De Novo characterization of a Cephalotaxus hainanensis transcriptome and genes related to paclitaxel biosynthesis

Cephalotaxus hainanensis, an endangered plant, is known to contain several metabolites with anti-cancer activity. Despite its clinical impact, the alkaloid metabolism of this species has remained largely uncharacterized. The potential of Cephalotaxus for metabolic engineering of medically interestin...

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Bibliographic Details
Published in:PloS one 2014-09, Vol.9 (9), p.e106900
Main Authors: Qiao, Fei, Cong, Hanqing, Jiang, Xuefei, Wang, Rongxiang, Yin, Junmei, Qian, Dan, Wang, Zhunian, Nick, Peter
Format: Article
Language:English
Subjects:
Agriculture
Anticancer properties
Bioinformatics
Biology and Life Sciences
Biosynthesis
Biotechnology
Cancer
Cephalotaxus
Cephalotaxus - genetics
Cephalotaxus hainanensis
Crops
Endangered plants
Exploitation
Gene expression
Gene Expression Regulation, Plant - genetics
Genes
Genes, Plant - genetics
Genomics
Germplasm
High-performance liquid chromatography
Horticulture
Laboratories
Landscape architecture
Leaves
Leukemia
Liquid chromatography
Metabolic engineering
Metabolism
Metabolites
Molecular chains
Natural products
Paclitaxel
Paclitaxel - biosynthesis
Plant Leaves - genetics
Ribonucleic acid
RNA
Science
Sequence Analysis, DNA - methods
Sequences
Transcriptome - genetics
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Summary:Cephalotaxus hainanensis, an endangered plant, is known to contain several metabolites with anti-cancer activity. Despite its clinical impact, the alkaloid metabolism of this species has remained largely uncharacterized. The potential of Cephalotaxus for metabolic engineering of medically interesting compounds has, so far, not been exploited, due to the almost complete lack of molecular information. We have therefore performed a high throughput RNA-seq analysis and assembled the transcriptome de novo. Raw reads comprising 4.3 Gbp were assembled de novo into 39,416 unique sequences (unigenes) with a mean length of 1,089.8 bp and a total assembly size of 45.8 Mbp, which equals to more than 50 times the number of Cephalotaxaceae sequences currently deposited in the GenBank (as of August 2013). As proof of principle for medically interesting pathways, gene fragments related to paclitaxel biosynthesis were searched and detected. To verify their functionality, the metabolic product paclitaxel, and its precursor baccatin III, were identified in the leaves of C. hainanensis by HPLC, and shown to be induced by MeJA. This finding demonstrates exemplarily the potential of the annotated transcriptome as information resource for the biotechnological exploitation of plant secondary metabolism.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0106900