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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Morpho-histological, histochemical, and molecular evidences related to cellular reprogramming during somatic embryogenesis of the model grass Brachypodium distachyon
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Morpho-histological, histochemical, and molecular evidences related to cellular reprogramming during somatic embryogenesis of the model grass Brachypodium distachyon

机译：与细胞胚胎发生期间的细胞重编程相关的形态组织化学，组织化学和分子证据

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摘要

The wild grass species Brachypodium distachyon (L.) has been proposed as a new model for temperate grasses. Among the biotechnological tools already developed for the species, an efficient induction protocol of somatic embryogenesis (SE) using immature zygotic embryos has provided the basis for genetic transformation studies. However, a systematic work to better understanding the basic cellular and molecular mechanisms that underlie the SE process of this grass species is still missing. Here, we present new insights at the morpho-histological, histochemical, and molecular aspects of B. distachyon SE pathway. Somatic embryos arose from embryogenic callus formed by cells derived from the protodermal-dividing cells of the scutellum. These protodermal cells showed typical meristematic features and high protein accumulation which were interpreted as the first observable steps towards the acquisition of a competent state. Starch content decreased along embryogenic callus differentiation supporting the idea that carbohydrate reserves are essential to morphogenetic processes. Interestingly, starch accumulation was also observed at late stages of SE process. Searches in databanks revealed three sequences available annotated as BdSERK, being two copies corresponding to SERK1 and one showing greater identity to SERK2. In silico analysis confirmed the presence of characteristic domains in a B. distachyon Somatic Embryogenesis Receptor Kinase genes candidates (BdSERKs), which suggests SERK functions are conserved in B. distachyon. In situ hybridization demonstrated the presence of transcripts of BdSERK1 in all development since globular until scutellar stages. The results reported in this study convey important information about the morphogenetic events in the embryogenic pathway which has been lacking in B. distachyon. This study also demonstrates that B. distachyon provides a useful model system for investigating the genetic regulation of SE in grass species.
机译：野生草地颅烟灰囊（L.）已被提出为温带草的新模型。在已经为物种开发的生物技术工具中，使用未成熟的Zygotic胚胎的体细胞胚胎发生（Se）的有效感应方案为遗传转化研究提供了基础。然而，更好地理解基本细胞和分子机制的系统工作仍然缺失。在这里，我们对B. distachyons Se途径的形态学，组织化学和分子方面提出了新的见解。躯体胚胎从衍生自瘢痕疙瘩的Protodermal分裂细胞形成的细胞形成的胚胎胚胎。这些Protodermal细胞显示出典型的分娩特征和高蛋白质积累，其被解释为朝着获得主管国家的第一个可观察步骤。淀粉含量沿胚胎源性愈伤组织分化减少，支持碳水化合物储备对形态发生过程至关重要。有趣的是，在SE过程的后期也观察到淀粉积累。在DataBANK中搜索显示有三个可用的序列作为BDSERK，是与SERK1相对应的两个副本，并为SERK2表示更大的身份。在硅分析中，证实了B.Tistachyon体细胞胚胎发生受体激酶基因激酶基因（BDSErks）中的特征结构域的存在，这表明SERK功能在B. distachyon中保守。原位杂交证明了由于球肠直到瘢痕疙序的所有开发中BDSERK1的成绩单。本研究报告的结果传达了关于胚胎发生途径中的形态发生事件的重要信息，这缺乏B. distachyon。本研究还表明B.Tistachyon提供了一种用于研究SE在草地的遗传调节的有用模型系统。

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