Transient expression of semi-dominant Zm ChlI alleles in N. benthamina. A) Schematic of expression constructs, showing orientation of the 35S promoter (35S), maize chelatase I gene (ZmChlI), HA-epitope tag (HA) and 35S terminator (TER) sequences. Wild-type ZmChlI and a number of variants carrying polymorphisms present in semi-dominant mutant alleles were expressed in N. benthamiana. See text and Table 1 for details. B) Leaves of N. benthamiana 12 days post-infiltration. Each leaf was inoculated with two constructs. C) Immunoblot analysis of ZmCHLI accumulation following infiltration with pOY-1 to 5 (lanes 1–5, respectively) or from non-transformed (NT) tissues. Immunoblot was challenged with anti-HA antibodies. The predicted size of the mature CHLI-HA fusion is approximately 41 kDa. A cross-hybridizing protein recognized by the HA antibody at approximately 100 kDa was used as a loading control.