Optimization of field asymmetric ion mobility spectrometry-based assessment of Aphanomyces root rot in pea

Abstract

When plants are infected with pathogens, disease response can result in changes in the profiles of volatile organic compounds (VOC). These changes in volatile organic compounds (VOC) profiles can be utilized for disease detection and quantification. In this study, field asymmetric ion mobility spectrometry (FAIMS) was used to evaluate the VOC profile variability in a pea near isogenic line (Pisum sativum L.) inoculated with zoospores of Aphanomyces euteiches Drechs, which causes Aphanomyces root rot disease. Pots were filled with silica sand and six plants per pot were grown under controlled conditions in a randomized complete block design with four replications. Four treatments, namely non-inoculated, 1 × 10⁵, 1 × 10⁶, and 2.79 × 10⁶ zoospores ml⁻¹ were applied to plants at 5 and 7 days after emergence. FAIMS was used to collect volatile profiles at 2, 4, 7 and 9 days after inoculation. Specific regions of interest – extracted from the ion current intensity from the FAIMS spectra – were analyzed using ANOVA. Similarly, multiple regions of interest were evaluated using principal component analysis and k-means clustering. Ion current profiles and curvature profiles were incorporated into the analysis using k-means clustering. Other ground reference data such as root rot index and physiological parameters were also recorded. The results showed a biomarker in a specific region of interest demonstrating ample ability to quantify and differentiate treatment effects during non-destructive sampling at 14 DAE (7 DAI). Data from this region could be used for early and non-destructive quantification and differentiation of treatment effects based on zoospore inoculation levels. The k-means clustering of ion current and curvature profiles showed patterns based on the treatments. These findings demonstrated that FAIMS could be used as a tool to assess plant-pathogen interactions using volatile biomarkers to evaluate disease responses and severity under controlled conditions.