Blue-Green Fluorescence for Downy Mildew Phenotyping in Lettuce: Linking Transcriptional, Metabolic, and Optical...

Effective plant disease phenotyping is crucial for resistance breeding, but traditional visual assessment is often inaccurate and inefficient. This is particularly challenging when breeding lettuce (Lactuca sativa) for resistance to downy mildew, given the obligate biotrophic lifestyle of the causal pathogen Bremia lactucae. We discovered that B. lactucae-infected lettuce exhibits patches of increased blue-green fluorescence (BGF) under UV-A excitation from 6 days post-inoculation, preceding visible symptoms. Co-localization of BGF with hyphae, visualized with trypan blue, indicates that BGF is induced by downy mildew colonization. We therefore investigated its potential for non-invasive disease detection and quantification, as well as the underlying physiological changes. Using a custom imaging system, we demonstrate that BGF leaf area correlates with downy mildew severity and can be automatically quantified via a U-Net-based convolutional neural network, enabling early, objective disease assessment. Exploring transcriptomic and metabolomic changes associated with BGF, we found that induction of the phenylpropanoid pathway led to accumulation of caffeoylquinic acids, whose fluorescence spectra overlap with that of BGF tissue, supporting the hypothesis that these compounds contribute to the fluorescence signal. BGF imaging offers a powerful tool for phenotyping in lettuce breeding and for identifying quantitative resistance traits that support durable downy mildew resistance.