AGROBACTERIUM-MEDIATED TRANSFORMATION OF POPULUS X EURAMERICANA OGY USING THE CHIMERIC CAMV 35S PIN2 GENE FUSION

A hybrid poplar clone, ''Ogy'' (Populus x euramericana), was transform ed with the potato proteinase inhibitor II (PI II) gene (pin2) to test the importance of serine proteinases in host-pathogen and host-insect interactions. The gene product PI IT is a specific inhibitor of tryps in and chymotrypsin. Transformants were obtained using an Agrobacteriu m binary vector system with a disarmed Ti plasmid (EHA 101) and plasmi d pRT104 containing pin2 regulated by a CaMV 35S promoter and a select able gene-encoding neomycin phosphotransferase II (NPT II). Putative t ransformed shoots from co-cultured leaves were selected using three di fferent kanamycin toxicity assays. Ten plantlets, selected from 431 sh oots derived from co-culture, were selected for nucleic acid and prote in analysis. DNA analysis (Southern blots and PCR) and protein gel blo ts (Western blots) confirmed pin2 integration and expression in transf ormant number TR5.