Morley: Image Analysis and Evaluation of Statistically Significant Differences in Geometric Sizes of Crop Seedlings Responded to Biotic Stimulation

Image analysis is widely applied in plant science for phenotyping and monitoring botanic and agricultural species. Although a lot of software is available, tools integrating image analysis and statistical assessment of seedling growth in large groups of plants are limited or absent, and do not cover the needs of the researchers. In this study, we developed Morley, a free, open-source graphical user interface written in Python. Morley automates the following workflow: (1) group-wise analysis of a few thousand seedlings from multiple images; (2) recognition of seeds, shoots and roots in seedling images; (3) calculation of shoot and root lengths and surface areas, (4) evaluation of statistically significant differences between plant groups, (5) calculation of germination rates, (6) visualization and interpretation. Morley is designed for laboratory studies of biotic effects on seedling growth, when molecular mechanisms underlying morphometric changes are analyzed. Performance was tested using cultivars of T. aestivum, P. sativum on seedlings of up to 1 week old. Accuracy of the measured morphometric parameters was comparable with the ones obtained using ImageJ and manual measurements. Dose-dependent laboratory tests for germination affected by new bioactive compounds and fertilizers, assuming extraction of seedlings from a substrate and/or dissection are among the suggested applications.