TRANSFORMATION OF EASTER LILY VIA PARTICLE BOMBARDMENT

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Particle bombardment was investigated as a potential transformation method for Easter lily. Bulb scale explants from Lilium longiflorum Thunb. `Nellie White' were used as target material. The uidA (or gusA) reporter gene for ß- glucuronidase (GUS) expression was used in all particle bombardments to assess efficiency of gene delivery. Parameters examined to achieve optimal levels of transient GUS expression included gene promoter, helium pressure (particle velocity), and target distance. The highest level of transient GUS expression (as measured by number of indigo-stained cells/scale explant) was observed with the rice actin 1 (Act1) promoter, a helium pressure of 1500 psi, and target distances of 9 to 12 cm. Parameters considered for recovery of stable transform ants included the choice of selective agent (phosphinothricin or hygromycin) and their respective selectable resistance genes (phosphinothricin acetyltransferase and hygromycin phosphotransferase), and preculture time of scale explants prior to bombardment. Polymerase chain reaction analysis will be used to screen putative stable transformants, and from this data it will be determined which parameters yielded the highest transformation rates.

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