Effects of low-dose gamma irradiation on artemisinin content and amorpha-4,11-diene synthase activity in Artemisia annua L

Abstract

Purpose: This research work aimed to create in vitro plantlet variants of Artemisia annua through gamma irradiation. The obtained variants were then evaluated for the correlation between their artemisinin content and enzyme activity of amorpha-4,11-diene synthase (ADS), the first enzyme of the artemisinin pathway.

Materials and methods: Shoot tips from plantlets of A. annua were exposed to gamma rays at a low-dose range and transferred to hormone-free MS medium for in vitro cultivation. A dose-response curve and the value of 50% of lethal dose (LD₅₀) were then obtained. This LD₅₀ dose of gamma rays was used for treating another batch of shoot tips. The surviving plantlets after four subsequent subcultures were evaluated for their ability to accumulate artemisinin in correlation with the enzyme activity of ADS.

Results: The dose-response curve showed that the LD₅₀ value was at 8 Gray (Gy). The surviving irradiated plantlets from this dose treatment had artemisinin content ranging from 0.03–0.70% (w/w) of dry weight, comparing with only 0.18% present in the original non-irradiated samples. Their correlation coefficient between the ADS activity and the artemisinin content appeared to be R² = 0.090 for all the 18 samples tested, although, selectively, more than half of these (11 samples) showed their R² value of as high as 0.851.

Conclusions: There was no correlation between the artemisinin content and ADS activity found in the whole population, but the correlation was observed in the main subpopulation of the irradiated A. annua plantlets.

Keywords:

• Artemisia annua L.
• Asteraceae
• gamma irradiation
• artemisinin
• amorpha-4,11-diene synthase
• in vitro plantlets