title: The ambrosia symbiosis is specific in some species and promiscuous in others: evidence from community pyrosequencing.
abstract: Symbioses are increasingly seen as dynamic ecosystems with multiple associates and varying fidelity. Symbiont specificity remains elusive in one of the most ecologically successful and economically damaging eukaryotic symbioses: the ambrosia symbiosis of wood-boring beetles and fungi. We used multiplexed pyrosequencing of amplified internal transcribed spacer II (ITS2) ribosomal DNA (rDNA) libraries to document the communities of fungal associates and symbionts inside the mycangia (fungus transfer organ) of three ambrosia beetle species, Xyleborus affinis, Xyleborus ferrugineus and Xylosandrus crassiusculus. We processed 93 beetle samples from 5 locations across Florida, including reference communities. Fungal communities within mycangia included 14-20 fungus species, many more than reported by culture-based studies. We recovered previously known nutritional symbionts as members of the core community. We also detected several other fungal taxa that are equally frequent but whose function is unknown and many other transient species. The composition of fungal assemblages was significantly correlated with beetle species but not with locality. The type of mycangium appears to determine specificity: two Xyleborus with mandibular mycangia had multiple dominant associates with even abundances; Xylosandrus crassiusculus (mesonotal mycangium) communities were dominated by a single symbiont, Ambrosiella sp. Beetle mycangia also carried many fungi from the environment, including plant pathogens and endophytes. The ITS2 marker proved useful for ecological analyses, but the taxonomic resolution was limited to fungal genus or family, particularly in Ophiostomatales, which are under-represented in our amplicons as well as in public databases. This initial analysis of three beetle species suggests that each clade of ambrosia beetles and each mycangium type may support a functionally and taxonomically distinct symbiosis.
authors.affiliation: 1] School of Forest Resources and Conservation, University of Florida, Gainesville, FL, USA [2] Department of Genetics and Microbiology, Faculty of Science, Charles University in Prague, Praha 2, Czech Republic [3] Institute of Microbiology AS CR, Praha 4, Czech Republic.
authors.forename: Martin
authors.initials: M
authors.lastname: Kostovcik
authors.affiliation: Department of Entomology and Nematology, University of Florida, Gainesville, FL, USA.
authors.forename: Craig C
authors.initials: CC
authors.lastname: Bateman
authors.affiliation: 1] Institute of Microbiology AS CR, Praha 4, Czech Republic [2] Department of Botany, Faculty of Science, Charles University, Praha 2, Czech Republic.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Kolarik
authors.affiliation: Department of Entomology and Nematology, Citrus Research and Education Center, University of Florida, Lake Alfred, FL, USA.
authors.forename: Lukasz L
authors.initials: LL
authors.lastname: Stelinski
authors.affiliation: Natural History Museum, University Museum of Bergen, University of Bergen, Bergen, Norway.
authors.forename: Bjarte H
authors.initials: BH
authors.lastname: Jordal
authors.affiliation: 1] School of Forest Resources and Conservation, University of Florida, Gainesville, FL, USA [2] Department of Entomology and Nematology, University of Florida, Gainesville, FL, USA.
authors.forename: Jiri
authors.initials: J
authors.lastname: Hulcr
mesh_terms: D000818:Animals; D044822:Biodiversity; D001517:Coleoptera / Q000382:microbiology*; D017931:DNA Primers; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D005431:Florida; D005658:Fungi / Q000145:classification / Q000235:genetics*; D005819:Genetic Markers / Q000235:genetics; D016133:Polymerase Chain Reaction; D013559:Symbiosis
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer; D005819:Genetic Markers
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