High Frequency β-glucuronidase Reporter Gene Expression in Shoot Tip, Leaf, and Hypocotyl Explants of Jatropha curcas L.

Abstract

Development of a suitable genetic transformation system is the backbone of transgenic technology for efficient recovery of Jatropha curcas plants with desirable traits. Although in vitro plant regeneration protocols for shoot tip, leaf, and hypocotyl explants of J. curcas have been reported, these protocol have not been translated into efficient genetic transformation systems. The objective of this study was to standardize the parameters influencing transient β-glucuronidase reporter gene expression following Agrobacterium-mediated genetic transformation using shoot tip, leaf, and hypocotyl explants of Jatropha curcas. Disarmed Agrobacterium tumefaciens strain EHA105, harboring pCAMBIA2301 carrying neomycin phosphotransferase (nptII) as a selectable marker and the β-glucuronidase (GUS) as a reporter gene, were used for transformation. Frequencies of GUS gene expression of 86.7%, 78.3%, and 74.2% were obtained for leaf, hypocotyls, and shoot tip explants, respectively. The transgenic nature of the kanamycin-resistant explants was also confirmed by using polymerase chain reaction (PCR) assay. Because, to date, there is no report on Agrobacterium-mediated transformation of shoot tip, leaf, and hypocotyl explants of Jatropha curcas, our findings/protocol should be useful for recovering stable transgenic Jatropha curcas plants from these explants.