Phenotypes of the Wild Type, trx m124-2, crr2-2 trx m124-2, and pgr5 trx m124-2 Mutants.
Seedlings were grown on MS medium in growth chambers (50 µmol photons m–2 s–1; continuous light; 23°C) for 3 to 4 weeks.
(A) Photographs of the plants. WT, wild type. Bar = 10 mm.
(B) Fresh weight of seedlings. Each value is shown as the mean ± sd (n = 10 to 15 independent plants). Columns with the same letters are not significantly different between genotypes according to one-way ANOVA with Tukey–Kramer multiple comparison test, P < 0.05 (Supplemental Table 5). WT, wild type.
(C) Chlorophyll content of seedlings, per unit fresh weight. Each value is the mean ± sd (n = 4 to 6 independent plants). Columns with the same letters are not significantly different between genotypes according to one-way ANOVA with Tukey–Kramer multiple comparison test, P < 0.05 (Supplemental Table 6). WT, wild type.
(D) and (E) Immunoblot analysis in the wild type (WT), crr2-2, pgr5, trx m124-2, crr2-2 trx m124-2, pgr5 trx m124-2, and 35Spro:PGR5. Chloroplasts were fractionated into the stromal fractions and thylakoid membranes. Trxs m1, m2, and m4, Trx f1, SBPase, FBPase, NADP-MDH, and RbcS were immunodetected using specific antibodies in the stromal fractions (D). PGR5, PGRL1, NDF1, PsbO, PetC, PsaD, and ATPC1 were immunodetected using specific antibodies in the thylakoid membranes (E). Protein samples corresponding to 1.0 µg chlorophyll were loaded in each lane.
(F) to (M) Chloroplasts in the wild type (WT), trx m124-2, and pgr5 trx m124-2. Chloroplast ultrastructure of WT (F) and (G), trx m124-2 (see [H] to [K]), and pgr5 trx m124-2 (see [L] and [M]). Seedlings were observed using TEM. Bar in (F), (H), (K), and (L) = 5 µm; bar in (G), (I), (J), and (M) = 1 µm.