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Abstract
Artemisinin is frequently used in the artemisinin-based combination therapy to cure drug-resistant malaria in Asian subcontinent and large swath of Africa. The hairy root system, using the Agrobacterium rhizogenes LBA 9402 strain to enhance the production of artemisinin in Artemisia annua L., is developed in our laboratory. The transgenic nature of hairy root lines and the copy number of transgene (rol B) were confirmed using polymerase chain reaction and Southern Blot analyses, respectively. The effect of different concentrations of methyl jasmonate (MeJA), fungal elicitors (Alternaria alternate, Curvularia limata, Fusarium solani, and Piriformospora indica), farnesyl pyrophosphate, and miconazole on artemisinin production in hairy root cultures were evaluated. Among all the factors used individually for their effect on artemisinin production in hairy root culture system, the maximum enhancement was achieved with P. indica (1.97 times). Increment of 2.44 times in artemisinin concentration by this system was, however, obtained by combined addition of MeJA and cell homogenate of P. indica in the culture medium. The effects of these factors on artemisinin production were positively correlated with regulatory genes of MVA, MEP, and artemisinin biosynthetic pathways, viz. hmgr, ads, cyp71av1, aldh1, dxs, dxr, and dbr2 in hairy root cultures of A. annua L.
| Abbreviations | ||
| ACT | = | artemisinin-based combination therapy |
| ADS | = | amorpha-4,11-diene synthase |
| ALDH1 | = | aldehyde dehydrogenase 1 |
| B5 | = | Gamborg's medium |
| CYP71AV1 | = | cytochrome P450 monooxygenase |
| DBR2 | = | artemisinic aldehyde double bond reductase |
| DW | = | dry weight |
| DXS | = | 1-deoxy-d-xylulose 5-phosphate synthase |
| DXR | = | 1-deoxy-d-xylulose 5-phosphate reductoisomerase |
| HMGR | = | 3-hydroxy-3-methylglutaryl coenzyme A reductase |
| HPLC | = | high-performance liquid chromatography |
| MS | = | Murashige and Skoog |
| qPCR | = | quantitative polymerase chain reaction |
| Abbreviations | ||
| ACT | = | artemisinin-based combination therapy |
| ADS | = | amorpha-4,11-diene synthase |
| ALDH1 | = | aldehyde dehydrogenase 1 |
| B5 | = | Gamborg's medium |
| CYP71AV1 | = | cytochrome P450 monooxygenase |
| DBR2 | = | artemisinic aldehyde double bond reductase |
| DW | = | dry weight |
| DXS | = | 1-deoxy-d-xylulose 5-phosphate synthase |
| DXR | = | 1-deoxy-d-xylulose 5-phosphate reductoisomerase |
| HMGR | = | 3-hydroxy-3-methylglutaryl coenzyme A reductase |
| HPLC | = | high-performance liquid chromatography |
| MS | = | Murashige and Skoog |
| qPCR | = | quantitative polymerase chain reaction |
Acknowledgments
This study was supported by a research grant to Prof. M. Z. Abdin from the Life Sciences Research Board, DRDO, Government of India and UGC-SAP. Seema Ahlawat is thankful to Department of Science and Technology, Government of India, for providing WoS-A Fellowship.
