De Novo Transcriptome Assembly and Characterization of Lithospermum officinale to Discover Putative Genes Involved in Specialized Metabolites Biosynthesis

Amit Rai; Taiki Nakaya; Yohei Shimizu; Megha Rai; Michimi Nakamura; Hideyuki Suzuki; Kazuki Saito; Mami Yamazaki

doi:10.1055/a-0630-5925

Planta Medica, Inhaltsverzeichnis

Planta Med 2018; 84(12/13): 920-934
DOI: 10.1055/a-0630-5925

Natural Product Chemistry and Analytical Studies

Original Papers

Georg Thieme Verlag KG Stuttgart · New York

De Novo Transcriptome Assembly and Characterization of Lithospermum officinale to Discover Putative Genes Involved in Specialized Metabolites Biosynthesis^[*]

Amit Rai

¹Graduate School of Pharmaceutical Sciences, Chiba University, Chiba, Japan

,

Taiki Nakaya

¹Graduate School of Pharmaceutical Sciences, Chiba University, Chiba, Japan

,

Yohei Shimizu

¹Graduate School of Pharmaceutical Sciences, Chiba University, Chiba, Japan

,

Megha Rai

¹Graduate School of Pharmaceutical Sciences, Chiba University, Chiba, Japan

,

Michimi Nakamura

¹Graduate School of Pharmaceutical Sciences, Chiba University, Chiba, Japan

,

Hideyuki Suzuki

²Kazusa DNA Research Institute, Chiba, Japan

,

Kazuki Saito

¹Graduate School of Pharmaceutical Sciences, Chiba University, Chiba, Japan

,

Mami Yamazaki

¹Graduate School of Pharmaceutical Sciences, Chiba University, Chiba, Japan

› Institutsangaben

Abstract

Lithospermum officinale is a valuable source of bioactive metabolites with medicinal and industrial values. However, little is known about genes involved in the biosynthesis of these metabolites, primarily due to the lack of genome or transcriptome resources. This study presents the first effort to establish and characterize de novo transcriptome assembly resource for L. officinale and expression analysis for three of its tissues, namely leaf, stem, and root. Using over 4Gbps of RNA-sequencing datasets, we obtained de novo transcriptome assembly of L. officinale, consisting of 77,047 unigenes with assembly N50 value as 1524 bps. Based on transcriptome annotation and functional classification, 52,766 unigenes were assigned with putative genes functions, gene ontology terms, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. KEGG pathway and gene ontology enrichment analysis using highly expressed unigenes across three tissues and targeted metabolome analysis showed active secondary metabolic processes enriched specifically in the root of L. officinale. Using co-expression analysis, we also identified 20 and 48 unigenes representing different enzymes of lithospermic/chlorogenic acid and shikonin biosynthesis pathways, respectively. We further identified 15 candidate unigenes annotated as cytochrome P450 with the highest expression in the root of L. officinale as novel genes with a role in key biochemical reactions toward shikonin biosynthesis. Thus, through this study, we not only generated a high-quality genomic resource for L. officinale but also propose candidate genes to be involved in shikonin biosynthesis pathways for further functional characterization.

Key words

Lithospermum officinale - Boraginaceae - shikonin biosynthesis - chlorogenic acid - lithospermic acid - RNA-sequencing - deep transcriptome analysis - KEGG-pathways enrichment analysis

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Referenzen

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Supporting Information

De Novo Transcriptome Assembly and Characterization of Lithospermum officinale to Discover Putative Genes Involved in Specialized Metabolites Biosynthesis[*]

De Novo Transcriptome Assembly and Characterization of Lithospermum officinale to Discover Putative Genes Involved in Specialized Metabolites Biosynthesis^[*]