Roots are central to the function of natural and agricultural ecosystems by driving plant acquisition of soil resources and influencing the carbon cycle. Root characteristics like length, diameter, and volume are critical to measure to understand plant and soil functions. RhizoVision Explorer is an open-source software designed to enable researchers interested in roots by providing an easy-to-use interface, fast image processing, and reliable measurements. The default broken roots mode is intended for roots sampled from pots and soil cores, washed, and typically scanned on a flatbed scanner, and provides measurements like length, diameter, and volume. The optional whole root mode for complete root systems or root crowns provides additional measurements such as angles, root depth, and convex hull. Both modes support providing measurements grouped by defined diameter ranges, the inclusion of multiple regions of interest, and batch analysis. RhizoVision Explorer was successfully validated against ground truth data using a new copper wire image set. In comparison, the current reference software, the commercial WinRhizo TM, drastically underestimated volume when wires of different diameters were in the same image. Additionally, measurements were compared with WinRhizo TM and IJ_Rhizo using a simulated root image set, showing general agreement in software measurements, except for root volume. Finally, scanned root image sets acquired in different labs for the crop, herbaceous, and tree species were used to compare results from RhizoVision Explorer with WinRhizo TM. The two software showed general agreement, except that WinRhizo TM substantially underestimated root volume relative to RhizoVision Explorer. In the current context of rapidly growing interest in root science, RhizoVision Explorer intends to become a reference software, improve the overall accuracy and replicability of root trait measurements, and provide a foundation for collaborative improvement and reliable access to all.

Roots are central to the function of natural and agricultural ecosystems by driving plant acquisition of soil resources and influencing the carbon cycle. Root characteristics like length, diameter, and volume are critical to measure to understand plant and soil functions. RhizoVision Explorer is an open-source software designed to enable researchers interested in roots by providing an easy-to-use interface, fast image processing, and reliable measurements. The default broken roots mode is intended for roots sampled from pots or soil cores, washed, and typically scanned on a flatbed scanner, and provides measurements like length, diameter, and volume. The optional whole root mode for complete root systems or root crowns provides additional measurements such as angles, root depth, and convex hull. Both modes support providing measurements grouped by defined diameter ranges, the inclusion of multiple regions of interest, and batch analysis. RhizoVision Explorer was successfully validated against ground truth data using a novel copper wire image set. In comparison, the current reference software, the commercial WinRhizoTM, drastically underestimated volume when wires of different diameters were in the same image. Additionally, measurements were compared with WinRhizoTM and IJ_Rhizo using a simulated root image set, showing general agreement in software measurements, except for root volume. Finally, scanned root image sets acquired in different labs for the crop, herbaceous, and tree species were used to compare results from RhizoVision Explorer with WinRhizoTM. The two software showed general agreement, except that WinRhizoTM substantially underestimated root volume relative to RhizoVision Explorer. In the current context of rapidly growing interest in root science, RhizoVision Explorer intends to become a reference software, improve the overall accuracy and replicability of root trait measurements, and provide a foundation for collaborative improvement and reliable access to all.

The root economics space is a useful framework for plant ecology but is rarely considered for crop ecophysiology. In order to understand root trait integration in winter wheat, we combined functional phenomics with trait economic theory, utilizing genetic variation, highthroughput phenotyping, and multivariate analyses.We phenotyped a diversity panel of 276 genotypes for root respiration and architectural traits using a novel high-throughput method for CO 2 flux and the open-source software RHIZOVISION EXPLORER to analyze scanned images.We uncovered substantial variation in specific root respiration (SRR) and specific root length (SRL), which were primary indicators of root metabolic and structural costs. Multiple linear regression analysis indicated that lateral root tips had the greatest SRR, and the residuals from this model were used as a new trait. Specific root respiration was negatively correlated with plant mass. Network analysis, using a Gaussian graphical model, identified root weight, SRL, diameter, and SRR as hub traits. Univariate and multivariate genetic analyses identified genetic regions associated with SRR, SRL, and root branching frequency, and proposed gene candidates.Combining functional phenomics and root economics is a promising approach to improving our understanding of crop ecophysiology. We identified root traits and genomic regions that could be harnessed to breed more efficient crops for sustainable agroecosystems.

SummaryThe root economics space is a useful framework for plant ecology, but rarely considered for crop ecophysiology. In order to understand root trait integration in winter wheat, we combined functional phenomics with trait economic theory utilizing genetic variation, high-throughput phenotyping, and multivariate analyses.We phenotyped a diversity panel of 276 genotypes for root respiration and architectural traits using a novel high-throughput method for CO2 flux and the open-source software RhizoVision Explorer for analyzing scanned images.We uncovered substantial variation for specific root respiration (SRR) and specific root length (SRL), which were primary indicators of root metabolic and construction costs. Multiple linear regression estimated that lateral root tips had the greatest SRR, and the residuals of this model were used as a new trait. SRR was negatively correlated with plant mass. Network analysis using a Gaussian graphical model identified root weight, SRL, diameter, and SRR as hub traits. Univariate and multivariate genetic analyses identified genetic regions associated with aspects of the root economics space, with underlying gene candidates.Combining functional phenomics and root economics is a promising approach to understand crop ecophysiology. We identified root traits and genomic regions that could be harnessed to breed more efficient crops for sustainable agroecosystems.

Plant growth and development in response to nutrient and water availability is an important adaptation for abiotic stress tolerance. Roots need to intercept both passing nutrients and water while foraging into new soil layers for further resources. Substantial amounts of nitrate can be lost in the field when leaching into groundwater; yet, very little is known about how deep rooting affects this process. Here, we phenotyped root system traits and deep 15N nitrate capture across 1.5 m vertical profiles of solid-media using tall mesocosms in switchgrass (Panicum virgatum L.), a promising cellulosic bioenergy feedstock. Root and shoot biomass traits, photosynthesis and respiration measures, and nutrient uptake and accumulation traits were quantified in response to a water and nitrate stress factorial experiment for switchgrass upland (VS16) and lowland (AP13) ecotypes. The two switchgrass ecotypes shared common plastic abiotic responses to nitrogen (N) and water availability and yet had substantial genotypic variation for root and shoot traits. A significant interaction between nitrogen and water stress combination treatments for axial and lateral root traits represents a complex and shared root development strategy for stress mitigation. Deep root growth and 15N capture were found to be closely linked to aboveground growth. Together, these results represent the wide genetic pool of switchgrass and that deep rooting promotes nitrate capture, plant productivity, and sustainability.