Plant pathogens and pests
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Aphid effector Me10 interacts with tomato TFT7, a 14‐3‐3 isoform involved in aphid resistance - Chaudhary - 2019 - New Phytologist -

Aphid effector Me10 interacts with tomato TFT7, a 14‐3‐3 isoform involved in aphid resistance - Chaudhary - 2019 - New Phytologist - | Plant pathogens and pests | Scoop.it
We demonstrated previously that expression of Macrosiphum euphorbiae salivary protein Me10 enhanced aphid reproduction on its host tomato (Solanum lycopersicum). However, the mechanism of action of Me10 remained elusive.
To confirm the secretion of Me10 by the aphid into plant tissues, we produced Me10 polyclonal antibodies. To identify the plant targets of Me10, we developed a tomato immune induced complementary DNA yeast two‐hybrid library and screened it with Me10 as bait. Immunoprecipitation and bimolecular fluorescence complementation (BiFC) assays were performed to validate one of the interactions in planta, and virus‐induced gene silencing was used for functional characterization in tomato.
We demonstrated that Me10 is secreted into the plant tissues and interacts with tomato 14‐3‐3 isoform 7 (TFT7) in yeast. Immunoprecipitation assays confirmed that Me10 and its homologue in Aphis gossypii, Ag10k, interact with TFT7 in planta. Further, BiFC revealed that Me10 interaction with TFT7 occurs in the plant cell cytoplasm. While silencing of TFT7 in tomato leaves did not affect tomato susceptibility to M. euphorbiae, it enhanced longevity and fecundity of A. gossypii, the non‐host aphid.
Our results suggest the model whereby TFT7 plays a role in aphid resistance in tomato and effectors of the Me10/Ag10k family interfere with TFT7 function during aphid infestation.
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Plant pathogens and pests
Mainly dedicated to plant pathogens , mechanisms of pathogenicity, life cycles, epidemiology and plant breeding methods and results helping to prevent their propagation.  This site is complementary to the Plant immunity and legume symbiosis Scoop-It site :      https://www.scoop.it/t/plant-pathogen-interactions-by-christophe-jacquet
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Apoplastic effector proteins of plant-associated fungi and oomycetes -

Apoplastic effector proteins of plant-associated fungi and oomycetes - | Plant pathogens and pests | Scoop.it

The outcome of an interaction between a plant and a fungus or an oomycete, whether compatibility or incompatibility, is often determined in the hostile extracellular spaces and matrices of the apoplast. Indeed, for compatibility to occur, many plant-associated fungi and oomycetes must first neutralize the apoplast, which is both monitored by plant cell-surface immune receptors, and enriched in plant (and frequently, competitor)-derived antimicrobial compounds. Research is highlighting the diverse roles that fungal and oomycete effector proteins play in the apoplast to promote compatibility, with most recent progress made towards understanding the role of these proteins in evading chitin-triggered immunity. Research is also showcasing the ability of apoplastic effector proteins to bring about incompatibility upon recognition by diverse plant cell-surface immune receptors, and the use of effectoromics to rapidly identify apoplastic effector protein–cell-surface immune receptor interactions.

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Mangroves in the Leaves: Anatomy, Physiology, and Immunity of Epithemal Hydathodes | Annual Review of Phytopathology

Mangroves in the Leaves: Anatomy, Physiology, and Immunity of Epithemal Hydathodes | Annual Review of Phytopathology | Plant pathogens and pests | Scoop.it
Hydathodes are organs found on aerial parts of a wide range of plant species that provide almost direct access for several pathogenic microbes to the plant vascular system. Hydathodes are better known as the site of guttation, which is the release of droplets of plant apoplastic fluid to the outer leaf surface. Because these organs are only described through sporadic allusions in the literature, this review aims to provide a comprehensive view of hydathode development, physiology, and immunity by compiling a historic and contemporary bibliography. In particular, we refine the definition of hydathodes.We illustrate their important roles in the maintenance of plant osmotic balance, nutrient retrieval, and exclusion of deleterious chemicals from the xylem sap. Finally, we present our current understanding of the infection of hydathodes by adapted vascular pathogens and the associated plant immune responses.
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Plant pathogenic fungi Colletotrichum and Magnaporthe share a common G1 phase monitoring strategy for proper appressorium development - Fukada - 2019 - New Phytologist -

Plant pathogenic fungi Colletotrichum and Magnaporthe share a common G1 phase monitoring strategy for proper appressorium development - Fukada - 2019 - New Phytologist - | Plant pathogens and pests | Scoop.it
To breach the plant cuticle, many plant pathogenic fungi differentiate specialized infection structures (appressoria). In Colletotrichum orbiculare (cucumber anthracnose fungus), this differentiation requires unique proper G1/S phase progression, regulated by two‐component GTPase activating protein CoBub2/CoBfa1 and GTPase CoTem1. Since their homologues regulate mitotic exit, cytokinesis, or septum formation from yeasts to mammals, we asked whether the BUB2 function in G1/S progression is specific to plant pathogenic fungi.
Colletotrichum higginsianum and Magnaporthe oryzae were genetically analyzed to investigate conservation of BUB2 roles in cell cycle regulation, septum formation, and virulence. Expression profile of cobub2Δ was analyzed using a custom microarray.
In bub2 mutants of both fungi, S phase initiation was earlier, and septum formation coordinated with a septation initiation network protein and contractile actin ring was impaired. Earlier G1/S transition in cobub2Δ results in especially high expression of DNA replication genes and differing regulation of virulence‐associated genes that encode proteins such as carbohydrate‐active enzymes and small secreted proteins. The virulence of chbub2Δ and mobub2Δ was significantly reduced.
Our evidence shows that BUB2 regulation of G1/S transition and septum formation supports its specific requirement for appressorium development in plant pathogenic fungi.
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Autocrine pheromone signalling regulates community behaviour in the fungal pathogen Fusarium oxysporum

Autocrine pheromone signalling regulates community behaviour in the fungal pathogen Fusarium oxysporum | Plant pathogens and pests | Scoop.it
Autocrine self-signalling via secreted peptides and cognate receptors regulates cell development in eukaryotes and is conserved from protozoans to mammals1,2. In contrast, secreted peptides from higher fungi have been traditionally associated with paracrine non-self-signalling during sexual reproduction3. For example, cells of the model fungus Saccharomyces cerevisiae fall into two distinct mating types (MAT), which produce either a- or α-pheromone and the cognate receptors Ste2 or Ste3, respectively4. Inappropriate autocrine pheromone signalling (APS) during mating is prevented by downregulation of the self-pheromone receptor4,5 and by a-type cell-specific cleavage of α-pheromone through the protease Bar1 (refs. 6,7,8). While APS can be artificially induced by manipulation of the pheromone secrete-and-sense circuit7,9,10,11, its natural occurrence in ascomycete fungi has not been described. Here, we show that Fusarium oxysporum—a devastating plant pathogen that lacks a known sexual cycle12—co-expresses both pheromone–receptor pairs, resulting in autocrine regulation of developmental programmes other than mating. We found that unisexual populations of MAT1-1 cells (α-type idiomorphs13) secrete and sense both a- and α-pheromone, and that their perception requires the cognate receptors and conserved elements of the cell wall integrity mitogen-activated protein kinase cascade. We further show that F. oxysporum uses APS to regulate spore germination in a cell-density-dependent manner, whereby the α-Ste2 interaction leads to repression of conidial germination while the a-Ste3 interaction relieves repression. Our results reveal the existence of a regulatory function for peptide pheromones in the quorum-sensing-mediated control of fungal development.
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Frontiers | Manipulation of Phytohormone Pathways by Effectors of Filamentous Plant Pathogens | Plant Science

Frontiers | Manipulation of Phytohormone Pathways by Effectors of Filamentous Plant Pathogens | Plant Science | Plant pathogens and pests | Scoop.it
Phytohormones regulate a large variety of physiological processes in plants. In addition, salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) are responsible for primary defense responses against abiotic and biotic stresses, while plant growth regulators, such as auxins, brassinosteroids (BRs), cytokinins (CKs), abscisic acid (ABA), and gibberellins (GAs), also contribute to plant immunity. To successfully colonize plants, filamentous pathogens like fungi and oomycetes have evolved diverse strategies to interfere with phytohormone pathways with the help of secreted effectors. These include proteins, toxins, polysaccharides as well as phytohormones or phytohormone mimics. Such pathogen effectors manipulate phytohormone pathways by directly altering hormone levels, by interfering with phytohormone biosynthesis, or by altering or blocking important components of phytohormone signaling pathways. In this review, we outline the various strategies used by filamentous phytopathogens to manipulate phytohormone pathways to cause disease.
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Cucumber mosaic virus infection as a potential selective pressure on Arabidopsis thaliana populations

Cucumber mosaic virus infection as a potential selective pressure on Arabidopsis thaliana populations | Plant pathogens and pests | Scoop.it
Author summary Plant-virus coevolution has not been demonstrated in any wild system, and it has been proposed that viruses often would be mutualistic symbionts, rather than pathogens, in wild plant ecosystems. We analyse if viruses are virulent pathogens of plants in wild ecosystems and, consequently, plants have evolved defences against virus infection. To test this hypothesis, we studied the genetic diversity of Arabidopsis thaliana for two defence traits, resistance and tolerance, to Cucumber mosaic virus (CMV) at a regional scale in the Iberian Peninsula. Resistance and tolerance to CMV showed substantial genetic variation within and between host populations, and depended on the virus x host genotype interaction, two conditions for coevolution. Resistance and tolerance were independent traits that co-occurred at the population and regional scales, and that have evolved independently from other adaptive life-history traits. Analyses also indicated that resistance and tolerance are likely under selection, most likely due to virus infection. These results support a hypothesis of plant-virus coevolution and contribute to demonstrate that plant viruses may be virulent parasites of plants in in wild ecosystems.
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Genomic Plasticity Mediated by Transposable Elements in the Plant Pathogenic Fungus Colletotrichum higginsianum | Genome Biology and Evolution | Oxford Academic

Genomic Plasticity Mediated by Transposable Elements in the Plant Pathogenic Fungus Colletotrichum higginsianum | Genome Biology and Evolution | Oxford Academic | Plant pathogens and pests | Scoop.it
Phytopathogen genomes are under constant pressure to change, as pathogens are locked in an evolutionary arms race with their hosts, where pathogens evolve effector genes to manipulate their hosts, while the hosts evolve immune components to recognize the products of these genes. Colletotrichum higginsianum (Ch), a fungal pathogen with no known sexual morph, infects Brassicaceae plants including Arabidopsis thaliana. Previous studies revealed that Ch differs in its virulence towards various A. thaliana ecotypes, indicating the existence of coevolutionary selective pressures. However, between-strain genomic variations in Ch have not been studied. Here, we sequenced and assembled the genome of a Ch strain, resulting in a highly contiguous genome assembly, which was compared to the chromosome-level genome assembly of another strain to identify genomic variations between strains. We found that the two closely related strains vary in terms of large-scale rearrangements, the existence of strain-specific regions, and effector candidate gene sets and that these variations are frequently associated with transposable elements (TEs). Ch has a compartmentalized genome consisting of gene-sparse, TE-dense regions with more effector candidate genes and gene-dense, TE-sparse regions harboring conserved genes. Additionally, analysis of the conservation patterns and syntenic regions of effector candidate genes indicated that the two strains vary in their effector candidate gene sets because of de novo evolution, horizontal gene transfer, or gene loss after divergence. Our results reveal mechanisms for generating genomic diversity in this asexual pathogen, which are important for understanding its adaption to hosts.
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Interactions of Tomato and Botrytis cinerea Genetic Diversity: Parsing the Contributions of Host Differentiation, Domestication, and Pathogen Variation

Interactions of Tomato and Botrytis cinerea Genetic Diversity: Parsing the Contributions of Host Differentiation, Domestication, and Pathogen Variation | Plant pathogens and pests | Scoop.it
Although the impacts of crop domestication on specialist pathogens are well known, less is known about the interaction of crop variation and generalist pathogens. To study how genetic variation within a crop affects plant resistance to generalist pathogens, we infected a collection of wild and domesticated tomato accessions with a genetically diverse population of the generalist pathogen Botrytis cinerea. We quantified variation in lesion size of 97 B. cinerea genotypes (isolates) on six domesticated tomato genotypes (Solanum lycopersicum) and six wild tomato genotypes (Solanum pimpinellifolium). Lesion size was significantly affected by large effects of the host and pathogen’s genotype, with a much smaller contribution of domestication. This pathogen collection also enables genome-wide association mapping of B. cinerea. Genome-wide association mapping of the pathogen showed that virulence is highly polygenic and involves a diversity of mechanisms. Breeding against this pathogen would likely require the use of diverse isolates to capture all possible mechanisms. Critically, we identified a subset of B. cinerea genes where allelic variation was linked to altered virulence against wild versus domesticated tomato, as well as loci that could handle both groups. This generalist pathogen already has a large collection of allelic variation that must be considered when designing a breeding program.
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The potato cyst nematode effector RHA1B is a ubiquitin ligase and uses two distinct mechanisms to suppress plant immune signaling

The potato cyst nematode effector RHA1B is a ubiquitin ligase and uses two distinct mechanisms to suppress plant immune signaling | Plant pathogens and pests | Scoop.it
Plant pathogens, such as bacteria, fungi, oomycetes and nematodes, rely on wide range of virulent effectors delivered into host cells to suppress plant immunity. Although phytobacterial effectors have been intensively investigated, little is known about the function of effectors of plant-parasitic nematodes, such as Globodera pallida, a cyst nematode responsible for vast losses in the potato and tomato industries. Here, we demonstrate using in vivo and in vitro ubiquitination assays the potato cyst nematode (Globodera pallida) effector RHA1B is an E3 ubiquitin ligase that employs multiple host plant E2 ubiquitin conjugation enzymes to catalyze ubiquitination. RHA1B was able to suppress effector-triggered immunity (ETI), as manifested by suppression of hypersensitive response (HR) mediated by a broad range of nucleotide-binding leucine-rich repeat (NB-LRR) immune receptors, presumably via E3-dependent degradation of the NB-LRR receptors. RHA1B also blocked the flg22-triggered expression of Acre31 and WRKY22, marker genes of pathogen‐associated molecular pattern (PAMP)‐triggered immunity (PTI), but this did not require the E3 activity of RHA1B. Moreover, transgenic potato overexpressing the RHA1B transgene exhibited enhanced susceptibility to G. pallida. Thus, our data suggest RHA1B facilitates nematode parasitism not only by triggering degradation of NB-LRR immune receptors to block ETI signaling but also by suppressing PTI signaling via an as yet unknown E3-independent mechanism.
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Albugo candida race diversity, ploidy and host‐associated microbes revealed using DNA sequence capture on diseased plants in the field - Jouet - 2019 - New Phytologist -

Albugo candida race diversity, ploidy and host‐associated microbes revealed using DNA sequence capture on diseased plants in the field - Jouet - 2019 - New Phytologist - | Plant pathogens and pests | Scoop.it
Physiological races of the oomycete Albugo candida are biotrophic pathogens of diverse plant species, primarily the Brassicaceae, and cause infections that suppress host immunity to other pathogens. However, A. candida race diversity and the consequences of host immunosuppression are poorly understood in the field.
We report a method that enables sequencing of DNA of plant pathogens and plant‐associated microbes directly from field samples (Pathogen Enrichment Sequencing: PenSeq). We apply this method to explore race diversity in A. candida and to detect A. candida‐associated microbes in the field (91 A. candida‐infected plants).
We show with unprecedented resolution that each host plant species supports colonization by one of 17 distinct phylogenetic lineages, each with an unique repertoire of effector candidate alleles. These data reveal the crucial role of sexual and asexual reproduction, polyploidy and host domestication in A. candida specialization on distinct plant species. Our bait design also enabled phylogenetic assignment of DNA sequences from bacteria and fungi from plants in the field.
This paper shows that targeted sequencing has a great potential for the study of pathogen populations while they are colonizing their hosts. This method could be applied to other microbes, especially to those that cannot be cultured.
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A novel fungal effector from Puccinia graminis suppressing RNA silencing and plant defense responses - Yin - - New Phytologist -

A novel fungal effector from Puccinia graminis suppressing RNA silencing and plant defense responses - Yin - - New Phytologist - | Plant pathogens and pests | Scoop.it
Fungal plant pathogens, like rust‐causing biotrophic fungi, secrete hundreds of effectors into plant cells to subvert host immunity and promote pathogenicity on their host plants by manipulating specific physiological processes or signal pathways, but the actual function has been demonstrated for very few of these proteins.
Here, we show that the PgtSR1 effector proteins, encoded by two allelic genes (PgtSR1‐a and PgtSR1‐b), from the wheat stem rust pathogen Puccinia graminis f. sp. tritici (Pgt), suppress RNA silencing in plants and impede plant defenses by altering the abundance of small RNAs that serve as defense regulators.
Expression of the PgtSR1s in plants revealed that the PgtSR1s promote susceptibility to multiple pathogens and partially suppress cell death triggered by multiple R proteins.
Overall, our study provides the first evidence that the filamentous fungus P. graminis has evolved to produce fungal suppressors of RNA silencing and indicates that PgtSR1s suppress both basal defenses and effector triggered immunity.
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Establishment of a selection marker recycling system for sequential transformation of the plant‐pathogenic fungus Colletotrichum orbiculare - Kumakura - 2019 - Molecular Plant Pathology - Wiley Onl...

Establishment of a selection marker recycling system for sequential transformation of the plant‐pathogenic fungus Colletotrichum orbiculare - Kumakura - 2019 - Molecular Plant Pathology - Wiley Onl... | Plant pathogens and pests | Scoop.it
Genome sequencing of pathogenic fungi has revealed the presence of various effectors that aid pathogen invasion by the manipulation of plant immunity. Effectors are often individually dispensable because of duplication and functional redundancy as a result of the arms race between host plants and pathogens. To study effectors that have functional redundancy, multiple gene disruption is often required. However, the number of selection markers that can be used for gene targeting is limited. Here, we established a marker recycling system that allows the use of the same selection marker in successive transformations in the model fungal pathogen Colletotrichum orbiculare, a causal agent of anthracnose disease in plants belonging to the Cucurbitaceae. We identified two C. orbiculare homologues of yeast URA3/pyrG, designated as URA3A and URA3B, which can be used as selection markers on medium with no uridine. The gene can then be removed from the genome via homologous recombination when the fungus is grown in the presence of 5‐fluoroorotic acid (5‐FOA), a chemical that is converted into a toxin by URA3 activity. The ura3a/b double mutants showed auxotrophy for uridine and insensitivity to 5‐FOA. Using the ura3a/b mutants, transformation with the URA3B marker and its removal were successfully applied to disrupt the virulence‐related gene, PKS1. The pks1 mutants showed a reduction in virulence, demonstrating that the method can be used to study virulence‐related genes in C. orbiculare. The establishment of a URA3‐based marker recycling system in plant‐pathogenic fungi enables the genetic analysis of multiple genes that have redundant functions, including effector genes.
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The fungal ribonuclease-like effector protein CSEP0064/BEC1054 represses plant immunity and interferes with degradation of host ribosomal RNA

The fungal ribonuclease-like effector protein CSEP0064/BEC1054 represses plant immunity and interferes with degradation of host ribosomal RNA | Plant pathogens and pests | Scoop.it
The biotrophic fungal pathogen Blumeria graminis causes the powdery mildew disease of cereals and grasses. We present the first crystal structure of a B. graminis effector of pathogenicity (CSEP0064/BEC1054), demonstrating it has a ribonuclease (RNase)-like fold. This effector is part of a group of RNase-like proteins (termed RALPHs) which comprise the largest set of secreted effector candidates within the B. graminis genomes. Their exceptional abundance suggests they play crucial functions during pathogenesis. We show that transgenic expression of RALPH CSEP0064/BEC1054 increases susceptibility to infection in both monocotyledonous and dicotyledonous plants. CSEP0064/BEC1054 interacts in planta with the pathogenesis-related protein PR10. The effector protein associates with total RNA and weakly with DNA. Methyl jasmonate (MeJA) levels modulate susceptibility to aniline-induced host RNA fragmentation. In planta expression of CSEP0064/BEC1054 reduces the formation of this RNA fragment. We propose CSEP0064/BEC1054 is a pseudoenzyme that binds to host ribosomes, thereby inhibiting the action of plant ribosome-inactivating proteins (RIPs) that would otherwise lead to host cell death, an unviable interaction and demise of the fungus.
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Molecular basis for functional diversity among microbial Nep1-like proteins

Molecular basis for functional diversity among microbial Nep1-like proteins | Plant pathogens and pests | Scoop.it
Necrosis and ethylene-inducing peptide 1 (Nep1)-like proteins (NLPs) are secreted by several phytopathogenic microorganisms. They trigger necrosis in various eudicot plants upon binding to plant sphingolipid glycosylinositol phosphorylceramides (GIPC). Interestingly, HaNLP3 from the obligate biotroph oomycete Hyaloperonospora arabidopsidis does not induce necrosis. We determined the crystal structure of HaNLP3 and showed that it adopts the NLP fold. However, the conformations of the loops surrounding the GIPC headgroup-binding cavity differ from those of cytotoxic Pythium aphanidermatum NLPPya. Essential dynamics extracted from μs-long molecular dynamics (MD) simulations reveals a limited conformational plasticity of the GIPC-binding cavity in HaNLP3 relative to toxic NLPs. This likely precludes HaNLP3 binding to GIPCs, which is the underlying reason for the lack of toxicity. This study reveals that mutations at key protein regions cause a switch between non-toxic and toxic phenotypes within the same protein scaffold. Altogether, these data provide evidence that protein flexibility is a distinguishing trait of toxic NLPs and highlight structural determinants for a potential functional diversification of non-toxic NLPs utilized by biotrophic plant pathogens.
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A two genes – for – one gene interaction between Leptosphaeria maculans and Brassica napus - Petit‐Houdenot - 2019 - New Phytologist -

A two genes – for – one gene interaction between Leptosphaeria maculans and Brassica napus - Petit‐Houdenot - 2019 - New Phytologist - | Plant pathogens and pests | Scoop.it
Interactions between Leptosphaeria maculans, causal agent of stem canker of oilseed rape, and its Brassica hosts are models of choice to explore the multiplicity of ‘gene‐for‐gene’ complementarities and how they diversified to increased complexity in the course of plant–pathogen co‐evolution. Here, we support this postulate by investigating the AvrLm10 avirulence that induces a resistance response when recognized by the Brassica nigra resistance gene Rlm10.
Using genome‐assisted map‐based cloning, we identified and cloned two AvrLm10 candidates as two genes in opposite transcriptional orientation located in a subtelomeric repeat‐rich region of the genome. The AvrLm10 genes encode small secreted proteins and show expression profiles in planta similar to those of all L. maculans avirulence genes identified so far.
Complementation and silencing assays indicated that both genes are necessary to trigger Rlm10 resistance. Three assays for protein–protein interactions showed that the two AvrLm10 proteins interact physically in vitro and in planta.
Some avirulence genes are recognized by two distinct resistance genes and some avirulence genes hide the recognition specificities of another. Our L. maculans model illustrates an additional case where two genes located in opposite transcriptional orientation are necessary to induce resistance. Interestingly, orthologues exist for both L. maculans genes in other phytopathogenic species, with a similar genome organization, which may point to an important conserved effector function linked to heterodimerization of the two proteins.
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An expanded subfamily of G-protein-coupled receptor genes in Fusarium graminearum required for wheat infection

An expanded subfamily of G-protein-coupled receptor genes in Fusarium graminearum required for wheat infection | Plant pathogens and pests | Scoop.it
The cAMP–PKA and MAP kinase pathways are essential for plant infection in the wheat head blight fungus Fusarium graminearum. To identify upstream receptors of these well-conserved signalling pathways, we systematically characterized the 105 G-protein-coupled receptor (GPCR) genes. Although none were required for vegetative growth, five GPCR genes (GIV1–GIV5) significantly upregulated during plant infection were important for virulence. The giv1 mutant was defective in the formation of specialized infection structures known as infection cushions, which was suppressed by application of exogenous cAMP and dominant active FST7 MEK kinase. GIV1 was important for the stimulation of PKA and Gpmk1 MAP kinase by compounds in wheat spikelets. GIV2 and GIV3 were important for infectious growth after penetration. Invasive hyphae of the giv2 mutant were defective in cell-to-cell spreading and mainly grew intercellularly in rachis tissues. Interestingly, the GIV2–GIV5 genes form a phylogenetic cluster with GIV6, which had overlapping functions with GIV5 during pathogenesis. Furthermore, the GIV2–GIV6 cluster is part of a 22-member subfamily of GPCRs, with many of them having in planta-specific upregulation and a common promoter element; however, only three subfamily members are conserved in other fungi. Taken together, F. graminearum has an expanded subfamily of infection-related GPCRs for regulating various infection processes.
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Non-invasive plant disease diagnostics enabled by smartphone-based fingerprinting of leaf volatiles

Non-invasive plant disease diagnostics enabled by smartphone-based fingerprinting of leaf volatiles | Plant pathogens and pests | Scoop.it
Plant pathogen detection conventionally relies on molecular technology that is complicated, time-consuming and constrained to centralized laboratories. We developed a cost-effective smartphone-based volatile organic compound (VOC) fingerprinting platform that allows non-invasive diagnosis of late blight caused by Phytophthora infestans by monitoring characteristic leaf volatile emissions in the field. This handheld device integrates a disposable colourimetric sensor array consisting of plasmonic nanocolorants and chemo-responsive organic dyes to detect key plant volatiles at the ppm level within 1 min of reaction. We demonstrate the multiplexed detection and classification of ten individual plant volatiles with this field-portable VOC-sensing platform, which allows for early detection of tomato late blight 2 d after inoculation, and differentiation from other pathogens of tomato that lead to similar symptoms on tomato foliage. Furthermore, we demonstrate a detection accuracy of ≥95% in diagnosis of P. infestans in both laboratory-inoculated and field-collected tomato leaves in blind pilot tests. Finally, the sensor platform has been beta-tested for detection of P. infestans in symptomless tomato plants in the greenhouse setting.
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The p33 protein of Citrus tristeza virus affects viral pathogenicity by modulating a host immune response - Sun - 2019 - New Phytologist -

The p33 protein of Citrus tristeza virus affects viral pathogenicity by modulating a host immune response - Sun - 2019 - New Phytologist - | Plant pathogens and pests | Scoop.it
Accumulation of reactive oxygen species (ROS) is a general plant basal defense strategy against viruses. In this study, we show that infection by Citrus tristeza virus (CTV) triggered ROS burst in Nicotiana benthamiana and in the natural citrus host, the extent of which was virus‐dose dependent.
Using Agrobacterium‐mediated expression of CTV‐encoded proteins in N. benthamiana, we found that p33, a unique viral protein, contributed to the induction of ROS accumulation and programmed cell death. The role of p33 in CTV pathogenicity was assessed based on gene knockout and complementation in N. benthamiana.
In the citrus–CTV pathosystem, deletion of the p33 open reading frame in a CTV variant resulted in a significant decrease in ROS production, compared to that of the wild type CTV, which correlated with invasion of the mutant virus into the immature xylem tracheid cells and abnormal differentiation of the vascular system. By contrast, the wild type CTV exhibited phloem‐limited distribution with a minor effect on the vasculature.
We conclude that the p33 protein is a CTV effector that negatively affects virus pathogenicity and suggest that N. benthamiana recognizes p33 to activate the host immune response to restrict CTV into the phloem tissue and minimize the disease syndrome.
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The t-SNARE protein FgPep12, associated with FgVam7, is essential for ascospore discharge and plant infection by trafficking Ca2+ ATPase FgNeo1 between Golgi and endosome/vacuole in Fusarium gramin...

The t-SNARE protein FgPep12, associated with FgVam7, is essential for ascospore discharge and plant infection by trafficking Ca2+ ATPase FgNeo1 between Golgi and endosome/vacuole in Fusarium gramin... | Plant pathogens and pests | Scoop.it
Soluble N-ethylmaleimide-sensitive factor attachment receptors (SNAREs) play a crucial role in the development and virulence through mediation of membrane fusion and vesicle trafficking in pathogens. Our previous studies reported that the SNARE protein FgVam7 and its binding proteins FgVps39/41 are involved in vesicle trafficking and are important for vegetative growth, asexual/sexual development, deoxynivalenol production and virulence in the Fusarium head blight fungus Fusarium graminearum. Here, we identified and characterized another FgVam7 binding protein in F. graminearum, FgPep12, an ortholog of yeast t-SNARE Pep12 with both the SNARE and TM domains being essential for its localization and function. Deletion of FgPep12 caused defects in vegetative growth, conidiogenesis, perithecia formation and virulence. Cytological observation revealed that FgPep12 localizes to the Golgi apparatus, late endosomes and vacuoles, and is necessary for transport from the vacuole to prevacuolar compartment. Further investigation revealed that both FgPep12 and FgVam7 are essential for ascospore discharge through interaction with and trafficking of the Ca2+ ATPase FgNeo1 between the Golgi and endosomal/vacuolar system. FgNeo1 has similar biological roles to FgPep12 and is required for ascospore discharge in F. graminearum. Together, these results provide solid evidence to help unravel the mechanisms underlying the manipulation of ascospore discharge and plant infection by SNARE proteins in F. graminearum.
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Niche-specific metabolic adaptation in biotrophic and necrotrophic oomycetes is manifested in differential use of nutrients, variation in gene content, and enzyme evolution

Niche-specific metabolic adaptation in biotrophic and necrotrophic oomycetes is manifested in differential use of nutrients, variation in gene content, and enzyme evolution | Plant pathogens and pests | Scoop.it
The use of host nutrients to support pathogen growth is central to disease. We addressed the relationship between metabolism and trophic behavior by comparing metabolic gene expression during potato tuber colonization by two oomycetes, the hemibiotroph Phytophthora infestans and the necrotroph Pythium ultimum. Genes for several pathways including amino acid, nucleotide, and cofactor biosynthesis were expressed more by Ph. infestans during its biotrophic stage compared to Py. ultimum. In contrast, Py. ultimum had higher expression of genes for metabolizing compounds that are normally sequestered within plant cells but released to the pathogen upon plant cell lysis, such as starch and triacylglycerides. The transcription pattern of metabolic genes in Ph. infestans during late infection became more like that of Py. ultimum, consistent with the former's transition to necrotrophy. Interspecific variation in metabolic gene content was limited but included the presence of γ-amylase only in Py. ultimum. The pathogens were also found to employ strikingly distinct strategies for using nitrate. Measurements of mRNA, 15N labeling studies, enzyme assays, and immunoblotting indicated that the assimilation pathway in Ph. infestans was nitrate-insensitive but induced during amino acid and ammonium starvation. In contrast, the pathway was nitrate-induced but not amino acid-repressed in Py. ultimum. The lack of amino acid repression in Py. ultimum appears due to the absence of a transcription factor common to fungi and Phytophthora that acts as a nitrogen metabolite repressor. Evidence for functional diversification in nitrate reductase protein was also observed. Its temperature optimum was adapted to each organism's growth range, and its Km was much lower in Py. ultimum. In summary, we observed divergence in patterns of gene expression, gene content, and enzyme function which contribute to the fitness of each species in its niche.
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Advances in understanding obligate biotrophy in rust fungi - Lorrain - 2019 - New Phytologist -

Advances in understanding obligate biotrophy in rust fungi - Lorrain - 2019 - New Phytologist - | Plant pathogens and pests | Scoop.it
Rust fungi (Pucciniales) are the largest group of plant pathogens and represent one of the most devastating threats to agricultural crops worldwide. Despite the economic importance of these highly specialized pathogens, many aspects of their biology remain obscure, largely because rust fungi are obligate biotrophs. The rise of genomics and advances in high‐throughput sequencing technology have presented new options for identifying candidate effector genes involved in pathogenicity mechanisms of rust fungi. Transcriptome analysis and integrated bioinformatics tools have led to the identification of key genetic determinants of host susceptibility to infection by rusts. Thousands of genes encoding secreted proteins highly expressed during host infection have been reported for different rust species, which represents significant potential towards understanding rust effector function. Recent high‐throughput in planta expression screen approaches (effectoromics) have pushed the field ahead even further towards predicting high‐priority effectors and identifying avirulence genes. These new insights into rust effector biology promise to inform future research and spur the development of effective and sustainable strategies for managing rust diseases.
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Aphid effector Me10 interacts with tomato TFT7, a 14‐3‐3 isoform involved in aphid resistance - Chaudhary - 2019 - New Phytologist -

Aphid effector Me10 interacts with tomato TFT7, a 14‐3‐3 isoform involved in aphid resistance - Chaudhary - 2019 - New Phytologist - | Plant pathogens and pests | Scoop.it
We demonstrated previously that expression of Macrosiphum euphorbiae salivary protein Me10 enhanced aphid reproduction on its host tomato (Solanum lycopersicum). However, the mechanism of action of Me10 remained elusive.
To confirm the secretion of Me10 by the aphid into plant tissues, we produced Me10 polyclonal antibodies. To identify the plant targets of Me10, we developed a tomato immune induced complementary DNA yeast two‐hybrid library and screened it with Me10 as bait. Immunoprecipitation and bimolecular fluorescence complementation (BiFC) assays were performed to validate one of the interactions in planta, and virus‐induced gene silencing was used for functional characterization in tomato.
We demonstrated that Me10 is secreted into the plant tissues and interacts with tomato 14‐3‐3 isoform 7 (TFT7) in yeast. Immunoprecipitation assays confirmed that Me10 and its homologue in Aphis gossypii, Ag10k, interact with TFT7 in planta. Further, BiFC revealed that Me10 interaction with TFT7 occurs in the plant cell cytoplasm. While silencing of TFT7 in tomato leaves did not affect tomato susceptibility to M. euphorbiae, it enhanced longevity and fecundity of A. gossypii, the non‐host aphid.
Our results suggest the model whereby TFT7 plays a role in aphid resistance in tomato and effectors of the Me10/Ag10k family interfere with TFT7 function during aphid infestation.
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Pathogen enrichment sequencing (PenSeq) enables population genomic studies in oomycetes - Thilliez - 2019 - New Phytologist -

Pathogen enrichment sequencing (PenSeq) enables population genomic studies in oomycetes - Thilliez - 2019 - New Phytologist - | Plant pathogens and pests | Scoop.it
The oomycete pathogens Phytophthora infestans and P. capsici cause significant crop losses world‐wide, threatening food security. In each case, pathogenicity factors, called RXLR effectors, contribute to virulence. Some RXLRs are perceived by resistance proteins to trigger host immunity, but our understanding of the demographic processes and adaptive evolution of pathogen virulence remains poor.
Here, we describe PenSeq, a highly efficient enrichment sequencing approach for genes encoding pathogenicity determinants which, as shown for the infamous potato blight pathogen Phytophthora infestans, make up < 1% of the entire genome.
PenSeq facilitates the characterization of allelic diversity in pathogen effectors, enabling evolutionary and population genomic analyses of Phytophthora species. Furthermore, PenSeq enables the massively parallel identification of presence/absence variations and sequence polymorphisms in key pathogen genes, which is a prerequisite for the efficient deployment of host resistance genes.
PenSeq represents a cost‐effective alternative to whole‐genome sequencing and addresses crucial limitations of current plant pathogen population studies, which are often based on selectively neutral markers and consequently have limited utility in the analysis of adaptive evolution. The approach can be adapted to diverse microbes and pathogens.
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Pathogen-induced changes in floral scent may increase honeybee-mediated dispersal of Erwinia amylovora

Pathogen-induced changes in floral scent may increase honeybee-mediated dispersal of Erwinia amylovora | Plant pathogens and pests | Scoop.it
Honeybees are well recognised for their key role in plant reproduction as pollinators. On the other hand, their activity may vector some pathogens, such as the bacterium Erwinia amylovora, the causative agent of fire blight disease in pomaceous plants. In this research, we evaluated whether honeybees are able to discriminate between healthy and E. amylovora-infected flowers, thus altering the dispersal of the pathogen. For this reason, honeybees were previously trained to forage either on inoculated or healthy (control) apple flower. After the training, the two honeybee groups were equally exposed to inoculated and control flowering apple plants. To assess their preference, three independent methods were used: (1) direct count of visiting bees per time frame; (2) incidence on apple flowers of a marker bacterium (Pantoea agglomerans, strain P10c) carried by foragers; (3) quantification of E. amylovora populations in the collected pollen loads, proportional to the number of visits to infected flowers. The results show that both honeybee groups preferred control flowers over inoculated ones. The characterisation of volatile compounds released by flowers revealed a different emission of several bioactive compounds, providing an explanation for honeybee preference. As an unexpected ecological consequence, the influence of infection on floral scent increasing the visit rate on healthy flowers may promote a secondary bacterial spread.
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Marine fungi - ScienceDirect

Marine fungi - ScienceDirect | Plant pathogens and pests | Scoop.it
Fungi play a dominant role in terrestrial environments where they thrive in symbiotic associations with plants and animals and are integral to nutrient cycling in diverse ecosystems. Everywhere that moisture and a carbon source coexist in the terrestrial biosphere, fungi are expected to occur. We know that fungi can be devastating to agricultural crops, both in the field and during their storage, and cause mortality in immunocompromised patients in numbers that rival the deaths from malaria. Yet fungi can also be harnessed as sources of food, chemicals and biofuels when humans exploit fungal metabolism. Despite their central role in the health and disease of the terrestrial biosphere, much less is known about the function and potential of marine fungi. Are fungi ubiquitous in marine environments as they are on land? Do they play the same or similar roles in these ecosystems? Here we describe the state of knowledge about the abundance and functions of fungi in the marine environment with a goal to stimulate new inquiry in this very open area.
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