Within-plant heterogeneity in fecundity and herbivory induced by localized DNA hypomethylation in the perennial herb Helleborus foetidus

Am J Bot. 2019 Jun;106(6):798-806. doi: 10.1002/ajb2.1291. Epub 2019 Jun 3.


Premise: Phenotypic heterogeneity of reiterated, homologous structures produced by individual plants has ecological consequences for plants and their animal consumers. This paper examines experimentally the epigenetic mosaicism hypothesis, which postulates that within-plant variation in traits of reiterated structures may partly arise from different parts of the same genetic individual differing in patterns or extent of genomic DNA methylation.

Methods: Leaves of paired ramets borne by field-growing Helleborus foetidus plants were infiltrated periodically over the entire flowering period with either a water solution of the demethylating agent zebularine or just water as the control. The effects of the zebularine treatment were assessed by quantifying genome-wide DNA cytosine methylation in leaves and monitoring inflorescence growth and flower production, number of ovules per flower, pollination success, fruit set, seed set, seed size, and distribution of sap-feeding insects.

Results: Genomic DNA from leaves in zebularine-treated ramets was significantly less methylated than DNA from leaves in control ones. Inflorescences in treated ramets grew smaller and produced fewer flowers, with fewer ovules and lower follicle and seed set, but did not differ from inflorescences in untreated ramets in pollination success or seed size. The zebularine treatment influenced the within-plant distribution of sap-feeding insects.

Conclusions: Experimental manipulation of genomic DNA methylation level in leaves of wild-growing H. foetidus plants induced considerable within-plant heterogeneity in phenotypic (inflorescences, flowers, fecundity) and ecologically relevant traits (herbivore distribution), which supports the hypothesis that epigenetic mosaicism may partly account for within-plant variation.

Keywords: DNA methylation; Ranunculaceae; demethylating agent; epigenetic mosaicism; herbivory; inflorescence growth; leaf infiltration; seed production; subindividual variation; zebularine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cytosine / metabolism*
  • DNA Methylation*
  • Epigenesis, Genetic*
  • Fertility / genetics*
  • Flowers / genetics
  • Flowers / physiology
  • Helleborus / genetics
  • Helleborus / physiology*
  • Herbivory / physiology*
  • Inflorescence / genetics
  • Inflorescence / physiology
  • Mosaicism
  • Plant Leaves / physiology
  • Seeds / genetics
  • Seeds / physiology


  • Cytosine