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

Abstract

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 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 area; (4) evaluation of statistically significant differences between plant groups; (5) calculation of germination rates; and (6) visualization and interpretation. Morley is designed for laboratory studies of biotic effects on seedling growth, when the molecular mechanisms underlying the morphometric changes are analyzed. The performance was tested using cultivars of Triticum aestivum and Pisum sativum on seedlings of up to 1 week old. The accuracy of the measured morphometric parameters was comparable with that obtained using ImageJ and manual measurements. Possible applications of Morley include dose-dependent laboratory tests for germination affected by new bioactive compounds and fertilizers.