During somatic embryogenesis (SE), explant cells undergo changes in the direction of their differentiation, which lead to diverse cell phenotypes. Although the genetic bases of the SE have been extensively studied in Arabidopsis thaliana, little is known about the chemical characteristics of the wall of the explant cells, which undergo changes in the direction of differentiation. Thus, we examined the occurrence of selected pectic and AGP epitopes in explant cells that display different phenotypes during SE. Explants examinations have been supplemented with an analysis of the ultrastructure. The deposition of selected pectic and AGP epitopes in somatic embryos was determined. Compared to an explant at the initial stage, a/embryogenic/totipotent and meristematic/pluripotent cells were characterized by a decrease in the presence of AGP epitopes, b/the presence of AGP epitopes in differentiated cells was similar, and c/an increase of analyzed epitopes was detected in the callus cells. Totipotent cells could be distinguished from pluripotent cells by: 1/the presence of the LM2 epitope in the latest one, 2/the appearance of the JIM16 epitope in totipotent cells, and 3/the more abundant presence of the JIM7 epitope in the totipotent cells. The LM5 epitope characterized the wall of the cells that were localized within the mass of embryogenic domain. The JIM8, JIM13 and JIM16 AGP epitopes appeared to be the most specific for the callus cells. The results indicate a relationship between the developmental state of the explant cells and the chemical composition of the cell walls.
Transcriptome, metabolome and histological profiling were performed on normal and aberrant somatic embryo germinants of Norway spruce (Picea abies L. Karst) providing a simplistic systems biology description of conifer germination. Aberrant germinants (AGs) formed periderm-like tissue at the apical pole and lacked shoot growth above the cotyledons. Transcriptome profiling (RNA-Sequencing) revealed a total of 370 differentially expressed genes at ≥1 or ≤-1 log2-fold change, where 92% were down-regulated in AGs compared with normal germinants (NGs). Genes associated with shoot apical meristem formation were down-regulated in AGs, or not differentially expressed between AGs and NGs. Genes involved in hormone signaling and transport were also down-regulated. Metabolite profiling by gas chromatography-mass spectrometry (MS) and liquid chromatography-MS revealed biochemical difference between AGs and NGs, notably increased levels of sugars including glucose in AGs. Genes involved in glucose signaling were down-regulated and genes involved in starch biosynthesis were up-regulated, suggesting involvement of sugar signaling during late embryo development and germination. The overall results provide new data enabling further studies to confirm potential markers for a normal germination process in conifers.
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