Biofilms Benefiting Plants Exposed to ZnO and CuO Nanoparticles Studied with a Root-Mimetic Hollow Fiber Membrane.

Plants exist with a consortium of microbes that influence plant health, including responses to biotic and abiotic stress. While nanoparticle (NP)-plant interactions are increasingly studied, the effect of NPs on the plant microbiome is less researched. Here a root-mimetic hollow fiber membrane (HFM) is presented for generating biofilms of plant-associated microbes nurtured by artificial root exudates (AREs) to correlate exudate composition with biofilm formation and response to NPs. Two microbial isolates from field-grown wheat, a bacillus endophyte and a pseudomonad root surface colonizer, were examined on HFMs fed with AREs varying in N and C composition. Bacterial morphology and biofilm architecture were characterized using scanning electron microscopy (SEM) and atomic force microscopy (AFM) and responses to CuO and ZnO NP challenges of 300 mg/L evaluated. The bacillus isolate sparsely colonized the HFM. In contrast, the pseudomonad formed robust biofilms within 3 days. Dependent on nutrient sources, the biofilm cells produced extensive extracellular polymeric substances (EPS) and large intracellular granules. Pseudomonad biofilms were minimally affected by ZnO NPs. CuO NPs, when introduced before biofilm maturation, strongly reduced biofilm formation. The findings demonstrate the utility of the HFM root-mimetic to study rhizoexudate influence on biofilms of root-colonizing microbes but without active plant metabolism. The results will allow better understanding of how microbe-rhizoexudate-NP interactions affect microbial and plant health.