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Transcriptomics analysis investigates sesquiterpenoids accumulation pattern in different tissues of Atractylodes lancea (Thunb.) DC. plantlet

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Abstract

Atractylodes lancea: (Thunb.) DC. is an endangered traditional Chinese medicinal herb with broad pharmacological activity, reflected by an abundance of sesquiterpenoids. However, the biosynthesis of these sesquiterpenoids is largely unknown due to a lack of genetic information. To understand sesquiterpenoid distribution in A. lancea, tissue-specific sesquiterpenoids accumulation patterns during different growth periods of plantlets were analyzed via gas chromatography (n = 3). Transcriptomic analyses were furthermore performed via RNA-Seq in an attempt to reveal the molecular mechanism regulating sesquiterpenoids formation in leaves, stems and roots. 85,170 unigenes with an average sequence length of 768 bp were acquired. Via analysis of differentially expressed genes, 857 unigenes were identified to be involved in sesquiterpenoids formation. Consequently, a putative sesquiterpenoid biosynthetic pathway was proposed, including mevalonate, 2-C-methyl-d-erythritol-4-phosphate, sesquiterpene synthase, cytochrome P450, and the peroxisome pathway. Moreover, the relationship between gene expression and sesquiterpenoid production in different growth periods of leaves, stems, and roots was analyzed based on Quantitative Real-Time PCR (n = 3) and gas chromatography data. In this study, the sesquiterpenoid distribution in different tissues of sterile tissue culture plantlets of A. lancea was systematically investigated via metabolite quantitative and transcriptomics analysis. This study provides a central theoretical basis for the manipulation of sesquiterpenoid biosynthesis in A. lancea.

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Abbreviations

A. lancea :

Atractylodes lancea (Thunb.) DC.

GC:

Gas chromatography

MS:

Mass spectrometry

L. canum:

Leucosceptrum canum

MS:

Murashige and Skoog

NR:

Non-redundant

NT:

Nucleotide

COG:

Classification of Orthologous Group

KEGG:

Kyoto encyclopedia of genes and genomes

GO:

Gene ontology

FPKM:

Fragments per kilobase of transcript per million fragments mapped

DEGs:

Differentially expressed genes

FDR:

False discovery rate

A. annua :

Artemisia annua

qRT-PCR:

Quantitative Real-Time PCR

MVA:

Mevalonate

MEP:

2-C-methyl-d-erythritol-4-phosphate

IPP:

Isopentenyl diphosphate

DMAPP:

Dimethylallyl diphosphate

HMGCR:

3-Hydroxy-3-methylglutaryl CoA synthase

DXR:

1-Deoxy-d-xylulose-5-phosphate reductoisomerase

IDI:

Isopentenyl diphosphate isomerase

FDPS:

Farnesyl diphosphate synthase

FS:

Farnesene synthase

GAS:

Germacrene A synthase

GDS:

Germacrene D synthase

CPS:

β-Caryophyllene synthase

ROS:

Reactive oxygen species

CAT:

Catalase

SOD:

Superoxide dismutase

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Acknowledgements

This work was financially supported by National Natural Science Foundation of China (21406119 and 31070443), Natural Science Foundation of the Higher Education Institutions of Jiangsu Province (14KJB180009), Integration of Production and Research Projects of Nanjing Science and Technology Commission (201306019), and a project funded by the Priority Academic Program Development (PAPD) of Jiangsu Higher Education Institutions.

Author contributions

Chuan-Chao Dai and Fei Chen conceived and designed the experiments. Fei Chen, Yu-Xiao Wei, Jing-Min Zhang, Xiu-Mei Sang performed the experiments. Fei Chen analyzed the data and wrote the manuscript.

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Authors and Affiliations

  1. Jiangsu Key Laboratory for Microbes and Functional Genomics, Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources, College of Life Sciences, Nanjing Normal University, Nanjing, 210023, China

    Fei Chen, Yu-Xiao Wei, Jing-Min Zhang, Xiu-Mei Sang & Chuan-Chao Dai

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  1. Fei Chen
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  2. Yu-Xiao Wei
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Correspondence to Chuan-Chao Dai.

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Chen, F., Wei, YX., Zhang, JM. et al. Transcriptomics analysis investigates sesquiterpenoids accumulation pattern in different tissues of Atractylodes lancea (Thunb.) DC. plantlet. Plant Cell Tiss Organ Cult 130, 73–90 (2017). https://doi.org/10.1007/s11240-017-1205-8

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