Efficient genetic transformation of sour orange, Citrus aurantium L. using Agrobacterium tumefaciens containing the coat protein gene of Citrus tristeza virus

Transgenic Citrus aurantium plants have been obtained by Agrobacterium tumefaciens-mediated gene transfer using seedling-derived epicotyl segments as explants and Basta herbicide as the selective agent. In the present research, hypocotyl, and epicotyls parts of C. aurantium plant were inoculated with A. tumefaciens strain EHA-105. Then, explants were transferred into the regeneration medium and shoot growth containing various concentrations of kinetin and IAA growth regulators. In order to detect transgenic shoots, herbicide Basta (ammonium glufosinate) was added at a rate of 1 mg/L to the regeneration medium. Production buds happened in explants which Marker BAR gene resistant to Basta herbicides was received. After PCR, to approve the final transgenic plants, transformation efficiency was evaluated by 9.16%. Regeneration of explants treated with hormones showed a significant difference at 1% compared with control explants. The Maximum regeneration was showed in 0.5 mg/L kinetin treatment. These results indicated that the combination between kinetin and IAA could be effective in increasing regeneration.