ISSN: 2332-0737
Erum Dilshad, Hammad Ismail, Waqas Khan Kayani and Bushra Mirza
The excessive attention in the use of plants as medicine is credited to the occurrence of active principles whose pharmacological activities have been investigated. Due to the limited production and very less quantity of these important metabolites present in the plant cells, their genetic engineering and increased in vitro production is the point of focus for many years and can be achieved by in vitro transformation and propagation of desired plant. In the current study, we report transformation protocol for Artemisia carvifolia Buch with Agrobacterium tumefaciens C58C1 harboring β-glucuronidase as reporter and neomycin phosphotransferase as selectable marker gene. We have optimized simple regeneration conditions after transformation involving two different types of explants (leaf and stem) on different media formulations for direct organogenesis and best regeneration response. MS media with 2.5 g/L benzylaminopurine (BAP), 0.25 g/L naphthalene acetic acid (NAA), giving maximum number of shoots was selected. Rooting was obtained on ½ MS medium supplemented with NAA (0.1 mg/L). Transient expression of gus reporter gene was observed in the leaf and stem explants after 2 days of bacterial infection. Artemisia carvifolia Buch can be successfully transformed with Agrobacterium tumefaciens strain C58C1 by using controlled in vitro regeneration conditions. Findings of the current study would be useful for micro propagation and genetic transformation of Artemisia carvifolia Buch in future.