E-MTAB-10927
RNA-seq analysis of the fungal mycelium of the plant pathogen Pyricularia oryzae treated with and antimicrobial peptide analog of a Trichoderma peptaibol
We aimed at investigating the mode of action of an antimicrobial peptide analog of a Trichoderma peptaibol.  Conidia of P. oryzae (IT10 strain) were cultured in Complete Medium at a final concentration of 2x105 conidia per mL for 3 days at 30 °C under shaking at 150 rpm. After three days, 50 µM of the Peptide 4rink were added to samples containing actively growing mycelium. Untreated samples were used as control. Three h after peptide treatment, fungal mycelia were collected, filtered with 100 µm Cell Strainer (Corning®), rinsed with sterile water and stored at -80 °C for RNA extraction. Total RNA was extracted with the RNeasy Plant Mini kit (Qiagen) following the manufacturer's instructions, performing the DNase digestion directly on the column. The integrity and concentration of the RNA samples were checked with Qubit fluorimeter and Bioanalyzer. RNA samples with RIN values < 7 were discarded. Four biological replicates of each treatment were submitted to RNA sequencing and analysis at CRIBI Biotechnology Center (University of Padova). Two µg of total RNA were subjected to PolyA tail capture and rRNA depletion for selection of mRNA. Directional (stranded) RNA libraries were prepared, quantified and subjected to quality control. Sequencing was performed on Illumina Nova Seq 6000 platform. Twenty million reads were produced for each sample and checked for quality control. Based on our RNA-seq data, the molecular mechanisms of the fungicidal activity of these peptide analogs appear to be related to their ability to cause cell wall and membrane damages and to induce autophagic cell death in fungal cells. This is the first transcriptomic analysis on a fungus treated with peptaibol analogs.
We aimed at investigating the mode of action of an antimicrobial peptide analog of a Trichoderma peptaibol.  Conidia of P. oryzae (IT10 strain) were cultured in Complete Medium at a final concentration of 2x105 conidia per mL for 3 days at 30 °C under shaking at 150 rpm. After three days, 50 µM of the Peptide 4rink were added to samples containing actively growing mycelium. Untreated samples were used as control. Three h after peptide treatment, fungal mycelia were collected, filtered with 100 µm Cell Strainer (Corning®), rinsed with sterile water and stored at -80 °C for RNA extraction. Total RNA was extracted with the RNeasy Plant Mini kit (Qiagen) following the manufacturer's instructions, performing the DNase digestion directly on the column. The integrity and concentration of the RNA samples were checked with Qubit fluorimeter and Bioanalyzer. RNA samples with RIN values < 7 were discarded. Four biological replicates of each treatment were submitted to RNA sequencing and analysis at CRIBI Biotechnology Center (University of Padova). Two µg of total RNA were subjected to PolyA tail capture and rRNA depletion for selection of mRNA. Directional (stranded) RNA libraries were prepared, quantified and subjected to quality control. Sequencing was performed on Illumina Nova Seq 6000 platform. Twenty million reads were produced for each sample and checked for quality control. Based on our RNA-seq data, the molecular mechanisms of the fungicidal activity of these peptide analogs appear to be related to their ability to cause cell wall and membrane damages and to induce autophagic cell death in fungal cells. This is the first transcriptomic analysis on a fungus treated with peptaibol analogs.