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. 2021 Jan 25;185(4):1574–1594. doi: 10.1093/plphys/kiab010

Figure 3.

Figure 3

The effect of transient expression of C-terminal GFP and HA fusions of ascorbate biosynthesis enzymes on ascorbate concentration in N. benthamiana leaves. A, Strategy followed to clone and to overexpress ascorbate biosynthesis genes from Arabidopsis translationally fused to GFP and HA at the protein C-terminus in N. benthamiana leaves. B, Immunoblots (α-GFP) of fusion protein accumulation 2, 3, and 4 d after agroinfiltration. Expression was driven by the 35S promoter. C–H, Leaf ascorbate concentration 3 d after agroinfiltration with different combinations of C-terminal GFP ad HA fusions of ascorbate biosynthesis enzymes. In H, “Cytosolic” refers to the coexpression of GMP, GME, GGP, GPP, and L-GalDH genes; “Whole” refers to the coexpression of those mentioned cytosolic genes plus mitochondrial L-GalLDH. For further details, constructs used in each infiltration are shown in Supplementary Table S1. Data displayed correspond to mean±SD from two leaves of at least three N. benthamiana plants infiltrated, collected at 3 d after infiltration. Different letters denote statistically significant differences for ascorbate concentration (one-way ANOVA, α = 0.05, post hoc Tukey test).