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At least 127 records · Page 7

A starting guide to root ecology: strengthening ecological concepts and standardising root classification, sampling, processing and trait measurements

In the context of a recent massive increase in research on plant root functions and their impact on the environment, root ecologists currently face many important challenges to keep on generating cutting-edge, meaningful and integrated knowledge. Consideration of the below-ground components in plant and ecosystem studies has been consistently called for in recent decades, but methodology is disparate and sometimes inappropriate. This handbook, based on the collective effort of a large team of experts, will improve trait comparisons across studies and integration of information across databases by providing standardised methods and controlled vocabularies. It is meant to be used not only as starting point by students and scientists who desire working on below-ground ecosystems, but also by experts for consolidating and broadening their views on multiple aspects of root ecology. Beyond the classical compilation of measurement protocols, we have synthesised recommendations from the literature to provide key background knowledge useful for: (1) defining below-ground plant entities and giving keys for their meaningful dissection, classification and naming beyond the classical fine-root vs coarse-root approach; (2) considering the specificity of root research to produce sound laboratory and field data; (3) describing typical, but overlooked steps for studying roots (e.g. root handling, cleaning and storage); and (4) gathering metadata necessary for the interpretation of results and their reuse. Most importantly, all root traits have been introduced with some degree of ecological context that will be a foundation for understanding their ecological meaning, their typical use and uncertainties, and some methodological and conceptual perspectives for future research. Considering all of this, we urge readers not to solely extract protocol recommendations for trait measurements from this work, but to take a moment to read and reflect on the extensive information contained in this broader guide to root ecology, including sections I–VII and the many introductions to each section and root trait description. Finally, it is critical to understand that a major aim of this guide is to help break down barriers between the many subdisciplines of root ecology and ecophysiology, broaden researchers’ views on the multiple aspects of root study and create favourable conditions for the inception of comprehensive experiments on the role of roots in plant and ecosystem functioning.

59 BASIC BIOLOGICAL SCIENCES↗

Choreographing root architecture and rhizosphere interactions through synthetic biology

Abstract Climate change is driving extreme changes to the environment, posing substantial threats to global food security and bioenergy. Given the direct role of plant roots in mediating plant-environment interactions, engineering the form and function of root systems and their associated microbiota may mitigate these effects. Synthetic genetic circuits have enabled sophisticated control of gene expression in microbial systems for years and a surge of advances has heralded the extension of this approach to multicellular plant species. Targeting these tools to affect root structure, exudation, and microbe activity on root surfaces provide multiple strategies for the advancement of climate-ready crops.

59 BASIC BIOLOGICAL SCIENCES↗

Belowground allocation and dynamics of recently fixed plant carbon in a California annual grassland

Plant roots and the organisms that surround them are a primary source for stabilized soil organic carbon (SOC). While grassland soils have a large capacity to store organic carbon (C), few field-based studies have quantified the amount of plant-fixed C that moves into soil and persists belowground over multiple years. Yet this characteristic of the soil C cycle is critical to C storage, soil water holding capacity, nutrient provisions, and the management of soil health. Here, we tracked the fate of plant-fixed C following a five-day 13 CO 2 labeling of a Northern California annual grassland, measuring C pools starting at the end of the labeling period, at three days, four weeks, six months, one year, and two years. Soil organic carbon was fractionated using a density-based approach to separate the free-light fraction (FLF), occluded-light fraction (OLF), and heavy fraction (HF). Using isotope ratio mass spectrometry, we measured 13 C enrichment and total C content for plant shoots, roots, soil, soil dissolved organic carbon (DOC), and the FLF, OLF, and HF. The chemical nature of C in the HF was further analyzed by solid state 13 C nuclear magnetic resonance (NMR) spectroscopy.

13C-NMR↗

Molecular Genetics of Root Thigmoresponsiveness in Arabidopsis thaliana

The molecular mechanisms that allow plant roots to use gravity and touch as growth guides are investigated. We are using a molecular genetic strategy in Arabidopsis thaliana to study these processes. When Arabidopsis thaliana seedlings grow on tilted hard-agar surfaces, their roots develop a wavy pattern of growth which appears to derive from a succession of left-handed and right-handed circumnutation-like processes triggered by gravity and touch stimulation (Okada and Shimura, 1990; Rutherford et al., 1998; Rutherford and Masson, 1996). Interestingly, mutations that affect root waving on tilted hard-agar surfaces can be identified and characterized. Some of these mutations affect root gravitropism, while others appear to be responsible for the production of abnormal waves (no waves, compressed or square waves, coils) without affecting gravitropism. The specific objectives of this project were to functionally characterize two genes (WVD2 and WVD6) which are required for root waving on tilted agar surfaces, but not for root gravitropism. Specific objectives included a physiological and cytological analysis of the mutants, and molecular cloning and characterization of the corresponding genes. As summarized in this paper, we have reached these objectives. We have also identified and partially characterized other mutations that affect root skewing on hard-agar surfaces (sku5-1 and ago1), and have completed our work on the root-wave phenotype associated with mutations in genes of the tryptophan biosynthesis pathway (Lynn et al., 1999; Rutherford et al., 1998; Sedbrook et al., 2000, 2002). We briefly describe our progress on the cloning and characterization of WVD6, WVD2 and SKU5, and provide a list of papers (published, or in preparation) that derived from this grant. We also discuss the biological implications of our findings, with special emphasis on the analysis of WVD2.

Masson, Patrick H.↗

Development of a Self-calibrating Dissolved Oxygen Microsensor Array for the Monitoring and Control of Plant Growth in a Space Environment

Plant experiments in space will require active nutrient delivery concepts in which water and nutrients are replenished on a continuous basis for long-term growth. The goal of this study is to develop a novel microsensor array to provide information on the dissolved oxygen environment in the plant root zone for the optimum control of plant cultivation systems in the space environment. Control of water and oxygen is limited by the current state-of-the-art in sensor technology. Two capabilities of the new microsensor array were tested. First, a novel in situ self-diagnosis/self-calibration capability for the microsensor was explored by dynamically controlling the oxygen microenvironment in close proximity to an amperometric dissolved oxygen microsensors. A pair of integrated electrochemical actuator electrodes provided the microenvironments based on water electrolysis. Miniaturized thin film dissolved oxygen microsensors on a flexible polyimide (Kapton(Registered Trademark)? substrate were fabricated and their performances were tested. Secondly, measurements of dissolved oxygen in two representative plant growth systems were made, which had not been performed previously due to lack of proper sensing technology. The responses of the oxygen microsensor array on a flexible polymer substrate properly reflected the oxygen contents on the surface of a porous tube nutrient delivery system and within a particulate substrate system. Additionally, we demonstrated the feasibility of using a 4-point thin film microprobe for water contents measurements for both plant growth systems. mechanical flexibility, and self-diagnosis. The proposed technology is anticipated to provide a reliable sensor feedback plant growth nutrient delivery systems in both terrestrial environment and the microgravity environment during long term space missions. The unique features of the sensor include small size and volume, multiple-point sensing,

Kim, Chang-Soo↗

Eucalyptus grandis MYB‐Like and RAN‐Like Zinc Finger Proteins Display Dual Roles in Regulating Plant Immunity and Symbiosis Pathways

Plant roots live in constant contact with diverse microbes in the soil. Plant fitness, therefore, relies on signaling pathways that mount an effective immune response against pathogens while fostering mutualistic symbioses. Plant pathways, and specifically immune genes that may act as "switches," discriminating between pathogenic or mutualistic fungi, remain largely unknown. Using Eucalyptus grandis as a model system, we investigate alterations to the root transcriptomic landscape during pre-symbiosis with either the pathogen Armillaria luteobubalina or the mutualistic fungus Pisolithus microcarpus. Comparative analyses identified three strongly counter-regulated genes that may act as immune switches to accommodate or to repress fungal colonization. We characterized two of these, a MYB-like and RAN-like zinc finger protein, using a transgenic approach and demonstrated that they have bifunctional roles in the regulation of cell death and a hypersensitive-like response, depending on the lifestyle of the associated fungus. Using co-expression network analysis, we identified hypothetical pathways correlated to these genes. We functionally validated these predictions using plants with transgenic roots with increased or decreased transcription of these genes, thereby showing the power of co-expression networks as an a priori approach to identify key immune response pathways in plants. Overall, our results demonstrate that prior to physical contact with microbes, MYB-like and RAN-like zinc finger proteins are key regulators of plant immune signaling that respond to fungal signals and enable or repress symbiotic establishment.

mycorrhizal fungi↗

Linked nitrogen and carbon dynamics reveal distinct pools and patterns in a deep, weathered bedrock rhizosphere

Nitrogen is one of the most limiting nutrients to forest productivity worldwide. Recently, it has been established that diverse ecosystems source a substantial fraction of their water from weathered bedrock, leading to questions about whether root-driven nitrogen cycling extends into weathered bedrock as well. In this study, we specifically examined nitrogen dynamics using specialized instrumentation distributed across a 16 m weathered bedrock vadose zone (WBVZ) underlying an old growth forest in northern California where the rhizosphere—composed of plant roots and their associated microbiome—extends meters into rock. We documented total dissolved nitrogen (TDN), dissolved organic carbon (DOC), inorganic nitrogen (ammonium and nitrate), and CO 2 and O 2 gases every 1.5 m to 16 m depth for 2 y. We found that TDN concentrations increased with depth, were an order of magnitude greater at 15 m than in the upper 30 cm, and that the majority of TDN throughout the weathered bedrock vadose zone was organic. We also found that TDN concentrations are influenced by depth, season, and interannual precipitation patterns. Carbon isotope composition of the DOC suggests that dissolved organic matter in the WBVZ is primarily derived from plant sources, and not the nitrogen-rich bedrock. We conclude that nitrogen dynamics in the WBVZ may be driven, in part, by an active rhizosphere, meters below the base of soil, and we argue that weathered bedrock horizons may play a key role in C-N cycling in ecosystems with deep-rooted plants.

Science & Technology - Other Topics↗

Synchrotron-based diffraction-enhanced imaging and diffraction-enhanced imaging combined with CT X-ray imaging systems to image seeds at 30 keV

Utilized the upgraded Synchrotron-based non-destructive Diffraction-enhanced imaging and Diffraction-enhanced imaging coupled with CT X-ray imaging systems to image the chickpea seeds, to enhance the contrast in plant root architecture, visibility of fine structures of root architecture growth and some aspects of physiology at acceptable level. DEI-CT images were acquired with 30 keV synchrotron X-rays. A series of DEI-CT slices were assembled together, to form a 3D data set. DEI-CT images explored more structural information and morphology. Noticed detailed anatomical, physiological observations, and contrast mechanisms. Furthermore, with these systems, some of the complex plant traits, root morphology, growth of laterals and subsequent laterals can be visualized directly.

36 MATERIALS SCIENCE↗

Enrichment of root-associated Streptomyces strains in response to drought is driven by diverse functional traits and does not predict beneficial effects on plant growth

The genus Streptomyces has consistently been found enriched in drought-stressed plant root microbiomes, yet the ecological basis and functional variation underlying this enrichment at the strain and isolate level remain unclear. Using two 16S rRNA sequencing methods with different levels of taxonomic resolution, we confirmed drought-associated enrichment (DE) of Streptomyces in field-grown sorghum roots and identified five closely related but distinct amplicon sequence variants (ASVs) belonging to the genus with variable drought enrichment patterns. From a culture collection of sorghum root endophytes, we selected 12 Streptomyces isolates representing these ASVs for phenotypic and genomic characterization. Whole-genome sequencing revealed substantial variation in gene content, even among closely related isolates, and exometabolomic profiling showed distinct metabolic responses to media supplemented with drought- versus well-watered root tissue. Traits linked to drought survival, including osmotic stress tolerance, siderophore production, and carbon utilization, varied widely among isolates and were not phylogenetically conserved. Using a broader panel of 48 Streptomyces, we demonstrate that DE scores, determined through mono-association experiments in gnotobiotic sorghum systems, showed high variability and lacked correlation with plant growth promotion. Pangenome-wide association identified orthogroups involved in osmolyte transport (e.g., proP) and membrane biosynthesis (e.g., fabG) as positively associated with DE, though most associations lacked phylogenetic signal. Collectively, these results demonstrate that Streptomyces DE is not a conserved genus-level trait but is instead strain-specific and functionally heterogeneous. Furthermore, DE in the root microbiome was shown not to predict beneficial effects on plant growth. This work underscores the need to resolve functional traits at the strain level and highlights the complexity of microbe-host-environment interactions under abiotic stress.

Fonseca-Garcia, Citlali↗

Some karyological observations on plants grown in space

Experiments were conducted to assess whether cell division in a plant root would be affected by prolonged exposure to microgravity. Root materials from sunflower, oat, and mung bean plants grown on STS-2 and STS-3 were utilized for the experiments. It is found that all oat, sunflower, and mung seedlings showed a reduced number of cells in division as they went through their first cell division cycle on earth when compared to their ground controls. A significant number of oat, mung, and sunflower plantlets exhibited random root orientation and the lack of strictly orthotropic growth of their shoot systems in the flight samples. In addition, it is found that the mung roots were apparently least affected in terms of their cytology despite the fact that their roots were often randomly oriented.

Krikorian, A. D.↗

How deep should we go to understand roots at the top of the world?

Harsh environmental conditions and the short summers of northern, high-latitude biomes impose unique constraints on the plants that live in the arctic tundra and the boreal forest. To escape the harsh aboveground environment, plants in these habitats often allocate a large portion of their biomass belowground to facilitate nutrient acquisition. In turn, the proximity of living plant roots to vast stores of sequestered soil carbon in these biomes means that shifts in rooting depth distribution and the size of the root–soil interface could significantly contribute to ongoing climate change. Indeed, plant ‘priming’ of rhizosphere decomposition via root exudation, particularly from shallowly distributed roots, can lead to losses of carbon from tundra soils. While we have a hard-won understanding of the distribution of plant communities across the arctic tundra and the boreal forest from direct field observations scaled to the landscape level using climate-informed mapping techniques (i.e. the Circumpolar Arctic Vegetation Map (CAVM); Walker et al. , 2005), these vegetation maps are only the tip of the iceberg (Iversen et al. , 2015). Root form and function remain hidden beneath the land surface. In an article recently published in New Phytologist , Blume-Werry et al. (2023, 10.1111/nph.18998) asked whether rooting depth distribution, and ensuing carbon emissions, could be inferred from commonly used vegetation mapping classifications across the pan-Arctic. An important question to guide our understanding, mapping, and prediction of belowground characteristics and ecosystem feedbacks at the top of the world. Unfortunately, they found that the answer was ‘not quite’. While rooting depth distribution varied demonstrably, in turn causing substantial changes in modeled carbon emissions via rhizosphere ‘priming’, variation across rooting depth profiles did not correspond with vegetation mapping classes. If we are unable to predict belowground rooting depth distributions across large spatial scales by leveraging aboveground vegetation community distributions, how then should belowground researchers proceed?

59 BASIC BIOLOGICAL SCIENCES↗

Visualizing the Hidden Half: Plant-Microbe Interactions in the Rhizosphere

Plant roots and the associated rhizosphere constitute a dynamic environment that fosters numerous intra- and interkingdom interactions, including metabolite exchange between plants and soil mediated by root exudates and the rhizosphere microbiome. These interactions affect plant fitness and performance, soil health, and the belowground carbon budget.

59 BASIC BIOLOGICAL SCIENCES↗

Interactions with microbial consortia have variable effects in organic carbon and production of exometabolites among genotypes of Populus trichocarpa

Abstract Poplar is a short‐rotation woody crop frequently studied for its significance as a sustainable bioenergy source. The successful establishment of a poplar plantation partially depends on its rhizosphere—a dynamic zone governed by complex interactions between plant roots and a plethora of commensal, mutualistic, symbiotic, or pathogenic microbes that shape plant fitness. In an exploratory endeavor, we investigated the effects of a consortium consisting of ectomycorrhizal fungi and a beneficial Pseudomonas sp. strain GM41 on plant growth (including height, stem girth, leaf, and root growth) and as well as growth rate over time, across four Populus trichocarpa genotypes. Additionally, we compared the level of total organic carbon and plant exometabolite profiles across different poplar genotypes in the presence of the microbial consortium. These data revealed no significant difference in plant growth parameters between the treatments and the control across four different poplar genotypes at 7 weeks post‐inoculation. However, total organic carbon and exometabolite profiles were significantly different between the genotypes and the treatments. These findings suggest that this microbial consortium has the potential to trigger early signaling responses in poplar, influencing its metabolism in ways crucial for later developmental processes and stress tolerance.

59 BASIC BIOLOGICAL SCIENCES↗

Does mycorrhizal symbiosis determine the climate niche for Populus as a bioenergy feedstock?

This is the final technical and scientific report for DE-SC0016097, in which we investigated the role of symbiotic fungi in determining the climatic niche of trees. While microbes have long been viewed as agents of disease, recent explorations of the microbiome have led biologists to recognize that beneficial microbes play an equally vital role in maintaining the health of plants and animals. Perhaps the most ubiquitous form of beneficial interaction in terrestrial ecosystems occurs between fungi and plant roots. These fungus-root, or “mycorrhizal”, symbioses involve the reciprocal exchange of plant sugars for soil nutrients obtained by the fungus, such as nitrogen and phosphorous, which are critical for plant growth. It is known that mycorrhizal associations are widespread - occurring in approximately 90% of plant species - and diverse, with single plants able to associate simultaneously with over 100 species of fungi. Despite this, the ecological factors that control large-scale distribution and abundance of mycorrhizal symbioses are still poorly known, making it challenging to predict how mycorrhizal symbioses may change in future climate conditions or how these fungal communities might be manipulated to improve regional agriculture or forestry projects. In this research project we focused on a native North American tree with strong biofuel potential – Populus – to answer fundamental questions about the role of climate, soil environment, and mycorrhizal interactions in determining growth and competition in plant communities. Populus provides a unique opportunity for ecologically relevant experiments because of its widespread distribution across North America and its natural variability in mycorrhizal association types.

59 BASIC BIOLOGICAL SCIENCES↗

Localized basal area affects soil respiration temperature sensitivity in a coastal deciduous forest

Abstract. Soil respiration (Rs), the flow of CO2 from the soil surface to the atmosphere, is one of the largest carbon fluxes in the terrestrial biosphere. The spatial variability of Rs is both large and poorly understood, limiting our ability to robustly scale it in space. One factor in Rs spatial variability is the autotrophic contribution from plant roots, but it is uncertain how the presence of plants affects the magnitude and temperature sensitivity of Rs. This study used 1 year of Rs measurements to examine the effect of localized basal area on Rs in the growing and dormant seasons, as well as during moisture-limited times, in a temperate, coastal, deciduous forest in eastern Maryland, USA. In a linear mixed-effects model, tree basal area within a 5 m radius (BA5) exerted a significant positive effect on the temperature sensitivity of soil respiration. Soil moisture was the dominant control on Rs during the dry portions of the year, while soil moisture, temperature, and BA5 all exerted significant effects on Rs in wetter periods. Our results suggest that autotrophic respiration is more sensitive to temperature than heterotrophic respiration at these sites, although we did not measure these source fluxes directly, and that soil respiration is highly moisture sensitive, even in a record-rainfall year. The Rs flux magnitudes (0.46–15.0 µmol m-2 s-1) and variability (coefficient of variability 10 %–23 % across plots) observed in this study were comparable to values observed in similar forests. Six Rs observations would be required in order to estimate the mean across all study sites to within 50 %, and 518 would be required in order to estimate it to within 5 %, with 95 % confidence. A better understanding of the spatial interactions between plants and microbes, as well as the strength and speed of above- and belowground coupling, is necessary to link these processes with large-scale soil-to-atmosphere C fluxes.

54 ENVIRONMENTAL SCIENCES↗

Root Pulling Force Across Drought in Maize Reveals Genotype by Environment Interactions and Candidate Genes

High-throughput, field-based characterization of root systems for hundreds of genotypes in thousands of plots is necessary for breeding and identifying loci underlying variation in root traits and their plasticity. We designed a large-scale sampling of root pulling force, the vertical force required to extract the root system from the soil, in a maize diversity panel under differing irrigation levels for two growing seasons. We then characterized the root system architecture of the extracted root crowns. We found consistent patterns of phenotypic plasticity for root pulling force for a subset of genotypes under differential irrigation, suggesting that root plasticity is predictable. Using genome-wide association analysis, we identified 54 SNPs as statistically significant for six independent root pulling force measurements across two irrigation levels and four developmental timepoints. For every significant GWAS SNP for any trait in any treatment and timepoint we conducted post hoc tests for genotype-by-environment interaction, using a mixed model ANOVA. We found that 8 of the 54 SNPs showed significant GxE. Candidate genes underlying variation in root pulling force included those involved in nutrient transport. Although they are often treated separately, variation in the ability of plant roots to sense and respond to variation in environmental resources including water and nutrients may be linked by the genes and pathways underlying this variation. While functional validation of the identified genes is needed, our results expand the current knowledge of root phenotypic plasticity at the whole plant and gene levels, and further elucidate the complex genetic architecture of maize root systems.

Woods, Patrick↗

Hormonal regulation of wheat growth during hydroponic culture

Hormonal control of root growth has been explored as one means to alleviate the crowding of plant root systems experienced in prototype hydroponic biomass production chambers being developed by the CELSS Breadboard Project. Four plant hormones, or their chemical analogs, which have been reported to selectively inhibit root growth, were tested by adding them to the nutrient solutions on day 10 of a 25 day growth test using spring wheat in hydroponic cultures. Growth and morphological changes is both shoot and root systems were evaluated. In no case was it possible to inhibit root growth without a comparable inhibition of shoot growth. It was concluded that this approach is unlikely to prove useful for wheat.

Wetherell, Donald↗