Anti-PsbO | 33 kDa of the oxygen evolving complex (OEC) of PSII (anti-peptide)

Subcellular and suborganellar localization of ANU10. (A–F) Confocal micrographs of the subepidermal layer of palisade mesophyll cells from (A–C) Ler and (D–F) anu10-1 35Spro:ANU10:GFP transgenic plants. Micrographs show (A, D) the chlorophyll autofluorescence of the chloroplasts, (B, E) the GFP fluorescence, and (C, F) an overlay of the chlorophyll and GFP signals, showing their co-localization in (F). Pictures were taken from first-node leaves collected 16 das. Scale bars indicate 50 ?m. (G) Western blot analysis of the proteins in chloroplast, stroma, and thylakoid fractions isolated from anu10-1 35Spro:ANU10:GFP transgenic plants collected 16 das. Primary antibodies against GFP, the large Rubisco subunit (RbcL), and the PsbO subunit of PSII were used. Molecular mass markers are indicated on the right.

Altered protein accumulation and stability of the chloroplast ATP synthase in the cgl160 mutant visualized by immunoblotting.A. Immunoblots with antibodies against essential subunits of the photosynthetic protein complexes of wild-type (Col-4) Arabidopsis and the two cgl160 T-DNA insertion lines grown under long-day and short-day conditions. Isolated thylakoid membranes were used, and equal amounts of chlorophyll were loaded onto the SDS-PAGE gel. For approximate quantification, wild-type samples from long-day plants were diluted to 10%, 25% and 50%, respectively. Accumulation of PSII was probed with antibodies against PsbB and PSBO. Additionally, the PSBS protein involved in NPQ and the minor PSII antenna protein LHCB4 were probed. Accumulation of the cytochrome b6f complex was probed with antibodies against the essential subunits PetA (cytochrome f), PetB (cytochrome b6), and PETC (Rieske protein). Accumulation of PSI was probed with antibodies against the reaction center subunit PsaB and the stromal ridge subunit PsaD. ATP synthase accumulation was probed with antibodies against the CF1 subunits AtpA (CF1?), AtpB (CF1?) and AtpD (CF1?) and antibodies against the CF0 subunits AtpF (CF0b) and AtpI (CF0a). B. Loading difference estimation for immunoblotting CF1 between wild type and cgl160-1. To obtain similar immunoblotting signal three times more (15 ?g protein) was needed for cgl160-1 compared to wild type (5 ?g protein). C. Maintenance of CF1 was measured by incubating leaves from wild type and cgl160-1 in solution containing the plastid protein synthesis inhibitor chloramphenicol for the indicated time points. Protein extract was isolated and separated by SDS-PAGE, immunoblotted and probed with specific antibodies against CF1 and LHCB2.1. Three times more protein was loaded from the mutant to obtain equal level of CF1 immunoblotting signal, as specified in B.

O2 evolution and protein profile of photosynthetic apparatus in A. thaliana seedlings with generational exposure to Fe deficiency. (A) SHR-trap 1445 c0, pH 7.7 s1 pH 7.7 s2, pH 7.7 s1 c2 seedlings were grown for 11 days in control AIS medium and net O2 evolution (expressed as ?mol O2 evolved min-1 mg chlorophyll-1) was measured under illumination at either 100 or 800 ?E m-2 s-1. Values are mean ± SE of three biological replicas, each consisting of at least 15 seedlings. Letters represent statistical differences, according to Student’s t-test, with p < 0.05. (B) Western blot analysis with antibodies against PsaA, CP43, PsbO, D1, LHCII, and PsbE of thylakoid membranes purified from 11 days-old SHR-trap 1445 c0 seedlings germinated in -Fe AIS medium or from 11 days-old SHR-trap 1445 c0, pH 7.7 s1, pH 7.7 s1 pH 7.7 s2, pH 7.7 s1 c2 seedlings germinated in control (+Fe) AIS medium. Samples corresponding to 1 ?g chlorophyll were loaded on each lane.

Changes of PSII and PSI antenna and core protein levels. Proteins from control and Tl-treated white mustard leaves were separated by SDS-PAGE followed by immunodetection with antibodies against Lhcb1, Lhcb2, Lhca1 (antenna proteins) and D1, D2, CP43, PsbO, PsaA (core proteins). Samples were loaded on the equal amount of chlorophyll (0.25 ?g). Description of samples abbreviation as given in the legend to Fig. 3

Changes in the protein levels of photosynthetic components under extended dark exposure in wild-type and gamera-1. Immunoblot detection of photosynthetic proteins from leaves of Ws and gamera-1 plants incubated after dark adaptation for 0, 2, and 4 days was examined. Specifically, essentially thylakoid fractions were assayed to determine the content of the following proteins: ?-subunit of ATP synthase; the D1 protein, OEC17, OEC23, and OEC33 of PSII; Cyt f and Rieske protein of the cytochrome b6f complex; and the F and D subunits of PSI, after extended dark treatment. Proteins were resolved via SDS-PAGE gel based on equal microgram chlorophyll per lane loading and processed as described in Section “Materials and Methods”. The Large subunit of RuBisco and LHCII stained with either CBB or Ponceau red, respectively, are presented here as loading controls. DAD indicates days after dark adaptation.