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.
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. For instance, flat surfaces such as nutrient agar, enable the influence of nutrient supply on microorganisms to be investigated. In contrast microstructured surfaces, such as isolated leaf cuticles or reconstituted leaf waxes enable the influence of physicochemical properties to be investigated. However, interest in replica leaf surfaces as an artificial surface is growing. As replica surfaces offer an improved representation of the complex topography of leaf surfaces. The use of replica leaf surfaces has to date primarily been focused on replicating the superhydrophobic surfaces of leaves. Whereas in this paper, we investigate potential replica surface materials for phyllosphere microbiology studies. Using a test pattern, we investigated the resolution, the degradation characteristics in environmental conditions, surface energy, and bacterial survival characteristics for each potential replica material. Our results indicate that PDMS is the most suitable material for producing replica leaf surfaces. Due to the high resolution achieved through replica molding, extended stability, hydrophobic properties, and bacterial survival characteristics comparable to isolated leaf cuticles. Our experiments highlight the importance of considering the inherent material properties, when selecting a replica leaf surface for phyllosphere microbiology studies. A replica leaf produced in PDMS offers a control surface that can be used for investigating microbe-microbe and microbe-plant interactions in the phyllosphere. Thus, in turn enabling mitigation strategies against pathogens to either the plant host or humans to be developed.
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