Deep RNA sequencing improved the structural annotation of the Tuber melanosporum transcriptome

New Phytol. 2011 Feb;189(3):883-891. doi: 10.1111/j.1469-8137.2010.03597.x.

Abstract

• The functional complexity of the Tuber melanosporum transcriptome has not yet been fully elucidated. Here, we applied high-throughput Illumina RNA-sequencing (RNA-Seq) to the transcriptome of T. melanosporum at different major developmental stages, that is free-living mycelium, fruiting body and ectomycorrhiza. • Sequencing of cDNA libraries generated a total of c. 24 million sequence reads representing > 882 Mb of sequence data. To construct a coverage signal profile across the genome, all reads were then aligned to the reference genome assembly of T. melanosporum Mel28. • We were able to identify a substantial number of novel transcripts, antisense transcripts, new exons, untranslated regions (UTRs), alternative upstream initiation codons and upstream open reading frames. • This RNA-Seq analysis allowed us to improve the genome annotation. It also provided us with a genome-wide view of the transcriptional and post-transcriptional mechanisms generating an increased number of transcript isoforms during major developmental transitions in T. melanosporum.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ascomycota / genetics*
  • Base Sequence
  • Codon, Initiator
  • Exons
  • Fruiting Bodies, Fungal
  • Gene Library
  • Genome, Fungal*
  • High-Throughput Nucleotide Sequencing / methods
  • Mycelium
  • Mycorrhizae
  • Open Reading Frames
  • RNA, Fungal / analysis*
  • Sequence Analysis, RNA / methods*
  • Untranslated Regions

Substances

  • Codon, Initiator
  • RNA, Fungal
  • Untranslated Regions