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Localization of BEN1-LIKE protein and nuclear degradation during development of metaphloem sieve elements in Triticum aestivum L.

机译:普通小麦的上皮韧皮部筛元素发育过程中BEN1-LIKE蛋白的定位和核降解。

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摘要

Metaphloem sieve elements (MSEs) in the developing caryopsis of Triticum aestivum L. undergo a unique type of programmed cell death (PCD); cell organelles gradually degrade with the MSE differentiation while mature sieve elements keep active. This study focuses on locating BEN1-LIKE protein and nuclear degradation in differentiating MSEs of wheat. Transmission electron microscopy (TEM) showed that nuclei degraded in MSE development. First, the degradation started at 2–3 days after flowering (DAF). The degraded fragments were then swallowed by phagocytic vacuoles at 4 DAF. Finally, nuclei almost completely degraded at 5 DAF. We measured the BEN1-LIKE protein expression in differentiating MSEs. In situ hybridization showed that BEN1-LIKE mRNA was a more obvious hybridization signal at 3–4 DAF at the microscopic level. Immuno-electron microscopy further revealed that BEN1-LIKE protein was mainly localized in MSE nuclei. Furthermore, MSE differentiation was tested using a TSQ Zn fluorescence probe which showed that the dynamic change of Zn accumulation was similar to BEN1-LIKE protein expression. These results suggest that nucleus degradation in wheat MSEs is associated with BEN1-LIKE protein and that the expression of this protein may be regulated by Zn accumulation variation.
机译:普通小麦发育中的颖果中的上级韧皮部筛分元件(MSE)经历了独特类型的程序性细胞死亡(PCD)。细胞细胞器随着MSE的分化而逐渐降解,而成熟的筛子元素保持活跃。这项研究的重点是在区分小麦MSEs中定位BEN1-LIKE蛋白和核降解。透射电子显微镜(TEM)显示,核在MSE发育中降解。首先,降解开始于开花后的2–3天(DAF)。然后在4 DAF下通过吞噬液泡吞下降解的片段。最后,原子核在5 DAF时几乎完全降解。我们测量了差异化MSE中的BEN1-LIKE蛋白表达。原位杂交表明,在微观水平,BEN1-LIKE mRNA在3–4 DAF处是更明显的杂交信号。免疫电子显微镜进一步显示,BEN1-LIKE蛋白主要位于MSE核中。此外,使用TSQ Zn荧光探针测试了MSE分化,结果表明Zn积累的动态变化与BEN1-LIKE蛋白表达相似。这些结果表明,小麦MSEs中的细胞核降解与BEN1-LIKE蛋白有关,并且该蛋白的表达可能受锌积累变化的调控。

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