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Adv Pharm Bull. 2020;10(3): 464-471.
doi: 10.34172/apb.2020.057
  Abstract View: 62
  PDF Download: 56

Research Article

Transformation of Amorphadiene Synthase and Antisilencing P19 Genes Into Artemisia annua L. and Its Effect on Antimalarial Artemisinin Production

Elfahmi Elfahmi 1,2 * ORCID logo, Fany Mutia Cahyani 1, Tati Kristianti 3, Sony Suhandono 4

1 School of Pharmacy, Bandung Institute of Technology, Bandung, Indonesia.
2 Biosciences and Biotechnology Research Center, Bandung Institute of Technology, Bandung, Indonesia.
3 Institut Pendidikan Indonesia, Garut, West Java, Indonesia.
4 School of Life Sciences and Technology, Bandung Institute of Technology, Bandung, Indonesia.

Abstract

Purpose: The low content of artemisinin related to the biosynthetic pathway is influenced bythe role of certain enzymes in the formation of artemisinin. The regulation of genes involved inartemisinin biosynthesis through genetic engineering is a choice to enhance the content. Thisresearch aims to transform ads and p19 gene as an antisilencing into Artemisia annua and to seetheir effects on artemisinin production.Methods: The presence of p19 and ads genes was confirmed through polymerase chain reaction(PCR) products and sequencing analysis. The plasmids, which contain ads and/or p19 genes,were transformed into Agrobacterium tumefaciens, and then inserted into leaves and hairy rootsof A. annua by vacuum and syringe infiltration methods. The successful transformation waschecked through the GUS histochemical test and the PCR analysis. Artemisinin levels weremeasured using HPLC.Results: The percentages of the blue area on leaves by using vacuum and syringe infiltrationmethod and on hairy roots were up to 98, 92.55%, and 99.00% respectively. The ads-p19 samplecontained a higher level of artemisinin (0.18%) compared to other samples. Transformed hairyroot with co-transformation of ads-p19 contained 0.095% artemisinin, where no artemisininwas found in the control hairy root. The transformation of ads and p19 genes into A. annuaplant has been successfully done and could enhance the artemisinin content on the transformedleaves with ads-p19 up to 2.57 folds compared to the untransformed leaves, while for p19, cotransformedand ads were up to 2.25, 1.29, and 1.14 folds respectively.Conclusion: Antisilencing p19 gene could enhance the transformation efficiency of ads andartemisinin level in A. annua.
Keywords: Agrobacterium tumefaciens, Artemisinin, Amorphadiene synthase, Artemisia annua, Malaria, p19
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Submitted: 20 Aug 2019
Revision: 12 Jan 2020
Accepted: 27 Jan 2020
ePublished: 11 May 2020
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