Phyllosphere-inhabiting endophytic bacteria feature a stationary phase-like lifestyle

Velasquez, A. C.; Huguet-Tapia, J. C.; He, S. Y.

doi:10.1101/2021.05.10.443510

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bioRxiv · 10.1101/2021.05.10.443510

Phyllosphere-inhabiting endophytic bacteria feature a stationary phase-like lifestyle

Velasquez, A. C.·Huguet-Tapia, J. C.·He, S. Y.

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Abstract

Plants are in constant association with a variety of microbes, and although much is known about how symbiotic and pathogenic microbes interact with plants, less is known about the population dynamics, adaptive traits, and transcriptional features of endophytic commensal microbes that live inside leaves. In this study, we evaluated the long-term population and transcriptional dynamics of two bacterial microbiota endophytes and compared them to those of a commensal-simulating non-pathogenic mutant of the bacterial pathogen Pseudomonas syringae. We found that population densities of all three endophytic phyllosphere bacteria remained static over a long period of time, which was caused by a continual equilibrium between bacterial multiplication and death, as evidenced by treatment of plants with antibiotics that only targeted dividing bacteria or by in planta visualization of bacteria carrying a fluorescent division reporter. Population stasis could not be explained by a lack of resources, as Arabidopsis leaves could support population densities up to 100 times higher than the normal microbiota populations, nor was population stasis reversed by significantly quenching PAMP-triggered immunity. Long-term temporal in planta transcriptomic analysis of these three bacterial endophytes revealed up-regulation of protein translation, the generation of energy, and the response to stress, and interestingly, for the microbiota strains, the longer the bacteria remained inside plants, the greater the up-regulation of some of these processes. Further transcriptomic analysis of in planta populations of commensal-simulating Pseudomonas syringae revealed a remarkable resemblance to those of in vitro bacteria in stationary phase, a metabolically active physiological state in which the production of secondary metabolites and stress responses are induced. This study provides novel insight into how endophytic bacteria survive and thrive inside plant leaves, and reshapes our current understanding of what it means to be part of the endophytic microbiota in the phyllosphere.

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Velasquez, A. C., Huguet-Tapia, J. C., He, S. Y.. 2021-05-10. Phyllosphere-inhabiting endophytic bacteria feature a stationary phase-like lifestyle. https://doi.org/10.1101/2021.05.10.443510

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