College of Horticulture, South China Agricultural University, Guangzhou 510642, China.
Physiol Plant. 2011 Aug;142(4):372-89. doi: 10.1111/j.1399-3054.2011.01478.x. Epub 2011 May 24.
A better understanding of somatic embryogenesis in banana (Musa spp.) may provide a practical way to improve regeneration of banana plants. In this study, we applied scanning electron microscopy (SEM) and transmission electron microscopy (TEM) to visualize the ultrastructural changes during somatic embryogenesis of banana (Musa AAA cv. 'Yueyoukang 1'). We also used histological and immunohistochemical techniques with 16 monoclonal antibodies to study the spatial distribution and cellular/subcellular localization of different arabinogalactan protein (AGP) components of the cell wall during somatic embryogenesis. Histological study with periodic acid-Schiff staining documented diverse embryogenic stages from embryogenic cells (ECs) to the late embryos. SEM revealed a mesh-like structure on the surface of proembryos which represented an early structural marker of somatic embryogenesis. TEM showed that ECs were rich in juvenile mitochondria, endoplasmic reticulum and Golgi stacks. Cells in proembryos and early globular embryos resembled ECs, but they were more vacuolated, showed more regular nuclei and slightly more developed organelles. Immunocytochemical study revealed that the signal of most AGP epitopes was stronger in starch-rich cells when compared with typical ECs. The main AGP component in the extracellular matrix surface network of banana proembryos was the MAC204 epitope. Later, AGP immunolabelling patterns varied with the developmental stages of the embryos. These results about developmental regulation of AGP epitopes along with developmental changes in the ultrastructure of cells are providing new insights into the somatic embryogenesis of banana.
更好地理解香蕉体细胞胚胎发生(Musa spp.)可能为提高香蕉植物的再生提供一种实用的方法。在这项研究中,我们应用扫描电子显微镜(SEM)和透射电子显微镜(TEM)来观察香蕉(Musa AAA cv. 'Yueyoukang 1')体细胞胚胎发生过程中的超微结构变化。我们还使用组织学和免疫组织化学技术,用 16 种单克隆抗体研究细胞壁中不同阿拉伯半乳糖蛋白(AGP)成分在体细胞胚胎发生过程中的空间分布和细胞/亚细胞定位。过碘酸-雪夫(PAS)染色的组织学研究记录了从胚性细胞(ECs)到晚期胚胎的不同胚胎发生阶段。SEM 显示了原胚表面的网格状结构,这是体细胞胚胎发生的早期结构标志。TEM 显示 ECs 富含幼龄线粒体、内质网和高尔基体堆叠。原胚和早期球形胚胎中的细胞与 ECs 相似,但它们更有空泡化,核更规则,细胞器稍发达。免疫细胞化学研究表明,与典型的 ECs 相比,富含淀粉的细胞中大多数 AGP 表位的信号更强。香蕉原胚细胞外基质表面网络中的主要 AGP 成分是 MAC204 表位。后来,AGP 免疫标记模式随胚胎发育阶段的变化而变化。这些关于 AGP 表位发育调控以及细胞超微结构发育变化的结果,为香蕉体细胞胚胎发生提供了新的见解。