METHOD FOR GENETICALLY TRANSFORMING ROOTSTOCKS OF CITRUS SPECIES OF THE GENERA CITRUS, PONCIRUS AND HYBRIDS THEREOF BASED ON YOUNG PLANTS

摘要

The invention relates to a method for transforming transgenic citrus lines based on young plants, comprising the following steps: (a) providing seeds selected from the group formed by seeds of pure or hybrid citrus varieties of the genera Carrizo and Poncirus; (b) treating the seeds with a fungicide composition comprising the compounds methyl-1-(butylcarbamoyl)-2-benzimidazole carbamate and ethanethiol as an active agent; (c) disinfecting the seeds using H₂O₂, and then using NaOCI; (d) sowing the seeds with their testas in dark conditions in Murashige and Tucker (MT) medium until the epicotyls become etoliated; (e) transforming the epicotyls using cells of Agrobacterium tumefaciens comprising the plasmid vectors having at least one gene of interest; (f) selecting the transgenic lines in a selection medium comprising an antibiotic for selection purposes, 6-benzylaminopurine (BAP) phytohormone and an antibiotic for controlling the growth of Agrobacterium; (g) growing the selected transformed lines; (h) rooting the grown lines in an optimised rooting medium, with a composition comprising at least one phytohormone; and (i) propagating the transgenic lines in order to produce the transformed rootstock.