Functional characterization and discovery of modulators of SbMATE, the agronomically important aluminium tolerance transporter from Sorghum bicolor.

About 50% of the world's arable land is strongly acidic (pH ≤ 5). The low pH solubilizes root-toxic ionic aluminium (Al3+) species from clay minerals, driving the evolution of counteractive adaptations in cultivated crops. The food crop Sorghum bicolor upregulates the membrane-embedded transporter protein SbMATE in its roots. SbMATE mediates efflux of the anionic form of the organic acid, citrate, into the soil rhizosphere, chelating Al3+ ions and thereby imparting Al-resistance based on excluding Al+3 from the growing root tip. Here, we use electrophysiological, radiolabeled, and fluorescence-based transport assays in two heterologous expression systems to establish a broad substrate recognition profile of SbMATE, showing the proton and/or sodium-driven transport of 14C-citrate anion, as well as the organic monovalent cation, ethidium, but not its divalent analog, propidium. We further complement our transport assays by measuring substrate binding to detergent-purified SbMATE protein. Finally, we use the purified membrane protein as an antigen to discover native conformation-binding and transport function-altering nanobodies using an animal-free, mRNA/cDNA display technology. Our results demonstrate the utility of using Pichia pastoris as an efficient eukaryotic host to express large quantities of functional plant transporter proteins. The nanobody discovery approach is applicable to other non-immunogenic plant proteins

Explainable deep learning in plant phenotyping

The increasing human population and variable weather conditions, due to climate change, pose a threat to the world's food security. To improve global food security, we need to provide breeders with tools to develop crop cultivars that are more resilient to extreme weather conditions and provide growers with tools to more effectively manage biotic and abiotic stresses in their crops. Plant phenotyping, the measurement of a plant's structural and functional characteristics, has the potential to inform, improve and accelerate both breeders' selections and growers' management decisions. To improve the speed, reliability and scale of plant phenotyping procedures, many researchers have adopted deep learning methods to estimate phenotypic information from images of plants and crops. Despite the successful results of these image-based phenotyping studies, the representations learned by deep learning models remain difficult to interpret, understand, and explain. For this reason, deep learning models are still considered to be black boxes. Explainable AI (XAI) is a promising approach for opening the deep learning model's black box and providing plant scientists with image-based phenotypic information that is interpretable and trustworthy. Although various fields of study have adopted XAI to advance their understanding of deep learning models, it has yet to be well-studied in the context of plant phenotyping research. In this review article, we reviewed existing XAI studies in plant shoot phenotyping, as well as related domains, to help plant researchers understand the benefits of XAI and make it easier for them to integrate XAI into their future studies. An elucidation of the representations within a deep learning model can help researchers explain the model's decisions, relate the features detected by the model to the underlying plant physiology, and enhance the trustworthiness of image-based phenotypic information used in food production systems