Suppress Plant Defenses Research Articles

Trichoderma virens Bys1 may competitively inhibit its own effector protein Alt a 1 to stabilize the symbiotic relationship with plant-evidence from docking and simulation studies.

The online version contains supplementary material available at 10.1007/s13205-021-02652-8.

HaCRT1 of Heterodera avenae Is Required for the Pathogenicity of the Cereal Cyst Nematode.

Cereal cyst nematodes are sedentary biotrophic endoparasites that secrete effector proteins into plant tissues to transit normal cells into specialized feeding sites and suppress plant defenses. To understand the function of nematode effectors in Heterodera avenae, here, we identified a calreticulin protein HaCRT1, which could suppress the cell death induced by Bax when expressed in Nicotiana benthamiana. HaCRT1 is synthetized in the subventral gland cells of pre-parasitic second-stage nematodes. Real-time PCR assays indicated that the expression of HaCRT1 was highest in parasitic second-stage juveniles. The expression of an HaCRT1-RFP fusion in N. benthamiana revealed that it was localized in the endoplasmic reticulum of the plant cell. The ability of H. avenae infecting plants was significantly reduced when HaCRT1 was knocked down by RNA interference in vitro. Arabidopsis thaliana plants expressing HaCRT1 were more susceptible than wild-type plants to Pseudomonas syringae. The induction of defense-related genes, PAD4, WRKY33, FRK1, and WRKY29, after treatment with flg22 was suppressed in HaCRT1-transgenic plants. Also, the ROS accumulation induced by flg22 was reduced in the HaCRT1-transgenic plants compared to wild-type plants. HaCRT1 overexpression increased the cytosolic Ca2+ concentration in A. thaliana. These data suggested that HaCRT1 may contribute to the pathogenicity of H. avenae by suppressing host basal defense.

Magnaporthe oryzae nucleoside diphosphate kinase is required for metabolic homeostasis and redox-mediated host innate immunity suppression.

The fungus Magnaporthe oryzae causes blast, the most devastating disease of cultivated rice. After penetrating the leaf cuticle, M. oryzae grows as a biotroph in intimate contact with living rice epidermal cells before necrotic lesions develop. Biotrophic growth requires maintaining metabolic homeostasis while suppressing plant defenses, but the metabolic connections and requirements involved are largely unknown. Here, we characterized the M. oryzae nucleoside diphosphate kinase-encoding gene NDK1 and discovered it was essential for facilitating biotrophic growth by suppressing the host oxidative burst-the first line of plant defense. NDK enzymes reversibly transfer phosphate groups from tri- to diphosphate nucleosides. Correspondingly, intracellular nucleotide pools were perturbed in M. oryzae strains lacking NDK1 through targeted gene deletion, compared to WT. This affected metabolic homeostasis: TCA, purine and pyrimidine intermediates, and oxidized NADP+ , accumulated in Δndk1. cAMP and glutathione were depleted. ROS accumulated in Δndk1 hyphae. Functional appressoria developed on rice leaf sheath surfaces, but Δndk1 invasive hyphal growth was restricted and redox homeostasis was perturbed, resulting in unsuppressed host oxidative bursts that triggered immunity. We conclude Ndk1 modulates intracellular nucleotide pools to maintain redox balance via metabolic homeostasis, thus quenching the host oxidative burst and suppressing rice innate immunity during biotrophy.

Trichoderma virens Alt a 1 protein may target maize PR5/thaumatin-like protein to suppress plant defence: An in silico analysis

Trichoderma spp. are plant beneficial fungi that colonize roots, but unlike plant pathogens, impart defence. Recently, however, some small secreted cysteine-rich proteins have been identified in Trichoderma secretome, deletion of which improves induced defence. The mechanism of such “unusual” behaviour of a plant-beneficial fungus is not understood. In the present study, through in silico analysis, we attempted to explore if such proteins indeed suppress defence by interacting with plant proteins, much like pathogens, taking Alt a 1 protein of T. virens and maize PR5, a thaumatin-like defence protein as predicted interacting partners (with cue from some plant-pathogen interactions). Using computational analysis, we test the hypothesis that Trichoderma Alt a 1 inactivates maize PR5 to suppress plant defence in order to be able to colonize roots. The tertiary structures of both Alt a 1 and PR5 were modelled through homology as well as de novo modelling, followed by model validation through different quality check tools. Prior to protein-protein docking, MD (molecular dynamics) optimisation and refinement of computational models were carried-out. Docking results indicated that Alt a 1 exhibits strong binding affinity with PR5 suggesting that Alt a 1 and PR5 might interact much like in a plant-pathogen interactions resulting in suppression of plant defence to facilitate colonization.

The large, diverse, and robust arsenal of Ralstonia solanacearum type III effectors and their in planta functions.

The type III secretion system with its delivered type III effectors (T3Es) is one of the main virulence determinants of Ralstonia solanacearum, a worldwide devastating plant pathogenic bacterium affecting many crop species. The pan‐effectome of the R. solanacearum species complex has been exhaustively identified and is composed of more than 100 different T3Es. Among the reported strains, their content ranges from 45 to 76 T3Es. This considerably large and varied effectome could be considered one of the factors contributing to the wide host range of R. solanacearum. In order to understand how R. solanacearum uses its T3Es to subvert the host cellular processes, many functional studies have been conducted over the last three decades. It has been shown that R. solanacearum effectors, as those from other plant pathogens, can suppress plant defence mechanisms, modulate the host metabolism, or avoid bacterial recognition through a wide variety of molecular mechanisms. R. solanacearum T3Es can also be perceived by the plant and trigger immune responses. To date, the molecular mechanisms employed by R. solanacearum T3Es to modulate these host processes have been described for a growing number of T3Es, although they remain unknown for the majority of them. In this microreview, we summarize and discuss the current knowledge on the characterized R. solanacearum species complex T3Es.

Can the plant immune system be activated against animal parasites such as nematodes?

Summary Plants have an immune system that can be exerted by every cell against a vast array of pathogens and parasites. This immune system has several levels of specialisation. A first level induces a general basal defence, after which specific pathogen-associated molecular patterns (PAMPs) are recognised by plant receptors. Normally, this basal defence is overcome by specialised pests by secreting specific effectors that suppress this first line of defence. Plant-parasitic nematodes have been found to secrete several such effectors through amphid and cuticle secretions or directly into the cells through their stylets. Many studies carried out on endoparasitic sedentary nematodes (ESN), in particular, reported the suppression of plant defence, especially in terms of down-regulation of the genes involved in salicylic acid (SA)-mediated defence, in the very first stages of plant-nematode interaction. However, plants can be immunised or primed against nematode attacks. Immunised plants respond to nematode infection more rapidly and efficiently and let a minor number of invasive juveniles become sedentary, develop to gravid females and reproduce. Immunisation of plants can be achieved by pre-treatments with chemicals that are functional analogues of the phytohormones that mediate defence reactions, such as salicylic acid (SA). Moreover, plants provided with the beneficial microorganisms commonly present in a healthy rhizosphere, such as arbuscular mycorrhizal fungi or opportunistic symbiotic fungi, have been found to be immunised against ESN. Immunisation is mainly mediated by up-regulation of SA-dependent defence genes, such as pathogenesis-related genes (PR-genes). Immunisation is systemic and can also work against herbivorous insects.

A Bursaphelenchus xylophilus effector, Bx-FAR-1, suppresses plant defense and affects nematode infection of pine trees

Bursaphelenchus xylophilus is an economically important pathogen that has caused serious damage to pine forests. Fatty acid and retinol-binding proteins (FARs) are nematode-specific proteins. One FAR gene, Bx-FAR-1, was identified from the transcriptome of B. xylophilus as a putative effector. It was upregulated in both highly and weakly virulent isolates in early infection stages. Transient expression assays showed that Bx-FAR-1 inhibited BAX- and INF1-induced cell death in Nicotiana benthamiana when secreted into the intracellular site. Bx-FAR-1 was expressed in the glandular tissues, intestines, and seminal vesicles of nematodes by in situ hybridization. Silencing Bx-FAR-1 via in vitro RNA interference attenuated the reproductive ability and pathogenicity of B. xylophilus as well as increased the expression of the pathogenesis-related gene 6 and JA biosynthesis gene LOX-5 (lipoxygenase-5) in pine trees. These results suggest that Bx-FAR-1 suppresses the plant immune response in the N. benthamiana intracellular site and may contribute to the pathogenicity of B. xylophilus in the early infection stages.

Resin acids as inducible chemical defences of pine seedlings against chewing insects.

Inducibility of defences in response to biotic stimuli is considered an important trait in plant resistance. In conifers, previous research has mostly focused on the inducibility of the volatile fraction of the oleoresin (mono- and sesquiterpenes), leaving the inducibility of the non-volatile resin acids largely unexplored, particularly in response to real herbivory. Here we investigated the differences in the inducibility of resin acids in two pine species, one native from Europe (Pinus pinaster Ait.) and another from North America (Pinus radiata D. Don), in response to wounding by two European insects: a bark chewer, the pine weevil (Hylobius abietis L.), and a defoliator, the pine processionary caterpillar (Thaumetopoea pityocampa Schiff.). We quantified the constitutive (control) and induced concentrations of resin acids in the stem and needles of both pine species by gas chromatography techniques. Both pine species strongly increased the concentration of resin acids in the stem after pine weevil feeding, although the response was greater in P. pinaster than in P. radiata. However, systemic defensive responses in the needles were negligible in both pine species after pine weevil feeding in the stem. On the other hand, P. radiata locally reduced the resin acid concentration in the needles after pine caterpillar feeding, whereas in P. pinaster resin acid concentration was apparently unaffected. Nevertheless, systemic induction of resin acids was only observed in the stem of P. pinaster in response to pine caterpillar feeding. In summary, pine induced responses were found highly compartmentalized, and specific to herbivore identity. Particularly, plant defence suppression mechanisms by the pine caterpillar, and ontogenetic factors might be potentially affecting the induced response of resin acids in both pine species.

A novel effector CfEC92 of Colletotrichum fructicola contributes to glomerella leaf spot virulence by suppressing plant defences at the early infection phase.

The ascomycete fungus Colletotrichum fructicola causes diseases on a broad range of plant species. On susceptible cultivars of apple, it induces severe early defoliation and fruit spots, named glomerella leaf spot (GLS), but the mechanisms of pathogenicity have remained elusive. Phytopathogens exhibit small secreted effectors to advance host infection by manipulating host immune reactions. We report the identification and characterization of CfEC92, an effector required for C. fructicola virulence. CfEC92 is a Colletotrichum‐specific small secreted protein that suppresses BAX‐triggered cell death in Nicotiana benthamiana. Accumulation of the gene transcript was barely detectable in conidia or vegetative hyphae, but was highly up‐regulated in appressoria formed during early apple leaf infection. Gene deletion mutants were not affected in vegetative growth, appressorium formation, or appressorium‐mediated cellophane penetration. However, the mutants were significantly reduced in virulence toward apple leaves and fruits. Microscopic examination indicated that infection by the deletion mutants elicited elevated deposition of papillae at the penetration sites, and formation of infection vesicles and primary hyphae was retarded. Signal peptide activity, subcellular localization, and cell death‐suppressive activity (without signal peptide) assays suggest that CfEC92 could be secreted and perform virulence functions inside plant cells. RNA sequencing and quantitative reverse transcription PCR results confirmed that the deletion mutants triggered elevated host defence reactions. Our results strongly support the interpretation that CfEC92 contributes to C. fructicola virulence as a plant immunity suppressor at the early infection phase.

LysM Proteins Regulate Fungal Development and Contribute to Hyphal Protection and Biocontrol Traits in Clonostachys rosea.

Lysin motif (LysM) modules are approximately 50 amino acids long and bind to peptidoglycan, chitin and its derivatives. Certain LysM proteins in plant pathogenic and entomopathogenic fungi are shown to scavenge chitin oligosaccharides and thereby dampen host defense reactions. Other LysM proteins can protect the fungal cell wall against hydrolytic enzymes. In this study, we investigated the biological function of LysM proteins in the mycoparasitic fungus Clonostachys rosea. The C. rosea genome contained three genes coding for LysM-containing proteins and gene expression analysis revealed that lysm1 and lysm2 were induced during mycoparasitic interaction with Fusarium graminearum and during colonization of wheat roots. Lysm1 was suppressed in germinating conidia, while lysm2 was induced during growth in chitin or peptidoglycan-containing medium. Deletion of lysm1 and lysm2 resulted in mutants with increased levels of conidiation and conidial germination, but reduced ability to control plant diseases caused by F. graminearum and Botrytis cinerea. The Δlysm2 strain showed a distinct, accelerated mycelial disintegration phenotype accompanied by reduced biomass production and hyphal protection against hydrolytic enzymes including chitinases, suggesting a role of LYSM2 in hyphal protection against chitinases. The Δlysm2 and Δlysm1Δlysm2 strains displayed reduced ability to colonize wheat roots, while only Δlysm1Δlysm2 failed to suppress expression of the wheat defense response genes PR1 and PR4. Based on our data, we propose a role of LYSM1 as a regulator of fungal development and of LYSM2 in cell wall protection against endogenous hydrolytic enzymes, while both are required to suppress plant defense responses. Our findings expand the understanding of the role of LysM proteins in fungal-fungal interactions and biocontrol.

Tetranychus evansi spider mite populations suppress tomato defenses to varying degrees.

Plant defense suppression is an offensive strategy of herbivores, in which they manipulate plant physiological processes to increase their performance. Paradoxically, defense suppression does not always benefit the defense‐suppressing herbivores, because lowered plant defenses can also enhance the performance of competing herbivores and can expose herbivores to increased predation. Suppression of plant defense may therefore entail considerable ecological costs depending on the presence of competitors and natural enemies in a community. Hence, we hypothesize that the optimal magnitude of suppression differs among locations. To investigate this, we studied defense suppression across populations of Tetranychus evansi spider mites, a herbivore from South America that is an invasive pest of solanaceous plants including cultivated tomato, Solanum lycopersicum, in other parts of the world. We measured the level of expression of defense marker genes in tomato plants after infestation with mites from eleven different T. evansi populations. These populations were chosen across a range of native (South American) and non‐native (other continents) environments and from different host plant species. We found significant variation at three out of four defense marker genes, demonstrating that T. evansi populations suppress jasmonic acid‐ and salicylic acid‐dependent plant signaling pathways to varying degrees. While we found no indication that this variation in defense suppression was explained by differences in host plant species, invasive populations tended to suppress plant defense to a smaller extent than native populations. This may reflect either the genetic lineage of T. evansi—as all invasive populations we studied belong to one linage and both native populations to another—or the absence of specialized natural enemies in invasive T. evansi populations.

Sting nematodes modify metabolomic profiles of host plants

Plant-parasitic nematodes are devastating pathogens of many important agricultural crops. They have been successful in large part due to their ability to modify host plant metabolomes to their benefit. Both root-knot and cyst nematodes are endoparasites that have co-evolved to modify host plants to create sophisticated feeding cells and suppress plant defenses. In contrast, the ability of migratory ectoparasitic nematodes to modify host plants is unknown. Based on global metabolomic profiling of sting nematodes in African bermudagrass, ectoparasites can modify the global metabolome of host plants. Specifically, sting nematodes suppress amino acids in susceptible cultivars. Upregulation of compounds linked to plant defense have negative impacts on sting nematode population densities. Pipecolic acid, linked to systemic acquired resistance induction, seems to play a large role in protecting tolerant cultivars from sting nematode feeding and could be targeted in breeding programs.

An aphid effector promotes barley susceptibility through suppression of defence gene expression.

Aphids secrete diverse repertoires of effectors into their hosts to promote the infestation process. While 'omics' approaches facilitated the identification and comparison of effector repertoires from a number of aphid species, the functional characterization of these proteins has been limited to dicot (model) plants. The bird cherry-oat aphid Rhopalosiphum padi is a pest of cereal crops, including barley. Here, we extend efforts to characterize aphid effectors with regard to their role in promoting susceptibility to the R. padi-barley interaction. We selected three R. padi effectors based on sequence similarity to previously characterized Myzus persicae effectors and assessed their subcellular localization, expression, and role in promoting plant susceptibility. Expression of R. padi effectors RpC002 and Rp1 in transgenic barley lines enhanced plant susceptibility to R. padi but not M. persicae, for which barley is a poor host. Characterization of Rp1 transgenic barley lines revealed reduced gene expression of plant hormone signalling genes relevant to plant-aphid interactions, indicating that this effector enhances susceptibility by suppressing plant defences in barley. Our data suggest that some aphid effectors specifically function when expressed in host species, and feature activities that benefit their corresponding aphid species.

The Unfolded Protein Response Regulates Pathogenic Development of Ustilago maydis by Rok1-Dependent Inhibition of Mating-Type Signaling.

Fungal pathogens require the unfolded protein response (UPR) to maintain protein homeostasis of the endoplasmic reticulum (ER) during pathogenic development. In the corn smut fungus Ustilago maydis, pathogenic development is controlled by the a and b mating-type loci. The UPR is specifically activated after plant penetration and required for efficient secretion of effectors and suppression of the plant defense response. The interaction between the UPR regulator Cib1 and the central developmental regulator Clp1 modulates the pathogenic program and triggers fungal colonization of the host plant. By contrast, when activated before plant penetration, the UPR interferes with fungal virulence by reducing expression of bE and bW, the central regulators of pathogenic development encoded by the b mating-type locus. Here, we show that this inhibitory effect results from UPR-mediated suppression of the pheromone response pathway upstream of the b regulatory network. UPR activity prompts dephosphorylation of the pheromone-responsive mitogen-activated protein kinase (MAPK) Kpp2, reducing activity of the pheromone response factor Prf1 that regulates expression of bE and bW Deletion of the dual specificity phosphatase rok1 fully suppressed UPR-dependent inhibition of Kpp2 phosphorylation, formation of infectious filaments, and fungal virulence. Rok1 determines the activity of mating-type signaling pathways and thus the degree of fungal virulence. We propose that UPR-dependent regulation of Rok1 aligns ER physiology with fungal aggressiveness and effector gene expression during biotrophic growth of U. maydis in the host plant.IMPORTANCE The unfolded protein response (UPR) is crucial for endoplasmic reticulum (ER) homeostasis and disease development in fungal pathogens. In the plant-pathogenic fungus Ustilago maydis, the UPR supports fungal proliferation in planta and effector secretion for plant defense suppression. In this study, we uncovered that UPR activity, which is normally restricted to the biotrophic stage in planta, inhibits mating and the formation of infectious filaments by Rok1-dependent dephosphorylation of the pheromone responsive mitogen-activated protein kinase (MAPK) Kpp2. This observation is relevant for understanding how the fungal virulence program is regulated by cellular physiology. UPR-mediated control of mating-type signaling pathways predicts that effector gene expression and the virulence potential are controlled by ER stress levels.

Conditional gene expression reveals stage-specific functions of the unfolded protein response in the Ustilago maydis-maize pathosystem.

Summary Ustilago maydis is a model organism for the study of biotrophic plant–pathogen interactions. The sexual and pathogenic development of the fungus are tightly connected since fusion of compatible haploid sporidia is prerequisite for infection of the host plant, maize (Zea mays). After plant penetration, the unfolded protein response (UPR) is activated and required for biotrophic growth. The UPR is continuously active throughout all stages of pathogenic development in planta. However, since development of UPR deletion mutants stops directly after plant penetration, the role of an active UPR at later stages of development remained to be determined. Here, we established a gene expression system for U. maydis that uses endogenous, conditionally active promoters to either induce or repress expression of a gene of interest during different stages of plant infection. Integration of the expression constructs into the native genomic locus and removal of resistance cassettes were required to obtain a wild‐type‐like expression pattern. This indicates that genomic localization and chromatin structure are important for correct promoter activity and gene expression. By conditional expression of the central UPR regulator, Cib1, in U. maydis, we show that a functional UPR is required for continuous plant defence suppression after host infection and that U. maydis relies on a robust control system to prevent deleterious UPR hyperactivation.

Colorado potato beetle microsymbiont Enterobacter BC-8 inhibits defense mechanisms of potato plants using crosstalk between jasmonate- and salicylate-mediated signaling pathways

One of the main insect potato pests is the Colorado potato beetle (Leptinotarsa decemlineata Say). It contains some microbial associates which can affect diverse interactions between host plant and the herbivorous insect. Previously, the most common L. decemlineata microsymbiont isolated from anterior and posterior parts of beetle gut was defined as Enterobacter BC-8. The role of Enterobacter BC-8 in manipulating plant defenses was investigated using antibiotics-treated beetles and model system simulating beetles attacks (wounding plants treated with Enterobacter BC-8 suspension). We demonstrated that the symbiotic bacteria suppressed plant defenses such as hydrogen peroxide and phenolic compounds accumulation and activity of peroxidases and trypsin inhibitors. It is worth noting that the influence of the insect symbionts on potato plants stimulated salicylate-sensitive genes and the marker of salicylate signaling pathway. Transcription activities of jasmonate-sensitive genes which encode some defense proteins against herbivores, were suppressed. So, Enterobacter BC-8 plays the role in salicylate/jasmonate crosstalks manipulating to suppress plant defense mechanisms.

Eurydema oleracea negatively affects defenses in Arabidopsis by inducing salicylic acid-mediated signaling pathway

The study of defense mechanisms of plants against herbivorous insects can clarify the evolutionary mechanisms of these interactions and has significant implications for agriculture. The herbivore Eurydema oleracea is an invasive crop pest; however, knowledge on how it affects plant immune responses is lacking. In the present study, we demonstrate that insect feeding causes the induction of both salicylic acid (SA) and jasmonic acid (JA)-mediated signaling pathways in infested Arabidopsis plants. Using transgenic SA-deficient NahG, we show that the two phytohormones crosstalk antagonistically. In particular, the rapid and strong induction of SA-related gene expression partially suppresses the JA-dependent signaling pathway. This results in increased plant susceptibility to the herbivore, evidenced by an increase of leaf damage inflicted by the adult insects and increased development of the nymphs. Our findings suggest that E. oleracea manipulates hormone signaling components of Arabidopsis as a strategy of attack to suppress plant defense traits. Our work contributes to knowledge on the adaptation of insect pests to plant responses, and is useful for developing management strategies to combat harmful herbivores in agriculture.

Candidate Effectors From Uromyces appendiculatus, the Causal Agent of Rust on Common Bean, Can Be Discriminated Based on Suppression of Immune Responses.

Rust fungi are devastating pathogens for several important crop plants. The biotrophic lifestyle of rust fungi requires that they influence their host plants to create a favorable environment for growth and reproduction. Rust fungi secrete a variety of effector proteins that manipulate host target proteins to alter plant metabolism and suppress defense responses. Because of the obligate biotrophic lifestyle of rust fungi, direct evidence for effector function is difficult to obtain, and so suites of experiments utilizing expression in heterologous systems are necessary. Here, we present results from a yeast cell death suppression assay and assays for suppression of PAMP-triggered immunity (PTI) and effector triggered immunity (ETI) based on delivery of effectors through the bacterial type III secretion system. In addition, subcellular localization was tested using transient expression of GFP fusion proteins in Nicotiana benthamiana through Agrobacterium infiltration. We tested 31 representative effector candidates from the devastating common bean rust pathogen Uromyces appendiculatus. These effector candidates were selected based on features of their gene families, most important lineage specificity. We show that several of our effector candidates suppress plant defense. Some of them also belong to families of effector candidates that are present in multiple rust species where their homologs probably also have effector functions. In our analysis of candidate effector mRNA expression, some of those effector candidates that gave positive results in the other assays were not up-regulated during plant infection, indicating that either these proteins have functions at multiple life stages or that strong up-regulation of RNA level in planta may not be as important a criterion for identifying effectors as previously thought. Overall, our pipeline for selecting effector candidates based on sequence features followed by screening assays using heterologous expression systems was successful in discriminating effector candidates. This work lays the foundation for functional characterization of U. appendiculatus effectors, the identification of effector targets, and identification of novel sources for resistance in common bean.

Draft genome sequence of fastidious pathogen Ceratobasidium theobromae, which causes vascular-streak dieback in Theobroma cacao

BackgroundCeratobasidium theobromae, a member of the Ceratobasidiaceae family, is the causal agent of vascular-streak dieback (VSD) of cacao, a major threat to the chocolate industry in the South-East Asia. The fastidious pathogen is very hard to isolate and maintain in pure culture, which is a major bottleneck in the study of its genetic diversity and genome.ResultThis study describes for the first time, a 33.90 Mbp de novo assembled genome of a putative C. theobromae isolate from cacao. Ab initio gene prediction identified 9264 protein-coding genes, of which 800 are unique to C. theobromae when compared to Rhizoctonia spp., a closely related group. Transcriptome analysis using RNA isolated from 4 independent VSD symptomatic cacao stems identified 3550 transcriptionally active genes when compared to the assembled C. theobromae genome while transcripts for only 4 C. theobromae genes were detected in 2 asymptomatic stems. De novo assembly of the non-cacao associated reads from the VSD symptomatic stems uniformly produced genes with high identity to predicted genes in the C. theobromae genome as compared to Rhizoctonia spp. or genes found in Genbank. Further analysis of the predicted C. theobromae transcriptome was carried out identifying CAZy gene classes, KEGG-pathway associated genes, and 138 putative effector proteins.ConclusionThese findings put forth, for the first time, a predicted genome for the fastidious basidiomycete C. theobromae causing VSD on cacao providing a model for testing and comparison in the future. The C. theobromae genome predicts a pathogenesis model involving secreted effector proteins to suppress plant defense mechanisms and plant cell wall degrading enzymes.

The Heterodera glycines effector Hg16B09 is required for nematode parasitism and suppresses plant defense response

Soybean cyst nematode (Heterodera glycines Ichinohe) is a sedentary root endoparasite that causes serious yield losses on soybean (Glycine max) worldwide. H. glycines secrets effector proteins into host cells to facilitate the success of parasitism. Nowadays, a large number of candidate effectors were identified from the genome sequence of H. glycines. However, the precise functions of these effectors in the nematode-host plant interaction are unknown. Here, an effector gene of dorsal gland protein Hg16B09 from H. glycines was cloned and functionally characterized through generating the transgenic soybean hairy roots. In situ hybridization assay and qRT-PCR analysis indicated Hg16B09 is exclusively expressed in the dorsal esophageal cells and up-regulated in the parasitic-stage juveniles. The constitutive expression of Hg16B09 in soybean hairy roots caused an enhanced susceptibility to H. glycines. In contrast, in planta silencing of Hg16B09 exhibited that nematode reproduction in hairy roots was decreased compared to the empty vector control. In addition, Hg16B09 also suppressed the expression of soybean defense-related genes induced by the pathogen-associated molecular pattern flg22. These data indicate that the effector Hg16B09 might aid H. glycines parasitism through suppressing plant basal defenses in the early parasitic stages.