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首页> 外文期刊>Protoplasma: An International Journal of Cell Biology >Two distinct cell sources of H2O2 in the lignifying Zinnia elegans cell culture system
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Two distinct cell sources of H2O2 in the lignifying Zinnia elegans cell culture system

机译：木质化百日草细胞培养系统中H2O2的两种不同细胞来源

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The use of transdifferentiating Zinnia elegans mesophyll cells has proved useful in investigations of the process of xylem differentiation from cambial derivatives. Cultured mesophyll cells can be induced by external stimuli to proceed through temporally controlled developmental programs which conclude in the formation of single-cell-derived dead vascular tracheids and parenchyma-like elements. However, there is a gap in our knowledge concerning the role played by reactive oxygen species (O-2(-) and H2O2) in the development of these vascular elements. In this study, we show by the following four independent and highly selective methods that transdifferentiating Z. elegans mesophyll cells are capable of producing reactive oxygen species: the 2,3-bis(2-methoxy-4-nitro-5sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay, which monitors O-2(-) production, and the xylenol orange, 2,7-dichlorofluorescein diacetate, and CeCl3 assays, which monitor H2O2 production and localization. The joint use of these biochemical (XTT and xylenol orange) assays and cytochemical (2,7-dichlorofluorescein diacetate and CeCl3) probes revealed that transdifferentiating Z elegans mesophyll cells do not show an oxidative burst but live in a strongly oxidative state during the entire culture period. In this state, H2O2 is produced by both tracheary and parenchyma-like elements, the nonlignifying parenchyma-like cells acting quantitatively as the main source. The existence of these two sources of H2O2 in this in vitro cell culture system may be especially relevant during the later stages of tracheary cell wall lignification, in which lignifying tracheary elements become hollow. In the case of differentiating tracheary elements, H2O2 was located in the same place and at the same time as the onset of tracheary element lignification, i.e., at the primary cell wall during secondary thickening, supporting the view that the H2O2 produced by this in vitro culture system is destined for use during lignin biosynthesis.
机译：事实证明，使用转分化百日菊属叶肉细胞可用于研究从冈比亚衍生物木质部分化的过程。培养的叶肉细胞可以由外部刺激诱导，以通过时间控制的发育程序进行，该程序以形成单细胞来源的死血管微管和薄壁组织样成分为结论。但是，关于活性氧（O-2（-）和H2O2）在这些血管元素发育中所起的作用，我们的知识尚有不足。在这项研究中，我们通过以下四种独立且高度选择性的方法表明，使线虫Z. elegans的叶肉细胞能够分化产生活性氧：2,3-双（2-甲氧基-4-硝基-5磺基苯基）-2H-监测O-2（-）产生的5-唑甲酸四唑鎓（XTT）检测以及监测H2O2产生和定位的二甲酚橙，2,7-二氯荧光素二乙酸酯和CeCl3检测。这些生物化学（XTT和二甲苯酚橙）测定法和细胞化学（2,7-二氯荧光素二乙酸酯和CeCl3）探针的共同使用表明，转分化的线虫Z线虫叶肉细胞在整个培养过程中没有显示出氧化爆发，而是处于强氧化状态期。在这种状态下，H2O2由气管和薄壁样细胞产生，非木质化薄壁样细胞在数量上起主要作用。在体外细胞培养系统中，这两种H2O2的存在可能与气管细胞壁木质化的后期阶段特别相关，在该阶段中，木质化的气管元素变为空心。在区分气管元素的情况下，H2O2与气管元素木质化发生的时间相同，并且位于同一时间，即在二次增厚过程中位于主细胞壁，这支持以下观点：培养系统注定要在木质素生物合成中使用。

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