RGB and Spectral Root Imaging for Plant Phenotyping and Physiological Research: Experimental Setup and Imaging Protocols

Better understanding of plant root dynamics is essential to improve resource use efficiency of agricultural systems and increase the resistance of crop ...

University of Natural Resources and Life Sciences

Plant root systems have a key role in ecology and agronomy. In spite of fast increase in root studies, still there is no classification that allows distinguishing among distinctive characteristics within the diversity of rooting strategies. Our hypothesis is that a multivariate approach for "plant functional type" identification in ecology can be applied to the classification of root systems. The classification method presented is based on a data-defined statistical procedure without a priori decision on the classifiers. The study demonstrates that principal component based rooting types provide efficient and meaningful multi-trait classifiers. The classification method is exemplified with simulated root architectures and morphological field data. Simulated root architectures showed that morphological attributes with spatial distribution parameters capture most distinctive features within root system diversity. While developmental type (tap vs. shoot-borne systems) is a strong, but coarse classifier, topological traits provide the most detailed differentiation among distinctive groups. Adequacy of commonly available morphologic traits for classification is supported by field data. Rooting types emerging from measured data, mainly distinguished by diameter/weight and density dominated types. Similarity of root systems within distinctive groups was the joint result of phylogenetic relation and environmental as well as human selection pressure. We concluded that the data-define classification is appropriate for integration of knowledge obtained with different root measurement methods and at various scales. Currently root morphology is the most promising basis for classification due to widely used common measurement protocols. To capture details of root diversity efforts in architectural measurement techniques are essential.

A statistical approach to root system classification.

Trait-based breeding is essential to improve wheat yield, particularly when stress adaptation is targeted. A set of modern and underutilized wheat genotypes was examined in a 2-year field experiment with distinct seasonal water supply. Yield formation and drought response strategies were analyzed in relation to components of Passioura's yield-water framework based on phenological, morphological, physiological, and root characteristics. Limited water supply resulted in 60% yield loss and substantially lower water use (37%), water use efficiency (32.6%), and harvest index (14%). Phenology and root length density were key determinants of water use. Late flowering underutilized wheat species with large root system and swift ground coverage showed greatest water use. Leaf chlorophyll concentration and stomata conductance were higher in modern cultivars, supporting their high biomass growth and superior water use efficiency. While, lower chlorophyll concentration and stomata conductance of underutilized wheats indicated a water saving strategy with an intrinsic limitation of potential growth. Harvest index was strongly dependent on phenology and yield components. Optimized flowering time, reduced tillering, and strong grain sink of modern cultivars explained higher harvest index compared to underutilized wheats. Cluster analysis revealed the consistent differentiation of underutilized and modern wheats based on traits underlying Passioura's yield-water framework. We identified physiological and root traits within modern cultivars to be targeted for trait-based crop improvement under water-limited conditions. High capacity of water use in underutilized genetic resources is related to yield-limiting phenological and morphological traits, constraining their potential role for better drought resistance. Still some genetic resources provide adaptive features for stress resistance compatible with high yield as revealed by high harvest index under drought of Khorasan wheat.

Dissection of drought response of modern and underutilized wheat varieties according to Passioura's yield-water framework.

Modern imaging technology provides new approaches to plant phenotyping for traits relevant to crop yield and resource efficiency. Our objective was to investigate water use strategies at early growth stages in durum wheat genetic resources using shoot imaging at the ScreenHouse phenotyping facility combined with physiological measurements. Twelve durum landraces from different pedoclimatic backgrounds were compared to three modern check cultivars in a greenhouse pot experiment under well-watered (75% plant available water, PAW) and drought (25% PAW) conditions. Transpiration rate was analyzed for the underlying main morphological (leaf area duration) and physiological (stomata conductance) factors. Combining both morphological and physiological regulation of transpiration, four distinct water use types were identified. Most landraces had high transpiration rates either due to extensive leaf area (area types) or both large leaf areas together with high stomata conductance (spender types). All modern cultivars were distinguished by high stomata conductance with comparatively compact canopies (conductance types). Only few landraces were water saver types with both small canopy and low stomata conductance. During early growth, genotypes with large leaf area had high dry-matter accumulation under both well-watered and drought conditions compared to genotypes with compact stature. However, high stomata conductance was the basis to achieve high dry matter per unit leaf area, indicating high assimilation capacity as a key for productivity in modern cultivars. We conclude that the identified water use strategies based on early growth shoot phenotyping combined with stomata conductance provide an appropriate framework for targeted selection of distinct pre-breeding material adapted to different types of water limited environments.

Identification of Water Use Strategies at Early Growth Stages in Durum Wheat from Shoot Phenotyping and Physiological Measurements.

Root phenotyping provides trait information for plant breeding. A shortcoming of high-throughput root phenotyping is the limitation to seedling plants and failure to make inferences on mature root systems. We suggest root system architecture (RSA) models to predict mature root traits and overcome the inference problem. Sixteen pea genotypes were phenotyped in (i) seedling (Petri dishes) and (ii) mature (sand-filled columns) root phenotyping platforms. The RSA model RootBox was parameterized with seedling traits to simulate the fully developed root systems. Measured and modelled root length, first-order lateral number, and root distribution were compared to determine key traits for model-based prediction. No direct relationship in root traits (tap, lateral length, interbranch distance) was evident between phenotyping systems. RootBox significantly improved the inference over phenotyping platforms. Seedling plant tap and lateral root elongation rates and interbranch distance were sufficient model parameters to predict genotype ranking in total root length with an RSpearman of 0.83. Parameterization including uneven lateral spacing via a scaling function substantially improved the prediction of architectures underlying the differently sized root systems. We conclude that RSA models can solve the inference problem of seedling root phenotyping. RSA models should be included in the phenotyping pipeline to provide reliable information on mature root systems to breeding research.

Alireza Nakhforoosh

Division of Agronomy,
Department of Crop Sciences,
Division of Agronomy, Department of Crop Sciences

University of Natural Resources and Life Sciences

RGB and Spectral Root Imaging for Plant Phenotyping and Physiological Research: Experimental Setup and Imaging Protocols

Alireza Nakhforoosh

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Division of Agronomy,Department of Crop Sciences,Division of Agronomy, Department of Crop Sciences

University of Natural Resources and Life Sciences

RGB and Spectral Root Imaging for Plant Phenotyping and Physiological Research: Experimental Setup and Imaging Protocols

Division of Agronomy,
Department of Crop Sciences,
Division of Agronomy, Department of Crop Sciences