DETERMINATION OF A GENOTYPE INDEPENDENT METHOD OF MAIZE TRANSFORMATION

WE PROPOSE TO EXAMINE A METHOD OF GENOTYPE INDEPENDENT GENETIC TRANSFORMATION OF ZEA MAYS (MAIZE). THE DNA DELIVERY WILL BE ACCOMPLISHED USING HIGH-VELOCITY MICROPROJECTILES. THE FIRST STEP IN THE EXPERIMENT WILL BE TO TEST THE VARIOUS TISSUES OF MAIZE AS POTENTIAL TARGETS FOR THE INCORPORATION OF DNA. THE TISSUES CHOSEN WILL EACH BE CAPABLE OF TRANSMITTING INTEGRATED DNA TO THE NEXT GENERATION OF PLANTS AND WILL NOT REQUIRE ANY HIGHLY GENOTYPE-DEPENDENT TISSUE CULTURE. TO EVALUATE THEIR SUITABILITY, EACH TARGET TISSUE WILL BE BOMBARDED WITH MICROPROJECTILES CONTAINING A B-GLUCURONIDASE GENE SO THAT TRANSIENT EXPRESSION OF THE NEWLY DELIVERED DNA CAN BE MEASURED BY THE PRODUCTION OF BLUE COLOR. THESE TISSUES WILL THEN BE EVALUATED TO DETERMINE THE EFFECT OF DNA BOMBARDMENT ON THEIR NORMAL DEVELOPMENT. THE SECOND STEP IN THE EXPERIMENT WILL BE TO USE A COLOR MARKER AS A MEANS OF DETERMINING WHETHER STABLY TRANSFORMED TISSUES CAN BE OBTAINED FROM EACH TARGET TISSUE. THESE THREE PIECES OF INFORMATION (TRANSIENT EXPRESSION, TISSUE DEVELOPMENT, AND STABLE EXPRESSION) WILL ALLOW US TO DETERMINE THE OPTIMAL CONDITIONS FOR A GENOTYPE INDEPENDENT METHOD OF TRANSFORMATION OF MAIZE.