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Arabidopsis LEC1 and LEC2 Orthologous Genes Are Key Regulators of Somatic Embryogenesis in Cassava

机译:拟南芥LEC1和LEC2直系同源基因是木薯体胚发生的关键调控因子。

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摘要

High genotype-dependent variation in friable embryogenic callus (FEC) induction and subsequent somaclonal variation constitute bottlenecks for the application and scaling of genetic transformation (GT) technology to more farmer- and industry-preferred cassava varieties. The understanding and identification of molecular factors underlying embryogenic development in cassava may help to overcome these constraints. Here, we described the Arabidopsis thaliana LEAFY COTYLEDON (LEC) LEC1 and LEC2 orthologous genes in cassava, designated as MeLEC1 and MeLEC2, respectively. Expression analyses showed that both, MeLEC1 and MeLEC2, are expressed at higher levels in somatic embryogenic (SE) tissues in contrast with differentiated mature tissues. The rapid expression increase of MeLEC genes at early SE induction times strongly suggests that they are involved in the transition from a somatic to an embryonic state, and probably, in the competence acquisition for SE development in cassava. The independent overexpression of the MeLEC genes resulted in different regenerated events with embryogenic characteristics such as MeLEC1OE plants with cotyledon-like leaves and MeLEC2OE plants with somatic-like embryos that emerged over the surface of mature leaves. Transcript increases of other embryo-specific regulating factors were also detected in MeLECOE plants, supporting their mutual interaction in the embryo development coordination. The single overexpression of MeLEC2 was enough to reprogram the vegetative cells and induce direct somatic embryogenesis, which converts this gene into a tool that could improve the recovery of transformed plants of recalcitrant genotypes. The identification of MeLEC genes contributes not only to improve our understanding of SE process in cassava, but also provides viable alternatives to optimize GT and advance in gene editing in this crop, through the development of genotype-independent protocols.
机译:易碎的胚性愈伤组织(FEC)诱导中高基因型依赖性变异以及随后的体细胞无性变异构成了将遗传转化(GT)技术应用和扩展到更多农民和行业偏爱的木薯品种的瓶颈。对木薯胚发生发展的分子因素的理解和鉴定可能有助于克服这些限制。在这里,我们描述了木薯中拟南芥叶的子叶(LEC)LEC1和LEC2直系同源基因,分别称为MeLEC1和MeLEC2。表达分析表明,与分化的成熟组织相比,MeLEC1和MeLEC2在体细胞胚胎发生(SE)组织中均以较高的水平表达。 MeLEC基因在早期SE诱导时期的快速表达增加强烈表明它们参与了从体细胞状态到胚胎状态的转变,并可能参与了木薯中SE发育的能力获取。 MeLEC基因的独立过表达导致具有胚发生特性的不同再生事件,例如带有子叶状叶片的MeLEC1 OE 植物和具有体细胞样胚的MeLEC2 OE 植物出现在成熟的叶子表面。在MeLEC OE 植物中还检测到其他胚特异性调节因子的转录本增加,支持它们在胚胎发育协调中的相互作用。 MeLEC2的单次过表达足以重新编程营养细胞并诱导直接体细胞胚发生,从而将该基因转化为可以改善转化为顽固基因型植物的工具。 MeLEC基因的鉴定不仅有助于增进我们对木薯中SE过程的理解,而且还提供了可行的替代方案,可通过开发不依赖基因型的方案来优化GT并促进该作物的基因编辑。

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