PREMISE OF THE STUDY: The phenotype of an individual can be modified by the environment experienced by its predecessors, a phenomenon called transgenerational or maternal effects. These effects are studied mostly across sexual generations and are thought to be mediated also by epigenetic variation. However, we do not know how important transgenerational effects are across asexual generations of clonal plants. METHODS: We investigated the role of different drought intensities and durations experienced by parental plants of Trifolium repens on the growth of offspring ramets after transplantation of clonal cuttings to control conditions. We also treated half of the plants with 5-azacytidine, which is a demethylating agent, to test the potential role of DNA methylation on transgenerational effects. KEY RESULTS: Transgenerational effects were manifested as increased biomass of offspring ramets if parental plants experienced medium drought applied for a short period and decreased biomass of offspring ramets if parental plants experienced intense drought for a short period. These transgenerational effects were not observed for offspring of parents from the same treatments if these were treated with 5-azacytidine, whose application significantly decreased the amount of 5-methyl-2'-deoxycytidine in plants. CONCLUSIONS: Transgenerational effects might play an important role in the clonal plant Trifolium repens and are probably mediated by epigenetic variation. The growth and behavior of clonal plants might be affected not only by the ambient environment but also by environments that are no longer present at the time of clonal reproduction. This phenomenon can have yet unacknowledged ecological and evolutionary implications for clonal plants.

Institute of Botany Czech Academy of Sciences Zámek 1 CZ 252 43 Průhonice Czech Republic

Institute of Botany Czech Academy of Sciences Zámek 1 CZ 252 43 Průhonice Czech Republic Department of Botany Charles University Benátská 2 CZ 128 00 Praha Czech Republic

Institute of Botany Czech Academy of Sciences Zámek 1 CZ 252 43 Průhonice Czech Republic Herbarium and Department of Botany Charles University Benátská 2 CZ 128 00 Praha Czech Republic

Institute of Experimental Botany Czech Academy of Sciences Rozvojová 263 CZ 165 02 Praha Czech Republic

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