# Genome-wide DNA methylation analysis of Medicago sativa L. treated with plasma and plasma-activated water

## TL;DR

This study examines how plasma and plasma-activated water affect DNA methylation in Medicago sativa L., revealing significant changes in methylation patterns and gene expression.

## Contribution

The study provides a genome-wide analysis of DNA methylation changes in Medicago sativa L. following plasma treatment, identifying thousands of differentially methylated genes.

## Key findings

- Treatment with plasma and plasma-activated water significantly altered DNA methylation levels in Medicago sativa L.
- Thousands of differentially methylated regions were identified across CG, CHG, and CHH contexts.
- Methylation changes were associated with potential impacts on gene expression and plant survival rates.

## Abstract

To explore the effects of plasma and plasma-activated water on whole-genome DNA methylation, in this study we used the Medicago sativa L. cultivar, as the experimental material, and mutant plants were obtained after treatment and screening. Changes in whole-genome DNA methylation in Medicago sativa L. were analyzed before and after mutagenesis using whole-genome bisulfite sequencing (WGBS) technology. We found that the percentage of methylated cytosines varied depending on the local sequence context (CG [dinucleotide context]), CHG and CHH (non-CG contexts, H is A [adenine] or T [thymine] or C [cytosine]) and external treatment. Differential methylated region (DMR) analysis revealed 41067 (CG), 5379 (CHG), and 257 (CHH) differentially methylated genes. This study quantitatively measured methylation levels, methylation sites, differentially methylated regions (DMRs), distribution of methylation in the genome, and methylation-related genes and pathways, for further investigations of the mechanism of plasma-induced mutagenesis.

•After mutation treatment, the survival rate of seed can be reduced to as low as 9%.•There were also significant changes at the molecular level before and after treatment.•After treatment, the roots of the seedlings grow rapidly.

After mutation treatment, the survival rate of seed can be reduced to as low as 9%.

There were also significant changes at the molecular level before and after treatment.

After treatment, the roots of the seedlings grow rapidly.

Plant biology; Interaction of plants with organisms; Plant Genetics