Gylytė Brigita, Jurkonienė Sigita, Cimmperman Reda, Šveikauskas Vaidevutis, Manusadžianas Levonas
Institute of Botany, Nature Research Centre, Vilnius, Lithuania.
PeerJ. 2021 Feb 17;9:e10930. doi: 10.7717/peerj.10930. eCollection 2021.
Cells of characean algae are attractive for plant cell physiologists because of their large size and their close relation to higher plant cells. The objective of our study was to evaluate the purity of the compartments (cell wall, cytoplasm with plastids, mitochondria, nuclei and endomembrane system, and vacuole) separated mechanically from the internodal cells of using enzymatic markers. These included -mannosidase and malate dehydrogenase, vacuolar and cytoplasmic enzymes, respectively. The biomarkers applied revealed the degree of compartment contamination with the material from unwanted cell parts. The cell wall was contaminated slightly by vacuole and cytoplasm residuals, respectively by 12.3 and 1.96% of corresponding biomarker activities. Relatively high activity of vacuolar marker in the cell wall could be associated with the cell vacuoles in the multicellular structure of the nodes. The biomarkers confirmed highly purified vacuolar (99.5%) and cytoplasmic (86.7%) compartments. Purity estimation of the cell fractions enabled reevaluating nCuO related Cu concentrations in the compartments of charophyte cell. The internalisation of CuO nanoparticles in cell occurred already after 0.5h. In general, the approach seems to be useful for assessing the accumulation and distribution of various xenobiotics and/or metabolites within plant cell. All this justifies internodal cells as a model organism for modern studies in cell biology and nanotoxicology.
轮藻细胞因其体积大且与高等植物细胞关系密切，对植物细胞生理学家具有吸引力。我们研究的目的是使用酶标记物评估从轮藻节间细胞机械分离的各组分（细胞壁、含质体的细胞质、线粒体、细胞核和内膜系统以及液泡）的纯度。这些标记物分别包括α -甘露糖苷酶和苹果酸脱氢酶，即液泡酶和细胞质酶。所应用的生物标志物揭示了来自不需要的细胞部分的物质对各组分的污染程度。细胞壁分别被液泡和细胞质残余物轻微污染，相应生物标志物活性分别为12.3%和1.96%。细胞壁中液泡标记物的相对高活性可能与节的多细胞结构中的细胞液泡有关。生物标志物证实了液泡组分（99.5%）和细胞质组分（86.7%）的高度纯化。细胞组分的纯度评估使得能够重新评估轮藻细胞各组分中与nCuO相关的铜浓度。在轮藻细胞中，CuO纳米颗粒在0.5小时后就已发生内化。总体而言，该方法似乎有助于评估各种外源生物和/或代谢物在植物细胞内的积累和分布。所有这些都证明轮藻节间细胞是细胞生物学和纳米毒理学现代研究的模式生物。
