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Glucose-6-phosphate dehydrogenase plays critical role in artemisinin production of Artemisia annua under salt stress

  • Original paper
  • Published:
Biologia Plantarum

Abstract

Artemisinin, a natural sesquiterpenoid isolated from Artemisia annua L., is regarded as the most efficient drug against malaria in the world. Artemsinin production in NaCl-treated A. annua seedlings and its relationships with the glucose-6-phosphate dehydrogenase (G6PDH) activity and generation of H2O2 and nitric oxide (NO) were investigated. Results revealed that artemisinin content in the seedlings was increased by 79.3 % over the control after 1-month treatment with 68 mM NaCl. The G6PDH activity was enhanced in the presence of NaCl together with stimulated generation of H2O2 and NO. Application of 1.0 mM glucosamine (GlcN), an inhibitor of G6PDH, blocked the increase of NADPH oxidase and nitrate reductase (NR) activities, as well as H2O2 and NO production in A. annua seedlings under the salt stress. The induced H2O2 was found to be involved in the upgrading gene expression of two key enzymes in the later stage of artemisinin biosynthetic pathway: amorphadiene synthase (ADS) and amorpha-4,11-diene monooxygenase (CYP71AV1). The released NO being attributed mainly to the increase of NR activity, negatively interacted with H2O2 production and enhanced gene expression of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR). Inhibition of NO generation partly blocked NaCl-induced artemisinin accumulation, and NO donor strongly rescued the decreased content of artemisinin caused by GlcN. These results suggest that G6PDH could play a critical role in NaCl-induced responses and artemisinin biosynthesis in A. annua.

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Abbreviations

ADS:

amorphadiene synthase

BSA:

bovine serum albumin

cPTIO:

2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide

CYP71AV1:

amorpha-4,11-diene monooxygenase

DPI:

diphenylene iodonium

DXR:

1-deoxy-D-xylulose-5-phosphate reductoisomerase

EDTA:

ethylenediaminetetraacetic acid

FPP:

farnesyl diphosphate

G6PDH:

glucose-6-phosphate dehydrogenase

GlcN:

glucosamine

HEPES:

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

HMGR:

3-hydroxy-3-methylglutaryl coenzyme A reductase

HPLC:

high-performance liquid chromatography

IPP:

isopentenyl diphosphate

L-NAME:

Nω-nitro-L-arginine methyl ester

MEP:

2-C-methyl-D-erythritol-4-phosphate

MTT:

3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide

MVA:

mevalonate

NO:

nitric oxide

NOS:

NO synthase

NR:

nitrate reductase

OPPP:

oxidative pentose phosphate pathway

PES:

phenazine ethosulfate

PVDF:

polyvinylidene difluoride

qPCR:

quantitative PCR

ROS:

reactive oxygen species

SA:

salicylic acid

SOD:

superoxide dismutase

TCA:

trichloroacetic acid

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Authors and Affiliations

  1. College of Pharmaceutical Sciences, Soochow University, Suzhou, 215123, P.R. China

    J. W. Wang, H. Tian & X. Yu

  2. Institute of Medicinal Plants, Yunnan Academy of Agricultural Sciences, Kunming, 650205, P.R. China

    H. Tian

  3. Department of Horticulture, School of Architecture, Soochow University, Suzhou, 215123, P.R. China

    L. P. Zheng

Authors
  1. J. W. Wang
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  2. H. Tian
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  3. X. Yu
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  4. L. P. Zheng
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Correspondence to J. W. Wang.

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Acknowledgements: The authors are grateful to the Graduate Program of Higher Education in Jiangsu Province (No. CXLX13-841), the Suzhou Scholar Program (No. 14317363), and the projects sponsored by the NSFC (No. 81273487) for financial support of this work.

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Wang, J.W., Tian, H., Yu, X. et al. Glucose-6-phosphate dehydrogenase plays critical role in artemisinin production of Artemisia annua under salt stress. Biol Plant 61, 529–539 (2017). https://doi.org/10.1007/s10535-016-0674-7

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  • DOI: https://doi.org/10.1007/s10535-016-0674-7

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