Contact reflectance spectroscopy for rapid, accurate, and non-destructive Phytophthora infestans clonal lineage discrimination.

Isolates of the predominantly asexual Phytophthora infestans, the oomycete causal agent of potato late blight, population in the US are characterized by clonal lineage, or asexual descendants of a single genotype. Current tools for clonal lineage identification are time consuming and require laboratory equipment. We previously found that foliar spectroscopy can be used for high accuracy pre- and post-symptomatic detection of P. infestans infections caused by clonal lineages US-08 and US-23. In this work, we find that there are subtle but distinct differences in spectral responses of potato foliage infected by these clonal lineages in both controlled, time course (12 to 24-h intervals over 5 days) growth chambers experiments and naturally-infected samples from commercial production fields. In both settings we measured continuous visible to shortwave infrared reflectance (400-2500 nm) on leaves using a portable spectrometer with contact probe. We consistently discriminated between infections caused by the two clonal lineages across all stages of disease progression using partial least squares discriminant analysis (PLS-DA) with total accuracies ranging from 88 to 98%. Three-class random forest differentiation between control, US-08, and US-23 yielded total discrimination accuracy ranging from 68 to 76%. Differences were greatest during pre-symptomatic infection stages and progressed towards uniformity as symptoms advanced. Using PLS-regression trait models we found that total phenolics, sugar, and leaf mass per area were different between lineages. Shortwave infrared wavelengths (>1100 nm) were important for clonal lineage differentiation. This work provides a foundation for future use of hyperspectral sensing as a non-destructive tool for pathovar differentiation.