Two-way microscale interactions between immigrant bacteria and plant leaf microbiota as revealed by live imaging


The phyllosphere – the aerial parts of plants – is an important microbial habitat that is home to diverse microbial communities. The spatial organization of bacterial cells on leaf surfaces is non-random, and correlates with leaf microscopic features. Yet, the role of microscale interactions between bacterial cells therein is not well understood. Here, we ask how interactions between immigrant bacteria and resident microbiota affect the spatial organization of the combined community. By means of live imaging in a simplified in vitro system, we studied the spatial organization, at the micrometer scale, of the biocontrol agent Pseudomonas fluorescens A506 and the plant pathogen P. syringae B728a when introduced to pear and bean leaf microbiota (the corresponding native plants of these strains). We found significant co-localization of immigrant and resident microbial cells at distances of a few micrometers, for both strains. Interestingly, this co-localization was in part due to preferential attachment of microbiota cells near newly formed P. fluorescens aggregates. Our results indicate that two-way immigrant bacteria – resident microbiota interactions affect the microscale spatial organization of leaf microbiota, and possibly that of other surface-related microbial communities.

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Fig. 1: Experimental setup.
Fig. 2: Surface colonization dynamics without microbiota (leaf solution only).
Fig. 3: Extracted microbiota from bean and pear leaves.
Fig. 4: Surface colonization of immigrant bacteria introduced to resident microbiota.
Fig. 5: Spatial analysis of combined immigrant bacteria and resident microbiota based on Pair Cross-correlation and Nearest Neighbor methods.
Fig. 6: Two-way microscale interaction dynamics.


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We thank Y. Hadar for valuable comments on the manuscript. We thank S. Lindow for kindly providing bacterial strains, and R. Feuchtwanger from Gan Hasadeh and N. Shachar for providing fresh leaves for this study. JP acknowledges the Lady Davis Trust for a postdoctoral fellowship. MB acknowledges the Rudin MSc scholarship. This work was supported by a research grant to NK from the James S. McDonnell Foundation (Studying Complex Systems Scholar Award, Grant #220020475) and from the Israel Science Foundation (ISF #1396/19).

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Steinberg, S., Grinberg, M., Beitelman, M. et al. Two-way microscale interactions between immigrant bacteria and plant leaf microbiota as revealed by live imaging. ISME J (2020).

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