Efficient Method for Agrobacterium Mediated Transformation of Artemisia annua L.

Pravej      Alam; Anis      Mohammad; M.M.      Ahmad; Mather   Ali   Khan; Mohd.      Nadeem; Riyazuddeen      Khan; Mohd.      Akmal; Seema      Ahlawat; M.Z.      Abdin

Abstract

Artemisinin, a potent antimalarial natural products isolated from aerial parts of Artemisia annua L. Many patents have been reported that the demand for artemisinin is exponentially increasing year after year due to increased incidences of drug resistant malaria throughout the world. Leaf explants were used frequently as target tissue to generate transgenic of Artemisia. annua L. However, obtaining a large number of transgenic lines through out the year is a laborious and delicate process. To circumvent this, we have developed a highly efficient leaf explant based Agrobacterium mediated transformation of A. annua L. plant. The gus gene was used as screenable marker to assess and optimize the performance of T-DNA delivery. The age of explant, kind of bacterial inoculation, suspension duration, infection times and co-culture conditions were optimized. The co-culture was carried out with Agrobacterium tumefaciens strain EHA105 under desiccation condition in the dark at 25-28 0C for 2-4 days. Complete analysis of transgene insertion demonstrated that the optimized method of transformation from leaf explants of A. annua L. was efficient and highly reproducible.

Keywords: Artemisia annua L., T-DNA delivery β−glucuronidase, Genetic transformation.

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