Project description:Gene expression in plastids of higher plants is dependent on two different transcription machineries, a plastid-encoded bacterial-type RNA polymerase (PEP) and a nuclear-encoded phage-type RNA polymerase (NEP), which recognize distinct types of promoters. The division of labor between PEP and NEP during plastid development and in mature chloroplasts is unclear due to a lack of comprehensive information on promoter usage. Here we present a thorough investigation into the distribution of PEP and NEP promoters within the plastid genome of barley (Hordeum vulgare L). Using a novel differential RNA sequencing approach, which discriminates between primary and processed transcripts, we obtained a genome-wide map of transcription start sites in plastids of mature first leaves. PEP-lacking plastids of the albostrians mutant allowed for the unambiguous identifications of NEP promoters. We observed that the chloroplast genome contains many more promoters than genes. According to our data, most genes (including genes coding for photosynthesis proteins) have both PEP and NEP promoters. We also detected numerous transcription start sites within operons indicating transcriptional uncoupling of genes in polycistronic gene clusters. Moreover, we mapped many transcription start sites in intergenic regions, as well as opposite to annotated genes demonstrating the existence of numerous non-coding RNA candidates. dRNA-seq analysis of total RNA from green and white plastids of the barley mutant line albostrians
Project description:Gene expression in plastids of higher plants is dependent on two different transcription machineries, a plastid-encoded bacterial-type RNA polymerase (PEP) and a nuclear-encoded phage-type RNA polymerase (NEP), which recognize distinct types of promoters. The division of labor between PEP and NEP during plastid development and in mature chloroplasts is unclear due to a lack of comprehensive information on promoter usage. Here we present a thorough investigation into the distribution of PEP and NEP promoters within the plastid genome of barley (Hordeum vulgare L). Using a novel differential RNA sequencing approach, which discriminates between primary and processed transcripts, we obtained a genome-wide map of transcription start sites in plastids of mature first leaves. PEP-lacking plastids of the albostrians mutant allowed for the unambiguous identifications of NEP promoters. We observed that the chloroplast genome contains many more promoters than genes. According to our data, most genes (including genes coding for photosynthesis proteins) have both PEP and NEP promoters. We also detected numerous transcription start sites within operons indicating transcriptional uncoupling of genes in polycistronic gene clusters. Moreover, we mapped many transcription start sites in intergenic regions, as well as opposite to annotated genes demonstrating the existence of numerous non-coding RNA candidates.
Project description:Signals originating within plastids modulate organelle differentiation by transcriptionally regulating nuclear-encoded genes. These retrograde signals are also integral regulators of plant development, including leaf morphology. The clb5 mutant displays severe leaf morphology defects due to Apocarotenoid Signal 1 (ACS1) accumulation in the developmentally arrested plastid. Transcriptomic analysis of clb5 validates ACS1 as a true retrograde signal impacting expression of hundreds of nuclear genes, including suppression of most genes encoding plastid ribosomal proteins.
2021-06-02 | GSE152252 | GEO
Project description:Novel plastid genome characteristics in Fugacium kawagutii and accelerated evolution of plastid proteins in dinoflagellates
Project description:The unicellular, free-living, nonphotosynthetic chlorophycean alga Polytomella parva, closely related to Chlamydomonas reinhardtii and Volvox carteri, contains colorless, starch-storing plastids. The P. parva plastids lack all light-dependent processes but maintain crucial metabolic pathways. The colorless alga also lacks a plastid genome, meaning no transcription or translation should occur inside the organelle. Here, using an algal fraction enriched in plastids as well as publicly available transcriptome data, we provide a proteomic characterization of the P. parva plastid, ultimately identifying several plastid proteins, both by mass spectrometry and bioinformatic analyses. Altogether these results led us to propose a plastid proteome for P. parva, i.e., a set of proteins that participate in carbohydrate metabolism; in the synthesis and degradation of starch, amino acids and lipids; in the biosynthesis of terpenoids and tetrapyrroles; in solute transport and protein translocation; and in redox homeostasis. This is the first detailed plastid proteome from a unicellular, free-living colorless alga.
Project description:Plants adapt to environmental changes by adjusting growth and defense, and the role of epigenetic modifications in this process is unclear. Sensing and adjusting to environmental changes are more important in certain tissues such as epidermis, vasculature, meristem, and reproductive tissues. These tissues possess sensory plastids that are specialized in environmental sensing. We show perturbation of four sensory plastid proteins MSH1, PPD3, CUE1, and SAL1 induce gene expression and DNA methylation changes targeted to networks associated to environmental sensing, with significant overlap with hda6-induced CHG hypermethylated genes at 12-hr daylength. At 16-hr daylength, hda6 loses CHG hypermethylation in gene body, and sensory plastid mutants have weaker phenotypes and DNA methylation- and gene expression- associated gene networks. We show daylength-responsive epistatic interaction between sensory plastid mutants with hda6. We also show that hda6 mutation confers daylength memory and, with msh1, enhanced tolerance to heat stress and biotic stress. These results suggest that HDA6 mediates programmed adjustments in plant phenotype triggered by sensory plastid-to-nucleus retrograde signaling in response to daylength and environmental cues.
Project description:Plants adapt to environmental changes by adjusting growth and defense, and the role of epigenetic modifications in this process is unclear. Sensing and adjusting to environmental changes are more important in certain tissues such as epidermis, vasculature, meristem, and reproductive tissues. These tissues possess sensory plastids that are specialized in environmental sensing. We show perturbation of four sensory plastid proteins MSH1, PPD3, CUE1, and SAL1 induce gene expression and DNA methylation changes targeted to networks associated to environmental sensing, with significant overlap with hda6-induced CHG hypermethylated genes at 12-hr daylength. At 16-hr daylength, hda6 loses CHG hypermethylation in gene body, and sensory plastid mutants have weaker phenotypes and DNA methylation- and gene expression- associated gene networks. We show daylength-responsive epistatic interaction between sensory plastid mutants with hda6. We also show that hda6 mutation confers daylength memory and, with msh1, enhanced tolerance to heat stress and biotic stress. These results suggest that HDA6 mediates programmed adjustments in plant phenotype triggered by sensory plastid-to-nucleus retrograde signaling in response to daylength and environmental cues.
Project description:Plastids emit signals that broadly affect cellular processes. Based on previous genetic analyses, we propose that plastid signaling regulates the downstream components of a light signaling network and that these interactions coordinate chloroplast biogenesis with both the light environment and development by regulating gene expression. We tested these ideas by analyzing light-regulated and plastid-regulated transcriptomes. We found that the plastid is a major regulator of light signaling, attenuating the expression of more than half of all light-regulated genes in our dataset and changing the nature of light regulation for a smaller fraction of these light-regulated genes. Our analyses provide evidence that light and plastid signaling are interactive processes and are consistent with these interactions serving as major drivers of chloroplast biogenesis and function.
Project description:Plastids communicate with the nucleus by means of retrograde plastid signals. The far-red (FR) light insensitive Arabidopsis mutant laf6 disrupted in a plastid-localised ABC-like protein (atABC1) accumulates the plastid signal protoporphyrin IX (proto IX) and has attenuated nuclear gene expression (Moller et al.2001 Genes Dev. 15:90-103). Our data suggests that proto IX accumulation results in hypocotyl elongation in response to FR light and we have demonstrated that by inhibiting the plastid localised protoporphyrinogen IX oxidase (PPO) using flumioxazin wild-type plants phenocopy laf6 by accumulating proto IX with a concomitant loss of hypocotyl growth inhibition in a dose-dependent manner. It is at present unclear what effect increased proto IX has on nuclear gene expression and how this is integrated with photomorphogenic responses such as hypocotyl elongation.
Project description:Biogenesis of plastid ribosomes is facilitated by auxiliary factors which process and modify ribosomal RNAs (rRNA) or are involved in ribosome assembly. In comparison to their bacterial and mitochondrial counterparts, the biogenesis of plastid ribosomes is less well understood and few auxiliary factors have been described so far. In this study, we report the functional characterization of CGL20 (CONSERVED ONLY IN THE GREEN LINEAGE20) in Arabidopsis thaliana (AtCGL20) a small, proline-rich protein which is targeted to mitochondria and chloroplasts. In Arabidopsis, CGL20 is encoded by segmentally duplicated genes of high similarity (AtCGL20A and AtCGL20B), and inactivation of both in the atcgl20ab mutant leads to a visible virescent phenotype, and growth arrest at low temperature. The chloroplast proteome, thylakoid complex abundance and pigment composition are significantly affected in atcgl20ab mutants, resulting in a combined proton gradient regulation (pgr) and chlororespiratory reduction (crr) phenotype. Loss of AtCGL20 alters plastid rRNA ratios, perturbs the formation of a hidden break in 23S rRNA and causes abnormal accumulation of 50S ribosomal subunits in the high-molecular-mass fraction of chloroplast stromal extracts. Moreover, AtCGL20A-eGFP fusion proteins comigrate with 50S ribosomal subunits in sucrose density gradients, even after RNase treatment of stromal extracts. Therefore, we propose that AtCGL20 participates in late stages of the biogenesis of 50S ribosomal subunits in plastids – a role which presumably evolved in the green lineage as a consequence of structural divergence of plastid ribosomes.