Unravelling hybridization in Phytophthora using phylogenomics and genome size estimation
Kris Van Poucke,
Annelies Haegeman,
Thomas Goedefroit,
Fran Focquet,
Leen Leus,
Marília Horta Jung,
Corina Nave,
Miguel Angel Redondo,
Claude Husson,
Kaloyan Kostov,
Aneta Lyubenova,
Petya Christova,
Anne Chandelier,
Slavcho Slavov,
Arthur de Cock,
Peter Bonants,
Sabine Werres,
Jonàs Oliva Palau,
Benoit Marçais,
Thomas Jung,
Jan Stenlid,
Tom Ruttink,
Kurt Heungens
Affiliations
Kris Van Poucke
Flanders Research Institute for Agriculture, Fisheries and Food (ILVO), Plant Sciences Unit
Annelies Haegeman
Flanders Research Institute for Agriculture, Fisheries and Food (ILVO), Plant Sciences Unit
Thomas Goedefroit
Flanders Research Institute for Agriculture, Fisheries and Food (ILVO), Plant Sciences Unit
Fran Focquet
Flanders Research Institute for Agriculture, Fisheries and Food (ILVO), Plant Sciences Unit
Leen Leus
Flanders Research Institute for Agriculture, Fisheries and Food (ILVO), Plant Sciences Unit
Marília Horta Jung
Phytophthora Research Centre (PRC), Mendel University
Corina Nave
Julius Kühn Institute (JKI) – Federal Research Center for Cultivated Plants, Institute for Plant Protection in Horticulture and Forestry (GF)
Miguel Angel Redondo
Department of Forest Mycology and Plant Pathology, Swedish University of Agricultural Sciences (SLU)
Claude Husson
Ministère de l’agriculture et de l’alimentation, Direction générale de l’alimentation (DGAL), Sous Direction de la Qualité et de la Protection des Végétaux (SDQPV), Département de la Santé des Forêts
Kaloyan Kostov
AgroBioInstitute (ABI), Agricultural Academy
Aneta Lyubenova
AgroBioInstitute (ABI), Agricultural Academy
Petya Christova
AgroBioInstitute (ABI), Agricultural Academy
Anne Chandelier
Life Sciences Department, Walloon Agricultural Research Centre (CRAW)
Slavcho Slavov
AgroBioInstitute (ABI), Agricultural Academy
Arthur de Cock
Westerdijk Fungal Biodiversity Institute
Peter Bonants
Wageningen University & Research, Business Unit Biointeractions & Plant Health
Sabine Werres
Julius Kühn Institute (JKI) – Federal Research Center for Cultivated Plants, Institute for Plant Protection in Horticulture and Forestry (GF)
Jonàs Oliva Palau
Department of Forest Mycology and Plant Pathology, Swedish University of Agricultural Sciences (SLU)
Benoit Marçais
Université de Lorraine – Institut National de la Recherche Agronomique (INRAe), L’Unité Mixte de Recherche Interactions arbres/microorganismes (UMR IAM)
Thomas Jung
Phytophthora Research Centre (PRC), Mendel University
Jan Stenlid
Department of Forest Mycology and Plant Pathology, Swedish University of Agricultural Sciences (SLU)
Tom Ruttink
Flanders Research Institute for Agriculture, Fisheries and Food (ILVO), Plant Sciences Unit
Kurt Heungens
Flanders Research Institute for Agriculture, Fisheries and Food (ILVO), Plant Sciences Unit
Abstract The genus Phytophthora comprises many economically and ecologically important plant pathogens. Hybrid species have previously been identified in at least six of the 12 phylogenetic clades. These hybrids can potentially infect a wider host range and display enhanced vigour compared to their progenitors. Phytophthora hybrids therefore pose a serious threat to agriculture as well as to natural ecosystems. Early and correct identification of hybrids is therefore essential for adequate plant protection but this is hampered by the limitations of morphological and traditional molecular methods. Identification of hybrids is also important in evolutionary studies as the positioning of hybrids in a phylogenetic tree can lead to suboptimal topologies. To improve the identification of hybrids we have combined genotyping-by-sequencing (GBS) and genome size estimation on a genus-wide collection of 614 Phytophthora isolates. Analyses based on locus- and allele counts and especially on the combination of species-specific loci and genome size estimations allowed us to confirm and characterize 27 previously described hybrid species and discover 16 new hybrid species. Our method was also valuable for species identification at an unprecedented resolution and further allowed correct naming of misidentified isolates. We used both a concatenation- and a coalescent-based phylogenomic method to construct a reliable phylogeny using the GBS data of 140 non-hybrid Phytophthora isolates. Hybrid species were subsequently connected to their progenitors in this phylogenetic tree. In this study we demonstrate the application of two validated techniques (GBS and flow cytometry) for relatively low cost but high resolution identification of hybrids and their phylogenetic relations.