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Immunolocalization of ricin accumulation during castor bean (Ricinus communis L.) seed development

机译:蓖麻籽种子发育过程中蓖麻毒蛋白积累的免疫定位

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Ricin is a dimeric glycoprotein that accumulates in protein storage vacuoles of endosperm cells of Ricinus communis L. (castor bean). The proricin travels through the Golgi apparatus and co-localizes throughout its route to the storage vacuoles of developing castor bean endosperm. We report here the pattern of seed morphological and ultrastructural changes during various stages of seed development, associated with ricin accumulation. ELISA was used to compare the ricin content in mature seeds of four Brazilian commercial cultivars. ELISA and immunoelectron microscopy analysis were used to study ricin accumulation during seed development from 10 to 60 days after pollination (DAP). Results have shown that no ricin could be localized in the endosperm cells in the early development stages (before 20 DAP) and only a few localization points could be observed at 30 DAP. However, a significant ricin localization signal was observed at 40 DAP in the matrix of the protein storage vacuoles. The signal increased significantly from 50 to 60 DAP, when ricin was observed in both the matrix and crystalloids of the protein storage vacuoles. Understanding ricin expression at the cellular level is fundamental for the development of strategies for gene suppression using molecular breeding approaches.
机译:蓖麻毒素是一种二聚体糖蛋白,积聚在蓖麻(蓖麻子)的胚乳细胞的蛋白储存液泡中。 proricin穿过高尔基体,并在整个路径中共定位到发育中的蓖麻籽胚乳的液泡中。我们在这里报告了种子发育各个阶段与蓖麻毒蛋白积累有关的种子形态和超微结构变化的模式。 ELISA用于比较四个巴西商业品种成熟种子中蓖麻毒素的含量。 ELISA和免疫电子显微镜分析用于研究授粉后(DAP)10到60天种子生长过程中蓖麻毒素的积累。结果表明,在生长的早期阶段(20 DAP之前),蓖麻毒素不能定位在胚乳细胞中,而在30 DAP时只能观察到几个定位点。但是,在蛋白质储存液泡的基质中,在40 DAP观察到明显的蓖麻毒蛋白定位信号。当在蛋白质储存液泡的基质和晶体中都观察到蓖麻毒蛋白时,信号从50 DAP显着增加到60 DAP。在细胞水平上了解蓖麻毒蛋白表达对于使用分子育种方法开发基因抑制策略至关重要。

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