Agrobacterium tumefaciens-Mediated Genetic Transformation of Eclipta alba

Abstract

Eclipta alba (Linn.) Hassk. (Asteraceae) is a high value medicinal plant which possesses diverse medicinal properties. It is an important herb for the treatment of various disorders, and is primarily used as a hepatoprotectant. Its major biochemical constituents include wedelolactone and dimethyl-wedelolactone (coumestans), which possess anti-hepatotoxic properties. Due to its numerous medicinal properties, it is in high demand by the pharmaceutical industry and therefore requires urgent biotechnological interventions for its improvement. Therefore, the present study was constructed with the aim of developing an efficient genetic transformation protocol for E. alba, which will help in the mass production of the active compounds found in E. alba. Agrobacterium tumefaciens strain LBA 4404, containing vector pBI121, was used to genetically transform the plant, and the effect of various factors such as infection type, light cycle effect, effect of pH, among others, on the genetic transformation efficiency was analyzed. Regenerated transformed shoots were confirmed using the standard Polymerase Chain Reaction PCR method. Kanamycin-resistant and beta- glucurosidaseGUS-positive shoots indicated the development of transgenic shoots in E. alba. Amplification of nptll and uidA genes confirmed the integration of t-DNA transgenic shoots. In conclusion, various factors affecting the transformation efficiency were analyzed, and a reliable A. tumefaciens-mediated genetic transformation protocol was developed.