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Early molecular events involved in Pinus pinaster Ait. somatic embryo development under reduced water availability: transcriptomic and proteomic analyses
Author(s) -
Morel Alexandre,
Teyssier Caroline,
Trontin JeanFrançois,
Eliášová Kateřina,
Pešek Bedřich,
Beaufour Martine,
Morabito Domenico,
Boizot Nathalie,
Le Metté Claire,
BelalBessai Leila,
Reymond Isabelle,
Harvengt Luc,
Cadene Martine,
Corbineau Françoise,
Vágner Martin,
Label Philippe,
LeluWalter MarieAnne
Publication year - 2014
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/ppl.12158
Subject(s) - somatic embryogenesis , transcriptome , proteome , biology , biochemistry , gel electrophoresis , gellan gum , embryo , embryogenesis , microbiology and biotechnology , botany , chemistry , gene expression , gene
Maritime pine somatic embryos ( SE s) require a reduction in water availability (high gellan gum concentration in the maturation medium) to reach the cotyledonary stage. This key switch, reported specifically for pine species, is not yet well understood. To facilitate the use of somatic embryogenesis for mass propagation of conifers, we need a better understanding of embryo development. Comparison of both transcriptome (Illumina RNA sequencing) and proteome [two‐dimensional sodium dodecyl sulfate‐polyacrylamide gel electrophoresis with mass spectrometry ( MS ) identification] of immature SE s, cultured on either high ( 9G ) or low ( 4G ) gellan gum concentration, was performed, together with analysis of water content, fresh and dry mass, endogenous abscisic acid ( ABA ; gas chromatography‐ MS ), soluble sugars (high‐pressure liquid chromatography), starch and confocal laser microscope observations. This multiscale, integrated analysis was used to unravel early molecular and physiological events involved in SE development. Under unfavorable conditions ( 4G ), the glycolytic pathway was enhanced, possibly in relation to cell proliferation that may be antagonistic to SE development. Under favorable conditions ( 9G ), SE s adapted to culture constraint by activating specific protective pathways, and ABA ‐mediated molecular and physiological responses promoting embryo development. Our results suggest that on 9G , germin‐like protein and ubiquitin‐protein ligase could be used as predictive markers of SE development, whereas protein phosphatase 2C could be a biomarker for culture adaptive responses. This is the first characterization of early molecular mechanisms involved in the development of pine SE s following an increase in gellan gum concentration in the maturation medium, and it is also the first report on somatic embryogenesis in conifers combining transcriptomic and proteomic datasets.