Establishment of an efficient transformation system is a prerequisite for genetic improvement of Jatropha curcas, a promising biodiesel feedstock plant, by transgenic approach. In this study an efficient Agrobacterium-mediated transformation protocol using cotyledon explants from J. curcas seeds was developed. The integration and expression of the transgenes in the putatively transformed J. curcas plants was confirmed by polymerase chain reaction (PCR), Southern blot analysis and β-glucuronidase (GUS) activity staining. In the protocol used, the critical step for successful transformation is that the selective agent kanamycin (20 mg/L) is not included in the callus-inducing medium within the first 4 weeks after co-cultivation with Agrobacterium.
Keywords: Agrobacterium-mediated transformation, biofuel, Jatropha curcas, kanamycin, physic nut, plant regeneration, transgenic plants.
BA, 6-Benzylaminopurine; CaMV, cauliflower mosaic virus; GUS,β-glucuronidase; IBA, indole-3-butyric acid; MS, Murashige and Skoog; NPT II,neomycin phosphotransferase II; PCR, polymerase chain reaction; YEP, yeast extract peptone; CIM, callus-inducing medium; SIM, shoot-inducing medium; RM,rooting medium; CTAB, cetyl trimethylammonium bromide.
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