Extended Data Fig. 1: Spectroscopy characterization of the wild type (WT), the pbf8 mutant, and pbf8 com, and identification of the pbf8 mutant.
From: Uncovering the photosystem I assembly pathway in land plants
a, Chlorophyll contents in WT, pbf8, and pbf8 com. Chlorophyll a (green) and chlorophyll b (yellow) contents were normalized to fresh weight (FW), with the chlorophyll a/b ratio above each bar. Statistical significance was determined by one-way ANOVA. P-values were adjusted by Tukey’s post-hoc test; **P < 0.01; ns, not significantly different. Data are means ± SD (n = 3 independent biological replicates). b, 77 K chlorophyll a fluorescence emission spectra of thylakoid membranes isolated from WT, pbf8, and pbf8 com upon excitation at 436 nm. Signals were normalized to the PSII emission maximum at 687 nm. c, Chlorophyll a fluorescence induction kinetics in WT, pbf8, and pbf8 com. AL, actinic light; FR, far-red light; SL, saturating light; Fm, maximal fluorescence; \({F}_{{\rm{m}}}^{{\prime} }\), maximal fluorescence in the light-adapted state; Fo, minimal fluorescence; \({F}_{{\rm{o}}}^{{\prime} }\), minimal fluorescence in the light-adapted state; Fv/Fm, maximum efficiency of PSII photochemistry. Data are means ± SD (n = 3 independent biological replicates). d, Chloroplast ultrastructure. (i–iii), Chloroplast ultrastructure of WT, pbf8, and pbf8 com. (iv–vi), Magnified images of the regions indicated by red frames in (i–iii). GT, grana thylakoids; SL, stromal lamellae. Bars, 1 μm (i–iii), 0.2 μm (iv–vi). e, Diagram of the T-DNA insertion in PBF8 (At3g56010). The T-DNA is inserted into the second exon. f, Identification of homozygous pbf8 plants. Genomic DNA was used as a template for PCR analysis with specific primers. LP, left border primer; RP, right border primer; o8474, specific T-DNA primer for GABI-Kat lines. Primer sequences are listed in Supplementary Table 2. g, Reverse-transcription PCR (RT-PCR) analysis of PBF8 transcript levels in WT, pbf8, and pbf8 com plants using specific primers listed in Supplementary Table 2. ACTIN2 was used as a reference transcript. h, Immunoblot analysis of PBF8 protein levels in WT, pbf8, and pbf8 com plants. Equal amounts of total leaf protein (10 μg) extracted from plants were separated by SDS-PAGE and immunodetected using the anti-PBF8 antibody generated in this study. β-Actin was used as a loading control. In b–d, f–h, three independent biological replicates were performed, and a representative replicate is shown.
