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van de Ruitenbeek, S. J. S.

Publications and source records attributed to van de Ruitenbeek, S. J. S..

4 recordsLinked to original sources

Globodera pallida virulence on major potato resistance has a common genetic basis across Western Europe

The potato cyst nematode Globodera pallida poses a major threat to potato production in Western Europe. Current management strategies largely depend on the use of potato varieties carrying the genetic resistance GpaVvrn. However, reports from multiple West-European countries indicate a steady rise in virulence against GpaVvrn-containing potato varieties, raising serious concerns about G. pallida control. Although recent studies have resolved the genetic basis of virulence in two Dutch G. pallida populations, it remains unclear how conserved this genetic adaptation is in populations from different regions. To investigate this, we first selected eight Dutch G. pallida populations on the GpaVvrn-containing potato variety Seresta and confirmed a previously identified virulence locus. Second, by analysing the allele frequencies of four virulence-associated SNPs in Dutch, British, and French GpaVvrn-selected G. pallida populations, we found that the same allele is consistently selected by GpaVvrn across Western Europe. Third, we analysed the propagation of eight G. pallida populations on 26 GpaVvrn-containing potato varieties and showed that a populations allele frequency of a single SNP (T173N) accurately reflects its reproduction on GpaVvrn. Fourth, we developed an allele-specific quantitative PCR (AS-qPCR) assay to determine a populations alternative allele frequency (AAF) of T173N and showed that AS-qPCR-based AAFs reliably indicate virulence levels on GpaVvrn in Dutch and German G. pallida populations. Together, these findings suggest that a common allele is consistently selected by GpaVvrn in populations from different regions across Western Europe. The AS-qPCR assay developed in this study provides a practical tool to estimate G. pallida virulence on GpaVvrn in the field, enabling field-tailored and sustainable resistance management strategies for farmers.

genetics↗

Environmental sex determination in the cyst nematode Globodera pallida defaults to male development

Environmental sex determination (ESD) enables organisms to adjust their sexual fate in response to external cues. Fluctuating sex ratios have long suggested the presence of ESD in populations of plant-parasitic nematodes. We show that in the potato cyst nematode Globodera pallida sex is regulated by nutritional cues. By manipulating sucrose availability to the host plant, we could steer the sex determination of G. pallida. Whereas high-sucrose medium promotes female development, low-sucrose medium promotes male development. Transcriptome analyses on the early stages of parasitism reveal that female development requires extensive transcriptional activation and post-transcriptional regulation. We identify Gp-lin-29, a transcription factor homologous to lin-29 in Caenorhabditis elegans, as a potential regulator of ESD. Small RNA sequencing uncovered the male-biased expression of Gp-let-7, a putative repressor of Gp-lin-29, and the female-biased expression of four miRNAs, including Gp-miR-100, located at the same genomic locus as Gp-let-7. Target prediction and enrichment analyses suggest that these female-biased miRNAs actively suppress male developmental programs. Together, our findings support a model in which G. pallida juveniles follow a default male developmental trajectory unless redirected by favourable environmental cues to become female. This study provides mechanistic insight into ESD in cyst nematodes and positions G. pallida as a tractable system for exploring epigenetic regulation of developmental plasticity.

developmental biology↗

The potato cyst nematode Globodera pallida overcomes major potato resistance through selection on standing variation at a single locus

Globodera pallida poses a major threat to potato production, with management strategies primarily relying on genetic resistance. However, reports from multiple locations in Western Europe indicate a steady increase in virulence levels among field populations, raising major concerns about G. pallida control. The evolutionary mechanisms driving this rise in virulence are poorly understood. To investigate this, we analysed the propagation of thirteen recently isolated field populations on thirty commercial potato varieties over four independent PCN resistance tests. Our findings indicate that (1) the genetic basis of resistance in potatoes is small, with the major resistance conferred by GpaV from Solanum vernei, and (2) the wide application of GpaVvrn has led to continuous selection on standing genetic variation in G. pallida field populations. To map virulence, we propagated two field populations on a GpaVvrn-resistant variety for four consecutive generations. High-coverage whole-genome sequencing of each generation revealed that GpaVvrn-mediated selection acted on a single locus of a newly assembled G. pallida Rookmaker reference genome. Examination of this virulence-associated locus identified Gp-pat-1 as a candidate gene. Silencing Gp-pat-1 increased G. pallida virulence on a GpaVvrn-resistant potato variety but had no effect on nematode virulence on a susceptible variety. Thereby classifying Gp-pat-1 as an avirulence gene and confirming its role in the breakdown of GpaVvrn-resistance. These findings demonstrate that negative selection on the Gp-pat-1 avirulence allele by GpaVvrn-mediated resistance is driving the emergence of virulence in the G. pallida field populations used in our selection experiments. Our results therefore strongly suggest that selection on Gp-pat-1 is likely underlying the current outbreak of GpaVvrn-resistance breaking populations. Our results provide a foundation for the development of molecular diagnostic tools to monitor virulence in field populations, to enhance understanding of resistance breakdown, and to inform the sustainable deployment of resistances in the field. SIGNIFICANCE STATEMENTIn Western Europe, management of the potato cyst nematode Globodera pallida primarily relies on genetic resistances in potato plants. However, resistance-breaking populations are emerging across Western Europe. Here, we identify a single resistance locus shared by all tested resistant potato varieties. We also identify a corresponding virulence locus on a newly assembled G. pallida reference genome and identify a gene within this locus that contributes to virulence. Our findings provide critical insights into the selective pressure acting on this plant-parasitic nematode, the emergence of virulence over time, and the molecular mechanism underlying resistance breakdown. These results provide much needed insights into a highly adapted soil-borne plant-pathogen.

genetics↗

A Chromosome-scale Reference Genome of Meloidogyne hapla reveals localized recombination hotspots enriched with Effector Proteins

Root-knot nematodes (Meloidogyne spp.) are among the most destructive agricultural pests that cause significant yield losses across a wide range of crops. Meloidogyne hapla, a diploid species, is a valuable model for studying root-knot nematodes due to its parasitic diversity, small genome, and a reproductive strategy that facilitates genetic analysis. Here, we present a high-quality chromosome-scale assembly of M. hapla, generated using multiple sequencing platforms-PacBio HiFi, ONT, Illumina and HiC. The 59 Mb assembly comprises 16 chromosome-length scaffolds, notably lacking canonical telomeric repeats. Instead, we identified a tandem 16-mer repeat mainly present at scaffold ends, suggesting an alternative system for chromosome-end maintenance. Genetic linkage analysis of F2 populations derived from crosses between M. hapla strains validated the assembly but also revealed anomalies indicating chromosome structure differences between parental isolates such as fissions, fusions, and rearrangements. This analysis also revealed sharply delineated zones of high recombination on most chromosome arms. We also identified 1,258 genes encoding putative secreted proteins (PSP), which should be enriched in genes involved in host interaction and pathogenicity. Most of the PSP genes had orthologs in other plant parasitic nematode species, and the majority were pioneers, lacking known functional domains. Notably, we found that PSPs are significantly enriched in high-recombination zones, possibly facilitating their rapid evolution. Overall, our study provides new insights into the genome structure of diploid root-knot nematodes and highlights the interplay between genome architecture, recombination, and parasitism. These findings raise new questions about how genetic and genomic adaptations drive the success of rootknot nematodes as plant parasites.

plant biology↗