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Short term UV-B radiation-mediated transcriptional responses and altered secondary metabolism of in vitro propagated plantlets of Artemisia annua L.

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Abstract

In vitro propagated Artemisia annua plantlets were exposed to low dose (2.8 W m−2) of UV-B (280–315 nm) radiation for different short-term (1, 2, 3 and 4 h) durations that resulted in 103 and 100 % enhanced artemisinin and flavonoid yield respectively after 3 h of UV-B irradiation. UV-B irradiation resulted in reduced chlorophyll a, b and carotenoids while the concentrations of UV-B-absorbing anthocyanins and phenolics were induced. Under UV-B radiation, superoxide radical formation increased in a time dependent manner and was maximum at 4 h. This lead to differential responses of enzymatic antioxidants (superoxide dismutase, catalase, peroxidases and glutathione reductase) to withstand the UV-B radiation-induced reactive oxygen species burst. The transcript analysis through RT-PCR revealed the significant up-regulation of HMGR, DXR, IPPi, FPS, ADS, CYP71AV1 and RED1 gene transcripts leading to higher artemisinin accumulation. Differential expression of stress regulated genes (AOX1a, NDB2, UPOX, PAL, LOX) helped the plant towards better adaptability at least up to 3 h UV-B irradiation period. Simultaneous clustering of genes and UV-B treatments using complete linkage and a euclidean distance matrix revealed that 3 h irradiation period is optimum for high artemisinin yield with better tolerance. Histological studies revealed the larger trichome size in UV-B treated plants as compared to untreated plants. The present study conclude that short term UV-B treatment to in vitro raised A. annua may be a safe approach for continuous supply of high artemisinin producing plantlets with simultaneous action of stress- regulated genes to keep the plant healthy.

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Abbreviations

BAP:

6-Benzylaminopurine

NAA:

1-Nephthaleneacetic acid

SOD:

Superoxide dismutase

POX:

Peroxidase

GR:

Glutathione reductase

ROS:

Reactive oxygen species

HMGR:

3-Hydroxyl-3-methyglutaryl CoA reductase

DXS:

1-Deoxy-d-xylulose-5-phosphate synthase

DXR:

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

IPPi:

Isopentenyl pyrophosphate isomerase

FPS:

Fernasyl diphoaphate synthase

GGPRS:

Geranylgeranyl pyrophosphate synthase

ADS:

Amorpha-4,11-diene synthase

CYP71AV1:

Cytochrome P450 dependent monooxygenase/hydroxylase

RED1:

Dihydroartemisinic aldehyde reductase

GAS:

Germacrene A synthase

ECS:

8-Epicedrol synthase

QHS:

β-Caryophyllene synthase

BFS:

β-Farnesene synthase

AOX1a:

Alternative oxidase 1a

NDB2:

NAD(P)H dehydrogenase B2

UPOX:

Gene at locus At2g21640 (up-regulated under oxidative stress)

FNR:

Ferredoxin NADP reductase

PAL:

Phenylalanine ammonia lyase

LOX:

Lipoxygenase

Chl:

Chlorophyll

T.S.:

Transverse section

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Acknowledgments

The authors are thankful to CSIR (council of scientific and industrial research), New Delhi, India for financial assistance, Dr. Priyanka Pandey for her assistance in heatmap preparation and Prof. R. P. Sinha for critical reading of the manuscript. Neha Pandey is thankful to UGC (University grant commission), New Delhi, India for financial assistance in the form of RFSMS (Research fellowship in science for meritorious students).

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  1. Laboratory of Morphogenesis, Department of Botany, Faculty of Science, Banaras Hindu University, Varanasi, India

    Neha Pandey & Shashi Pandey-Rai

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  1. Neha Pandey
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Correspondence to Shashi Pandey-Rai.

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Pandey, N., Pandey-Rai, S. Short term UV-B radiation-mediated transcriptional responses and altered secondary metabolism of in vitro propagated plantlets of Artemisia annua L.. Plant Cell Tiss Organ Cult 116, 371–385 (2014). https://doi.org/10.1007/s11240-013-0413-0

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