Plant phenotypic plasticity in response to nutrient and water availability is an important adaptation for abiotic stress tolerance. Roots intercept water and nutrients while foraging through soil searching for further resources. Substantial amounts of nitrate can leach into groundwater; yet, little is known about how deep rooting affects this process. Here, we phenotyped root system traits and deep 15N nitrate capture across 1.5 m profiles of solid-media using tall mesocosms in switchgrass (Panicum virgatum L.), a cellulosic bioenergy feedstock. Root and shoot biomass, photosynthesis and respiration, and nutrient uptake traits were quantified in response to a water and nitrate stress factorial experiment in the greenhouse for switchgrass upland (VS16) and lowland (AP13) ecotypes. The two switchgrass ecotypes shared common plastic abiotic responses to nitrogen (N) and water availability and yet showed genotypic differences for root and shoot traits. A significant interaction between nitrogen and water stress for axial and lateral root traits represents a complex and shared root development strategy for stress mitigation. Deep root growth and 15N capture were found to be closely linked to aboveground growth. Together, these results represent the wide genetic pool of switchgrass and that deep rooting promotes nitrate capture, plant productivity, and sustainability. Highlight Two main ecotypes of switchgrass have both shared and different root responses to varying water and nitrogen conditions, with deep rooting shown to be closely linked to aboveground growth.

/pdf/interactions-among-rooting-traits-for-deep-water-and-1kpm2sg185.pdf

Interactions among rooting traits for deep water and nitrogen uptake in upland and lowland ecotypes of switchgrass (Panicum virgatum L.)

Abstract Sustainable biofuel cropping systems aim to address climate change while meeting energy needs. Understanding how soil and plant‐associated microbes respond to these different cropping systems is key to promoting agriculture sustainability and evaluating changes in ecosystem functions. Here, we leverage a long‐term biofuel cropping system field experiment to dissect soil and root microbiome changes across a soil‐depth gradient in poplar, restored prairie and switchgrass to understand their effects on the microbial communities. High throughput amplicon sequencing of the fungal internal transcribed spacer (ITS) and prokaryotic 16S DNA regions showed a common trend of root and soil microbial community richness decreasing and evenness increasing with depth. Ecological niche (root vs. soil) had the strongest effect on community structure, followed by depth, then crop. Stochastic processes dominated the structuring of fungal communities in deeper soil layers while operational taxonomic units (OTUs) in surface soil layers were more likely to co‐occur and to be enriched by plant hosts. Prokaryotic communities were dispersal limited at deeper depths. Microbial networks showed a higher density, connectedness, average degree and module size in deeper soils. We observed a decrease in fungal‐fungal links and an increase of bacteria–bacteria links with increasing depth in all crops, particularly in the root microbiome.

https://onlinelibrary.wiley.com/doi/pdfdirect/10.1111/1462-2920.16269

Stochastic and deterministic processes shape bioenergy crop microbiomes along a vertical soil niche

Crop root segmentation models developed through deep learning have increased the throughput of in situ crop phenotyping studies. However, models trained to identify roots in one image dataset may not accurately identify roots in another dataset, especially when the new dataset contains known differences, called domain shifts. The objective of this study was to quantify how model performance changes when models are used to segment image datasets that contain domain shifts and evaluate approaches to reduce error associated with domain shifts. We collected maize root images at two growth stages (V7 and R2) in a field experiment and manually segmented images to measure total root length (TRL). We developed five segmentation models and evaluated each model's ability to handle a temporal (growth‐stage) domain shift. For the V7 growth stage, a growth‐stage‐specific model trained only on images captured at the V7 growth stage was best suited for measuring TRL. At the R2 growth stage, combining images from both growth stages into a single dataset to train a model resulted in the most accurate TRL measurements. We applied two of the field models to images from a greenhouse experiment to evaluate how model performance changed when exposed to a cross‐site domain shift. Field models were less accurate than models trained only on the greenhouse images even when crop growth stage was identical. Although models may perform well for one experiment, model error increases when applied to images from different experiments even when crop species, growth stage, and soil type are similar.

Toward improved image‐based root phenotyping: Handling temporal and cross‐site domain shifts in crop root segmentation models

Bioenergy could help limit global warming to 2°C above pre‐industrial levels while supplying almost a fourth of the world's renewable energy needs by 2050. However, the deployment of bioenergy raises concerns that adoption at meaningful scales may lead to unintended negative environmental consequences. Meanwhile, the full consolidation of a bioenergy industry is currently challenged by a sufficient, resilient, and resource‐efficient biomass supply and an effective conversion process. Here, we provide a comprehensive analysis of how stable isotope approaches have accelerated the development of a robust bioeconomy by advancing knowledge about environmental sustainability, feedstock development, and biological conversion. We show that advances in stable isotope research have generated crucial information to (1) gain mechanistic insight into the potential of bioenergy crops to mitigate climate change as well as their impact on water and nutrient cycling; (2) develop high‐yielding, resilient feedstocks that produce high‐value bioproducts in planta; and (3) engineer microbes to enhance feedstock conversion to bioenergy products. Further, we highlight knowledge gaps that could benefit from future research facilitated by stable isotope approaches. We conclude that advances in mechanistic knowledge and innovations within the field of stable isotopes in cross‐disciplinary research actions will greatly contribute to breaking down the barriers to establishing a robust bioeconomy.

https://onlinelibrary.wiley.com/doi/pdfdirect/10.1111/gcbb.13056

Accelerating the development of a sustainable bioenergy portfolio through stable isotopes

Phytotechnology has traditionally been considered as a tool to remediate contaminated soils. While phytotechnology has been generally defined as the application of science and engineering to study problems and provide solutions involving plants, the practical applications go far beyond restoring contaminated land. This review aims to broaden the way we think about phytotechnologies while highlighting how these living technologies can restore, conserve and regenerate the multiple functions and ecosystem services provided by the soil, particularly in the context of agroecosystems. At first, the main problems of soil degradation in agroecosystems are shortly underlined. Subsequently, the importance of plants and their living roots as engines of restoration are reviewed. This paper demonstrates the importance of root traits and functions for soil restoration. It also demonstrates that plant and root diversity together with perenniality are key component of an efficient soil restoration process. Then, a phytotechnology toolbox which includes three pillars for agroecosystems restoration is presented. The three pillars are agricultural practices and land management (1), rhizosphere engineering (2) and ecological intensification (3). This paper also highlights the importance of developing targeted phytotechnology-based restoration strategies developed from root functions and knowledge of rhizosphere processes. More work is needed to evaluate the potential benefits of incorporating phytotechnology-based restoration strategies in the context of grain or vegetable crop productions as most of the studies for agroecosystem restoration strategies were intended to mimic natural prairies.

Restoring Soil Functions and Agroecosystem Services Through Phytotechnologies

Bioenergy production often focuses on the aboveground feedstock production for conversion to fuel and other materials. However, the belowground component is crucial for soil carbon sequestration, greenhouse gas fluxes, and ecosystem function. Roots maximize feedstock production on marginal lands by acquiring soil resources and mediating soil ecosystem processes through interactions with the microbial community. This belowground world is challenging to observe and quantify; however, there are unprecedented opportunities using current methodologies to bring roots, microbes, and soil into focus. These opportunities allow not only breeding for increased feedstock production but breeding for increased soil health and carbon sequestration as well. A recent workshop hosted by the USDOE Bioenergy Research Centers highlighted these challenges and opportunities while creating a roadmap for increased collaboration and data interoperability through standardization of methodologies and data using F.A.I.R. principles. This article provides a background on the need for belowground research in bioenergy cropping systems, a primer on root system properties of major U.S. bioenergy crops, and an overview of the roles of root chemistry, exudation, and microbial interactions on sustainability. Crucially, we outline how to adopt standardized measures and databases to meet the most pressing methodological needs to accelerate root, soil, and microbial research to meet the pressing societal challenges of the century.

https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/ppj2.20028

Bioenergy Underground: Challenges and opportunities for phenotyping roots and the microbiome for sustainable bioenergy crop production

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Accelerating the development of a sustainable bioenergy portfolio through stable isotopes

Bioenergy Underground: Challenges and opportunities for phenotyping roots and the microbiome for sustainable bioenergy crop production