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Vol 36(2020) N 3 p. 34-45; DOI 10.21519/0234-2758-2020-36-3-34-45
A.P. Firsov1*, T.Yu. Mitiuchkina1, A.S. Pushin1, A.K. Tzareva2, A.M. Vainsteinl, S.V. Dolgov1

Agrobacteium Transformation of Tobacco with a Genetic Module of Antimalarial Agent Artemisinin Biosynthesis

1Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Pushchino Branch, Russian Academy of Sciences, Pushchino, Moscow Oblast, 142290 Russia
2Faculty of Biotechnology, Lomonosov Moscow State University, Moscow, 119991 Russia
3Hebrew University of Jerusalem, Rehovot, 76100 Israel

*aleksey_firsov@mail.ru
Received - 19.12.2019; Accepted - 19.05.2020

References

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 All(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.

artemisinin, malaria, metabolic engineering, tobacco, transgenic plants

This work was supported by a Grant from the Russian Science Foundation no. 19-14-00190.



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