Transcriptome Analysis of Taxus cuspidata Needles Based on 454 Pyrosequencing

Abstract

Taxus species are highly valued as renewable resources for the production of Taxol. Despite the commercial and medicinal importance of Taxus, little genomic information is available for yew species, and Taxol biosynthesis still needs to be fully elucidated. In this study, 454 pyrosequencing technology was employed to produce an expressed sequence tag (EST) from the needles of Taxus cuspidata. In all, 81 148 high-quality reads from the needles of T. cuspidata were produced using Roche GS FLX Titanium. A total of 20 557 unique sequences were obtained, including 12 975 singletons and 7582 contigs. Approximately 14 095 unique sequences were annotated by a similarity search against five public databases. Gene ontology revealed 11 220 unique sequences that could be assigned to 45 vocabularies. In the Kyoto Encyclopedia of Genes and Genomes mapping, 2403 transcripts were established as associated with 3821 biochemical pathways. Enzymes in the plastidial 2-C-methyl-D-erythritol 4-phosphate pathway were well represented. Candidates of the putative genes of Taxol biosynthesis were revealed, including those in the remaining steps. In total, 291 transcripts were identified, representing putative homologues of transcription factors. Furthermore, 753 simple sequence repeat motifs, which are potential molecular markers for genetic application, were identified. These results provide the largest EST collections in Taxus and will contribute to biosynthetic and biochemical studies that lead to drug improvement.

Key words

transcriptome analysis - Taxus cuspidata - Taxaceae - 454 pyrosequencing - Taxol biosynthesis - transcription factor

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