Dive into the world of cutting-edge plant research in the CAPITALISE project at the PSI Research Center in the Czech Republic to see how we study our crops’ photosynthetic and growth performance under drought stress conditions using high-throughput image-based phenotyping. We applied an integrative phenotyping protocol to get insights into the performance and resilience to drought stress in barley genotypes as part of Tier 3 phenotyping experiments. In collaboration with Dr. Eyal Fridman at Volcani Center (ARO), we screened spring barley (Hordeum vulgare) genotypes selected from the CMPP (Cytoplasmic multi-parental DH population) and HEB (Halle Exotic Barley) families, including wild (Hs/Hs) and cultivated (Hv/Hv) alleles.
Plant cultivation: Seedlings were transplanted into 3L pots and grown under a short photoperiod in walk-in chambers until the fourth leaf stage. They were then moved to a PlantScreenTM phenotyping system at PSI Research Center for automated scoring in the greenhouse under long photoperiod (16 h) and were grown until full maturation.
Stress application: Drought stress treatment was induced at the tillering stage, 24 days after transplanting, with soil relative water content maintained at 55% for control plants and 25% for stressed plants, reducing to 20% at the flowering stage (65 days after transplanting). Daily watering and weighing ensured consistent pot weight and monitored water consumption.
Cultivation and growth of barley plants under controlled conditions at PSI research center
Integrative Phenotyping: Growth dynamics and physiological responses were assessed by measuring and calculating 145 traits using multiple imaging sensors over 70 time points. Plants were phenotyped three times a week using chlorophyll fluorescence for photosynthetic efficiency, RGB imaging for growth rate and color analysis, and thermal imaging for canopy temperature. Additionally, hyperspectral imaging was conducted once a week to detect early senescence from leaf reflectance indices.
Multiple imaging sensors were used to assess morphological and physiological responses from side and top-view images
Yield-Related Traits Assessment: At maturity, the above-ground biomass, including leaves, tillers, and spikes, was harvested and the dry weight was measured. To improve spike assessment, we developed SpikeAPP, a deep-learning platform for efficient and reproducible image-based spike detection during large-scale phenotyping. We also assessed grain-related traits using X-ray imaging to measure grain number, length, and width collected from five spikes per plant, followed by ground truth validation.
By combining comprehensive metadata analysis with an integrative phenotyping approach, we were able to rank the genotypes based on their stress resilience and overall performance, including yield.
Yield-related trait assessment from RGB and X-ray images followed by ground truth validation
References:
Tiwari, L.D., Bdolach, E., Prusty, M.R.,Bodenheimer, S., Be’ery, A., Faigenboim-Doron, A. et al. (2024) Cytonuclear interactions modulate the plasticity of photosynthetic rhythmicity and growth in wild barley.Physiologia Plantarum, 176(1), e14192. https://doi.org/10.1111/ppl.14192
Merchuk-Ovnat, L., Silberman, R., Laiba, E., Maurer, A., Pillen, K., Faigenboim, A., & Fridman, E. (2018). Genome scan identifies flowering-independent effects of barley HsDry2. 2 locus on yield traits under water deficit. Journal of Experimental Botany, 69(7), 1765-1779. https://doi.org/10.1093/jxb/ery016
