Abstract

Artificial surfaces are routinely used instead of leaves to enable a reductionist approach in phyllosphere microbiology, the study of microorganisms residing on plant leaf surfaces. Commonly used artificial surfaces include, flat surfaces, such as metal and nutrient agar, and microstructured surfaces, such as isolate leaf cuticles or reconstituted leaf waxes. However, interest in replica leaf surfaces as an artificial surface is growing, as replica surfaces provide an improved representation of the complex topography of leaf surfaces. To date, leaf surfaces have predominantly been replicated for their superhydrophobic properties. In contrast, in this paper we investigated the potential of agarose, the elastomer polydimethylsiloxane (PDMS), and gelatin as replica leaf surface materials for phyllosphere microbiology studies. Using a test pattern of pillars, we investigated the ability to replicate microstructures into the materials, as well as the degradation characteristics of the materials in environmental conditions. Pillars produced in PDMS were measured to be within 10% of the mold master and remained stable throughout the degradation experiments. In agarose and gelatin the pillars deviated by more than 10% and degraded considerably within 48 hours in environmental conditions. Furthermore, we investigated the surface energy of the materials, an important property of a leaf surface, which influences resource availability and microorganism attachment. We found that the surface energy and bacterial viability on PDMS was comparable to isolated Citrus × aurantium and Populus × canescens leaf cuticles. Hence indicating that PDMS is the most suitable material for replica leaf surfaces. In summary, our experiments highlight the importance of considering the inherent material properties when selecting a replica leaf surface for phyllosphere microbiology studies. As demonstrated, a PDMS replica leaf offers a control surface that can be used for investigating microbe-microbe and microbe-plant interactions in the phyllosphere, which will enable mitigation strategies against pathogens to be developed.

Highlights

  • Our work has demonstrated the potential of PDMS as a replica leaf material for phyllosphere microbiology

  • Our results highlighted the drawbacks of agarose and gelatin through comparing optical resolution, degradation characteristics, hydrophobic properties, and bacterial survival to PDMS

  • We demonstrated that agarose and gelatin replicas would not provide topography comparable to that of a living leaf

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Summary

Introduction

Microorganisms contribute to the health of the plant host. The rhizosphere is the microbial habitat in the vicinity of the plant roots, which is influenced by the products secreted by the roots.[6, 7] recently phyllosphere microbiology, the study of microorganisms which reside on plant leaf surfaces, has gained increasing interest. This increase is attributed to growing interest in the role that microorganisms in the phyllosphere, have on the health of the plant host. The cuticle prevents water, ion, and nutrient loss.[9]

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