Correlation data.

Roots are major contributors to nutrient acquisition, water absorption, and plant anchoring and stability. However, little is known about the root system of industrial hemp (Cannabis sativa L.), an increasingly important crop worth $16 billion annually. Hemp is commonly cultivated for grain as an oilseed, stalk biomass for fiber and industrial materials, but has also had growing interest for its carbon sequestration potential due to its reported deep rooting profile. The objectives of this research were to (1) phenotype a panel of 46 industrially-relevant hemp genotypes, (2) quantify the phenotypic differences of shoot and root traits through 2D image analysis, (3) and to investigate genotype grouping strategies and gene targets that could be useful for crop improvement. To phenotype the root system architecture of multiple hemp genotypes representative of production hemp, a large format raised-bed was developed in a greenhouse in which hemp was planted in rows. Root and shoot traits varied across genotypes, with a difference of 175% in total root length between the largest and smallest genotype, and heritability values ranging from 0.51 to 0.88 for key root traits. A strong positive correlation was found between root and shoot biomass (R = 0.93) suggests coordinated resource allocation strategies across genotypes. Of the 46 genotypes studied, two genotypes consistently showed the greatest differences across most of the traits analyzed in the panel. A root-to-shoot quadrant framework was applied to classify hemp ideotypes based on biomass allocation and architectural traits. In addition, comparative genomic analysis identified 74 candidate root architecture genes in hemp that are orthologous to known regulators in maize, rice, and Arabidopsis. These findings highlight substantial phenotypic diversity in hemp root systems and provide a foundation for developing genotype grouping strategies and selecting breeding targets for mapping populations.