ChIP-seq for identifying molecules associated with sound vibration in plant

Plant can perceive and respond natural sound vibration (SV). Artificial SV also served as a novel trigger of induced resistance, although approaches for activating such plant innate immunity intensively studied on the use of biological and chemical agents (BCA). Artificial SV pre-treatment protected Arabidopsis thaliana seedlings against insect pests and fungal pathogens. However, SV-mediated epigenetic modulation remains unexplored while CBA-mediated induced resistance is known as a complicated process involving epigenetic regulation. Here, we performed a ChIP-seq analysis to understand the role of 10 kHz SV-mediated epigenetic modification in induced resistance against a soil-borne pathogenic bacterium Ralstonia solanacearum. Overall design: A group of plants was prepared to be expose to sound vibration (SV, 10 kHz) at 90 dB every 3 h for 10 days. The other group of plants with non-treatment were also prepared as a control group. Then, all samples were drenched with a pathogen (R. solanacearum). ChIP-seq analysis was performed on 36 arabidopsis samples, which were obtained at 0d and 1d after dreching pathogen. Cells were fixed using 1% formaldehyde, lysed and sonicated using a Bioruptor (Diagenode Inc.). Immunoprecipitation was performed with anti-H3k36ac(Millipore, 07-540) or anti-H3k27me3(07-449, Milipore). 250-400 bp genomic libraries were generated from the input and immunoprecipitated DNA and sequenced using Illumina HiSeq2500 to generate 100 bp paired-end reads.