Increasing labor market insecurities among young people? Labor market entry process in Hungary since the early 1980s

Pairwise SNVs for All Strain

Photosynthetic parameters in wild-type plants and the two <i>cgl160</i> T-DNA-insertion lines were determined for plants grown under long-day and plants grown under short-day conditions.

<p>Chlorophyll <i>a</i>/<i>b</i> ratio, chlorophyll content per leaf area, maximum quantum efficiency of PSII in the dark-adapted state, and maximum linear electron flux capacity, as determined from the yield of PSII, were determined on intact leaves. Photosynthetic complex contents were determined in isolated thylakoids, and re-normalized to a leaf area basis. The values represent averages of four to eight plants, the standard deviations are indicated. Data were subjected to a one-way analysis of variance (ANOVA) using a pair-wise multiple comparison procedure (Holm-Sidak method). Statistically significant differences, relative to the wild type grown under the corresponding growth regime, are shown in italics.</p><p>Photosynthetic parameters in wild-type plants and the two <i>cgl160</i> T-DNA-insertion lines were determined for plants grown under long-day and plants grown under short-day conditions.</p

The Thylakoid Membrane Protein CGL160 Supports CF₁CF₀ ATP Synthase Accumulation in <i>Arabidopsis thaliana</i>

<div><p>The biogenesis of the major thylakoid protein complexes of the photosynthetic apparatus requires auxiliary proteins supporting individual assembly steps. Here, we identify a plant lineage specific gene, <i>CGL160</i>, whose homolog, <i>atp1</i>, co-occurs with ATP synthase subunits in an operon-like arrangement in many cyanobacteria. <i>Arabidopsis thaliana</i> T-DNA insertion mutants, which no longer accumulate the nucleus-encoded CGL160 protein, accumulate less than 25% of wild-type levels of the chloroplast ATP synthase. Severe cosmetic or growth phenotypes result under either short day or fluctuating light growth conditions, respectively, but this is ameliorated under long day constant light growth conditions where the growth, ATP synthase activity and photosynthetic electron transport of the mutants are less affected. Accumulation of other photosynthetic complexes is largely unaffected in <i>cgl160</i> mutants, suggesting that CGL160 is a specific assembly or stability factor for the CF₁CF₀ complex. CGL160 is not found in the mature assembled complex but it does interact specifically with subunits of ATP synthase, predominantly those in the extrinsic CF₁ sub-complex. We suggest therefore that it may facilitate the assembly of CF₁ into the holocomplex.</p></div

Organization of the ATP synthase operon of cyanobacteria and co-expression of genes of the ATP synthase operon with CGL160 in Synechocystis sp PCC 6803.

<p>A. The genes encoding the ATP synthase are organized in a single operon together with atp1 that is related to CGL160 (previously CGLD22 in Chlamydomonas). The genes are depicted as arrows, with the orientation indicated by the direction of the arrow. This information was obtained from The SEED viewer (<a href="http://pubseed.theseed.org" target="_blank">http://pubseed.theseed.org</a>) B. The genes of the ATP synthase are co-expressed with atp1 as described in [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0121658#pone.0121658.ref041" target="_blank">41</a>]. The atp1 gene encodes the putative ortholog of CGL160; the curve showing the expression profile of atp1 is in red. The microarray data used to generate the expression curves were obtained from the Gene Expression Omnibus. Microarray values normalized against the median of the ratio of each sample against the reference, and log-transformed. The plotted data include 24 experiments. C-D. Affymetrix (ATH1 Gene Chip) Gene expression data in different Arabidopsis organs for CGL160 and for select nucleus-encoded genes. Genes for chloroplast localized ATP synthase subunits C, or D, mitochondrial ATP synthase. Data were retrieved from the Arabidopsis Electronic Fluorescent Pictograph (eFP) Browser. E. Expression profile of CGL160 and mitochondrial ATP synthase genes in maize (Zea mays) (ATP3, RMZM2G321725; ATP5, GRMZM2G156068) and chloroplast ATP synthase genes (ATPC, GRMZM2G048907; ATPD: GRMZM2G025171) during maize leaf development. Representative images for CGL160 and ATP synthase encoding genes are taken from <a href="http://bar.utoronto.ca/efp_maize/cgi-bin/efpWeb.cgi" target="_blank">http://bar.utoronto.ca/efp_maize/cgi-bin/efpWeb.cgi</a>. Proteins that are involved in photosynthesis typically exhibit peak expression in mature chloroplasts corresponding to leaf sections 9 and 14.</p

Isolation and characterization of Arabidopsis <i>cgl160</i> mutants.

<p>A. DNA insertion sites in CGL160 gene of Arabidopsis. DNA insertion sites (black triangles) are shown in relation to the CGL160 gene structure. The two <i>cgl160</i> alleles analyzed in this study are denoted as <i>cgl160-1</i> and <i>cgl160-2</i>. The CGL160 coding region is indicated by the translational start (ATG). The CGL160 genomic locus contains nine exons but only the first four are shown in the fig (grey boxes), shown are also the first four introns (black thin connecting lines). Before ATG is the promoter region in light gray. The region used for CGL160 specific antibody is shown as antigen. B. Characterization of CGL160 amount in Arabidopsis <i>cgl160-1</i> mutant from isolated chloroplasts. The CGL160 antibody was used for immunoblotting and 10 μg protein was loaded in each lane. The LHCB1 antibody was used as a loading control. C. Characterization of CGL160 amount in Arabidopsis <i>cgl160-2</i> mutant from isolated chloroplasts. The CGL160 antibody was used for immunoblotting and 10 μg protein was loaded in each lane. The LHCB1 antibody was used as a loading control.</p

Altered protein accumulation and stability of the chloroplast ATP synthase in the <i>cgl160</i> mutant visualized by immunoblotting.

<p>A. Immunoblots with antibodies against essential subunits of the photosynthetic protein complexes of wild-type (Col-4) Arabidopsis and the two <i>cgl160</i> 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 <i>b</i>₆<i>f</i> complex was probed with antibodies against the essential subunits PetA (cytochrome <i>f</i>), PetB (cytochrome <i>b</i>₆), 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 (CF₁α), AtpB (CF₁β) and AtpD (CF₁δ) and antibodies against the CF₀ subunits AtpF (CF₀b) and AtpI (CF₀a). B. Loading difference estimation for immunoblotting CF₁ between wild type and <i>cgl160-1</i>. To obtain similar immunoblotting signal three times more (15 μg protein) was needed for <i>cgl160-1</i> compared to wild type (5 μg protein). C. Maintenance of CF₁ was measured by incubating leaves from wild type and <i>cgl160-1</i> 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.</p

Total genes RNA-seq

This spreadsheet shows all the genes found by RNA-seq and their counts (RPKM) in all four samples, WT-mock, WT-Rose Bengal, sak1-mock, sak1-Rose Bengal. Each column shows results from a single sequencing lane, each sample was loaded on two lanes

Phenotypic characterization of Arabidopsis <i>cgl160</i> mutant plants.

<p>A. Top panel shows, from left to right, a wild type plant, <i>cgl160-1</i> and wild type, <i>cgl160-2</i> plants grown for 3 weeks under normal light conditions (120 μmol m^-2 s^-1) at long day (16 hours light and 8 hours dark). B. Wild type, <i>cgl160-1</i> and <i>cgl160-2</i> plants grown under short day conditions (120 μmol m^-2 s^-1; 8 hours light and 16 hours dark) for 4 weeks. C. Shows, from left to right, a wild type plant, <i>cgl160-1</i> and wild type, <i>cgl160-2</i> plants grown for 1 week under normal light conditions (120 μmol m^-2 s^-1) at long day (16 hours light and 8 hours dark) and then shifted to fluctuating low/normal light conditions (5 min 120 μmol m^-2 s^-1, 5 min 20 μmol m^-2 s^-1 changing every 5 min for 16 hours light and then 8 hours dark) for two weeks. D. Bar graph of Fv/Fm as a measure of photosynthetic performance under the indicated conditions.</p

2D native / SDS-PAGE gel immunoblotting and crosslinking indicate interaction between CGL160 and the chloroplast ATP synthase complex.

<p>A. Blue-native gel electrophoresis analysis of thylakoidal protein complexes in wild type and mutant plants. PSII-SC photosystem II supercomplex, PSII-D photosystem II dimer, PSII-M photosystem II monomer, LHCII-T light harvesting complex II trimer. The Blue-native gel was used for immunoblotting against D1, CF₁ and CGL160 antibodies as indicated. B. Solubilized and cross-linked thylakoid membranes were separated by SDS-PAGE and probed with specific antibodies against CGL160 and CF₁. C. Detection of CGL160 and ATP synthase subcomplexes by immunoblot analyses of 2D BN/SDS gels as in panel A, employing antibodies specific for CGL160 and individual ATPase subunits as indicated in the figure.</p

Linear electron transport, non-photochemical quenching (qN) and redox state of the PSII acceptor side are affected in <i>cgl160</i> mutants both under long- and short-day conditions.

<p>Measurements were done in long (A-C)—and short-day (D-F) adapted plants (16 hours light and 8 hours light, respectively). A and D. Light response curve of linear electron flux as calculated from PSII yield measurements. In both <i>cgl160</i> T-DNA insertion lines, linear electron flux capacity is reduced. B and E. Light response curves of non-photochemical quenching (qN). In both T-DNA insertion lines, the induction of qN is shifted towards lower light intensities, but the maximum light-saturated qN is unaltered C and F. Light response curves of the redox state of the PSII acceptor side, determined as qL. When qL is one, Q_A is fully oxidized; when qL is zero, Q_A is fully reduced. With increasing light intensity, the PSII acceptor side becomes more rapidly reduced in both T-DNA insertion lines.</p