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Design and demonstration of a low‐field magnetic resonance imaging rhizotron for in‐field imaging of energy sorghum roots

Abstract Root phenotyping provides critical information to plant breeders for developing varieties with improved drought tolerance, greater root biomass, and greater nutrient use efficiency. Phenotyping roots in the natural environment is important for understanding the effect of the soil environment on root genotypic expressions. The goal of this work was to design and test a field‐scale mobile low‐field magnetic resonance imaging (LF‐MRI) Rhizotron that produces actionable root phenotyping data. We demonstrated this novel technology for root visualization and quantification using a LF‐MRI Rhizotron operating at 47 mT with two soil types. The LF‐MRI Rhizotron weights 453 kg, with a height of 90 cm, a diameter of 28 cm and an imaging field of view of 28 cm × 28 cm. The unit was operated in a Belk clay (Entic Hapluderts) and Weswood silt loam (Udifluventic Halustepts) generating 2‐D and 3‐D image data sets. The 2‐D image data had a collection time of 16.5 min per image at an image resolution of 2.2 mm per pixel. The 3‐D data had a collection time of 13 h per image with a 2.2 × 2.2 × 2.2 mm voxel resolution. Low‐field magnetic resonance imaging worked well for visualizing roots in moderate to high clay soils, demonstrating the potential for this technology; however, the broad application of this platform is hampered due to the prohibitively long scanning time to obtain 3‐D images. By increasing the field strength, and therefore the signal‐to‐noise ratio, faster scan times can enable a more useful system for root phenotyping.

Bagnall, G. Cody↗

Quantitative phenotyping of crop roots with spectral electrical impedance tomography: a rhizotron study with optimized measurement design

Background: Root systems are key contributors to plant health, resilience, and, ultimately, yield of agricultural crops. To optimize plant performance, phenotyping trials are conducted to breed plants with diverse root traits. However, traditional analysis methods are often labour-intensive and invasive to the root system, therefore limiting high-throughput phenotyping. Spectral electrical impedance tomography (sEIT) could help as a non-invasive and cost-efficient alternative to optical root analysis, potentially providing 2D or 3D spatio-temporal information on root development and activity. Although impedance measurements have been shown to be sensitive to root biomass, nutrient status, and diurnal activity, only few attempts have been made to employ tomographic algorithms to recover spatially resolved information on root systems. In this study, we aim to establish relationships between tomographic electrical polarization signatures and root traits of different fine root systems (maize, pinto bean, black bean, and soy bean) under hydroponic conditions. Results: Our results show that, with the use of an optimized data acquisition scheme, sEIT is capable of providing spatially resolved information on root biomass and root surface area for all investigated root systems. We found strong correlations between the total polarization strength and the root biomass (R 2 = 0.82) and root surface area (R 2 = 0.8). Our findings suggest that the captured polarization signature is dominated by cell-scale polarization processes. Additionally, we demonstrate that the resolution characteristics of the measurement scheme can have a significant impact on the tomographic reconstruction of root traits. Conclusion: Our findings showcase that sEIT is a promising tool for the tomographic reconstruction of root traits in high-throughput root phenotyping trials and should be evaluated as a substitute for traditional, often time-consuming, root characterization methods.

59 BASIC BIOLOGICAL SCIENCES↗

A Field-Deployable Magnetic Resonance Imaging Rhizotron for Modeling and Enhancing Root Growth and Biogeochemical Function

A collaborative team from Texas A&M AgriLife Research, ABQMR Inc., the Soil Health Institute, the Athinoula A. Martinos Center for Biomedical Imaging, and NIST developed low-field magnetic resonance imaging (LF-MRI) instrumentation capable of imaging intact soil-root systems. The system measured root biomass, architecture, 3D mass distribution, and growth rates, providing a non-destructive means to evaluate ideal plant characteristics based on root metrics. It also successfully generated three-dimensional images of soil water content, a key property influencing root growth and exploration. Operating much like an MRI used in a medical setting, the system functioned in field conditions without damaging plants, overcoming the limitations of traditional methods such as trenching, soil coring, and root excavation. Over the course of the project, the team designed and built three functional prototype systems. These prototypes provided new insights into root–water–soil interactions that drive processes such as nutrient uptake, water use, and carbon management. This information contributed to efforts to optimize plants for carbon sequestration without sacrificing economic yield. The project also supported the identification of desirable traits for energy sorghum, including high root growth rates, more vertical root angles, and enhanced drought resilience under water-limiting conditions.

09 BIOMASS FUELS↗

Root image collection from a genotype by environment (GxE) root architecture study of the bioenergy species, Populus trichocarpa

Here we present a curated dataset; raw images of fresh roots collected from a genotype by environment (growth matrix) greenhouse root architecture study of the bioenergy species, Populus trichocarpa. Five separate experiments were performed to generate the dataset. Biological replicates (ramets, vegetative propagants) of Populus trichocarpa genotypes BESC 24 and BESC 375 were used for all experiments. The first two experiments were an eight-week and a six-week pot experiment where plants with grown in 4" X 14" sized pots were filled with Farfard 3B potting soil (Fafard Sungro #3B), and watered twice daily. Average greenhouse humidity and growth temperature were around 50-60% and 23°C, respectively. Ramets of P. trichocarpa genotypes BESC 24 and BESC 375 were used in various experiments. The source of this material was dormant field cuttings grown in greenhouse. The second experiment was a Microcosm root architecture experiment. This was done in a microcosm built with double-decker tissue culture Magenta box. Tissue culture plants with new root initials were transferred to microcosm with pro choice rapid dry growth matrix with 1x Hoagland’s solution pH5.7 and grown for six weeks until harvest. Temperature was held at ~22°C. The closed container trapped moisture. The source material was tissue culture stock plants established from shoot tips of greenhouse stock plants (obtained from same set of dormant field cuttings as rest of the study). For the remaining two experiments, greenwood stem cuttings from greenhouse grown plants were grown in six-week rhizotron and two-week hydroponics experiments in the greenhouse. For rhizotron experiments, plants were grown in 2-D rhizotron with black panels on both sides (31cm H x 30cm L x 1.2cm W) and watered daily. For hydroponics experiment, plants were grown in a 15ml falcon tube with distilled water and covered in foil and shaken a few times a day. Greenhouse temperature was held at ~ 23°C and humidity was between 50-60%. Stems were cut off and the root system was separated, gently washed to rid of matrix, immersed in a water tray and scanned for obtaining raw root images reported here. All scans of roots floating in a water tray were performed using an Epson Perfection V850 Pro Photo Scanner. Additional meta data is embedded in the raw image files.

59 BASIC BIOLOGICAL SCIENCES↗

Specialized Plant Growth Chamber Designs to Study Complex Rhizosphere Interactions

The rhizosphere is a dynamic ecosystem shaped by complex interactions between plant roots, soil, microbial communities and other micro- and macro-fauna. Although studied for decades, critical gaps exist in the study of plant roots, the rhizosphere microbiome and the soil system surrounding roots, partly due to the challenges associated with measuring and parsing these spatiotemporal interactions in complex heterogeneous systems such as soil. To overcome the challenges associated with in situ study of rhizosphere interactions, specialized plant growth chamber systems have been developed that mimic the natural growth environment. This review discusses the currently available lab-based systems ranging from widely known rhizotrons to other emerging devices designed to allow continuous monitoring and non-destructive sampling of the rhizosphere ecosystems in real-time throughout the developmental stages of a plant. We categorize them based on the major rhizosphere processes it addresses and identify their unique challenges as well as advantages. We find that while some design elements are shared among different systems (e.g., size exclusion membranes), most of the systems are bespoke and speaks to the intricacies and specialization involved in unraveling the details of rhizosphere processes. We also discuss what we describe as the next generation of growth chamber employing the latest technology as well as the current barriers they face. We conclude with a perspective on the current knowledge gaps in the rhizosphere which can be filled by innovative chamber designs.

59 BASIC BIOLOGICAL SCIENCES↗