Imaging O2 dynamics and microenvironments in the seagrass leaf phyllosphere with magnetic optical sensor nanoparticles

Abstract

Eutrophication leads to epiphyte blooms on seagrass leaves that strongly affect plant health, yet the actual mechanisms of such epiphyte-induced plant stress remain poorly understood. We used magnetic optical sensor nanoparticles in combination with luminescence lifetime imaging to map the O2 concentration and dynamics in the heterogeneous seagrass phyllosphere under changing light conditions. By incorporating magnetite into the sensor nanoparticles, it was possible to image the spatial O2 distribution under flow over seagrass leaf segments in the presence of a strong magnetic field. Local microniches with low leaf surface O2 concentrations were found under thick epiphytic biofilms, often leading to anoxic microhabitats in darkness. High irradiance led to O2 supersaturation across most of the seagrass phyllosphere, whereas leaf microenvironments with reduced O2 conditions were found under epiphytic biofilms at low irradiance, probably driven by self-shading. Horizontal micro-profiles extracted from the O2 images revealed pronounced heterogeneities in local O2 concentration over the base of the epiphytic biofilm, with up to 52% reduction in O2 concentrations in areas with relatively thick (>2 mm), compared with thin (≤1 mm), epiphyte layers in darkness. We also present evidence of enhanced relative internal O2 transport within leaves with epiphyte overgrowth, compared with bare seagrass leaves, in light as a result of limited mass transfer across thick outward diffusion pathways. The local availability of O2 was still markedly reduced in the epiphyte-covered leaves, however. The leaf phyllosphere is thus characterized by a complex microlandscape of O2 availability that strongly affects microbial processes occurring within the epiphytic biofilm, which may have implications for seagrass health, as anoxic microhabitats have been shown to promote the microbiological production of reduced toxic compounds, such as nitric oxide.