Agrobacteium transformation of tobacco with a genetic module of antimalarial agent artemisinin biosynthesis

A. P. Firsov*, T. Yu Mitiushkina, A. S. Pushin, A. K. Tzareva, A. M. Vainstein, S. V. Dolgov

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Artemisinin-based medicines are the most effective treatment for malaria. To date, the wormwood plants (Artemisia annua L.) are the main source of artemisinin. Due to the limited nature of this source, considerable efforts are directed towards the development of methods for artemisinin production via heterologous expression systems. We used in this study agrobacterial transformation to transfer the genetic module of the artemisinin biosynthesis pathway into plants and then analyzed its transcription in a heterologous host. Tobacco plants were transformed with the artemisinin biosynthesis genes encoding amorpha-4,11-diene synthase, artemisin-aldehyde Δ11(13) reductase, amorpha-4,11-diene monooxygenase, cytochrome P450 reductase from A. annua and yeast 3-hydroxy-3-methylglutaryl-coenzyme A reductase cloned in the pArtemC vector; farnesyl diphosphate synthase and aldehyde dehydrogenase were used to transform the plants as parts of vector p2356. As a result of transformation with the pArtemC and p2356 vectors, in twos transgenic lines with all target genes were obtained. Five genes of artemisinin biosynthesis and two genes of biosynthesis of its precursors were successfully transferred into the genome of transgenic tobacco lines as a result of the co-transformation with abovementioned vectors. Thus, the entire artemisinin biosynthesis pathway was first reconstructed in heterologous plants: the transcription of the artemisinin biosynthesis genes in the tobacco plants was shown via RT-PCR. The obtained results will be used in further research on expression systems for the production of artemisinin and other non-protein substances in heterologous host plants.

Original languageAmerican English
Pages (from-to)34-45
Number of pages12
JournalBiotekhnologiya
Volume36
Issue number3
DOIs
StatePublished - 2020

Bibliographical note

Publisher Copyright:
© 2020 State Research Institute for Genetics and Selection of Industrial Microorganisms.

Keywords

  • Artemisinin
  • Malaria
  • Metabolic engineering
  • Tobacco
  • Transgenic plants

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