Project description:Pentatricopeptide repeat (PPR) proteins are RNA binding proteins that function in posttranscriptional regulation as gene-specific regulators of RNA metabolism in plant organelles. Plant PPR proteins are divided into four classes: P, PLS, E and DYW. The E- and DYW-class proteins are mainly implicated in RNA editing, whereas most of the P-class proteins predominantly participate in RNA cleavage, splicing and stabilization. In contrast, the functions of PLS-class proteins still remain obscure. Here, we report the function of PLS-class PpPPR_31 and PpPPR_9 in Physcomitrella patens. The knockout (KO) mutants of PpPPR_31 and PpPPR_9 exhibited slower protonema growth compared to the wild type. The PpPPR_31 KO mutants showed a considerable reduction in the splicing of nad5 intron 3 and atp9 intron 1. The PpPPR_9 KO mutants displayed severely reduced splicing of cox1 intron 3. An RNA electrophoresis mobility shift assay showed that the recombinant PpPPR_31 protein bound to the 5' region of nad5 exon 4 and the bulged-A region in domain VI of atp9 group II intron 1 while the recombinant PpPPR_9 bound to the translated region of ORF622 in cox1 intron 3. These results suggest that PLS-class PPR proteins may influence the splicing efficiency of mitochondrial group II introns.
2020-08-01 | GSE145050 | GEO
Project description:Editing factor knockout complementation in Physcomitrium patens
Project description:To investigate transcriptomic profiles when auxin-dependent transcription factor ARFC2 from Physcomitrium patens is expressed, we established an inducible overexpressor line for PpARFC2. We then perform bulk RNA-seq on protonemata tissues that have been mock-treated or induced by beta-estradiol for 16 h.
Project description:Physcomitrium (Physcomitrella) patens Gransden wild type strain and four mutants Δdek1 (Perroud et al., 2014); Δloop (Demko et al., 2014); Δlg3 (Johansen et al., 2016) and oex1 (this work) were sequenced for time series analysis.
Project description:RAPID ALKALINIZATION FACTOR (RALF) peptides are shown to regulate multiple physiological processes in plants, including modulation of immune response in angiosperms. This family of cysteine-rich peptide hormones has considerably expanded during land plant evolution, but the ancestral roles of RALF peptides in stress response remains poorly understood. Here, we used the moss Physcomitrium patens as a model to gain insight into the function of PpRALF peptides in response to abiotic and biotic stress factors in non-vascular plants. The quantitative proteomic analysis revealed cooperative down-regulation of M6 metalloproteases and membrane proteins, including those involved in stress response, in PpRALF1, 2 and 3 knockout (KO) lines.
Project description:Here, we provide an isobaric tag for relative and absolute quantitation (iTRAQ)-based quantitative proteomic dataset of the moss Physcomitrium patens smg1 knockout line. The kinase SMG1 is one of the key components of the NMD system in many organisms, including plants. Several lines with a deleted SMG1 in a land Physcomitrium patens subsp. patens (“Gransden 2004”, Frieburg) were produced by James P. B. Lloyd [doi.org/10.1111/tpj.12329]. One of these lines, SMG1 KO mutant line 2, was used for this study. Protonemata of WT and mutant lines were grown in200 ml liquid BCD medium supplemented with 5 mM ammonium tartrate (BCDAT) during a 16-h photoperiod at 250C for 8 days [https://doi.org/10.1101/pdb.prot5136]. All experiments were performed in three biological replicates.
Project description:At dawn of a scorching summer day, land plants must anticipate upcoming extreme midday temperatures by timely establishing molecular defenses that can maintain heat-labile membranes and proteins in a functional state. A gradual morning pre-exposure to increasing sub-damaging temperatures induces the accumulation of heat-shock proteins (HSPs) that are central to the onset of plant acquired thermotolerance (AT). In an attempt to gain knowledge on the mechanisms of AT in the model land plant Physcomitrium patens, we used label-free LC-MS/MS proteomics to quantify the accumulated and depleted proteins before and following a mild heat-priming treatment.
