Cytological, Biochemical and Molecular Events of the Embryogenic State in Douglas-fir (Pseudotsuga menziesii [Mirb.])

0502551 - ÚEB 2019 RIV CH eng J - Journal Article
Gautier, F. - Label, P. - Eliášová, Kateřina - Leple, J.C. - Motyka, Václav - Boizot, N. - Vondráková, Zuzana - Malbeck, Jiří - Trávníčková, Alena - Le Metté, C. - Lesage-Descauses, M. C. - Lomenech, A. M. - Trontin, J.F. - Costa, G. - Lelu-Walter, M.A. - Teyssier, C.
Cytological, Biochemical and Molecular Events of the Embryogenic State in Douglas-fir (Pseudotsuga menziesii [Mirb.]).
Frontiers in Plant Science. Roč. 10, FEB 28 (2019), č. článku 118. ISSN 1664-462X. E-ISSN 1664-462X
R&D Projects: GA ČR(CZ) GA16-14649S; GA MŠMT(CZ) EF16_019/0000738; GA MŠMT(CZ) LTC17030
Institutional support: RVO:61389030
Keywords : comparative proteomic analysis * transcription factor family * early somatic embryogenesis * abscisic-acid * nonembryogenic tissues * sugar transporters * genome sequence * cell fate * auxin * expression * embryonal mass * non-embryogenic callus * histo-cytology * soluble carbohydrate * phytohormone * proteomic * network * transcriptomic
OECD category: Biochemical research methods
Impact factor: 4.402, year: 2019

Somatic embryogenesis techniques have been developed for most coniferous species, but only using very juvenile material. To extend the techniques' scope, better integrated understanding of the key biological, physiological and molecular characteristics of embryogenic state is required. Therefore, embryonal masses (EMs) and non-embryogenic calli (NECs) have been compared during proliferation at multiple levels. EMs and NECs originating from a single somatic embryo (isogenic lines) of each of three unrelated genotypes were used in the analyses, which included comparison of the lines' anatomy by transmission light microscopy, transcriptomes by RNAseq Illumina sequencing, proteomes by free-gel analysis, contents of endogenous phytohormones (indole-3-acetic acid, cytokinins and ABA) by LC-MS analysis, and soluble sugar contents by HPLC. EMs were characterized by upregulation (relative to levels in NECs) of transcripts, proteins, transcription factors and active cytokinins associated with cell differentiation accompanied by histological, carbohydrate content and genetic markers of cell division. In contrast, NECs were characterized by upregulation (relative to levels in EMs) of transcripts, proteins and products associated with responses to stimuli (ABA, degradation forms of cytokinins, phenols), oxidative stress (reactive oxygen species) and carbohydrate storage (starch). Sub-Network Enrichment Analyses that highlighted functions and interactions of transcripts and proteins that significantly differed between EMs and NECs corroborated these findings. The study shows the utility of a novel approach involving integrated multi-scale transcriptomic, proteomic, biochemical, histological and anatomical analyses to obtain insights into molecular events associated with embryogenesis and more specifically to the embryogenic state of cell in Douglas-fir.
Permanent Link: http://hdl.handle.net/11104/0294457