Variation in DNA methylation and response to short-term herbivory in Thlaspi arvense

We investigated the effect of experimental demethylation and herbivory treatments following a 2x3 factorial design. First, half the seeds were 28 submerged in a water solution of the demethylating agent 5-azacytidine and the other half 29 only in water, as controls. Then, we assigned control and demethylated plants to three herbivory categories (i) insect herbivory, (ii) artificial herbivory, and (iii) undamaged plants. The effects of the demethylation and herbivory treatments were assessed by quantifying genome-wide global DNA cytosine methylation and differentially methylated cytosines using epiGBS