Towards automated detection of hyperhydricity in plant in vitro culture

Abstract

Hyperhydricity (HH) is one of the most important physiological disorders that negatively affects various plant tissue culture techniques. The objective of this study was to characterize optical features to allow an automated detection of HH. For this purpose, HH was induced in two plant species, apple and Arabidopsis thaliana, and the severity was quantified based on visual scoring and determination of apoplastic liquid volume. The comparison between the HH score and the apoplastic liquid volume revealed a significant correlation, but different response dynamics. Corresponding leaf reflectance spectra were collected and different approaches of spectral analyses were evaluated for their ability to identify HH-specific wavelengths. Statistical analysis of raw spectra showed significantly lower reflection of hyperhydric leaves in the VIS, NIR and SWIR region. Application of the continuum removal hull method to raw spectra identified HH-specific absorption features over time and major absorption peaks at 980 nm, 1150 nm, 1400 nm, 1520 nm, 1780 nm and 1930 nm for the various conducted experiments. Machine learning (ML) model spot checking specified the support vector machine to be most suited for classification of hyperhydric explants, with a test accuracy of 85% outperforming traditional classification via vegetation index with 63% test accuracy and the other ML models tested. Investigations on the predictor importance revealed 1950 nm, 1445 nm in SWIR region and 415 nm in the VIS region to be most important for classification. The validity of the developed spectral classifier was tested on an available hyperspectral image acquisition in the SWIR-region.