Simultaneous imaging of Pseudomonas fluorescens WCS365 populations expressing three different autofluorescent proteins in the rhizosphere:: New perspectives for studying microbial communities
Simultaneous imaging of Pseudomonas fluorescens WCS365 populations expressing three different autofluorescent proteins in the rhizosphere:: New perspectives for studying microbial communities
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DOI：
10.1094/mpmi.2000.13.11.1170
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发表时间：
2000-11-01
影响因子：
3.5
通讯作者：
Lugtenberg, BJJ
中科院分区：
文献类型：
--
作者：
Bloemberg, GV;Wijfjes, AHM;Lugtenberg, BJJ
为了在单细胞水平上以非侵入性的方式同时观察根际中不同的假单胞菌群体，构建了一组四个根际稳定的质粒，其表达三种不同的绿色荧光蛋白（GFP）衍生物，即增强青色（ECFP）、增强绿色（EGFP）、增强黄色（EYFP）和最近发表的红色荧光蛋白（RFP; DsRed）。在番茄幼苗接种荧光假单胞菌WCS 365群体（每个群体表达不同的自体荧光蛋白）后，植物生长5天，使用共聚焦激光扫描显微镜检查根际。我们能够同时可视化并清楚地区分多达三种不同的细菌种群。在根系的基部经常观察到由混合种群组成的微菌落，而进一步向根尖的微菌落主要由单一种群组成，表明微菌落随时间的动态行为。由于克隆载体pME 6010对革兰氏阴性菌具有广泛的宿主范围，因此构建的质粒可用于多种目的。特别是，它们对于微生物群落的分析具有重要价值，例如在生物控制、生物施肥、生物刺激、生态位竞争、定殖和生物膜形成等过程中。
To visualize simultaneously different populations of pseudomonads in the rhizosphere at the single cell level in a noninvasive way, a set of four rhizosphere-stable plasmids was constructed expressing three different derivatives of the green fluorescent protein (GFP), namely enhanced cyan (ECFP), enhanced green (EGFP), enhanced yellow (EYFP), and the recently published red fluorescent protein (RFP; DsRed), Upon tomato seedling inoculation with Pseudomonas fluorescens WCS365 populations, each expressing a different autofluorescent protein followed by plant growth for 5 days, the rhizosphere was inspected using confocal laser scanning microscopy. We were able to visualize simultaneously and clearly distinguish from each other up to three different bacterial populations. Micro colonies consisting of mixed populations were frequently observed at the base of the root system, whereas microcolonies further toward the root tip predominantly consisted of a single population, suggesting a dynamic behavior of microcolonies over time. Since the cloning vector pME6010 has a broad host range for gram-negative bacteria, the constructed plasmids can be used for many purposes. In particular, they will be of great value for the analysis of microbial communities, for example in processes such as biocontrol, biofertilization, biostimulation, competition for niches, colonization, and biofilm formation.