Automatic root hair measurement to quantify abiotic stresses in
microscopy images
Abstract
Improving nutrient and water uptake in crops is one of the major
challenges to sustain a fast-growing population that faces increasingly
nutrient limited soils. Root hairs, which are specialized epidermal
cells, are important drivers of nutrient and water uptake from the soil.
Microscopy provides a mean to record root hairs as digital images.
However, due to their geometry and complex spatial arrangements
quantifying root hairs in microscopy images manually remains a
bottleneck. Manual selection of representative root hairs can result in
inaccurate estimations of root hair traits and misrepresentation of root
hair functions. We present a method to quantify phenotypes automatically
by measuring all individual root hairs in digital microscopy images. Our
method uses random forests classification to separate root hair from the
parent root and the image background. We define metrics to evaluate
segments of root hairs that intersect or form blobs of two or more root
hairs. Using simulated annealing for combinatorial optimization, we
reconstruct individual root hairs by resolving intersections in a
globally optimal way. As a result, we measure root hair length, its
distribution, and root hair density in each image. We validate our
method on examples of three maize cultivars under phosphorus, nitrogen,
and potassium stress. Results show that our measurements of root hair
traits strongly correlate with manually measured validation data in mean
root hair length (Pearson-correlation: 0.74 to 0.88, p<.001),
as well as in root hair density (Pearson-correlation: 0.65 to 0.84,
p<.001). We show that our method distinguishes subtle
differences between genotypes and treatments based on the extracted
traits and believe that our study paves a way towards identifying the
genetic control of root hair traits and increased agricultural
production.