Cardinale Massimiliano
Institute of Plant Sciences, University of Graz Graz, Austria ; Institute of Environmental Biotechnology, Graz University of Technology Graz, Austria.
Front Microbiol. 2014 Mar 7;5:94. doi: 10.3389/fmicb.2014.00094. eCollection 2014.
No plant or cryptogam exists in nature without microorganisms associated with its tissues. Plants as microbial hosts are puzzles of different microhabitats, each of them colonized by specifically adapted microbiomes. The interactions with such microorganisms have drastic effects on the host fitness. Since the last 20 years, the combination of microscopic tools and molecular approaches contributed to new insights into microbe-host interactions. Particularly, confocal laser scanning microscopy (CLSM) facilitated the exploration of microbial habitats and allowed the observation of host-associated microorganisms in situ with an unprecedented accuracy. Here I present an overview of the progresses made in the study of the interactions between microorganisms and plants or plant-like organisms, focusing on the role of CLSM for the understanding of their significance. I critically discuss risks of misinterpretation when procedures of CLSM are not properly optimized. I also review approaches for quantitative and statistical analyses of CLSM images, the combination with other molecular and microscopic methods, and suggest the re-evaluation of natural autofluorescence. In this review, technical aspects were coupled with scientific outcomes, to facilitate the readers in identifying possible CLSM applications in their research or to expand their existing potential. The scope of this review is to highlight the importance of confocal microscopy in the study of plant-microbe interactions and also to be an inspiration for integrating microscopy with molecular techniques in future researches of microbial ecology.
自然界中不存在没有与其组织相关联的微生物的植物或隐花植物。植物作为微生物宿主是不同微生境的集合，每个微生境都被特定适应的微生物群落所定殖。与这些微生物的相互作用对宿主适应性有巨大影响。自过去20年来，显微镜工具和分子方法的结合为微生物与宿主相互作用的新见解做出了贡献。特别是，共聚焦激光扫描显微镜（CLSM）促进了对微生物栖息地的探索，并以前所未有的精度实现了对宿主相关微生物的原位观察。在此，我概述了微生物与植物或类植物生物相互作用研究的进展，重点关注CLSM在理解其重要性方面的作用。我批判性地讨论了CLSM程序未得到适当优化时的误读风险。我还回顾了CLSM图像的定量和统计分析方法、与其他分子和显微镜方法的结合，并建议重新评估自然自发荧光。在本综述中，技术方面与科学成果相结合，以方便读者在其研究中识别可能的CLSM应用或扩展其现有潜力。本综述的范围是强调共聚焦显微镜在植物 - 微生物相互作用研究中的重要性，并为未来微生物生态学研究中将显微镜与分子技术相结合提供启发。
