Abstract

The ability to nondestructively image and automatically phenotype complex root systems, like those of rice (Oryza sativa), is fundamental to identifying genes underlying root system architecture (RSA). Although root systems are central to plant fitness, identifying genes responsible for RSA remains an underexplored opportunity for crop improvement. Here we describe a nondestructive imaging and analysis system for automated phenotyping and trait ranking of RSA. Using this system, we image rice roots from 12 genotypes. We automatically estimate RSA traits previously identified as important to plant function. In addition, we expand the suite of features examined for RSA to include traits that more comprehensively describe monocot RSA but that are difficult to measure with traditional methods. Using 16 automatically acquired phenotypic traits for 2,297 images from 118 individuals, we observe (1) wide variation in phenotypes among the genotypes surveyed; and (2) greater intergenotype variance of RSA features than variance within a genotype. RSA trait values are integrated into a computational pipeline that utilizes supervised learning methods to determine which traits best separate two genotypes, and then ranks the traits according to their contribution to each pairwise comparison. This trait-ranking step identifies candidate traits for subsequent quantitative trait loci analysis and demonstrates that depth and average radius are key contributors to differences in rice RSA within our set of genotypes. Our results suggest a strong genetic component underlying rice RSA. This work enables the automatic phenotyping of RSA of individuals within mapping populations, providing an integrative framework for quantitative trait loci analysis of RSA.

Highlights

  • Root systems are complex three-dimensional (3D) structures that provide functions central to plant fitness, such as water and nutrient acquisition

  • Only plants that had germinated within 1 d of each other were used for analysis

  • Of the 66 possible pairwise comparisons between 12 genotypes, 64 had at least one trait that varied significantly between a pair of genotypes even when accounting for the effect of performing multiple comparisons. This was true even when analyzing plants that had germinated at slightly different times. These results suggest a strong genetic component underlying rice varietal mean root system architecture (RSA) traits

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Summary

Introduction

Root systems are complex three-dimensional (3D) structures that provide functions central to plant fitness, such as water and nutrient acquisition. Using our gel-based platform, we generated approximately 2,300 images of multiple individuals of 12 different rice genotypes at the 14th d after planting (dap; Fig. 1). This analysis system is comprised of a pipeline that preprocesses each image, calculates root features for each image, and combines all of the phenotyping information into a comprehensive trait-ranking step (Supplemental Fig. S1).

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