Advances in automated image analysis using open-source computer vision tools, such as PlantCV, have greatly increased the throughput of aboveground phenotyping in a variety of crop species. However, PlantCV was largely optimized to analyze images collected under controlled laboratory conditions, and has seldom been used to analyze images collected under field conditions. Further, there are no known applications of PlantCV for analyzing images collected belowground, such as those obtained from minirhizotron imaging devices. In this study, we demonstrated applications of PlantCV for extracting plant trait information from aboveground and belowground images collected in two perennial crop mapping populations. The first population was composed of nearly 1,200 individuals of a potential perennial oilseed crop (Silphium Integrifolium x Perfoliatum ), and the second population was composed of nearly 1,700 individuals of a perennial cover crop (Trifolium ambiguum , Kura Clover). We designed and used a field-based imaging cart to collect overhead and profile images of individuals from both populations in August and October, which improved the efficiency of field-based image capture. Around the time of aboveground image collection, belowground images of root networks were collected using minirhizotron imaging devices. We then assessed the application of PlantCV for measuring aboveground traits (crop canopy area, height, leaf color and growth rates) and belowground traits (root length and growth rates), and we explored future directions of PlantCV for field-based image analysis of aboveground and belowground crop tissues.

Nitrogen inputs can be an important cost consideration for farmers in terms of economic profit as well as environmental impact. Elucidating genetic regions that are associated with plant phenotype response to nitrogen stress can help in facilitating breeding approaches that can mitigate these costs. A diverse population of 272 maize lines was planted at a field site in Champaign, IL in two consecutive years in reduced nitrogen conditions. 302 phenotypes were recorded including: seed ionomic content, root structural traits derived from 2-dimensional images as well as a 3-dimensional representation generated from x-ray computed tomography (XRT) scans, root traits extrapolated from mini-rhizotron systems, drone images across the growing season and end of season agronomic traits such as biomass and yield. BLUP models were fit to obtain estimates of single year genotypic values as well as across years values both of which took into account year specific spatial variation. Individual years and combined years BLUP values were used as response variables in genome wide association studies (GWAS) to identify loci significantly associated with each set of values. Significant associations were identified for all phenotype categories.