AGROBACTERIUM-MEDIATED TRANSFORMATION OF CITRUS USING TWO BINARY VECTORS

Abstract

In the present study, epicotyl explants of 'Midsweet' sweet orange (Citrus sinensis L. Osb.) were transformed using a mixture of two binary vectors individually introduced into Agrobacterium strain AGL1. One binary Ti vector was assembled from pCAMBIA2301 with GUS as the reporter gene while the other binary Ti vector was assembled from pGreen0029 with GFP as the reporter gene. Two gene constructs containing specific gene fragments from Poncirus were used. The epicotyl explants were transformed by use of 1:1 mixture of AGL1 carrying the binary vectors separately. Shoots regenerated from transformed epicotyl explants on selective medium were micrografted onto 'Carrizo' seedlings. The GFP expression and the GUS activity in the leaves were also determined. Further confirmation by molecular analyses is currently underway.