Anti-Lhca1 | PSI type I chlorophyll a/b-binding protein

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

Immunoblot analysis of photosynthetic complex accumulation in wild-type tobacco and the two ?psaI lines grown under low, intermediate, and high-light conditions. Because the accumulation of most tested proteins was highest under high-light conditions, lanes one to three contain samples diluted to 25%, 50%, and a 100% sample of wild-type tobacco grown under high-light conditions, to allow for semi-quantitative determination of changes in protein abundance. Lanes four and five contain the two transplastomic lines grown at 1000 µE m?2 s?1. Lanes six to eight contain wild-type tobacco and the mutants grown at intermediate light intensities, and lanes nine to eleven contain samples grown at low light intensities. For PSII, the accumulation of the essential subunits PsbB (CP43) and PsbD (D2) and the LHCB1 antenna protein were determined, while for the cytochrome b6f complex, the accumulation of the essential redox-active subunits PetA (cytochrome f), PetB (cytochrome b6), and PETC (Rieske FeS protein) was tested. AtpB was probed as an essential subunit of the chloroplast ATP. For PSI, in addition to the three essential plastome-encoded subunits PsaA, PsaB, and PsaC, the accumulation of the nuclear-encoded subunits PSAD, PSAH, PSAK, PSAL, and PSAN and of the four LHCI proteins (LHCA1, LHCA2, LHCA3, LHCA4) was determined. Finally, we examined the accumulation of Ycf4, the chloroplast-encoded PSI-biogenesis factor encoded in the same operon as PsaI, and the nuclear-encoded assembly factor Y3IP1.

Lack of CP12-1 leads to decreased levels of PRK protein. (A) Equal crude protein samples from mature rosettes of the indicated genotypes were analysed by immunoblotting using the antibodies shown. PRK (phosphoribulokinase), GAPDH (glyceraldehyde-3-phosphate dehydrogenase), TK (transketolase), and LhcA (light-harvesting complex). Each sample represents a pool of 2–4 different plants. (B) qRT-PCR analysis of PRK gene expression in CP12 mutants. Relative levels of expression of the PRK gene in the CP12-T-DNA mutant collection and cp12-1/2/3RNAi plants with respect to wild-type (WT) expression levels. The results are the mean from 3–4 biological replicates, and the error bars indicate the SE. Actin2 (AT3G18780), the elongation factor gene (AT1G07940.1), and Cyclophilin (AT2G29960) were used as internal standards (mean CV, 0.1869; mean M value, 0.4668).

Assessment of the purity of isolated elaioplasts using immunoblots.Different fractions of the sucrose gradient (Band 1 to Band 4) are compared to peel proteins using antibodies for plastid stroma large Rubisco subunit (RbcL), cytosolic UGPase, vacuolar (v)-ATPase, mitochondrial voltage-dependent anion-selective channel protein 1 (VDAC1), and photosynthesis Light-harvesting complex (Lhca1 and Lhca4); b Coomassie blue protein profiles of purified elaioplasts (Band 1 to Band 4) from kumquat peel, as compared with purified chromoplast (Chro.) from sweet orange flesh and total kumquat peel proteins

Characterization of the CTE-disrupted pcya1-1 mutant. (A) Schematic diagram of an AphVIII insertion cassette into the PCYA1 locus in the pcya1-1 mutant. Black solid line represents 5?UTR and 3?UTR; black dotted line symbolizes intron; black box stands for exon and black arrows indicate the targeting sites of PCR primers. Purple, red, blue, and orange solid lines represent TP, NTE, FDBR and CTE regions of PCYA1, respectively. (B) PCR confirmation of the AphVIII insertion in pcya1-1. Genomic DNA was extracted from 4A+, HS211 and pcya1-1, respectively. PCR were performed using primers depicted in (A). HS211, the parental strain of pcya1-1. (C) Detection of CrPCYA1 expression in the dark or under light (?160 ?mol photons m-2s-1). Upper, immunoblot analysis of CrPCYA1 protein accumulation using antibody against CrPCYA1 mature protein; lower, Coomassie blue stain as an equal loading control. (D) Comparison of phototrophic growth phenotypes of pcya1-1 with 4A+, hmox1 and HS211 under constant low light (?60 ?mol photons m-2s-1), high light (?700 ?mol photons m-2s-1) and 12 h dark/12 h light cycle. (E) Accumulation of the photosystem I related proteins in pcya1-1, 4A+, HS211 and hmox1 in the dark or under light (?160 ?mol photons m-2s-1). Immunoblot analyses were performed with LHCA1 and PSAD antibodies. Coomassie blue stain (CB) as an equal loading control. Approximately 30 ?g total proteins were loaded in each lane.

PCYA1 knockdown by artificial microRNA. (A) Analysis of CrPCYA1 protein accumulation in artificial microRNA-mediated PCYA1 knockdown lines. (B) Phototrophic growth of PCYA1 knockdown mutants were compared with 4A+ and hmox1 under constant low light, high light and dark/light cycle. Low light, ?60 ?mol photons m-2s-1; High light, ?700 ?mol photons m-2s-1. (C) Accumulation of representative photosystem I related proteins in PCYA1 knockdown mutants, 4A+ and hmox1 in the dark or under light (?160 ?mol photons m-2s-1). LHCA1 and PSAD antibodies were used for immunoblotting analyses. Around 30 ?g total proteins were loaded in each lane.