We present RootPainter, a GUI-based software tool for the rapid training of deep neural networks for use in biological image analysis. RootPainter facilitates both fully-automatic and semiautomatic image segmentation. We investigate the effectiveness of RootPainter using three plant image datasets, evaluating its potential for root length extraction from chicory roots in soil, biopore counting and root nodule counting from scanned roots. We also use RootPainter to compare dense annotations to corrective ones which are added during the training based on the weaknesses of the current model. Deep Learning | GUI | Segmentation | Phenotyping | Biopore | Rhizotron | Root nodule | Interactive segmentation Correspondence: ags@di.ku.dk Fig. 1. RootPainter corrective annotation concept. (a) Roots in soil. (b) AI root predictions. (c) Human corrections. (d) AI learns from corrections. Smith et al. | bioRχiv | April 16, 2020 | 1-16