Quantifying aluminum toxicity effects on corn phenotype using advanced imaging technologies.

Soil acidity (pH <5.5) limits agricultural production due to aluminum (Al) toxicity. The primary target of Al toxicity is the plant root. However, symptoms can be observed on the shoots. This study aims to determine the potential use of chlorophyll fluorescence imaging, multispectral imaging, and 3D multispectral scanning technology to quantify the effects of Al toxicity on corn. Corn seedlings were grown for 13 days in nutrient solutions (pH 4.0) with four Al treatments: 50, 100, 200, and 400 µM and a control (0 µM AlCl3 L-1). During the experiment, four measurements were performed: four (MT1), six (MT2), 11 (MT3), and 13 (MT4) days after the application of Al treatments. The most sensitive traits affected by Al toxicity were the reduction of plant growth and increased reflectance in the visible wavelength (affected at MT1). The reflectance of red wavelengths increased more significantly compared to near-infrared and green wavelengths, leading to a decrease in the normalized difference vegetation index and the Green Leaf Index. The most sensitive chlorophyll fluorescence traits, effective quantum yield of PSII, and photochemical quenching coefficient were affected after prolonged Al exposure (MT3). This study demonstrates the usability of selected phenotypic traits in remote sensing studies to map Al-toxic soils and in high-throughput phenotyping studies to screen Al-tolerant genotypes.

Monitoring Drought Stress in Common Bean Using Chlorophyll Fluorescence and Multispectral Imaging.

Drought is a significant constraint in bean production. In this study, we used high-throughput phenotyping methods (chlorophyll fluorescence imaging, multispectral imaging, 3D multispectral scanning) to monitor the development of drought-induced morphological and physiological symptoms at an early stage of development of the common bean. This study aimed to select the plant phenotypic traits which were most sensitive to drought. Plants were grown in an irrigated control (C) and under three drought treatments: D70, D50, and D30 (irrigated with 70, 50, and 30 mL distilled water, respectively). Measurements were performed on five consecutive days, starting on the first day after the onset of treatments (1 DAT-5 DAT), with an additional measurement taken on the eighth day (8 DAT) after the onset of treatments. Earliest detected changes were found at 3 DAT when compared to the control. D30 caused a decrease in leaf area index (of 40%), total leaf area (28%), reflectance in specific green (13%), saturation (9%), and green leaf index (9%), and an increase in the anthocyanin index (23%) and reflectance in blue (7%). The selected phenotypic traits could be used to monitor drought stress and to screen for tolerant genotypes in breeding programs.

Classification of high-throughput phenotyping data for differentiation among nutrient deficiency in common bean.

The development of automated, image-based, high-throughput plant phenotyping enabled the simultaneous measurement of many plant traits. Big and complex phenotypic datasets require advanced statistical methods which enable the extraction of the most valuable traits when combined with other measurements, interpretation, and understanding of their (eco)physiological background. Nutrient deficiency in plants causes specific symptoms that can be easily detected by multispectral imaging, 3D scanning, and chlorophyll fluorescence measurements. Screening of numerous image-based phenotypic traits of common bean plants grown in nutrient-deficient solutions was conducted to optimize phenotyping and select the most valuable phenotypic traits related to the specific nutrient deficit. Discriminant analysis was used to compare the efficiency of groups of traits obtained by high-throughput phenotyping techniques (chlorophyll fluorescence, multispectral traits, and morphological traits) in discrimination between nutrients [nitrogen (N), phosphorus (P), potassium (K), magnesium (Mg), and iron (Fe)] at early and prolonged deficiency. Furthermore, a recursive partitioning analysis was used to select variables within each group of traits that show the highest accuracy for assigning plants to the respective nutrient deficit treatment. Using the entire set of measured traits, the highest classification success by discriminant function was achieved using multispectral traits. In the subsequent measurements, chlorophyll fluorescence and multispectral traits achieved comparably high classification success. Recursive partitioning analysis was able to intrinsically identify variables within each group of traits and their threshold values that best separate the observations from different nutrient deficiency groups. Again, the highest success in assigning plants into their respective groups was achieved based on selected multispectral traits. Selected chlorophyll fluorescence traits also showed high accuracy for assigning plants into control, Fe, Mg, and P deficit but could not correctly assign K and N deficit plants. This study has shown the usefulness of combining high-throughput phenotyping techniques with advanced data analysis to determine and differentiate nutrient deficiency stress.