Figure 4.

Validation of chloroplast targeting peptide for ZmLOX6. Precise cleavage site of the signal peptide was determined by N‐terminal sequencing of the immunopurified mature ZmLOX6 protein from maize leaves. DNA encoding 1–62 amino acids long target peptide was ligated to a green fluorescent protein (AcGFP) gene and transformed into maize seedling leaves for transient expression (a‐d). Guard cell pair of a single stomate is visible when autofluorescence is overlaid with chlorophyll fluorescence (a); (b) chlorophyll fluorescence alone; (c) AcGFP fluorescence alone; (d) chlorophyll fluorescence overlaid with AcGFP fluorescence; (e) chlorophyll and AcGFP fluorescence together with a 60 amino acids long signal peptide fused to AcGFP and (f) AcGFP fluorescence alone with the 60 amino acids long signal peptide. AcGFP was targeted to cytoplasm when the signal peptide length was truncated by 2 amino acids to 60 amino acids (e and f).