An agrobacterium-mediatesd transie4nt gene expression system for intact leaves

An efficient and reproducible AgrohLlc~teriut,l-mediated transient gene expression system for intact leaf tissue was developed. A high level of transient expression was observed when bacteria. which were pretreated in C?Y gene-induc- ing conditions. were infiltrated into complete leaf tissue. Histochemical /I-glucuronidase assays showed large transgene-expressing sectors comprising of up to 90 percent of the leaf area. As a consequence of infiltration, the induced bacteria entered into the intercellular spaces, thus enabling T-DNA transfer in all cell layers of the leaf. The protocol was optimized for Phasrolus wlgaris leaves. but similar results were obtained with other plant species. such as P/mroh~.s uc,utifdim. poplar, and tobacco. A /I-glucuronidase chimeric gene interrupted by an intron was used as a marker for histological detection of the sectors. Copyright :CJ 1997 Elsevier Science Ireland Ltd.