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NEP-TC a rRNA Methyltransferase Involved on Somatic Embryogenesis of Tamarillo (Solanum betaceum Cav.)

机译:NEP-TC a rRNA甲基转移酶参与番茄的体细胞胚发生。

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摘要

Somatic embryogenesis (SE) is an important biotechnological tool for large-scale clonal propagation and for embryogenesis research. Moreover, genetic transformation and cryopreservation procedures in many species rely on efficient SE protocols. We have been studying different aspects related to SE induction and somatic embryo development in tamarillo (Solanum betaceum Cav.), a small tree from the Solanaceae family. Previous proteomic analyses identified a protein (NEP-TC, 26.5 kDa) consistently present in non-embryogenic calluses of tamarillo, but absent in the embryogenic ones. In this work, the role of NEP-TC during SE was assessed by gene expression analysis and immunolocalization. The results obtained demonstrated that NEP-TC is a putative member of the SpoU rRNA methylase family. This protein, present in the cytoplasm and nucleus, is expressed in non-embryogenic cells and not expressed in embryogenic cells. Slightly enhanced SE induction levels in tamarillo plants with NEP-TC down-regulated levels also supports the role of this protein on SE induction. Heterologous expression was used to confirm NEP-TC rRNA methyltransferase activity, with enhanced activity levels when rRNA was used as a substrate. These data relate a putative member of the SpoU methylase family with plant morphogenesis, in particular with SE induction.
机译:体细胞胚胎发生(SE)是大规模克隆繁殖和胚胎发生研究的重要生物技术工具。此外,许多物种的遗传转化和冷冻保存程序都依赖有效的SE方案。我们一直在研究番茄(茄属科的小树)番茄中SE诱导和体细胞胚发育的不同方面。先前的蛋白质组学分析鉴定出一种蛋白质(NEP-TC,26.5 kDa)始终存在于番茄(tamarillo)的非胚性愈伤组织中,而在胚发生性愈伤组织中却不存在。在这项工作中,通过基因表达分析和免疫定位评估了NEP-TC在SE中的作用。获得的结果表明,NEP-TC是SpoU rRNA甲基化酶家族的假定成员。存在于细胞质和细胞核中的这种蛋白质在非胚性细胞中表达,而在胚性细胞中不表达。 NEP-TC下调水平的番茄植物中SE诱导水平略有提高,也支持该蛋白在SE诱导中的作用。当rRNA用作底物时,异源表达用于确认NEP-TC rRNA甲基转移酶活性,并具有增强的活性水平。这些数据将SpoU甲基化酶家族的推定成员与植物形态发生,特别是与SE诱导相关。

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