通过免疫和分子遗传技术结合高分辨率显微工具和3D图像分析,通过原位鉴定,对寄主植物栖息地中细菌的空间定位和功能进行分析,大大有助于更好地理解微生物的功能相互作用。植物。在分子遗传学方法中,16S-rRNA基因对于重建新分离的细菌的系统发育并将其原位定位至关重要。然而,它们通常不允许解析属以下的系统发育隶属关系。尤其是,从植物有益菌株中分离机会性人类病原体,目前分配给同一物种,需要基于基因组的解析技术。整个细菌基因组序列允许区分系统发育密切相关的菌株。此外,完整的基因组序列使得能够对生物技术相关菌株进行菌株特异性监测。在这篇小型综述中,我们提出了高分辨率的方法来分析植物微生物群的组成和关键功能,关注重氮营养植物生长促进细菌的相互作用,像巴西螺旋藻一样,非豆类寄主植物。将高分辨率显微分析与特异性免疫学检测方法和分子遗传学工具相结合,包括特别是细菌和植物伙伴的转录组分析,使新的见解有益菌-植物相互作用的关键特征在整个系统。
Analyses of the spatial localization and the functions of bacteria in host plant habitats through in situ identification by immunological and molecular genetic techniques combined with high resolving microscopic tools and 3D-image analysis contributed substantially to a better understanding of the functional interplay of the microbiota in plants. Among the molecular genetic methods, 16S-rRNA genes were of central importance to reconstruct the phylogeny of newly isolated bacteria and to localize them in situ. However, they usually do not allow resolution for phylogenetic affiliations below genus level. Especially, the separation of opportunistic human pathogens from plant beneficial strains, currently allocated to the same species, needs genome-based resolving techniques. Whole bacterial genome sequences allow to discriminate phylogenetically closely related strains. In addition, complete genome sequences enable strain-specific monitoring for biotechnologically relevant strains. In this mini-
review we present high resolving approaches for analysis of the composition and key functions of plant microbiota, focusing on interactions of diazotrophic plant growth promoting bacteria, like Azospirillum brasilense, with non-legume host plants. Combining high resolving microscopic analyses with specific immunological detection methods and molecular genetic tools, including especially transcriptome analyses of both the bacterial and plant partners, enables new insights into key traits of beneficial bacteria-plant interactions in holobiontic systems.