title: Seed response to strigolactone is controlled by abscisic acid-independent DNA methylation in the obligate root parasitic plant, Phelipanche ramosa L. Pomel.
abstract: Seed dormancy release of the obligate root parasitic plant, Phelipanche ramosa, requires a minimum 4-day conditioning period followed by stimulation by host-derived germination stimulants, such as strigolactones. Germination is then mediated by germination stimulant-dependent activation of PrCYP707A1, an abscisic acid catabolic gene. The molecular mechanisms occurring during the conditioning period that silence PrCYP707A1 expression and regulate germination stimulant response are almost unknown. Here, global DNA methylation quantification associated with pharmacological approaches and cytosine methylation analysis of the PrCYP707A1 promoter were used to investigate the modulation and possible role of DNA methylation during the conditioning period and in the PrCYP707A1 response to GR24, a synthetic strigolactone analogue. Active global DNA demethylation occurs during the conditioning period and is required for PrCYP707A1 activation by GR24 and for subsequent seed germination. Treatment with 5-azacytidine, a DNA-hypomethylating molecule, reduces the length of the conditioning period. Conversely, hydroxyurea, a hypermethylating agent, inhibits PrCYP707A1 expression and seed germination. Methylated DNA immunoprecipitation followed by PCR experiments and bisulfite sequencing revealed that DNA demethylation particularly impacts a 78-nucleotide sequence in the PrCYP707A1 promoter. The results here demonstrate that the DNA methylation status during the conditioning period plays a crucial role independently of abscisic acid in the regulation of P. ramosa seed germination by controlling the strigolactone-dependent expression of PrCYP707A1.
authors.affiliation: Laboratoire de Biologie et Pathologie Végétales, SFR 4207 QUASAV, Université de Nantes, 2 rue de la Houssinière, BP 92208, 44322 Nantes Cedex 3, France.
authors.forename: Marc-Marie
authors.initials: MM
authors.lastname: Lechat
authors.affiliation: Laboratoire de Biologie et Pathologie Végétales, SFR 4207 QUASAV, Université de Nantes, 2 rue de la Houssinière, BP 92208, 44322 Nantes Cedex 3, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Brun
authors.affiliation: Laboratoire de Biologie et Pathologie Végétales, SFR 4207 QUASAV, Université de Nantes, 2 rue de la Houssinière, BP 92208, 44322 Nantes Cedex 3, France.
authors.forename: Grégory
authors.initials: G
authors.lastname: Montiel
authors.affiliation: Laboratoire de Biologie et Pathologie Végétales, SFR 4207 QUASAV, Université de Nantes, 2 rue de la Houssinière, BP 92208, 44322 Nantes Cedex 3, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Véronési
authors.affiliation: Laboratoire de Biologie et Pathologie Végétales, SFR 4207 QUASAV, Université de Nantes, 2 rue de la Houssinière, BP 92208, 44322 Nantes Cedex 3, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Simier
authors.affiliation: Laboratoire de Biologie et Pathologie Végétales, SFR 4207 QUASAV, Université de Nantes, 2 rue de la Houssinière, BP 92208, 44322 Nantes Cedex 3, France.
authors.forename: Séverine
authors.initials: S
authors.lastname: Thoiron
authors.affiliation: Laboratoire de Biologie et Pathologie Végétales, SFR 4207 QUASAV, Université de Nantes, 2 rue de la Houssinière, BP 92208, 44322 Nantes Cedex 3, France.
authors.forename: Jean-Bernard
authors.initials: JB
authors.lastname: Pouvreau
authors.affiliation: Laboratoire de Biologie et Pathologie Végétales, SFR 4207 QUASAV, Université de Nantes, 2 rue de la Houssinière, BP 92208, 44322 Nantes Cedex 3, France philippe.delavault@univ-nantes.fr.
authors.forename: Philippe
authors.identifier: http://orcid.org/0000-0002-0189-4830
authors.initials: P
authors.lastname: Delavault
mesh_terms: D000040:Abscisic Acid / Q000378:metabolism; D001374:Azacitidine / Q000494:pharmacology; D001483:Base Sequence; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D019175:DNA Methylation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D044127:Epigenesis, Genetic; D018525:Germination / Q000187:drug effects; D006918:Hydroxyurea / Q000494:pharmacology; D007783:Lactones / Q000494:pharmacology*; D008969:Molecular Sequence Data; D031672:Orobanche / Q000187:drug effects / Q000502:physiology*; D057445:Plant Dormancy / Q000187:drug effects; D010937:Plant Growth Regulators / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000469:parasitology; D012639:Seeds / Q000187:drug effects / Q000502:physiology*; D017422:Sequence Analysis, DNA
chemical_list: D004791:Enzyme Inhibitors; D007783:Lactones; D010937:Plant Growth Regulators; D010940:Plant Proteins; D000040:Abscisic Acid; C483518:strigol; D003577:Cytochrome P-450 Enzyme System; C107203:abscisic acid 8'-hydroxylase; D001374:Azacitidine; D006918:Hydroxyurea
keywords: Bisulfite sequencing; DNA methylation; branched broomrape (Phelipanche ramosa); dormancy release; epigenetic regulation; parasitic plant; seed germination; strigolactone.
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