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At least 217 records · Page 12

Cultivation of novel Atribacterota from oil well provides new insight into their diversity, ecology, and evolution in anoxic, carbon-rich environments

Background: The Atribacterota are widely distributed in the subsurface biosphere. Recently, the first Atribacterota isolate was described and the number of Atribacterota genome sequences retrieved from environmental samples has increased significantly; however, their diversity, physiology, ecology, and evolution remain poorly understood. Results: We report the isolation of the second member of Atribacterota, Thermatribacter velox gen. nov., sp. nov., within a new family Thermatribacteraceae fam. nov., and the short-term laboratory cultivation of a member of the JS1 lineage, Phoenicimicrobium oleiphilum HX-OS.bin.34 TS , both from a terrestrial oil reservoir. Physiological and metatranscriptomics analyses showed that Thermatribacter velox B11 T and Phoenicimicrobium oleiphilum HX-OS.bin.34 TS ferment sugars and n-alkanes, respectively, producing H 2 , CO 2 , and acetate as common products. Comparative genomics showed that all members of the Atribacterota lack a complete Wood-Ljungdahl Pathway (WLP), but that the Reductive Glycine Pathway (RGP) is widespread, indicating that the RGP, rather than WLP, is a central hub in Atribacterota metabolism. Ancestral character state reconstructions and phylogenetic analyses showed that key genes encoding the RGP (fdhA, fhs, folD, glyA, gcvT, gcvPAB, pdhD) and other central functions were gained independently in the two classes, Atribacteria (OP9) and Phoenicimicrobiia (JS1), after which they were inherited vertically; these genes included fumarate-adding enzymes (faeA; Phoenicimicrobiia only), the CODH/ACS complex (acsABCDE), and diverse hydrogenases (NiFe group 3b, 4b and FeFe group A3, C). Finally, we present genome-resolved community metabolic models showing the central roles of Atribacteria (OP9) and Phoenicimicrobiia (JS1) in acetate- and hydrocarbon-rich environments. Conclusion: Our findings expand the knowledge of the diversity, physiology, ecology, and evolution of the phylum Atribacterota. This study is a starting point for promoting more incisive studies of their syntrophic biology and may guide the rational design of strategies to cultivate them in the laboratory.

59 BASIC BIOLOGICAL SCIENCES↗

Multicellular magnetotactic bacteria are genetically heterogeneous consortia with metabolically differentiated cells

Consortia of multicellular magnetotactic bacteria (MMB) are currently the only known example of bacteria without a unicellular stage in their life cycle. Because of their recalcitrance to cultivation, most previous studies of MMB have been limited to microscopic observations. To study the biology of these unique organisms in more detail, we use multiple culture-independent approaches to analyze the genomics and physiology of MMB consortia at single-cell resolution. We separately sequenced the metagenomes of 22 individual MMB consortia, representing 8 new species, and quantified the genetic diversity within each MMB consortium. This revealed that, counter to conventional views, cells within MMB consortia are not clonal. Single consortia metagenomes were then used to reconstruct the species-specific metabolic potential and infer the physiological capabilities of MMB. To validate genomic predictions, we performed stable isotope probing (SIP) experiments and interrogated MMB consortia using fluorescence in situ hybridization (FISH) combined with nanoscale secondary ion mass spectrometry (NanoSIMS). By coupling FISH with bioorthogonal noncanonical amino acid tagging (BONCAT), we explored their in situ activity as well as variation of protein synthesis within cells. We demonstrate that MMB consortia are mixotrophic sulfate reducers and that they exhibit metabolic differentiation between individual cells, suggesting that MMB consortia are more complex than previously thought. These findings expand our understanding of MMB diversity, ecology, genomics, and physiology, as well as offer insights into the mechanisms underpinning the multicellular nature of their unique lifestyle.

59 BASIC BIOLOGICAL SCIENCES↗

Rhizosphere Soil Biogeochemical Data and Photosynthetic Data of Vicia Faba in a Rhizobox

Here we share the data in column format via csv files for pH, redox, and dissolved oxygen collected at hourly resolution from microelectrodes. Dissolved organic carbon concentrations collected from TOC are also provided in a similar format but are composited samples from hourly microdialysis collection. This provided resolution of diel rhizosphere dynamics belowground. Plant physiological data was also collected at every 5 min for 24 hr cycles in order to capture diel dynamics aboveground. This data was used to parameterize the reaction transport model eSTOMP-ROOTS, which examines the rhizosphere biogeochemistry of a growing Vicia faba plant. The aim was to investigate plant activity and belowground biogeochemical processes, particularly their impact on mineral-organic associations in the rhizosphere. We combined in-situ rhizosphere microsensor and plant physiological measurements with a 3-D plant-soil reactive transport model to explore the behavior of dissolved organic carbon (DOC) in the rhizosphere. Over several days, microdialysis probes placed at the root-soil interface in live soil showed distinct daily patterns of DOC concentration in the pore water. Spikes in DOC concentrations during the day aligned with peaks in leaf-level photosynthesis, accompanied by decreasing redox potential and dissolved oxygen levels, and increasing pH in the rhizosphere. This new mechanistic modeling framework, which integrates aboveground plant physiological data with non-destructive, high-resolution monitoring of rhizosphere processes, offers significant potential for studying the factors that control carbon storage in soils.

54 ENVIRONMENTAL SCIENCES↗

Rays for Roots - Integrating Backscatter X-Ray Phenotyping, Modeling and Genetics to Increase Carbon Sequestration and Switchgrass Resource Use (Final Report)

To increase carbon (C) deposition in the soil and enhance crop resource use efficiency, characterizing root form and function is essential. Several root and soil traits have been linked to increased root-to-soil C transfer. Technology that could provide high-resolution characterization of many of these traits in field conditions would revolutionize our ability to study and understand how to increase C sequestration. In this effort, we developed an initial early prototype backscatter X-ray system for non-destructive imaging of root traits. We collected initial backscatter X-ray data in field and lab settings and carried out early analysis of these data. Along with this prototype, we also developed a suite of root phenotyping approaches including advanced minirhizotron image analysis, soil core imaging, and mesocosm imaging. Minirhizotron (MR) tubes are clear tubes inserted into the soil in the field and used to image roots and the surrounding soil. Our team has developed deep learning-based methods that can segment roots from soil that can learn from imprecise image-level labels. The ability to learn or fine-tune our deep learning algorithms from image-level labels allows easier and faster application of these approaches to new locations and new plant species. We have successfully implemented and applied our MR analysis approaches to thousands of switchgrass MR images collected across geographical regions. An advantage of MR imaging is the ability to collect root and soil images over time. Our soil core analysis included collecting hundreds of soil core samples from harvested switchgrass fields and imaging these cores with both X-ray CT and backscatter X-ray imaging. Initial segmentation approaches for the X-ray CT images of these cores have been developed and applied. An advantage of soil core analysis is that it preserves the three-dimensional structures of the roots and soil in the core collected. Our group also developed photogrammetry-based mesocosm root imaging and phenotyping approaches. In this approach, a plant was grown in a large mesocosm with a three-dimensional grid of thin supporting lines inserted throughout the mesocosm. After the plant (and, correspondingly, the root architecture is grown and established) the soil media was removed and the supporting lines approximately preserved the three-dimensional root architecture. Then, we applied photogrammetry techniques to create a three-dimensional digital representation of the root architecture for which we developed analysis algorithms including skeletonization. We carried out our phenotyping development with powerful switchgrass resources and physiological and agroecosystem modeling to deliver novel technology. This project contributes to multiple ARPA-E missions including reduction of foreign imports of energy, reduction of energy-related emissions including greenhouse gases, and ensuring that the United States maintains a technological lead in developing and deploying advanced energy technology. Furthermore, the developed tools could transform public and private plant breeding and could be broadly applicable to other crops and, potentially, other application areas. Our team of engineers, plant and soil scientists, and modelers i) developed an early prototype backscatter X-ray platform that can operate in field conditions; ii) developed a suite of root phenotyping and characterization approaches as described above; iii) developed and carried out plant biology and physiology roots studies and; iv) developed and implemented mechanistic physiological modeling.

42 ENGINEERING↗

Growth-associated polyhydroxybutyrate accumulation in Azospira suillum PS during aerobic and perchlorate respiration

Polyhydroxyalkanoates (PHAs) are widespread microbial storage polymers increasingly recognized for roles beyond carbon and energy storage, including redox homeostasis and stress physiology. While PHA accumulation is classically associated with stationary-phase metabolism under severe nutrient imbalance, comparatively little is known about growth-associated PHA synthesis in facultative anaerobes with unusual respiratory strategies. Here, we investigated polyhydroxybutyrate (PHB) metabolism in Azospira suillum PS, a genetically tractable perchlorate-reducing bacterium capable of both aerobic and anaerobic respiration. Using physiological growth experiments, targeted gene deletions, PHB extractions, and intracellular redox measurements, we examined PHB accumulation under varying respiratory and nutrient conditions. We demonstrate that PHB accumulates during exponential growth under moderately nitrogen-limited conditions, both aerobically and during perchlorate respiration, representing a rare example of growth-associated PHB synthesis under anaerobic conditions. Genomic analysis revealed four phaC homologs, one of which could not be deleted under the experimental conditions tested and co-localized with phaB and phaR. Redox profiling further revealed a strong positive correlation between PHB accumulation and intracellular NADPH/NADP+ ratios. Together, these findings expand the physiological contexts in which PHB synthesis is known to occur and highlight perchlorate-respiring bacteria as underexplored model systems for studying growth-integrated carbon and redox storage strategies.

Meier, David A O↗

Stoichiometrically coupled carbon and nitrogen cycling in the MIcrobial-MIneral Carbon Stabilization model version 1.0 (MIMICS-CN v1.0)

Explicit consideration of microbial physiology in soil biogeochemical models that represent coupled carbon–nitrogen dynamics presents opportunities to deepen understanding of ecosystem responses to environmental change. The MIcrobial-MIneral Carbon Stabilization (MIMICS) model explicitly represents microbial physiology and physicochemical stabilization of soil carbon (C) on regional and global scales. Here we present a new version of MIMICS with coupled C and nitrogen (N) cycling through litter, microbial, and soil organic matter (SOM) pools. The model was parameterized and validated against C and N data from the Long-Term Inter-site Decomposition Experiment Team (LIDET; six litter types, 10 years of observations, and 13 sites across North America). The model simulates C and N losses from litterbags in the LIDET study with reasonable accuracy (C: R 2 =0.63; N: R 2 =0.29), which is comparable with simulations from the DAYCENT model that implicitly represents microbial activity (C: R 2 =0.67; N: R 2 =0.30). Subsequently, we evaluated equilibrium values of stocks (total soil C and N, microbial biomass C and N, inorganic N) and microbial process rates (soil heterotrophic respiration, N mineralization) simulated by MIMICS-CN across the 13 simulated LIDET sites against published observations from other continent-wide datasets. We found that MIMICS-CN produces equilibrium values in line with measured values, showing that the model generates plausible estimates of ecosystem soil biogeochemical dynamics across continental-scale gradients. MIMICS-CN provides a platform for coupling C and N projections in a microbially explicit model, but experiments still need to identify the physiological and stoichiometric characteristics of soil microbes, especially under environmental change scenarios.

58 GEOSCIENCES↗

Miniaturized, automated in-vitro tissue bioreactor

In one embodiment, a system includes a bioreactor coupled to a substrate. The bioreactor includes: a plurality of walls defining a reservoir; a plurality of fluidic channels in at least some of the walls; a fluidic inlet in fluidic communication with the reservoir via, the fluidic channels; a fluidic outlet in fluidic communication with the reservoir via the fluidic channels; and one or more sensors coupled to the reservoir, each sensor being configured to detect one or more of: environmental conditions in the reservoir; and physiological conditions of one or more cells optionally present in the reservoir. In another embodiment, a method includes delivering media to a reservoir of a bioreactor; delivering a plurality of cells to the reservoir, controlling a reservoir temperature and a reservoir gas composition; using one or more of the sensors to monitor environmental and physiological conditions; and reporting the environmental and/or physiological conditions.

42 ENGINEERING↗

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↗

Salinity exposure affects lower-canopy specific leaf area of upland trees in a coastal deciduous forest

Sea level rise and increasing storm surges are likely to affect the canopy physiology, ecology, and structure of coastal forests, even well in advance of tree mortality. Laboratory and greenhouse studies have documented that saltwater exposure can trigger changes in leaf-level physiology and morphology, but few in situ studies have examined how tree-specific leaf area (SLA), the ratio of leaf area to mass and a crucial trait and model parameter, is affected by saline soils. We conducted an observational study of SLA in a mid-Atlantic (USA) coastal deciduous forest, taking advantage of a natural gradient in salinity along a tidal creek. Measured SLA of the 239 trees and seven species sampled ranged from Carya glabra (N = 6 trees, mean SLA = 277.9±36.3cm 2 /g) to Fagus grandifolia (N=60, 321.9±62.9cm 2 g); as expected, trees species and canopy position (sun versus shade) significantly affected SLA. For trees (N=100) directly exposed to the tidal creek, salinity was highly significant after accounting for species (P<0.001), with trees in the lower reaches of the creek having lower SLA. Leaf area index (LAI), computed from SLA and litter traps, ranged from 4.8 to 15.8 and was inversely related to salinity exposure; the spatial variability in leaf litter production contributed much more to LAI uncertainty than did SLA variability. These in situ results are correlative but consistent with the hypothesis, based on previous greenhouse studies, that the stress of chronic salinity exposure changes species’ leaf morphology. Our findings are useful for understanding the growing effects of saltwater intrusion into upland forests, as well as parameterizing and testing ecosystem-scale models simulating forest stressors and disturbances at the terrestrial-aquatic interface.

54 ENVIRONMENTAL SCIENCES↗

Exploring thermal state in mixed immersive virtual environments

Combining immersive virtual environment (IVE) with a controlled environment is a potential solution for analyzing human thermal experience during building design. Existing studies in this field have not adequately analyzed scenarios involving stabilized comfortable and uncomfortable temperature conditions using both thermal state votes and physiological responses, or the influence of the seasons. By combining IVE with a climate chamber, called mixed IVE (MIVE) in this study, experiments were conducted to test the hypothesis that participants' virtual experience did not significantly alter their thermal experience compared to their in-situ experience. Response variables were the control temperature distribution, the thermal state vote (at temperature steps 18.3 °C, 23.8 °C, and 29.4 °C), and physiological responses (heart rate and skin temperature). The results show that the first two response variables were not significantly different between the MIVE and in-situ settings (except for one case). Due to the heat development of the head mounted display device, the mean forehead skin temperature in the MIVE experiments was significantly higher than that in the in-situ experiments in most cases. Furthermore, such difference in skin temperature did not seem to affect general thermal state votes. In addition, significant skin temperature differences at some locations were also observed between the MIVE and in-situ settings.

42 ENGINEERING↗

Three-dimensional spheroid culture induces apical-basal polarity and the original characteristics of immortalized human renal proximal tubule epithelial cells

Highlights: • Immortalized proximal tubule cells (HK-2 cells) formed a tubule-like structure. • The 3D spheroid culture enhanced apical-basal polarity of HK-2 cells. • The 3D spheroid culture induced Na transport function of HK-2 cells. • The 3D spheroid culture promoted up-regulation of kidney development-related genes. The proximal tubules, which are part of the kidney, maintain blood homeostasis by absorbing amino acids, glucose, water, and ions such as sodium (Na), potassium, and bicarbonate. Proximal tubule dysfunction is associated with the pathogenesis of many kidney diseases. Renal proximal tubular epithelial cells (RPTECs) are responsible for the main functions of the proximal tubules. Therefore, in vitro experiments using RPTECs would greatly enhance our understanding of nephron physiology and pathobiology. It is preferable to use immortalized cell lines, such as human kidney-2 (HK-2) cells, because they are derived from humans and maintain growth indefinitely. However, tissue-specific RPTEC phenotypes, including apical-basal polarization, are frequently lost in conventional two-dimensional culture methods in part due to microenvironmental deficiencies. To overcome this limitation, we developed a three-dimensional (3D) spheroid culture method for HK-2 cells using an extracellular matrix. HK-2 spheroids in 3D culture formed a tubule-like architecture with cellular polarity and showed markedly restored Na transport function. 3D culture of HK-2 cells also increased expression of kidney development–related genes, including WNT9B. Models of human renal tubules using HK-2 spheroids will greatly improve our understanding of the physiology and pathobiology of the kidney.

60 APPLIED LIFE SCIENCES↗

Seasonal trends in leaf-level photosynthetic capacity and water use efficiency in a North American Eastern deciduous forest and their impact on canopy-scale gas exchange

Vegetative transpiration (E) and photosynthetic carbon assimilation (A) are known to be seasonally dynamic, with changes in their ratio determining the marginal water use efficiency (WUE). Despite an understanding that stomata play a mechanistic role in regulating WUE, it is still unclear how stomatal and nonstomatal processes influence change in WUE over the course of the growing season. As a result, limited understanding of the primary physiological drivers of seasonal dynamics of canopy WUE remains one of the largest uncertainties in earth system model projections of carbon and water exchange in temperate deciduous forest ecosystems. Here, we investigated seasonal patterns in leaf-level physiological, hydraulic, and anatomical properties, including the seasonal progress of the stomatal slope parameter (g 1 ; inversely proportional to WUE) and the maximum carboxylation rate (V cmax ). V cmax and g 1 were seasonally variable; however, their patterns were not temporally synchronized. g 1 generally showed an increasing trend until late in the season, while V cmax peaked during the midsummer months. Seasonal progression of V cmax was primarily driven by changes in leaf structural, and anatomical characteristics, while seasonal changes in g 1 were most strongly related to changes in V cmax and leaf hydraulics. Using a seasonally variable V cmax and g 1 to parameterize a canopy-scale gas exchange model increased seasonally aggregated A and E by 3% and 16%, respectively.

54 ENVIRONMENTAL SCIENCES↗

Quantitative measurements of reactive oxygen species partitioning in electron transfer flavoenzyme magnetic field sensing

Biological magnetic field sensing that gives rise to physiological responses is of considerable importance in quantum biology. The radical pair mechanism (RPM) is a fundamental quantum process that can explain some of the observed biological magnetic effects. In magnetically sensitive radical pair (RP) reactions, coherent spin dynamics between singlet and triplet pairs are modulated by weak magnetic fields. The resulting singlet and triplet reaction products lead to distinct biological signaling channels and cellular outcomes. A prevalent RP in biology is between flavin semiquinone and superoxide (O 2 •–) in the biological activation of molecular oxygen. This RP can result in a partitioning of reactive oxygen species (ROS) products to form either O 2 •– or hydrogen peroxide (H 2 O 2 ). Here, we examine magnetic sensing of recombinant human electron transfer flavoenzyme (ETF) reoxidation by selectively measuring O 2 •– and H 2 O 2 product distributions. ROS partitioning was observed between two static magnetic fields at 20 nT and 50 μT, with a 13% decrease in H 2 O 2 singlet products and a 10% increase in O 2 •– triplet products relative to 50 µT. RPM product yields were calculated for a realistic flavin/superoxide RP across the range of static magnetic fields, in agreement with experimental results. For a triplet born RP, the RPM also predicts about three times more O 2 •– than H 2 O 2 , with experimental results exhibiting about four time more O 2 •– produced by ETF. The method presented here illustrates the potential of a novel magnetic flavoprotein biological sensor that is directly linked to mitochondria bioenergetics and can be used as a target to study cell physiology.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

A Printed Microscopic Universal Gradient Interface for Super Stretchable Strain‐Insensitive Bioelectronics

Abstract Stretchable electronics capable of conforming to nonplanar and dynamic human body surfaces are central for creating implantable and on‐skin devices for high‐fidelity monitoring of diverse physiological signals. While various strategies have been developed to produce stretchable devices, the signals collected from such devices are often highly sensitive to local strain, resulting in inevitable convolution with surface strain‐induced motion artifacts that are difficult to distinguish from intrinsic physiological signals. Here all‐printed super stretchable strain‐insensitive bioelectronics using a unique universal gradient interface (UGI) are reported to bridge the gap between soft biomaterials and stiff electronic materials. Leveraging a versatile aerosol‐based multi‐materials printing technique that allows precise spatial control over the local stiffnesses with submicron resolution, the UGI enables strain‐insensitive electronic devices with negligible resistivity changes under a 180% uniaxial stretch ratio. Various stretchable devices are directly printed on the UGI for on‐skin health monitoring with high signal quality and near‐perfect immunity to motion artifacts, including semiconductor‐based photodetectors for sensing blood oxygen saturation levels and metal‐based temperature sensors. The concept in this work will significantly simplify the fabrication and accelerate the development of a broad range of wearable and implantable bioelectronics for real‐time health monitoring and personalized therapeutics.

Song, Kaidong [Department of Aerospace and Mechani↗

Cell-Type-Specific Proteomics Analysis of a Small Number of Plant Cells by Integrating Laser Capture Microdissection with a Nanodroplet Sample Processing Platform

Plant organs and tissues contain multiple cell types, which are well organized in 3-dimensional structure to efficiently perform physiological functions such as homeostasis, response to environmental perturbation, pathogen infection. It is critically important to perform molecular measurements at the cell-type-specific level to discover mechanisms and unique features of cell populations that govern differentiation and respond to external perturbations. Although mass spectrometry-based proteomics has been demonstrated as an enabling discovery tool to study plant physiology, conventional approaches require millions of cells to generate robust biological conclusions. Such requirements mask the cell-to-cell heterogeneities and limit the comprehensive profiling of plant proteins at spatially resolved and cell-type-specific resolutions. This protocol describes a recently-developed proteomics workflow for studying a small number of plant cells by integrating laser capture microdissection, microfluidic nanodroplet-based sample preparation, with ultrasensitive liquid chromatography-mass spectrometry. Using poplar as a model tree species, we provide detailed protocols, including plant tissue harvest, tissue preparation, cryosectioning, laser microdissection, protein digestion, mass spectrometry measurement, and data analysis. We show the workflow enables the precise identification and quantification of thousands of proteins from hundreds of isolated plant root and leaf cells.

59 BASIC BIOLOGICAL SCIENCES↗

Hyperspectral leaf reflectance of grasses varies with evolutionary lineage more than with site

Abstract To predict ecological responses at broad environmental scales, grass species are commonly grouped into two broad functional types based on photosynthetic pathway. However, closely related species may have distinctive anatomical and physiological attributes that influence ecological responses, beyond those related to photosynthetic pathway alone. Hyperspectral leaf reflectance can provide an integrated measure of covarying leaf traits that may result from phylogenetic trait conservatism and/or environmental conditions. Understanding whether spectra‐trait relationships are lineage specific or reflect environmental variation across sites is necessary for using hyperspectral reflectance to predict plant responses to environmental changes across spatial scales. We measured hyperspectral leaf reflectance (400–2400 nm) and 12 structural, biochemical, and physiological leaf traits from five grass‐dominated sites spanning the Great Plains of North America. We assessed if variation in leaf reflectance spectra among grass species is explained more by evolutionary lineage (as captured by tribes or subfamilies), photosynthetic pathway (C 3 or C 4 ), or site differences. We then determined whether leaf spectra can be used to predict leaf traits within and across lineages. Our results using redundancy analysis ordination (RDA) show that grass tribe identity explained more variation in leaf spectra (adjusted R 2 = 0.12) than photosynthetic pathway, which explained little variation in leaf spectra (adjusted R 2 = 0.00). Furthermore, leaf reflectance from the same tribe across multiple sites was more similar than leaf reflectance from the same site across tribes (adjusted R 2 = 0.12 and 0.08, respectively). Across all sites and species, trait predictions based on spectra ranged considerably in predictive accuracies ( R 2 = 0.65 to <0.01), but R 2 was >0.80 for certain lineages and sites. The relationship between Vc max , a measure of photosynthetic capacity, and spectra was particularly promising. Chloridoideae, a lineage more common at drier sites, appears to have distinct spectra‐trait relationships compared with other lineages. Overall, our results show that evolutionary relatedness explains more variation in grass leaf spectra than photosynthetic pathway or site, but consideration of lineage‐ and site‐specific trait relationships is needed to interpret spectral variation across large environmental gradients.

Pau, Stephanie [Department of Geography University↗

Morphophysiological Plant Phenotyping for the Development of Plant Breeding Under Drought and Heat Conditions: A Practical Approach

ABSTRACT Currently, the breeding programs focus their efforts on identifying and developing tolerant genotypes to adverse conditions, such as drought and high temperatures. In this context, the physiological approach, which involves phenotyping several traits, is useful for breeding programs. Leaf photosynthetic traits have become one of the main objectives to be evaluated for breeders due to their relationship with improving grain yield and biomass production. Gas exchange ( Ge ) and chlorophyll “a” fluorescence ( Chf ) are the main tools to characterize the photosynthetic activity in real time at the leaf level. Consequently, several association studies using proximal and nonproximal sensing (e.g., RGB, thermography) have been developed. However, for the correct application of this breeding approach, it is essential to have a basic knowledge of both the physiological principles involved in the readings and the limitations of phenotyping due to the characteristics of the devices available on the market. This revision also covers other traits, such as the morphological and anatomical characteristics of leaves and roots, and the use of isotopes complementing Ge and Chf measurements.

Estrada, Félix [Instituto de Investigaciones Agrop↗

Factors influencing pregnancy, litter size, and reproductive parameters of invasive wild pigs

Abstract Reproduction is the most energetically expensive life stage with the demands of productivity representing a balance between physiological requirements and environmental conditions. Wild pigs ( Sus scrofa ) throughout most of North America are genetic hybrids of feral domestic pigs and wild boar and have the highest reproductive potential of any wild ungulate. The phenology of reproduction, extent of multiple reproductive events per year, how individual and extrinsic factors contribute to variability in productivity, and impact of genetic lineage on these parameters is not well understood in wild pigs. We quantified reproductive parameters in wild pigs relative to a suite of individual and environmental attributes across seasons and multiple years in South Carolina, USA, from March 2017 and May 2020. We hypothesized that individual attributes (mass, age class, number of teats, rump fat, relative genetic association to wild boar vs. domestic pigs) and extrinsic factors (mast availability) would influence probability of pregnancy and fetal litter size. Wild pigs produced offspring throughout all months with peaks in conception corresponding to a seasonal pulse in food availability. The likelihood of pregnancy was influenced by female mass and nutritional condition and was greatest during years with abundant resources. Similarly, litter size increased with female mass and age, implying larger and older females represent the most important group for population recruitment. In evaluating the relationship between reproductive output and ancestral associations to domestic pigs versus wild boar, the proportion of wild boar ancestry was not an important influence on productivity in our population. We determined juveniles reach a physiological threshold of sexual maturity at approximately 30 kg. Average litter size was comparable to other populations, and wild pigs maintain an average fetal litter size of 5.43 offspring despite 13.6% embryonic mortality. A thorough understanding of biotic and extrinsic factors influencing reproduction are important for realistic population models, which are necessary for identifying areas to focus management needs and implementation.

59 BASIC BIOLOGICAL SCIENCES↗