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Minou Nowrousian, Jason E. Stajich, Meiling Chu, Ines Engh, Eric Espagne, Karen Halliday, Jens Kamerewerd, Frank Kempken, Birgit Knab, Hsiao-Che Kuo, Heinz D. Osiewacz, Stefanie Pöggeler, Nick D. Read, Stephan Seiler, Kristina M. Smith, Denise Zickler, Ulrich Kück, Michael Freitag

• Filamentous fungi are of great importance in ecology, agriculture, medicine, and biotechnology. Thus, it is not surprising that genomes for more than 100 filamentous fungi have been sequenced, most of them by Sanger sequencing. While next-generation sequencing techniques have revolutionized genome resequencing, e.g. for strain comparisons, genetic mapping, or transcriptome and ChIP analyses, \(\textit {de novo}\) assembly of eukaryotic genomes still presents significant hurdles, because of their large size and stretches of repetitive sequences. Filamentous fungi contain few repetitive regions in their 30–90 Mb genomes and thus are suitable candidates to test \(\textit {de novo}\) genome assembly from short sequence reads. Here, we present a high-quality draft sequence of the \(\textit {Sordaria macrospora}\) genome that was obtained by a combination of Illumina/Solexa and Roche/454 sequencing. Paired-end Solexa sequencing of genomic DNA to 85-fold coverage and an additional 10-fold coverage by single-end 454 sequencing resulted in ~4 Gb of DNA sequence. Reads were assembled to a 40 Mb draft version (N50 of 117 kb) with the Velvet assembler. Comparative analysis with \(\it Neurospora\) genomes increased the N50 to 498 kb. The \(\textit {S.macrospora}\) genome contains even fewer repeat regions than its closest sequenced relative, \(\textit {Neurospora crassa.}\) Comparison with genomes of other fungi showed that \(\textit {S. macrospora}\), a model organism for morphogenesis and meiosis, harbors duplications of several genes involved in self/nonself-recognition. Furthermore, \(\textit {S. macrospora}\) contains more polyketide biosynthesis genes than \(\textit {N. crassa}\). Phylogenetic analyses suggest that some of these genes may have been acquired by horizontal gene transfer from a distantly related ascomycete group. Our study shows that, for typical filamentous fungi, \(\textit {de novo}\) assembly of genomes from short sequence reads alone is feasible, that a mixture of Solexa and 454 sequencing substantially improves the assembly, and that the resulting data can be used for comparative studies to address basic questions of fungal biology.