A Low-Cost Method for Phenotyping Wilting and Recovery of Wheat Leaves under Heat Stress Using Semi-Automated Image Analysis

Abstract

Leaf wilting is the most common symptom of dehydration stress. Methods to analyze this phenomenon are particularly relevant to evaluate crop agronomic performance, to genetically dissect out the wilting process, and for functional analysis of genetically modified plants. In this study, a low-cost, semi-automated method to quantify leaf folding of wilting plants is described that can replace visual analysis. Standardized heat-stress conditions were applied with a thermostatic drier, on plantlets or excised leaves from three wheat genotypes (Trinakria, Cappelli, and a Water-mutant of Trinakria). The best time–temperature binomial to record both the leaf wilting and recovery phases was identified using a free time-lapse application, by a smartphone camera. The quantitative description of the wilting phenomenon was obtained through the Kinovea software, which automatically tracked the leaf angle changes over time, computed various kinematic data (angular velocity, centripetal acceleration, total degrees of displacement) and constructed the graphs. The possibility of applying standardized heat-stress conditions and quantitatively describe the leaf folding kinematics means that this instrumentation and its use represents a very low cost tool for objective phenotyping of the degree of the heat-stress tolerance of wheat and of morphologically similar species.