Abstract

Bacteria are social organisms that interact extensively within and between species while responding to external stimuli from their environments. Designing synthetic microbial communities can enable efficient and beneficial microbiome implementation in many areas. However, in order to design an efficient community, one must consider the interactions between their members. Using a reductionist approach, we examined pairwise interactions of three related Pseudomonas species in various microenvironments including plant roots and inert surfaces. Our results show that the step between monoculture and co-culture is already very complex. Monoculture root colonization patterns demonstrate that each isolate occupied a particular location on wheat roots, such as root tip, distance from the tip, or scattered along the root. However, pairwise colonization outcomes on the root did not follow the bacterial behavior in monoculture, suggesting various interaction patterns. In addition, we show that interspecies interactions on a microscale on inert surface take part in co-culture colonization and that the interactions are affected by the presence of root extracts and depend on its source. The understanding of interrelationships on the root may contribute to future attempts to manipulate and improve bacterial colonization and to intervene with root microbiomes to construct and design effective synthetic microbial consortia.

Highlights

  • One of the frontlines of synthetic biology is engineering of synthetic microbial communities (SMCs) aiming at efficient and beneficial microbiome implementation in many areas, including biosynthesis (Santoyo et al, 2012), biodegradation of complex organic matter (McCarty and Ledesma-Amaro, 2019), and improving crop quality and health (Rainey, 1999; Lugtenberg and Kamilova, 2009)

  • We examined whether monoculture colonization patterns of these three related Pseudomonas strains (Tovi et al, 2019) were sufficient to predict pairwise colonization and interaction outcomes in liquid culture, on glass surface, and in roots

  • Based on confocal laser scanning microscopy (CLSM), quantitative polymerase chain reaction, and live imaging fluorescent microscopy, we demonstrated that a colonization pattern by a single isolate will not necessarily predict its behavior when introduced as a pair

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Summary

Introduction

One of the frontlines of synthetic biology is engineering of synthetic microbial communities (SMCs) aiming at efficient and beneficial microbiome implementation in many areas, including biosynthesis (Santoyo et al, 2012), biodegradation of complex organic matter (McCarty and Ledesma-Amaro, 2019), and improving crop quality and health (Rainey, 1999; Lugtenberg and Kamilova, 2009). Many studies have shown the positive effects of inoculation with a single strain of beneficial soil microbe, in particular, Pseudomonas species, on crop yields, health, and quality (Loper, 1988; Capper and Higgins, 1993; Weller, 2007). Recent studies address the question of multispecies inoculum, aiming at engineering SMC to improve plant development, health, and nutrition (Kong et al, 2018; McCarty and Ledesma-Amaro, 2019; Zafarul-Hye et al, 2020).

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