Figure 3

PCR-based mutagenesis of Zm ChlI. Full-length ZmChlI was amplified using a mutagenic PCR procedure and fragments cloned into the pGEM T-Easy. Plasmids were electroporated into E. coli and DNA from several clones sequenced. Fragments were then subcloned into the pBIN61 binary vector and transformed into Agrobacterium. Clones were grown overnight and expressed in N. benthamiana using agroinfiltration. Following infiltration, plants were screened for the development of chlorosis indicating the generation of novel dominant mutant variants.