Plasmopara viticola (P. viticola, Berk. & M. A. Curtis; Berl. & De Toni), is a global threat to Eurasian wine grapes Vitis vinifera. Although resistant grapevine varieties are becoming more accessible, P. viticola populations are rapidly evolving to overcome these resistances. We aimed to uncover avirulence genes related to Rpv3.1-mediated grapevine resistance. We sequenced the genomes and characterized the development of 136 P. viticola strains on resistant and sensitive grapevine cultivars. A genome-wide association study was conducted to identify genomic variations associated with resistant-breaking phenotypes. We identified a genomic region associated with the breakdown of Rpv3.1 grapevine resistance (avrRpv3.1 locus). A diploid-aware reassembly of the P. viticola INRA-Pv221 genome revealed structural variations in this locus, including a 30 kbp deletion. Virulent P. viticola strains displayed multiple deletions on both haplotypes at the avrRpv3.1 locus. These deletions involve two paralog genes coding for proteins with 800-900 amino acids and signal peptides. These proteins exhibited a structure featuring LWY-fold structural modules, common among oomycete effectors. When transiently expressed, these proteins induced cell death in grapevines carrying Rpv3.1 resistance, confirming their avirulence nature. This discovery sheds light on the genetic mechanisms enabling P. viticola to adapt to grapevine resistance, laying a foundation for developing strategies to manage this destructive crop pathogen.


Subject(s)
Disease Resistance , Plant Diseases , Vitis , Vitis/genetics , Vitis/microbiology , Plant Diseases/microbiology , Plant Diseases/genetics , Plant Diseases/immunology , Disease Resistance/genetics , Oomycetes/pathogenicity , Genome-Wide Association Study , Sequence Deletion , Genes, Plant , Haplotypes/genetics , Gene Deletion , Phenotype
11.
J Exp Bot ; 2024 Jul 19.
Article in English | MEDLINE | ID: mdl-39028677

ABSTRACT

Both filamentous pathogens' hyphae and pollen tube penetrate the host's outer layer and involve growth within the host tissues. Early epidermal responses are decisive for the outcome of these two-cell interaction processes. We identified a single cell type, the papilla of Arabidospis thaliana's stigma, as a tool to conduct a comprehensive comparative analysis on how an epidermal cell responds to the invasion of an unwanted pathogen or a welcomed pollen tube. We showed that Phytophtora parasitica, a root oomycete, effectively breaches the stigmatic cell wall and develops as a biotroph within the papilla cytoplasm. These invasive features resemble the behaviour exhibited by the pathogen within its natural host cells, but diverge from the manner in which the pollen tube progresses, being engulfed within the papilla cell wall. Quantitative analysis revealed that both invaders trigger reorganisation of the stigmatic endomembrane system and the actin cytoskeleton. While some remodelling processes are shared between the two interactions, others appear more specific towards the respective invader. These findings underscore the remarkable ability of an epidermal cell to differentiate between two types of invaders, thereby enabling it to trigger the most suitable response during the onset of invasion.

12.
J Invertebr Pathol ; 206: 108159, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38925366

ABSTRACT

The oomycete Aphanomyces astaci is the causative agent of crayfish plague, a disease threatening susceptible freshwater crayfish species in Europe. To detect its spatiotemporal occurrence in Switzerland, we reviewed (1) the literature regarding occurrence of crayfish plague and North American crayfish carrier species and (2) the necropsy report archive of the Institute for Fish and Wildlife Health (FIWI) from 1968 to 2020. In the past, crayfish plague was diagnosed through several methods: conventional PCR, culture, and histology. When available, we re-evaluated archived Bouin's or formalin-fixed, paraffin-embedded samples collected during necropsies (1991-2020) with a recently published quantitative PCR. Literature research revealed putative reports of crayfish plague in Switzerland between the 1870s and 1910s and the first occurrence of three North American crayfish species between the late 1970s and 1990s. Finally, 54 (28.1%) cases were classified as positive and 9 (4.7%) cases as suspicious. The total number of positive cases increased by 14 (14.7%) after re-evaluation of samples. The earliest diagnosis of crayfish plague was performed in 1980 and the earliest biomolecular confirmation of A. astaci DNA dated 1991. Between 1980-1990, 1991-2000 and 2001-2010 crayfish plague spread from one to two and finally three catchment basins, respectively. Similar to other European countries, crayfish plague has occurred in Switzerland in two waves: the first at the end of the 19th and the second at the end of the 20th century in association with the first occurrence of North American crayfish species. The spread from one catchment basin to another suggests a human-mediated pathogen dispersal.


Subject(s)
Aphanomyces , Astacoidea , Animals , Astacoidea/microbiology , Astacoidea/parasitology , Switzerland/epidemiology
13.
Pestic Biochem Physiol ; 204: 106085, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39277398

ABSTRACT

Fluoxapiprolin, a novel piperidinyl thiazole isoxazoline fungicide, was developed by Bayer Crop Science in 2012. Despite its well-documented inhibitory activity against plant pathogenic oomycetes such as Phytophthora capsici and Phytophthora infestans, limited information regarding its antifungal spectrum and protective and curative activity is available. Fluoxapiprolin exhibited strong inhibitory activity against Phytophthora spp. and several Pythium spp., with EC50 values ranging from 2.12 × 10-4 to 2.92 µg/mL. It was much more effective against P. capsici in inhibiting mycelial growth, sporangium production, and cystospore germination than at reducing zoospore release. Moreover, fluoxapiprolin displayed both protective and curative activity against P. capsici infection in pepper plants under greenhouse conditions, with systemic translocation capability confirmed by High Performance Liquid Chromatography (HPLC) analysis. The results demonstrated the strong inhibitory activity of fluoxapiprolin against economically important plant oomycete pathogens, including Phytophthora spp. and several Pythium spp., and its certain translocation activity in pepper plants.


Subject(s)
Capsicum , Fungicides, Industrial , Phytophthora , Plant Diseases , Fungicides, Industrial/pharmacology , Phytophthora/drug effects , Plant Diseases/microbiology , Plant Diseases/prevention & control , Capsicum/microbiology , Capsicum/drug effects , Oomycetes/drug effects , Pythium/drug effects
14.
Plant Dis ; 108(9): 2771-2777, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38720542

ABSTRACT

A real-time loop-mediated isothermal amplification (LAMP) assay for the detection of Bremia lactucae, the causal pathogen of lettuce downy mildew, was developed and validated to aid in-field detection of airborne inoculum. Assay specificity was confirmed against a range of other pathogenic oomycete and fungal spp., and sensitivity of the assay for the detection of DNA extracted from sporangia was evaluated. The B. lactucae LAMP assay reliably detected DNA equivalent to 1 spore/reaction (16.7 pg DNA/reaction). Following extraction of DNA from Rotorod air samplers, to which sporangial suspensions were added, the assay reliably detected 25 sporangia/Rotorod. Detection of airborne inoculum of B. lactucae collected through the season from air samplers deployed in-field in plots infected with B. lactucae and in commercial lettuce fields in Scotland over two growing seasons was assessed. The method can be deployed on samples collected from commercial lettuce production to inform disease management strategies and limit the use of unnecessary prophylactic pesticide applications.


Subject(s)
Lactuca , Nucleic Acid Amplification Techniques , Oomycetes , Plant Diseases , Nucleic Acid Amplification Techniques/methods , Lactuca/microbiology , Plant Diseases/microbiology , Oomycetes/genetics , Oomycetes/isolation & purification , Sensitivity and Specificity , Molecular Diagnostic Techniques/methods
15.
Plant Dis ; 108(7): 2081-2089, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38386301

ABSTRACT

Houttuynia cordata is a prevalent vegetable in several Asian countries and is commonly used as a traditional Chinese medicinal herb for treating various diseases in China. Unfortunately, its yield and quality are adversely affected by root rot. However, the pathogen responsible for the losses remains unidentified, and effective fungicides for its management have not been thoroughly explored. In this work, we demonstrate the first report of Globisporangium spinosum as the causative agent causing root rot of H. cordata. Moreover, we evaluated the efficacy of hymexazol to manage the disease, which displayed remarkable inhibitory effects against mycelial growth of G. spinosum in vitro, with EC50 values as low as 1.336 µg/ml. Furthermore, hymexazol completely inhibited sporangia in G. spinosum at a concentration of 0.3125 µg/ml. Specifically, we observed that hymexazol was highly efficacious in reducing the incidence of H. cordata root rot caused by G. spinosum in a greenhouse setting. These findings offer a potential management tool for utilization of hymexazol in controlling H. cordata root rot in field production.


Subject(s)
Fungicides, Industrial , Houttuynia , Plant Diseases , Plant Roots , Houttuynia/microbiology , Plant Diseases/microbiology , Plant Diseases/prevention & control , Plant Roots/microbiology , Fungicides, Industrial/pharmacology
16.
J Asian Nat Prod Res ; 26(10): 1166-1174, 2024 Oct.
Article in English | MEDLINE | ID: mdl-38859556

ABSTRACT

Twenty 3-acyloxymaltol/ethyl maltol derivatives (7a-j and 8a-j) were synthesized and evaluated in vitro for their anti-oomycete activity against Phytophthora capsici, respectively. Among all of twenty derivatives, more than half of the compounds 7f, 7h, 8a-h and 8j had anti-oomycete activity higher than the positive control zoxamide (EC50 = 22.23 mg/L), and the EC50 values of 18.66, 20.32, 12.80, 16.18, 10.59, 14.98, 16.80, 10.36, 15.32, 12.64, and 13.59 mg/L, respectively. Especially, compounds 8c and 8f exhibited the best anti-oomycete activity against P. capsici with EC50 values of 10.59 and 10.36 mg/L, respectively. Overall, hydroxyl group of maltol/ethyl maltol is important active modification site.


Subject(s)
Phytophthora , Molecular Structure , Phytophthora/drug effects , Pyrones/pharmacology , Pyrones/chemistry , Pyrones/chemical synthesis , Structure-Activity Relationship , Drug Design
17.
Vet Radiol Ultrasound ; 65(2): 87-98, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38192159

ABSTRACT

Pythium insidiosum is an aquatic oomycete that causes granulomatous infection in dogs, most commonly cutaneous and gastrointestinal. Ultrasonographic characteristics of gastrointestinal pythiosis have been described; occasionally, CT is utilized in the clinical setting, and CT features of pythiosis have not been published. The purpose of this retrospective, multicenter, descriptive study is to describe CT characteristics of noncutaneous canine pythiosis. The following CT parameters were recorded: lesion anatomic location, number, shape, margination, size, attenuation pre- and postcontrast, enhancement pattern, lymph nodes affected, other lesions identified, and presence of peritoneal effusion or steatitis. Descriptive statistics demonstrating the frequency of lesion appearances were performed. Twenty-five dogs with noncutaneous pythiosis lesions that underwent CT were included; 19 had primarily gastrointestinal infections, four primarily arterial infections, one intrathoracic and intra-abdominal infection, and one primary pulmonary infection. In dogs with primary gastrointestinal infection, lesions were most common at the ileocolic junction and were most frequently focal, well-defined, moderate to marked circumferential wall thickening that was homogeneous and smoothly marginated precontrast, with moderate heterogeneous contrast enhancement. Most dogs had involvement of multiple gastrointestinal regions. Of four dogs with primary arterial involvement, three had large aneurysmal dilatations of the cranial mesenteric artery with severe mural thickening. All dogs had regional lymphadenopathy, which was variable but generally mild. Nine dogs had peritoneal effusion; six dogs had steatitis. CT features of pythiosis can overlap with neoplasia, but pythiosis should be considered as a differential, especially in young dogs. Findings supported using CT as an adjunct imaging test for increasing clinical suspicion of noncutaneous pythiosis.


Subject(s)
Dog Diseases , Gastrointestinal Diseases , Pythiosis , Steatitis , Dogs , Animals , Pythiosis/diagnostic imaging , Retrospective Studies , Gastrointestinal Diseases/veterinary , Tomography, X-Ray Computed/veterinary , Dog Diseases/diagnostic imaging , Dog Diseases/pathology
18.
Mycoscience ; 65(3): 133-137, 2024.
Article in English | MEDLINE | ID: mdl-39233756

ABSTRACT

Pythium amaminum sp. nov. was isolated from river and reservoir water on Amami island, Kagoshima Prefecture, Japan. The species can grow at temperatures between 10 °C and 35 °C. At the optimum temperature of 25 °C, the radial growth rate is 22.5 mm per day. Pythium amaminum produces filamentous sporangia consisting of branched, lobulate or digitate elements forming large complexes. Zoospores form inside the vesicle, which is discharged through a long tube at least 320 µm. Globose oogonia are ornamented with conical blunt spines. Oospores are aplerotic and globose. Antheridia twine around the oogonia or stick to them. These features having a both of the long discharge tube from sporangium and oogonia with spines are not observed in any other species of the genus Pythium, and thus we conclude that P. amaminum is a new Pythium species.

19.
Mol Plant Microbe Interact ; 36(1): 26-46, 2023 Jan.
Article in English | MEDLINE | ID: mdl-36306437

ABSTRACT

Phytophthora species are oomycete plant pathogens that cause great economic and ecological impacts. The Phytophthora genus includes over 180 known species, infecting a wide range of plant hosts, including crops, trees, and ornamentals. We sequenced the genomes of 31 individual Phytophthora species and 24 individual transcriptomes to study genetic relationships across the genus. De novo genome assemblies revealed variation in genome sizes, numbers of predicted genes, and in repetitive element content across the Phytophthora genus. A genus-wide comparison evaluated orthologous groups of genes. Predicted effector gene counts varied across Phytophthora species by effector family, genome size, and plant host range. Predicted numbers of apoplastic effectors increased as the host range of Phytophthora species increased. Predicted numbers of cytoplasmic effectors also increased with host range but leveled off or decreased in Phytophthora species that have enormous host ranges. With extensive sequencing across the Phytophthora genus, we now have the genomic resources to evaluate horizontal gene transfer events across the oomycetes. Using a machine-learning approach to identify horizontally transferred genes with bacterial or fungal origin, we identified 44 candidates over 36 Phytophthora species genomes. Phylogenetic reconstruction indicates that the transfers of most of these 44 candidates happened in parallel to major advances in the evolution of the oomycetes and Phytophthora spp. We conclude that the 31 genomes presented here are essential for investigating genus-wide genomic associations in genus Phytophthora. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


Subject(s)
Phytophthora , Phytophthora/genetics , Phylogeny , Gene Transfer, Horizontal , Genome , Genomics , Plants/genetics
20.
Mol Plant Microbe Interact ; 36(6): 313-314, 2023 Jun.
Article in English | MEDLINE | ID: mdl-37516252

ABSTRACT

The plant pathogen Phytophthora palmivora causes rot disease in several monocots and dicots. The plant 14-3-3 proteins are targets of different types of effector molecules secreted by the pathogens. An RXLR-type effector FIRE (14-3-3 interacting RXLR effector) and its target 14-3-3 proteins that localize to haustoria have been identified, pointing to a potential site of interaction. The pathogen hijacks the host 14-3-3 proteins through FIRE-mediated interaction and lowers the immunity for disease progression. The effector FIRE and 14-3-3 interaction deciphered in this study could pave the way for genetic modification of plants with altered 14-3-3 protein for broad host resistance. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


Subject(s)
14-3-3 Proteins , Phytophthora infestans , 14-3-3 Proteins/genetics , Amino Acid Motifs , Plants , Plant Diseases
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