Members of the B family of membrane-bound ATP-binding cassette (ABC) transporters represent key components of the auxin efflux machinery in plants. Over the last two decades, experimental studies have shown that modifying ATP-binding cassette sub-family B (ABCB) expression affects auxin distribution and plant phenotypes. However, precisely how ABCB proteins transport auxin in conjunction with the more widely studied family of PIN-formed (PIN) auxin efflux transporters is unclear, and studies using heterologous systems have produced conflicting results. Here, we integrate ABCB localization data into a multicellular model of auxin transport in the Arabidopsis thaliana root tip to predict how ABCB-mediated auxin transport impacts organ-scale auxin distribution. We use our model to test five potential ABCB–PIN regulatory interactions, simulating the auxin dynamics for each interaction and quantitatively comparing the predictions with experimental images of the DII-VENUS auxin reporter in wild-type and abcb single and double loss-of-function mutants. Only specific ABCB–PIN regulatory interactions result in predictions that recreate the experimentally observed DII-VENUS distributions and long-distance auxin transport. Our results suggest that ABCBs enable auxin efflux independently of PINs; however, PIN-mediated auxin efflux is predominantly through a co-dependent efflux where co-localized with ABCBs.

The spatial arrangement of the root system, termed root system architecture, is important for resource acquisition as it directly affects the soil zone explored. Methods for phenotyping roots are mostly destructive, which prevents analysis of roots over time as they grow. Here, we used X‐ray microcomputed tomography (μCT) to non‐invasively characterize wheat (Triticum aestivum L.) seedling root development across time under high and low nitrate nutrition. Roots were imaged multiple times with the 3D models co‐aligned and timestamped in the architectural plant model OpenSimRoot for subsequent root growth and nitrate uptake simulations. Through 4D imaging, we found that lateral root traits were highly responsive to nitrate limitation in soil with greater lateral root length under low N. The root growth model using all μCT root scans was comparable to a parameterized model using only the final root scan in the series. In a second μCT experiment, root growth and nitrate uptake simulations of candidate wheat genotypes found significant root growth and uptake differences between lines. A high nitrate uptake wheat line selected from field data had a greater lateral root count and length at seedling growth stage compared with a low uptake line.