A novel imaging and software platform was developed for the high-throughput phenotyping of three-dimensional root traits during seedling development. To demonstrate the platform's capacity, plants of two rice (Oryza sativa) genotypes, Azucena and IR64, were grown in a transparent gellan gum system and imaged daily for 10 d. Rotational image sequences consisting of 40 two-dimensional images were captured using an optically corrected digital imaging system. Three-dimensional root reconstructions were generated and analyzed using a custom-designed software, RootReader3D. Using the automated and interactive capabilities of RootReader3D, five rice root types were classified and 27 phenotypic root traits were measured to characterize these two genotypes. Where possible, measurements from the three-dimensional platform were validated and were highly correlated with conventional two-dimensional measurements. When comparing gellan gum-grown plants with those grown under hydroponic and sand culture, significant differences were detected in morphological root traits (P , 0.05). This highly flexible platform provides the capacity to measure root traits with a high degree of spatial and temporal resolution and will facilitate novel investigations into the development of entire root systems or selected components of root systems. In combination with the extensive genetic resources that are now available, this platform will be a powerful resource to further explore the molecular and genetic determinants of root system architecture.
The beetle Hemisphaerota cyanea (Chrysomelidae; Cassidinae) responds to disturbance by activating a tarsal adhesion mechanism by which it secures a hold on the substrate. Its tarsi are oversized and collectively bear some 60,000 adhesive bristles, each with two terminal pads. While walking, the beetle commits but a small fraction of the bristles to contact with the substrate. But when assaulted, it presses its tarsi flatly down, thereby touching ground with all or nearly all of the bristles. Once so adhered, it can withstand pulling forces of up to 0.8 g (Ϸ60 times its body mass) for 2 min, and of higher magnitudes, up to >3 g, for shorter periods. Adhesion is secured by a liquid, most probably an oil. By adhering, the beetle is able to thwart attacking ants, given that it is able to cling more persistently than the ant persists in its assault. One predator, the reduviid Arilus cristatus, is able to feed on the beetle, possibly because by injecting venom it prevents the beetle from maintaining its tarsal hold.
High-throughput phenotyping of root systems requires a combination of specialized techniques and adaptable plant growth, root imaging and software tools. A custom phenotyping platform was designed to capture images of whole root systems, and novel software tools were developed to process and analyse these images. The platform and its components are adaptable to a wide range root phenotyping studies using diverse growth systems (hydroponics, paper pouches, gel and soil) involving several plant species, including, but not limited to, rice, maize, sorghum, tomato and Arabidopsis. The RootReader2D software tool is free and publicly available and was designed with both user-guided and automated features that increase flexibility and enhance efficiency when measuring root growth traits from specific roots or entire root systems during large-scale phenotyping studies. To demonstrate the unique capabilities and high-throughput capacity of this phenotyping platform for studying root systems, genome-wide association studies on rice (Oryza sativa) and maize (Zea mays) root growth were performed and root traits related to aluminium (Al) tolerance were analysed on the parents of the maize nested association mapping (NAM) population.
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