We outline a novel method of experimental plant DNA demethylation for ecological epigenetic experiments. While the traditionally-used approach causes underdeveloped root systems and high mortality of treated plants, this new-one overcomes the unwanted effects while maintaining the demethylation efficiency. Also, we demonstrate its application for ecological epigenetic experiments, testing transgenerational effects of plant–plant competition. This novel method could be better suited for experimental studies seeking valuable insights into ecological epigenetics. As this method is based on periodical spraying of azacytidine on established plants, it can be suitable for clonal species reproducing asexually, and it opens the possibility of community-level experimental demethylation of plants.
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