Vol. 79 No: 3

Title:
Transient expression and stable integration of chimeric Gus gene in watermelon following electroporation

Authors:
M.V. HEMA, D THEERTHA PRASAD and A VANI

pp: 364-369

Abstract:
A novel technique for introducing foreign genes into intact plant organs has been demonstrated in watermelon (Citrullus lanatus L.) following electroporation of zygotic embryos and nodal buds using the chimeric β-glucuronidase (gus) gene. DNA transfer was done under in vitro and in vivo conditions using plasmid constructs harbouring the gus reporter gene under the control of the constitutive promoter CaMV35S and nos terminator. The parameters for electroporation were optimized to study the influence of different plasmid constructs on the uptake, transient expression and stable integration of the transgene in the electroporated tissues. Maximum GUS expression was observed by electroporating the tissues with five electric pulses of 400 V cm^-1 discharged from a 960 μF capacitor in the presence of 200 fig DNA ml^-1. Microscopic observation of the thin sections of paraffin-embedded embryos clearly showed that the β-glucuronidase expression was exclusively located inside the treated cells. Further, we confirmed the stable integration of chimeric gene in R₁ progeny obtained from in vivo electroporated watermelon plants by dot blot and Southern blot analysis.

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