Tissue Culture of Stone Fruit Plants Basis for Their Genetic Engineering

Genetic engineering of stone fruit species like apricot, plum, peach and cherry are hampered by the inefficient and low-level regeneration processes in tissue culture. The first transgenic stone fruit species have emerged from transformed hypocotyls. These great achievements were applauded by the scientific community contrary the fact that hypocotyl derived transgenic plants have no real brooding value. Tissue culture of different organs of valuable cultivars are recorded with an extremely low-level of regeneration in the literature. To improve the tissue culture basis of stone fruit plants an extensive tissue culture programme were launched and dozens of different media were compared including a series of hormone concentration in the tissue culture systems. Our continuous efforts were crowned by a very efficient method for achieving up to 30-40% regenerable petioles. Usually on a single petiole several well-separated meristems were induced. After 3-4 weeks of cultivation shoots were developed. The basic media $K_2$ were supplemented with 10g/L saccharose, 10g/L glucose and 10g/L maltose. The following plant hormones were used BAP 1mg/L, TDZ 1mg/L, 2-iP 1mg/L and IAA 0,1 mg/L concentrations. The Petri dishes were kept for 3 weeks in dark at a temperature 22$^{\circ}C$ for 8 hours and 22-24$^{\circ}C$ for 16 hours. The Petri dishes were sealed with Parafilm. The regeneration of the petioles were genotype independent and we were able to regenerate different plum cultivars with almost the same efficiency.