title: A Laser Dissection-RNAseq Analysis Highlights the Activation of Cytokinin Pathways by Nod Factors in the Medicago truncatula Root Epidermis.
abstract: Nod factors (NFs) are lipochitooligosaccharidic signal molecules produced by rhizobia, which play a key role in the rhizobium-legume symbiotic interaction. In this study, we analyzed the gene expression reprogramming induced by purified NF (4 and 24 h of treatment) in the root epidermis of the model legume Medicago truncatula Tissue-specific transcriptome analysis was achieved by laser-capture microdissection coupled to high-depth RNA sequencing. The expression of 17,191 genes was detected in the epidermis, among which 1,070 were found to be regulated by NF addition, including previously characterized NF-induced marker genes. Many genes exhibited strong levels of transcriptional activation, sometimes only transiently at 4 h, indicating highly dynamic regulation. Expression reprogramming affected a variety of cellular processes, including perception, signaling, regulation of gene expression, as well as cell wall, cytoskeleton, transport, metabolism, and defense, with numerous NF-induced genes never identified before. Strikingly, early epidermal activation of cytokinin (CK) pathways was indicated, based on the induction of CK metabolic and signaling genes, including the CRE1 receptor essential to promote nodulation. These transcriptional activations were independently validated using promoter:β-glucuronidase fusions with the MtCRE1 CK receptor gene and a CK response reporter (TWO COMPONENT SIGNALING SENSOR NEW). A CK pretreatment reduced the NF induction of the EARLY NODULIN11 (ENOD11) symbiotic marker, while a CK-degrading enzyme (CYTOKININ OXIDASE/DEHYDROGENASE3) ectopically expressed in the root epidermis led to increased NF induction of ENOD11 and nodulation. Therefore, CK may play both positive and negative roles in M. truncatula nodulation.
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.).
authors.forename: Marie-Françoise
authors.initials: MF
authors.lastname: Jardinaud
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.).
authors.forename: Stéphane
authors.identifier: 0000-0002-4084-2312
authors.initials: S
authors.lastname: Boivin
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.).
authors.forename: Nathalie
authors.identifier: 0000-0003-3361-4730
authors.initials: N
authors.lastname: Rodde
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.).
authors.forename: Olivier
authors.identifier: 0000-0002-5820-1660
authors.initials: O
authors.lastname: Catrice
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.).
authors.forename: Anna
authors.identifier: 0000-0002-4918-7836
authors.initials: A
authors.lastname: Kisiala
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.).
authors.forename: Agnes
authors.initials: A
authors.lastname: Lepage
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.).
authors.forename: Sandra
authors.initials: S
authors.lastname: Moreau
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.).
authors.forename: Brice
authors.identifier: 0000-0001-6923-0902
authors.initials: B
authors.lastname: Roux
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.).
authors.forename: Ludovic
authors.initials: L
authors.lastname: Cottret
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.).
authors.forename: Erika
authors.identifier: 0000-0001-6062-5043
authors.initials: E
authors.lastname: Sallet
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.).
authors.forename: Mathias
authors.initials: M
authors.lastname: Brault
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.).
authors.forename: R J Neil
authors.identifier: 0000-0003-1304-6430
authors.initials: RJ
authors.lastname: Emery
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.).
authors.forename: Jérôme
authors.initials: J
authors.lastname: Gouzy
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.).
authors.forename: Florian
authors.initials: F
authors.lastname: Frugier
authors.affiliation: LIPM, Université de Toulouse, Institut National de la Recherche Agronomique, Centre National de la Recherche Scientifique, 31326 Castanet-Tolosan, France (M.-F.J., N.R., O.C., A.L., S.M., B.R., L.C., E.S., J.G., P.G.);INPT-Université de Toulouse, ENSAT, 31326 Castanet-Tolosan, France (M.-F.J.);Institute of Plant Sciences-Paris Saclay University, Centre National de la Recherche Scientifique/Institut National de la Recherche Agronomique/Universités Paris-Sud/Paris-Diderot/d'Evry, 91190 Gif-sur-Yvette, France (S.B., M.B., F.F.);Biology Department, Trent University, Peterborough, Ontario, Canada K9J 7B8 (A.K., R.J.N.E.); andDepartment of Plant Genetics, Physiology, and Biotechnology, University of Technology and Life Sciences, 85-789 Bydgoszcz, Poland (A.K.) pascal.gamas@toulouse.inra.fr.
authors.forename: Pascal
authors.identifier: 0000-0002-6253-4249
authors.initials: P
authors.lastname: Gamas
mesh_terms: D003583:Cytokinins / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D007834:Lasers; D008070:Lipopolysaccharides / Q000378:metabolism* / Q000494:pharmacology; D046913:Medicago truncatula / Q000235:genetics / Q000378:metabolism*; D019441:Plant Epidermis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D053204:Root Nodules, Plant / Q000235:genetics / Q000378:metabolism; D017423:Sequence Analysis, RNA / Q000379:methods; D015398:Signal Transduction
chemical_list: D003583:Cytokinins; C438018:ENOD11 protein, Medicago truncatula; D008070:Lipopolysaccharides; D010940:Plant Proteins