Exploring the Genome Diversity of Mycorrhizal Fungi – A JGI Sequencing Project Update Annegret Kohler1, Joshua Herr1,2, Emmanuelle Morin1, Claude Murat1, Annick Brun1, Claire Venault-Fourrey1, David Hibbett3, Igor Grigoriev4,, and Francis Martin1 1 Tree-Microbe Interactions Research Group, INRA-Nancy, UMR 1136, Champenoux, FRANCE 2 Schatz Center for Tree Molecular Genetics, Pennsylvania State University, University Park, PA 16802 USA, 3 Biology Department, Clark University, Worcester, MA 01610 USA 4 Eukaryotic Genomics Group, Joint Genome Institute, Walnut Creek, CA 94598 USA Abstract Mycorrhizal fungi are responsible for plant and ecosystem health by providing an array of benefits. Mycorrhizal fungi help plants (including woody trees and shrubs, forbs, grasses, etc.) to acquire nutrients - notably nitrogen and phosphorus – and aid in water acquisition. Additionally, mycorrhizal fungi provide both a physical barrier to pathogens and initiate gene expression to aid in responses to be biotic and abiotic stresses. In ecosystems, mycorrhizal fungi contribute to nutrient cycling and may possess novel enzymes for the breakdown of a wide array of natural and synthetic chemicals. The goal of this sequencing initiative is to identify and compare genome determinants shaping evolution across the disparate fungal taxa in symbiosis with plants. Sequencing mycorrhizal fungal genomes will contribute to, which includes but is not limited to, the better understanding of interactions with host-trees; the basis for understanding phenotypic differences in fruit body structure and development; the description and understanding of organic secondary metabolites typical of antibiotics and volatile compounds; the comparative genomics of additional fungal parasites and saprobes; and the development of molecular markers to be used for the tractability of fungi in various environments. Sequencing the Genomes of Differing Status of Mycorrhizal Fungal Genome Sequencing Mycorrhizal Basidiomycetes in the Types of Mycorrhizal Fungi • Along with completed genomes and those in the sequencing, assembly, and Joint Genome Institute’s Fungal annotation stages (red and orange boxes, respectively), two tiers of genome • Although Smith & Read (2008) describe seven types of sequencing efforts have been planned through JGI for mycorrhizal fungi. Genomics Program mycorrhizae, mycorrhizal fungi can be described in terms of • The sequencing of mycorrhizal genomes by JGI is part of the ecto- and endomycorrhizae, which may form an infection • These sequencing efforts, planned for the next two years, will yield upwards of larger Fungal Genome Project (JGI-FGP). gradient from plant to plant. Describing mycorrhizal fungi in 30 fungal genomes from all fungal phyla and sub-phyla exhibiting the this way is an artificial classification that probably owes as mycorrhizal habit. These fungal genomes will contribute to our knowledge of • Basidiomycete representatives of both mycorrhizal and much to the plant host as the fungal symbiont. biochemical evolution and adaptation as well as ecosystem functioning. saprophytic genomes will be compared to elucidate how ecological roles have shaped genome architecture in the • Ectomycorrhizae interact with a plant symbiont by forming a Species Name Taxonomic Position Mycorrhizal Status Basidiomycetes (Martin & Nehls 2009). Type sheath around a plant root but do not penetrate the plant Laccaria bicolor Basidiomycota, Agaricales, Ecto Completed (Martin et al 2008), Version 2.0 apoplast. This form of mycorrhizae is common with trees and S238N-H82 Hydnangiaceae soon to be released scrubs and accounts for approximately 10% of plant species, Tuber Ascomycota, Pezizales, Ecto Completed (Martin et al 2010), Version 1.0 which cover large temperate and tropical ecosystems. Genomes melansporum Tuberaceae Completed • Endomycorrhizae are Cenococcum Ascomycota, Dothideomycetes Ecto In Sequencing Queue characterized by the geophilum 1.58 penetration of the fungal Glomus Glomerulomycota, Glomales Endo Illumina, 454 & Fosmid Sequencing In Progress intraradices tissue into the plant Hebeloma Basidiomycota, Agaricales, Ecto Illumina, 454 & Fosmid Sequencing Complete, apoplast, but typically do cylindrosporum h7 Cortinariaceae Assembly Near Completion not contribute a large Paxillus involutus Basidiomycota, Boletales Ecto Illumina, 454 & Fosmid Sequencing Complete, amount of fungal hyphae Assembly In Progress Pisolithus Basidiomycota, Boletales Ecto Illumina, 454 & Fosmid Sequencing Complete, to cover a plant root. microcarpus 441 Assembly In Progress Mycorrhizal types such as Pisolithus tinctorius Basidiomycota, Boletales Ecto On Hold Pending Pisolithus microcarpus 441 Ericoid and Orchid Marx 270 Assembly mycorrhizae, for the sake Rhizopogon Basdiomycota, Boletales, Ecto Illumina, 454 & Fosmid Sequencing Complete, salebrosus Suillineae (?) Assembly In Progress plant root of simplicity, can be put in Sequencing, Genomes Tuber aestivum Ascomycota, Pezizales, Ecto Sequencing started by Genoscope Genome Assembly, and Annotation. Image from into this category. Tuberaceae Institute, France Bonfante & Amanita muscaria Basidiomycota, Agaricales, Ecto In Sequencing Queue Genre 2010 BX008 Aminitaceae Laccaria Basidiomycota, Agaricales, Ecto 454 Sequencing completed through INRA amethystina Hydnangiaceae The Laccaria Pan-Genome Lactarius quietus Basidiomycota, Russulales Ecto Soon to be shipped to JGI Phylogenetic distribution of Basidiomycete representatives of the • Laccaria is a cosmopolitan genus of mycorrhizal fungi found Paxillus Basidiomycota, Boletales, Ecto Soon to be shipped to JGI JGI-FGP with in progress or pending genome sequencing. Green in temperate regions throughout the world. rubicundulus Paxilineae Piloderma croceum Basidiomycota, Atheliales Ecto Illumina & Fosmids Completed, 454 In Progress text represents mycorrhizal forming fungi and black text • Laccaria are model mycorrhizal taxa because, unlike many F1598 represents saprophytic or parasitic fungi. The box with ‘other’ Suillus luteus Basidiomycota, Boletales, Ecto Soon to be shipped to JGI Fungal Genomes taxa represent completed Basidiomycete genomes sequenced by fungi, they can be cultured and manipulated in the laboratory. Suillineae Additionally, copious amounts of ecosystem information JGI or other organizations which will be used for the purposes of Thelephora Basidiomycota, Thelephorales Ecto Soon to be shipped to JGI comparative genomics of the Basidiomycota and Eukaryotes. concerning their natural history is available. terrestris Sebacina vermifera Basidiomycota, Sebacinales Endo/Orchid Sequencing In-Progress, No Fosmids • Numerous Laccaria species are being sequenced using the MAFF 305830 Illumina GA platform for both intra-genus and population Meliniomyces Ascomycota, Helotiales Ecto/Ericoid Soon to be shipped to JGI A Tale of Two Symbionts: Laccaria bicolor genomic studies. The Laccaria bicolor S238N-H82 genome Mycorrhizal Oidiodendron Ascomycota, Leotiomycetales Endo Illumina Completed, INRA 454 Completed, JGI bicolor vs. Tuber melanosporum will be used for genome assembly (Martin et al 2008). – maius Zn 454 In Progress • Sequencing the genomes of Laccaria bicolor (Martin et al • Sequencing these genomes will allow the comparison of Rhyzoscyphus Ascomycota, Helotiales Ecto/Endo Soon to be shipped to JGI 2008) and Tuber melanosporum (Martin et al 2010) have very adaptive mutations to influences of environment, geography, Scheduled for Completion in 2011. ericeae different genomes, but exhibit a very similar suite of symbiosis- Tier 1 Terfezia boudieri Ascomycota, Pezizales Ecto/Endo DNA ready to be shipped to JGI and host specificity that have been noted across the genus. induced nutrient cycling enzymes (Plett & Martin 2011). Boletus edulis Basidiomycota, Boletales, Ecto DNA ready to be shipped to JGI • The analysis will consist of ‘ingroup’ L. bicolor (annotation Boleineae Laccaria bicolor 2.0) and L. amethystina (draft genome) with ‘outgroup’ L. Cantharellus Basidiomycota, Cantharellales Ecto DNA in production • Laccaria bicolor appears to be laccata and L. proxima. cibarius less dependent on the host and • The analysis will Coltricia Basidiomycota, Ecto DNA in production more active at acquiring carbon also consist of 20 cinnamomea Hymenochaetales Cortinarius Basidiomycota, Agaricales, Ecto DNA in production from the soil substrates and, as European, glaucopus Cortinariaceae a result, may act as a weak American, and Gymnomyces Basidiomycota, Russulales Ecto DNA in production saprotroph in the environment Fungal Genomes Genomes Fungal xanthosporus Asian strains of L. Ramaria formosa Basidiomycota, Gomphales Ecto DNA in production (Martin et al 2008). bicolor and L. amethystina for the Scleroderma Basidiomycota, Boletales, Ecto DNA in production Tuber melanosporum • Tuber melansporum is more purpose of citrinum Sclerodermataceae aggressive in its colonization of determining Tomentella Basidiomycota, Thelephorales Ecto DNA in production plant roots and does not appear geographic, sublilacina Mycorrhizal Tricholoma Basidiomycota, Ecto DNA in production to be able to acquire carbon Evolutionary relationships environmental, and – matsutake Tricholomataceae from the soil and therefore is within the host influence on Tulasnella Basidiomycota, Cantharellales, Endo/Orchid DNA in production calospora Tulasnellaceae more dependent on the host for genus Laccaria genome in 2012. Completion for Scheduled Meliniomyces Ascomycota, Helotiales Ericoid, DNA in production its survival (Martin et al