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Danchin, E. G. J.

Publications and source records attributed to Danchin, E. G. J..

9 recordsLinked to original sources

The host range paradox of Meloidogyne incognita: a physiological and transcriptomic analysis of nine susceptible interactions across six plant orders

The plant-parasitic nematode Meloidogyne incognita is the pathogen with the broadest host range among all known biotrophic interactions. This species is also the single most damaging of a group of agriculturally important plant-parasites, which together are estimated to contribute to losses in excess of $170 billion/year to world agriculture. Understanding how M. incognita is able to infect representatives from most orders of flowering plants, covering more than 3000 species, addresses a fundamentally important question of how pathogens adapt to their host-environment, and may inform control of a pathogen which threatens global food security. Here, we analyse the plant-nematode infection phenotype, and cross-kingdom transcriptome, of nine interactions across six orders of flowering plants at 25 days post infection. At this stage, majority of nematodes found within roots were immature and mature females (49.2% - 91.8%). Our data show that the phylogenetic distribution of hosts does not explain the phenotypic distribution of parasitism. Interestingly, though, M. incognita do have distinct transcriptional responses to different groups of hosts, but in a pattern which is independent of host phylogenetic. Three distinct nematode "transcriptional programmes" - Group 1, 2, and 3 - are evident, and we find that effectors are neither uniformly deployed across hosts, nor across groups of hosts. Importantly, we show that this differential deployment of effectors can have profound consequences for host specificity. Finally, we show that there is essentially no widespread core gall transcriptome at 25 days post infection, prompting the proposal of a model best described as "all roads lead to Rome".

plant biology↗

DNA-protein interactions in Meloidogyne incognita

BackgroundThe root-knot nematode Meloidogyne incognita, is a highly destructive parasite that manipulates host plant processes through effector proteins, affecting agriculture globally. Despite advances in genomic and transcriptomic studies, the regulatory mechanisms controlling effector gene expression, especially at the chromatin level, are still poorly understood. Gene regulation studies in plant-parasitic nematodes (PPN) face several challenges, including the absence of transformation systems and technical barriers in chromatin preparation, particularly for transcription factors (TFs) expressed in secretory gland cells. Conventional methods like Chromatin Immunoprecipitation (ChIP) are limited in PPN due to low chromatin yields, the impermeability of nematode cuticles, and difficulties in producing antibodies for low-abundance TFs. These issues call for alternative approaches, such as dCas9-based CAPTURE (CRISPR Affinity Purification in siTU of Regulatory Elements) that allows studying chromatin interactions by using a catalytically inactive dCas9 protein to target specific genomic loci without relying on antibodies. ResultsThis study presents an optimized in vitro dCas9-based CAPTURE for M. incognita that addresses key challenges in chromatin extraction and stability. The protocol focuses on the promoter region of the effector gene 6F06, a critical gene for parasitism. Several optimizations were made, including improvements in nematode disruption, chromatin extraction, and protein-DNA complex stability. This method successfully isolated chromatin-protein complexes and identified four putative chromatin-associated proteins, including BANF1, linked to chromatin remodelling complexes like SWI/SNF. ConclusionThe optimized in vitro dCas9-based CAPTURE protocol offers a new tool for investigating chromatin dynamics and regulatory proteins in non-transformable nematodes. This method expands the scope of effector gene regulation research and provides new insights into parasitism in M. incognita. Future research will aim to validate these regulatory proteins and extend the method to other effector loci, potentially guiding the development of novel nematode control strategies.

molecular biology↗

Genomic rearrangements promote diversification of a facultative meiotic parthenogenetic nematode pest (Meloidogyne graminicola)

Meloidogyne graminicola (Mg), commonly known as the rice root-knot nematode, is a highly destructive pest that inflicts significant damage on rice crops worldwide. Mg is thought to reproduce primarily by meiotic parthenogenesis, but its success across diverse habitats and hosts raises important questions about its adaptation mechanisms, particularly those driving the evolution of its virulence. Documenting the origin of the pathogen, its reproductive strategies and other evolutionary processes shaping its genome are thus crucial to understand its recent, rapid expansion. In this study, we first improved gene annotations of the Mg genome to enhance identification of potential secreted parasitism genes. Next, comparative genomics analyses of 13 Mg isolates from diverse geographic locations revealed evolutionary changes in the genome, including single nucleotide variations (SNVs), loss of heterozygosity (LoH), and copy number variations (CNVs). These events affected a substantial number of genes, including those coding for secreted proteins, suggesting their roles in nematode adaptation. LoH, the reduction of linkage disequilibrium between SNPs with distance as well as the 4-gamete test all provide evidence for meiotic recombination, supporting some sexual reproduction in Mg. Clustering of populations, based on LoH profiles and SNVs, allowed the definition of groups of isolates not correlating with current geographic distribution. The low sequence divergence at the genome level and the lack of clear phylogeographic structure among isolates support the hypothesis of a recent, widespread dissemination of the parasite, especially across Southeast Asia. Overall, our study supports a dual reproductive mode (sexual/asexual) in Mg, which offers an evolutionary advantage by balancing clonal proliferation in favorable conditions with occasional sexual reproduction allowing generation of new allelic combinations in adverse environments.

genomics↗

Identification and characterization of specific motifs in effector proteins of plant parasites using MOnSTER.

Plant pathogens cause billions of dollars of crop loss every year and are a major threat to global food security. Identifying and characterizing pathogens effectors is crucial towards their improved control. Because of their poor sequence conservation, effector identification in protein sequences predicted from genomes is challenging and current methods generate too many candidates without indication for prioritizing further experimental studies. In most phyla, effectors contain specific sequence motifs which influence their localization and targets in the plant. Although bacterial, fungal and oomycetes effectors have been studied extensively and conserved characteristic motifs have been identified, research on plant-parasitic nematode effectors (PPN) identified some enriched degenerate motifs in only one species so far. The different lifestyles of PPNs might reflect effectors with different functions according to the nematodes specific needs, thus presenting a high variety of characteristic motifs. To circumvent these limitations, we have developed MOnSTER a novel tool that identifies clusters of motifs of protein sequences (CLUMPs). MOnSTER can be fed with motifs identified by de novo tools or from databases such as Pfam and InterProScan. The advantage of MOnSTER is the reduction of motif redundancy by clustering them and associating a score. This score encompasses the physicochemical properties of AAs and the motif occurrences. We built up our method to identify discriminant CLUMPs in candidate parasitism proteins of plant-pathogenic oomycetes. We showed the reliability of MOnSTER by identifying five CLUMPs that correspond to the known motifs: RxLR, -dEER and LxLFLAK-HVLVxxP. Consequently, we applied MOnSTER on PPN candidate parasitism proteins and identified peculiar motifs in their sequences. We identified six CLUMPs in about 60% of the known nematode candidate parasitism proteins. Furthermore, we found that specific co-occurrences of at least two CLUMPs are present in PPN candidate parasitism protein sequences bearing protein domains important for invasion and pathogenicity. The potentiality of this tool goes beyond the candidate parasitism proteins and can be used to easily cluster motifs and calculate the CLUMP-score on any set of protein sequences. Authors summaryPopulation growth, environmental degradation and climate change are already bringing harm to human communities and the natural world that needs to be addressed rapidly. Ensuring food security for a population that will exceed 9 billion people by 2050 while preserving the environment and biodiversity is a major challenge. Agricultural pathogens, to cause the infection, secrete effector proteins that promote colonization of the host plant. Identifying and characterizing pathogens effectors is crucial towards understanding how they manipulate the plant and better combat them. Because of their poor sequence conservation, effector identification in protein sequences predicted from genomes is challenging and current methods generate too many candidates without indication for prioritizing further experimental studies. To address these challenges, we have developed a novel tool called MOnSTER, that identifies and score clusters of motifs of protein sequences (CLUMPs). MOnSTER is an easy to use tool that can be included in any pipeline needing motif calling and will be of great use to accelerate both computational and experimental studies relating to protein motif discovery. Altogether our findings provide improvements in the understanding of the mechanisms set up by the pathogens to infect the plant and can elucidate important signatures to block the development of plant-pathogen interactions and allow to engineer of durable disease resistance.

bioinformatics↗

Ionizing radiation responses are incidental to desiccation responses in the bdelloid rotifer Adineta vaga

BackgroundThe remarkable resistance to ionizing radiation found in anhydrobiotic organisms, such as some bacteria, tardigrades, and bdelloid rotifers has been hypothesized to be incidental to the desiccation resistance. Both stresses produce reactive oxygen species and cause damage to DNA and other macromolecules. However, this hypothesis has only been investigated in a few species. ResultsIn this study, we analyzed the transcriptomic response of the bdelloid rotifer Adineta vaga to desiccation and to low- (X-rays) and high- (Fe) LET radiation to highlight the molecular and genetic mechanisms triggered by both stresses. We identified numerous genes encoding antioxidants, but also chaperones, that are constitutively highly expressed, which may contribute to the protection of proteins against oxidative stress during desiccation and ionizing radiation. We also detected a transcriptomic response common to desiccation and ionizing radiation with the over-expression of genes mainly involved in DNA repair and protein modifications but also genes with unknown functions being bdelloid-specific. A distinct transcriptomic response specific to rehydration was also found, with the over-expression of genes mainly encoding Late Embryogenesis Abundant proteins, specific Heat Shock Proteins, and glucose repressive proteins. ConclusionsThese results suggest that the extreme resistance of bdelloid rotifers to radiation might indeed be a consequence of their capacity to resist complete desiccation. This study paves the way to functional genetic experiments on A. vaga targeting promising candidate proteins playing central roles in radiation and desiccation resistance.

molecular biology↗

AvP: a software package for automatic phylogenetic detection of candidate horizontal gene transfers.

Horizontal gene transfer (HGT) is the transfer of genes between species outside the transmission from parent to offspring. Due to their impact on the genome and biology of various species, HGTs have gained broader attention, but high-throughput methods to robustly identify them are lacking. One rapid method to identify HGT candidates is to calculate the difference in similarity between the most similar gene in closely related species and the most similar gene in distantly related species. Although metrics on similarity associated with taxonomic information can rapidly detect putative HGTs, these methods are hampered by false positives that are difficult to track. Furthermore, they do not inform on the evolutionary trajectory and events such as duplications. Hence, phylogenetic analysis is necessary to confirm HGT candidates and provide a more comprehensive view of their origin and evolutionary history. However, phylogenetic reconstruction requires several time-consuming manual steps to retrieve the homologous sequences, produce a multiple alignment, construct the phylogeny and analyze the topology to assess whether it supports the HGT hypothesis. Here, we present AvP which automatically performs all these steps and detects candidate HGTs within a phylogenetic framework. Availability and implementationAvP is written in Python and is available under GNU General Public License v3.0 at (https://github.com/GDKO/AvP).

bioinformatics↗

Improvement of eukaryotic proteins prediction from soil metagenomes

BackgroundDuring the last decades, shotgun metagenomics and metabarcoding have highlighted the diversity of microorganisms from environmental or host-associated samples. Most assembled metagenome public repositories use annotation pipelines tailored for prokaryotes regardless of the taxonomic origin of contigs and metagenome-assembled genomes (MAGs). Consequently, eukaryotic contigs and MAGs, with intrinsically different gene features, are not optimally annotated, resulting in an incorrect representation of the eukaryotic component of biodiversity, despite their biological relevance. ResultsUsing an automated analysis pipeline, we have filtered 7.9 billion of contigs from 6,873 soil metagenomes in the IMG/M database of the Joint Genome Institute to identify eukaryotic contigs. We have re-annotated genes using eukaryote-tailored methods, yielding 8 million eukaryotic proteins. Of these, 5.6 million could be traced back to non-chimeric higher confidence eukaryotic contigs. Our pipeline improves eukaryotic proteins completeness, contiguity and quality. Moreover, the better quality of eukaryotic proteins combined with a more comprehensive assignment method improves the taxonomic annotation as well. ConclusionsUsing public soil metagenomic data, we provide a dataset of eukaryotic soil proteins with improved completeness and quality as well as a more reliable taxonomic annotation. This unique resource is of interest for any scientist aiming at studying the composition, biological functions and gene flux in soil communities involving eukaryotes. O_TEXTBOXKey Points O_LIEukaryotic micro-organisms have key roles in soil microbial communities and ecosystems. C_LIO_LIEukaryotic genes and proteins are incorrectly represented in metagenomic libraries due to inappropriate methods dedicated to prokaryotes. C_LIO_LIWe have improved the quality and completeness of the de novo proteins prediction and taxonomic annotation of eukaryotic organisms from soil or plant-associated microbiome. C_LI C_TEXTBOX

genomics↗

The genome and lifestage-specific transcriptomes of a plant-parasitic nematode and its host reveal susceptibility genes involved in trans-kingdom synthesis of vitamin B.

Plant-parasitic nematodes are a major, and in some cases a dominant, threat to crop production in all agricultural systems. The relative scarcity of classical resistance genes highlights a pressing need to identify new ways to develop nematode-resistant germplasm. Here, we sequence and assemble a high-quality genome of the model cyst nematode Heterodera schachtii to provide a platform for the first system-wide dual analysis of host and parasite gene expression over time, covering all major stages of the interaction. This novel approach enabled the analysis of the hologenome of the infection site, to identify metabolic pathways that were incomplete in the parasite but complemented by the host. Using a combination of bioinformatic, genetic, and biochemical approaches, we show that the highly atypical completion of vitamin B5 biosynthesis by the parasitic animal, putatively enabled by a horizontal gene transfer from a bacterium, is critically important for parasitism. Knockout of either the plant-encoded or the now nematode-encoded steps in the pathway blocks parasitism. Our experiments establish a reference for cyst nematodes, use this platform to further our fundamental understanding of the evolution of plant-parasitism by nematodes, and show that understanding congruent differential expression of metabolic pathways represents a new way to find nematode susceptibility genes, and thereby, targets for future genome editing-mediated generation of nematode-resistant crops.

genetics↗

Chromatin landscape dynamics in development of the plant parasitic nematode Meloidogyne incognita

In model organisms, epigenome dynamics underlies a plethora of biological processes. The role of epigenetic modifications in development and parasitism in nematode pests remains unknown. The root-knot nematode Meloidogyne incognita adapts rapidly to unfavorable conditions, despite its asexual reproduction. However, the mechanisms underlying this remarkable plasticity and their potential impact on gene expression remain unknown. This study provides the first insight into contribution of epigenetic mechanisms to this plasticity, by studying histone modifications in M. incognita. The distribution of five histone modifications revealed the existence of strong epigenetic signatures, similar to those found in the model nematode Caenorhabditis elegans. We investigated their impact on chromatin structure and their distribution relative to transposable elements (TE) loci. We assessed the influence of the chromatin landscape on gene expression at two developmental stages: eggs, and pre-parasitic juveniles. H3K4me3 histone modification was strongly correlated with high levels of expression for protein-coding genes implicated in stage-specific processes during M. incognita development. We provided new insights in the dynamic regulation of parasitism genes kept under histone modifications silencing. In this pioneering study, we establish a comprehensive framework for the importance of epigenetic mechanisms in the regulation of the genome expression and its stability in plant-parasitic nematodes. Author summaryThe nematode Meloidogyne incognita is one of the most destructive plant parasites worldwide. Its ability to infect a wide range of hosts and its high adaptability contribute to its parasitic success. We investigated the role of epigenetic mechanisms -- specifically post-translational histone modifications -- in the parasitic life cycle. We showed these modifications are linked to gene expression regulation and likely contribute to nematode development and pathogenicity.

genomics↗