RGB imaging-based evaluation of waterlogging tolerance in cultivated and wild chrysanthemums

Waterlogging is a major stress that impacts the chrysanthemum industry. Large-scale germplasm screening for identifying waterlogging-tolerant resources in a quick and accurate manner is essential for developing new cultivars with improved waterlogging tolerance. To overcome this phenotyping bottleneck, consumer-grade digital cameras have been used to acquire the red-green-blue (RGB) images of 180 chrysanthemum cultivars and their wild relatives under waterlogging stress and well-watered conditions. A total of 103 image-based digital traits (i-traits), including 10 morphological i-traits and 93 texture i-traits, were extracted and systematically analyzed. Most of these i-traits presented high coefficients of variation (CVs) and broad-sense heritability (H ² ), with an average CV of 34.04 ​% and an average H ² of 0.93. We identified several novel texture i-traits associated with the hue (H) component, which strongly correlated with the traditional waterlogging tolerance index, the membership function value of waterlogging (MFVW) (R ​= ​0.63-0.77). We further employed the random forest (RF) and gradient boosting tree (GBT) machine learning algorithms to predict aboveground biomass and MFVW on the basis of different i-trait datasets. The RF model achieved superior predictive performance, with a coefficient of determination (R ² ) of up to 0.88 for shoot weight and 0.86 for MFVW. Moreover, a subset of the top 13 most important i-traits could accurately predict MFVW (R ²  ​> ​0.80) via the cross-validation method. A total of 10 highly tolerant resources were selected by traditional and RGB-based evaluation, and 50 ​% belonged to Artemisia. Our findings confirmed that RGB-based technology provides a promising novel approach for quantifying waterlogging response that contributes to future breeding programs and genetic dissection for waterlogging tolerance.