Embrapa Recursos Genéticos e Biotecnologia, Brasília, DF, Brazil.
Programa de Pós-Graduação em Botânica, Universidade de Brasília, Brasília-DF, Brazil.
PLoS One. 2018 Dec 31;13(12):e0209445. doi: 10.1371/journal.pone.0209445. eCollection 2018.
To understand the molecular processes triggered during the different steps of somatic embryogenesis (SE) in oil palm, the expression of 19 genes associated to SE identified in proteomic and transcriptomic studies was investigated by qRT-PCR. To evaluate the differential expression of these genes, two interspecific hybrid genotypes (Elaeis oleifera x Elaeis guineensis) contrasting for the acquisition of embryogenic competence were used. Aclorophyllated leaves of both hybrids, one responsive (B351733) and the other non-responsive (B352933) to SE were submitted to callus induction and collected at different time points: 0 (before induction), 14, 30, 90 and 150 days of callus induction (doi). The results obtained showed that all evaluated genes were downregulated at 14 doi in the responsive genotype when compared to the non-responsive. It was also possible to observe that most of the genes changed their expression behavior at 30 doi and were upregulated thereafter until 150 doi, with the exception of the pathogenesis-related PRB1-3-like (PRB1-3) gene, which did not show differential expression at 30 doi and was downregulated at 90 and 150 doi when compared to the non-responsive hybrid. These results indicate that 30 doi is a turning point in gene expression, probably associated to embryogenic competence acquisition. We also show that the expression behavior of the responsive genotype is more stable than that of the non-responsive when the different induction time points are compared to 0 doi (before induction). Moreover, the results obtained in this study corroborate our hypothesis that the regulation of genes involved in the control of oxidative stress and energy metabolism are crucial for the acquisition of embryogenic competence in oil palm.
为了了解油棕体胚发生(SE)不同阶段触发的分子过程,通过 qRT-PCR 研究了蛋白质组学和转录组学研究中鉴定的与 SE 相关的 19 个基因的表达。为了评估这些基因的差异表达,使用了两个种间杂种基因型(Elaeis oleifera x Elaeis guineensis),它们在获得胚胎发生能力方面表现出不同。将两个杂种的去叶绿素化叶片(对 SE 有反应的 B351733 和无反应的 B352933)进行愈伤组织诱导,并在不同时间点(0 天(诱导前)、14、30、90 和 150 天)收集:0 天(诱导前)、14、30、90 和 150 天。结果表明,与无反应的基因型相比,所有评估的基因在有反应的基因型中在 14 天的 doi 时均下调。还可以观察到,大多数基因在 30 天的 doi 时改变了它们的表达行为,并在此后一直上调,直到 150 天的 doi,除了与发病相关的 PRB1-3 样(PRB1-3)基因外,该基因在 30 天的 doi 时没有表现出差异表达,与无反应的杂种相比,在 90 天和 150 天的 doi 时下调。这些结果表明,30 天的 doi 是基因表达的一个转折点,可能与胚胎发生能力的获得有关。我们还表明,与无反应的基因型相比,当比较不同的诱导时间点与 0 天(诱导前)时,有反应的基因型的表达行为更加稳定。此外,本研究的结果证实了我们的假设,即参与氧化应激和能量代谢控制的基因的调节对于油棕获得胚胎发生能力至关重要。