Project description:To gain insight into the basic mechanism of Cd detoxification in the methylotrophic yeast, H. polymorpha, we analyzed temporal changes in transcriptional profiles in response to Cd exposure. We used H. polymorpha whole-genome cDNA microarrays, which contain 98% of all H. polymorpha ORFs in duplicate or triplicate. Keywords: time course, cadmium
Project description:Transcriptome profiling analysis of the Hansenula polymorpha MET4 gene deletion strain have been carried out to obtain comprehensive information on the HpMet4p-mediated regulatory networks in association with the cadmium (Cd) detoxification and sulfur regulation in H. polymorpha. Total RNA samples were collected from H. polymorpha wild type, and HpMET4 deletion strain, under sulfur starvation or Cd (0.6 mM) stress conditions. The differential fluorescence intensities of each RNA sample were measured after labeling with Cy3 or Cy5. For all analyses, we performed dye swapping experiments to avoid dye bias. Thus, four intensity values were generated for each ORF and averaged for analysis.
Project description:To gain insight into the basic mechanism of Hydrogen peroxide detoxification in the methylotrophic yeast, H. polymorpha, we analyzed changes in transcriptional profiles in response to hydrogen peroxide exposure. Total RNA samples were collected from H. polymorpha cells after 30 min incubation with 0.5mM hydrogen peroxide. Using the RNA sample obtained prior to hydrogen peroxide addition as a reference, the differential fluorescence intensities of each RNA sample prepared at the indicated time was measured after labeling with Cy3 or Cy5 fluorochromes. For all analyses, we performed dye swapping experiments to avoid dye bias.
Project description:Transcriptome profiling analysis of the Hansenula polymorpha MET4 gene deletion strain have been carried out to obtain comprehensive information on the HpMet4p-mediated regulatory networks in association with the cadmium (Cd) detoxification and sulfur regulation in H. polymorpha.
Project description:In the present study, Marchantia polymorpha Mppcs loss of function mutants were generated through CRISPR/cas9 mediated genome-editing. To assess whether the knockout of MpPCS gene affects the transcription of M. polymorpha nuclear genes in unstressed condition, the Mppcs-2 knockout mutant and Cam2 wild-type transcriptomes were compared by RNA-Seq.
Project description:In order to elucidate the role of the single Marchantia B-GATA ortholog in response to high light intensities, a transcriptomic analysis of Marchantia polymorpha BoGa, Mpb-gata1 mutants and MpB-GATA1ox under high-ligh stress conditions was performed.
Project description:Pattern-recognition receptor (PRR)-triggered immunity (PTI) plays a pivotal role in plant immunity to ward off a wide range of pathogenic microbes. The model liverwort Marchantia polymorpha is gaining popularity in investigating the evolution of plant-microbe interactions. The M. polymorpha is capable of triggering defense-related gene expression by sensing components in bacterial and fungal extracts, suggesting existence of PTI in this plant model. However, the molecular components that would form PTI in M. polymorpha have not yet been described. We show that, in M. polymorpha, lysin motif (LysM) receptor-like kinase (LYK) MpLYK1 and LYK-related (LYR) MpLYR, among four LysM receptor homologs, are required for sensing chitin and peptidoglycan (PGN) fragments and thereby triggering a series of immune responses. Phosphoproteomic analysis of M. polymorpha in response to chitin treatment comprehensively identified regulatory proteins that would shape LysM-mediated PTI. The identified proteins covered homologs of well-described PTI components in angiosperms as well as proteins whose roles in PTI are not yet determined including the blue-light receptor phototropin MpPHOT. We revealed that MpPHOT is required for a negative feedback of defense-related gene expression during PTI. Taken together, this study provides the basic framework of LysM-mediated PTI in M. polymorpha and demonstrates the utility of M. polymorpha as a plant model for discovering novel or fundamental molecular mechanisms underlying PRR-triggered immune signaling in plants.
Project description:Pattern-recognition receptor (PRR)-triggered immunity (PTI) plays a pivotal role in plant immunity to ward off a wide range of pathogenic microbes. The model liverwort Marchantia polymorpha is gaining popularity in investigating the evolution of plant-microbe interactions. The M. polymorpha is capable of triggering defense-related gene expression by sensing components in bacterial and fungal extracts, suggesting existence of PTI in this plant model. However, the molecular components that would form PTI in M. polymorpha have not yet been described. We show that, in M. polymorpha, lysin motif (LysM) receptor-like kinase (LYK) MpLYK1 and LYK-related (LYR) MpLYR, among four LysM receptor homologs, are required for sensing chitin and peptidoglycan (PGN) fragments and thereby triggering a series of immune responses. Phosphoproteomic analysis of M. polymorpha in response to chitin treatment comprehensively identified regulatory proteins that would shape LysM-mediated PTI. The identified proteins covered homologs of well-described PTI components in angiosperms as well as proteins whose roles in PTI are not yet determined including the blue-light receptor phototropin MpPHOT. We revealed that MpPHOT is required for a negative feedback of defense-related gene expression during PTI. Taken together, this study provides the basic framework of LysM-mediated PTI in M. polymorpha and demonstrates the utility of M. polymorpha as a plant model for discovering novel or fundamental molecular mechanisms underlying PRR-triggered immune signaling in plants.