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Overexpression of allene oxide cyclase promoted tanshinone/phenolic acid production in Salvia miltiorrhiza

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Abstract

Key message

This study provides a desirable candidate gene resource (SmAOC) to increase the content of valuable natural products via appropriate JA pathway genetic engineering.

Abstract

Jasmonates (JAs) are important signal molecules in plants. They regulate transcripts of defense and secondary biosynthetic metabolite genes in response to environmental stresses. Currently, JAs are widely used as elicitors to improve the content of useful secondary metabolism in plants. Synthesis of the naturally occurring enantiomer of various jasmonates is catalyzed by allene oxide cyclase (AOC, EC 5.3.99.6). Here, we cloned and characterized the AOC gene (SmAOC) from Salvia miltiorrhiza. As expected, SmAOC expression was induced by abiotic stimuli such as methyl jasmonate (MeJA), ultraviolet radiation (UV) and low temperature (4 °C) in S. miltiorrhiza plantlets. To demonstrate whether the engineered internal JAs pool by overexpressing AOC gene could promote secondary metabolism production, the SmAOC was incorporated into S. miltiorrhiza hairy roots. The results revealed that SmAOC overexpression significant enhanced the yields of tanshinone IIA, rosmarinic acid (RA) and lithospermic acid B (LAB) in S. miltiorrhiza hairy roots. In addition, expression levels for key genes involved in the biosynthetic pathway of diterpenes and phenolic acids were also altered. These suggest that genetic manipulation of AOC would be helpful for improving the production of valuable secondary metabolites by regulating the biosynthesis of JAs.

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Abbreviations

4CL:

4-Coumarate coenzyme A ligase

AOC:

Allene oxide cyclase

AOS:

Allene oxide synthase

BV:

Blank vector

CMK:

Cytidine monophosphate kinase

CTAB:

Cetyl trimethyl ammonium bromide

DXR:

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

HMGR:

3-Hydroxy-3-methylglutaryl-CoA reductase

HPPR:

4-Hydroxyphenylpyruvate reductase

hpt:

Hygromycin phosphotransferase gene

IPTG:

Isopropyl thiob-d-galactoside

JAs:

Jasmonates

JMT:

Jasmonic acid caboxyl methyltransferase

LAB:

Lithospermic acid B

LOX:

Lipoxygenase

MeJA:

Methyl jasmonate

MRM:

Multiple reactions monitoring

ORF:

Open reading frame

PAL:

Phenylalanine ammonialyase

RA:

Rosmarinic acid

RT-qPCR:

Real-time quantitative PCR

TFs:

Transcription factors

WT:

Wild type

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Acknowledgments

This work was supported by grants from National Natural Science Foundation of China (30900786, 31100221 and 31160059).

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Authors and Affiliations

  1. Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai, 200003, People’s Republic of China

    Xiao-Ce Gu, Jun-Feng Chen, Ying Xiao, Peng Di, Hong-Jiao Xuan, Xun Zhou & Wan-Sheng Chen

  2. Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, Shanghai, 200433, People’s Republic of China

    Lei Zhang

  3. Department of Plant Sciences, University of California, Davis, CA, 95616, USA

    Lei Zhang

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  1. Xiao-Ce Gu
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  2. Jun-Feng Chen
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  4. Peng Di
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  5. Hong-Jiao Xuan
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  6. Xun Zhou
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  7. Lei Zhang
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Correspondence to Lei Zhang or Wan-Sheng Chen.

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Communicated by E. Benvenuto.

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Gu, XC., Chen, JF., Xiao, Y. et al. Overexpression of allene oxide cyclase promoted tanshinone/phenolic acid production in Salvia miltiorrhiza . Plant Cell Rep 31, 2247–2259 (2012). https://doi.org/10.1007/s00299-012-1334-9

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  • DOI: https://doi.org/10.1007/s00299-012-1334-9

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