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Molecules 2017, 22(12), 2155; doi:10.3390/molecules22122155

De Novo RNA Sequencing and Expression Analysis of Aconitum carmichaelii to Analyze Key Genes Involved in the Biosynthesis of Diterpene Alkaloids

1
Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675, Japan
2
Tsukuba Division, Research Center for Medicinal Plant Resources, National Institutes of Biomedical Innovation, Health and Nutrition, Tsukuba 305-0843, Japan
3
Medical Mycology Research Center, Chiba University, Chiba 260-8673, Japan
4
Kazusa DNA Research Institute, Chiba 292-0818, Japan
*
Authors to whom correspondence should be addressed.
Received: 16 November 2017 / Revised: 30 November 2017 / Accepted: 1 December 2017 / Published: 5 December 2017
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Abstract

Aconitum carmichaelii is an important medicinal herb used widely in China, Japan, India, Korea, and other Asian countries. While extensive research on the characterization of metabolic extracts of A. carmichaelii has shown accumulation of numerous bioactive metabolites including aconitine and aconitine-type diterpene alkaloids, its biosynthetic pathway remains largely unknown. Biosynthesis of these secondary metabolites is tightly controlled and mostly occurs in a tissue-specific manner; therefore, transcriptome analysis across multiple tissues is an attractive method to identify the molecular components involved for further functional characterization. In order to understand the biosynthesis of secondary metabolites, Illumina-based deep transcriptome profiling and analysis was performed for four tissues (flower, bud, leaf, and root) of A. carmichaelii, resulting in 5.5 Gbps clean RNA-seq reads assembled into 128,183 unigenes. Unigenes annotated as possible rate-determining steps of an aconitine-type biosynthetic pathway were highly expressed in the root, in accordance with previous reports describing the root as the accumulation site for these metabolites. We also identified 21 unigenes annotated as cytochrome P450s and highly expressed in roots, which represent candidate unigenes involved in the diversification of secondary metabolites. Comparative transcriptome analysis of A. carmichaelii with A. heterophyllum identified 20,232 orthogroups, representing 30,633 unigenes of A. carmichaelii, gene ontology enrichment analysis of which revealed essential biological process together with a secondary metabolic process to be highly enriched. Unigenes identified in this study are strong candidates for aconitine-type diterpene alkaloid biosynthesis, and will serve as useful resources for further validation studies. View Full-Text
Keywords: Aconitum; RNA sequencing; transcript abundance; diterpene alkaloids; aconitine; KEGG pathway enrichment analysis Aconitum; RNA sequencing; transcript abundance; diterpene alkaloids; aconitine; KEGG pathway enrichment analysis
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MDPI and ACS Style

Rai, M.; Rai, A.; Kawano, N.; Yoshimatsu, K.; Takahashi, H.; Suzuki, H.; Kawahara, N.; Saito, K.; Yamazaki, M. De Novo RNA Sequencing and Expression Analysis of Aconitum carmichaelii to Analyze Key Genes Involved in the Biosynthesis of Diterpene Alkaloids. Molecules 2017, 22, 2155.

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