PDF(Pubmed)
Abstract：
Although some budding yeasts have proved tractable and intensely studied models, others are more recalcitrant. Debaryomyces hansenii, an important yeast species in food and biotechnological industries with curious physiological characteristics, has proved difficult to manipulate genetically and remains poorly defined. To remedy this, we have combined live cell fluorescent dyes with high-resolution imaging techniques to define the sub-cellular features of D. hansenii, such as the mitochondria, nuclei, vacuoles and the cell wall. Using these tools, we define biological processes like the cell cycle, organelle inheritance and various membrane trafficking pathways of D. hansenii for the first time. Beyond this, reagents designed to study Saccharomyces cerevisiae proteins were used to access proteomic information about D. hansenii. Finally, we optimised the use of label-free holotomography to image yeast, defining the physical parameters and visualising sub-cellular features like membranes and vacuoles. Not only does this work shed light on D. hansenii but this combinatorial approach serves as a template for how other cell biological systems, which are not amenable to standard genetic procedures, can be studied.
摘要：
尽管一些发芽酵母已被证明是易于处理和深入研究的模型，其他人更顽固。汉森德巴酵母,一种在食品和生物技术行业中具有奇特生理特征的重要酵母，已证明难以操纵基因，并且定义不清。为了补救这一点，我们将活细胞荧光染料与高分辨率成像技术相结合，以定义D.hansenii的亚细胞特征，比如线粒体，原子核,液泡和细胞壁。使用这些工具，我们定义了像细胞周期这样的生物过程，首次发现D.hansenii的细胞器遗传和各种膜运输途径。除此之外,设计用于研究酿酒酵母蛋白质的试剂用于获取有关D.hansenii的蛋白质组信息。最后,我们优化了无标签全断层扫描对酵母成像的使用，定义物理参数并可视化亚细胞特征，如膜和液泡。这项工作不仅揭示了D.hansenii，而且这种组合方法作为其他细胞生物系统的模板，不适合标准遗传程序，可以研究。
