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At least 1,297 records · Page 72

[Some peculiar features of liquid supply to the root medium of plants growing in microgravity]

Sixteen point probes monitored moisture level in the root medium of the wheat plants grown in greenhouse SVET on the MIR/NASA space science program. The article outlines types of water migration in the absence of gravity. Hydrophysical characteristics of perspective root media have been explored. Results of the water supply monitoring and control in the course of experiment are reported. The authors put forward porous root media to facilitate water migration and aeration.

NASA Experiment Number 284093 2/2↗

Verification Testing and Veg-05 Tomato Crop Production on the International Space Station

Production of fresh, nutritious, and tasty produce for astronauts during spaceflight may provide health-promoting, bioavailable nutrients and enhance the dietary experience as we move into longer-duration missions. Growing and caring for plants may also reduce the psychological stresses associated with spaceflight and enhance connections to Earth. Requirements to consistently grow a diversity of crops under spaceflight environmental conditions remain poorly defined. The VEG-05 experiment is part of a series of experiments with pick-and-eat salad crops to better define best practices for crop production and handling in space. VEG-05 and predecessor experiments VEG-04A and VEG-04B, use the Veggie vegetable production facilities on the International Space Station to grow salad crops under different spectral compositions. In VEG-04A and B, mizuna mustard was cultivated with two different red: blue light treatments, and in VEG-05 we are cultivating ‘Red Robin’ dwarf cherry tomatoes under the same light spectra. Light can impact the growth habit, yield, nutritional composition, microbial levels, and even flavor attributes within crops, and our team will assess these characteristics for this crop during VEG-05. Prior to launch and installation on ISS in late 2022, both a science verification test (SVT), and an experiment verification test (EVT) were conducted at Kennedy Space Center in ISS Environment Simulator Chambers. Science verification testing, and a previous fertilizer test, grew plants in both plant pillows and PONDS (Passive Orbital Nutrient Delivery System) units and tested two different fertilizer treatments in both sets of hardware, with each test under only one of the light conditions. Because of challenges validating the PONDS hardware on ISS, and with good crop production in plant pillows, the EVT moved forward using only plant pillows with the highest fertilizer composition tested during SVT, and two Veggie units were utilized. One Veggie had light settings consisting of equal levels of red: blue light (150 µmol/m2/s for each color) plus green light (30 µmol/m2/s) while the second Veggie had a 90:10 ratio of red: blue light (270 µmol/m2/s red and 30 µmol/m2/s blue) plus green, so each unit provided 330 µmol/m2/s of photosynthetically active radiation to the tomato crops on average. Our original plan, based on prior ground testing, was to grow the crop for 104 days and harvest at 80, 90 and 104 days after initiation. For SVT, under the equal red: blue light treatment, fruit ripening in plant pillows was delayed and fruit were not ripe by day 80, so actual harvest days were days 90, 97, and 104. For EVT we saw fruit ripening earlier, especially in the high red treatment, and so we harvested at days 83, 90, and 99 days after initiation. In SVT we had mostly daily watering, and this led to excess water in plant pillows, which leaked out. This excess water also caused fungus to grow on one leaf and a couple of plant stems. To reduce this excess moisture, we throttled back the watering for EVT, and used the root mat reservoir more frequently. This led to watering only every other day, reducing crew time needed for plant care, however, two wilting events occurred during this EVT, at days 51 and 75. Plants recovered from these wilting events, but these events may have influenced the rate of fruit ripening and flower formation. Regardless, more than 10 fruit were produced from each plant on average, with the high red treatment producing slightly heavier fruit. Microbial testing from fruit during SVT indicated that fruit were safe for consumption with microbial levels below detection limits. VEG-05 flight and ground operations are expected to run between December 2022 and March 2023. This research was co-funded by the Human Research Program and Space Biology (MTL#1075) in the ILSRA 2015 NRA call.

Gioia D. Massa↗

Center for Lignocellulose Structure and Formation (CLSF)

Lignocellulose is the carbon- and energy-rich material synthesized by plants when they make secondary cell walls, e.g. in woody tissues. Plants annually convert gigatons of atmospheric CO 2 into these complex and useful biomaterials that comprise over half of all biological carbon on Earth. This biological conversion helps to reduce atmospheric CO 2 that contributes to climate change while generating renewable plant cell walls that are used commercially on a massive scale in the paper, timber and textile industries. Recent technical developments open new possibilities for use of modified wood and cellulose fibers as large-scale alternatives to steel, plastics, and other nonrenewable materials with high carbon footprints and energy inputs, as well as for generation of biofuels and bio-hydrogen. Despite the utility of plant-based biomaterials, many physical and biological aspects of cell wall structure and formation remain obscure, and this limits the current scope for engineering these renewable resources for greater utility.

09 BIOMASS FUELS↗

IPD3, a master regulator of arbuscular mycorrhizal symbiosis, affects genes for immunity and metabolism of non-host Arabidopsis when restored long after its evolutionary loss

Abstract Arbuscular mycorrhizal symbiosis (AM) is a beneficial trait originating with the first land plants, which has subsequently been lost by species scattered throughout the radiation of plant diversity to the present day, including the model Arabidopsis thaliana . To explore if elements of this apparently beneficial trait are still present and could be reactivated we generated Arabidopsis plants expressing a constitutively active form of Interacting Protein of DMI3 , a key transcription factor that enables AM within the Common Symbiosis Pathway, which was lost from Arabidopsis along with the AM host trait. We characterize the transcriptomic effect of expressing IPD3 in Arabidopsis with and without exposure to the AM fungus (AMF) Rhizophagus irregularis , and compare these results to the AM model Lotus japonicus and its ipd3 knockout mutant cyclops-4 . Despite its long history as a non-AM species, restoring IPD3 in the form of its constitutively active DNA-binding domain to Arabidopsis altered expression of specific gene networks. Surprisingly, the effect of expressing IPD3 in Arabidopsis and knocking it out in Lotus was strongest in plants not exposed to AMF, which is revealed to be due to changes in IPD3 genotype causing a transcriptional state, which partially mimics AMF exposure in non-inoculated plants. Our results indicate that molecular connections to symbiosis machinery remain in place in this nonAM species, with implications for both basic science and the prospect of engineering this trait for agriculture.

59 BASIC BIOLOGICAL SCIENCES↗

Experiments on plants grown in space. Status and prospects

Space flight provides a unique capability to gravitational research by providing an environment in which gravity can be manipulated below the Earth norm of 1 to a state comparable to weightlessness. Although space experiments with seeds and plants have been conducted for almost 25 years, we are only now entering an era that promises the capability of routinely conducting controlled plant experiments in space. Results from experiments with higher plants and seeds flown thus far in both manned and unmanned spacecraft from both the USA and USSR are briefly summarized. An overview of the current status of space biology experimentation as it relates to plants is presented and present and future opportunities to conduct plant experiments under space flight conditions are discussed.

Flight Experiment↗

Arabidopsis plants expressing only the redox–regulated Rca–α isoform have constrained photosynthesis and plant growth

Rubisco activase (Rca) facilitates the release of sugar–phosphate inhibitors from the active sites of Rubisco and thereby plays a central role in initiating and sustaining Rubisco activation. In Arabidopsis, alternative splicing of a single Rca gene results in two Rca isoforms, Rca–α and Rca–β. Redox modulation of Rca–α regulates the function of Rca–α and Rca–β acting together to control Rubisco activation. Although Arabidopsis Rca–α alone less effectively activates Rubisco in vitro, it is not known how CO 2 assimilation and plant growth are impacted. Here, we show that two independent transgenic Arabidopsis lines expressing Rca–α in the absence of Rca–β (‘Rca–α only’ lines) grew more slowly in various light conditions, especially under low light or fluctuating light intensity, and in a short day photoperiod compared to wildtype. Photosynthetic induction was slower in the Rca–α only lines, and they maintained a lower rate of CO 2 assimilation during both photoperiod types. Here, our findings suggest Rca oligomers composed of Rca–α only are less effective in initiating and sustaining the activation of Rubisco than when Rca–β is also present. Currently there are no examples of any plant species that naturally express Rca–α only but numerous examples of species expressing Rca–β only. That Rca–α exists in most plant species, including many C3 and C4 food and bioenergy crops, implies its presence is adaptive under some circumstances.

59 BASIC BIOLOGICAL SCIENCES↗

Experiments to be flown in an Earth orbiting laboratory: The US experiments on the first international microgravity laboratory, from concept to flight

The current life cycle of NASA ARC-managed flight experiments is presented. The two main purposes are: (1) to bring to the attention of biologists, and in particular cell and plant biologists, some of the requirements for flying a life science experiment in space; and (2) to introduce the subject to biologists embarking on studies in the field and to delineate some of the specific requirements that will be encountered by an ARC-managed microgravity experiment. This is not intended to be an exhaustive encyclopedia of all techniques used to prepare an experiment to evaluate the effect of microgravity on plant and animal cells. However, many of the requirements are the same for all biological systems and for other NASA centers. Emphasis is on the principle investigator's (PI's) involvement in the activities required for successful completion of major reviews. The PI support required for activities other than these reviews is also discussed, as are the interactions between ARC and the PI that will be required as problems or questions arise throughout experiment and payload development. It is impossible to predict the extent of this activity because it varies according to the complexity of the experiment and the flight experience of the PI.

Winget, C. M.↗

Responses of vascular plant fine roots and associated microbial communities to whole-ecosystem warming and elevated CO 2 in northern peatlands

Warming and elevated CO 2 (eCO 2 ) are expected to facilitate vascular plant encroachment in peatlands. The rhizosphere, where microbial activity is fueled by root turnover and exudates, plays a crucial role in biogeochemical cycling, and will likely at least partially dictate the response of the belowground carbon cycle to climate changes. We leveraged the Spruce and Peatland Responses Under Changing Environments (SPRUCE) experiment, to explore the effects of a whole-ecosystem warming gradient (+0°C to 9°C) and eCO 2 on vascular plant fine roots and their associated microbes. We combined trait-based approaches with the profiling of fungal and prokaryote communities in plant roots and rhizospheres, through amplicon sequencing. Warming promoted self-reliance for resource uptake in trees and shrubs, while saprophytic fungi and putative chemoorganoheterotrophic bacteria utilizing plant-derived carbon substrates were favored in the root zone. Conversely, eCO 2 promoted associations between trees and ectomycorrhizal fungi. Trees mostly associated with short-distance exploration-type fungi that preferentially use labile soil N. Additionally, eCO 2 decreased the relative abundance of saprotrophs in tree roots. Our results indicate that plant fine-root trait variation is a crucial mechanism through which vascular plants in peatlands respond to climate change via their influence on microbial communities that regulate biogeochemical cycles.

59 BASIC BIOLOGICAL SCIENCES↗

A plant host, Nicotiana benthamiana, enables the production and study of fungal lignin-degrading enzymes

Abstract Lignin has significant potential as an abundant and renewable source for commodity chemicals yet remains vastly underutilized. Efforts towards engineering a biochemical route to the valorization of lignin are currently limited by the lack of a suitable heterologous host for the production of lignin-degrading enzymes. Here, we show that expression of fungal genes in Nicotiana benthamiana enables production of members from seven major classes of enzymes associated with lignin degradation (23 of 35 tested) in soluble form for direct use in lignin activity assays. We combinatorially characterized a subset of these enzymes in the context of model lignin dimer oxidation, revealing that fine-tuned coupling of peroxide-generators to peroxidases results in more extensive C-C bond cleavage compared to direct addition of peroxide. Comparison of peroxidase isoform activity revealed that the extent of C-C bond cleavage depends on peroxidase identity, suggesting that peroxidases are individually specialized in the context of lignin oxidation. We anticipate the use of N. benthamiana as a platform to rapidly produce a diverse array of fungal lignin-degrading enzymes will facilitate a better understanding of their concerted role in nature and unlock their potential for lignin valorization, including within the plant host itself.

59 BASIC BIOLOGICAL SCIENCES↗

Serendipita Fungi Modulate the Switchgrass Root Transcriptome to Circumvent Host Defenses and Establish a Symbiotic Relationship

The fungal family Serendipitaceae encompasses root-associated lineages with endophytic, ericoid, orchid, and ectomycorrhizal lifestyles. Switchgrass is an important bioenergy crop for cellulosic ethanol production owing to high biomass production on marginal soils otherwise unfit for food crop cultivation. The aim of this study was to investigate the host plant responses to Serendipita spp. colonization by characterizing the switchgrass root transcriptome during different stages of symbiosis in vitro. For this, we included a native switchgrass strain, Serendipita bescii, and a related strain, S. vermifera, isolated from Australian orchids. Serendipita colonization progresses from thin hyphae that grow between root cells to, finally, the production of large, bulbous hyphae that fill root cells during the later stages of colonization. We report that switchgrass seems to perceive both fungi prior to physical contact, leading to the activation of chemical and structural defense responses and putative host disease resistance genes. Subsequently, the host defense system appears to be quenched and carbohydrate metabolism adjusted, potentially to accommodate the fungal symbiont. In addition, prior to contact, switchgrass exhibited significant increases in root hair density and root surface area. Furthermore, genes involved in phytohormone metabolism such as gibberellin, jasmonic acid, and salicylic acid were activated during different stages of colonization. Both fungal strains induced plant gene expression in a similar manner, indicating a conserved plant response to members of this fungal order. Understanding plant responsiveness to Serendipita spp. will inform our efforts to integrate them into forages and row crops for optimal plant–microbe functioning, thus facilitating low-input, sustainable agricultural practices.

59 BASIC BIOLOGICAL SCIENCES↗

Disentangling the Effects of Vapor Pressure Deficit and Soil Water Availability on Canopy Conductance in a Seasonal Tropical Forest During the 2015 El Niño Drought

In land models used in Earth system models, vegetation water stress is heuristically related to soil water matric potential, and root water uptake in soil layers is simply proportional to root density to meet the transpiration demand. The efficiency of roots utilizing deeper soil water under water stress cannot be well represented, but it is of critical importance for transpiration regulation and carbon flux. To improve modeling of tropical forest response to drought, a plant hydrodynamic scheme (HYDRO), adapted from a trait-based model in the Functionally Assembled Terrestrial Ecosystem Simulator (FATES), has been implemented in the Energy Exascale Earth System Model (E3SM) Land Model (ELM) to represent the hydraulic dynamics and root water uptake strategy. The adapted model conforms to the big-leaf photosynthesis model in ELM. Vegetation water stress in the model is a function of prognostic leaf water potential and is used to regulate stomatal conductance. Point simulations were performed at two tropical forest sites in the Amazon and Panama. Model sensitivity to the numerical solver, plant hydraulic traits, soil properties, and atmospheric forcing has been evaluated. Our results showed that, even though water stress factors calculated at our study sites using HYDRO are comparable to that simulated by the heuristic water stress model, soil moisture prediction is improved with soil water redistributions through roots. Simulated results using HYDRO is not as sensitive to plant hydraulic traits measured for different tree species as to the soil properties and atmospheric forcing. Simplified numerical solver may cause inaccuracy as to where roots uptake water. This study suggests that plant hydrodynamic representation is critical for improved simulations of soil moisture under water stresses.

54 ENVIRONMENTAL SCIENCES↗

Quantifying Seasonal and Diurnal Cycles of Solar‐Induced Fluorescence With a Novel Hyperspectral Imager

Abstract Solar‐induced fluorescence (SIF) is a proxy of ecosystem photosynthesis that often scales linearly with gross primary productivity (GPP) at the canopy scale. However, the mechanistic relationship between GPP and SIF is still uncertain, especially at smaller temporal and spatial scales. We deployed a ultra‐hyperspectral imager over two grassland sites in California throughout a soil moisture dry down. The imager has high spatial resolution that limits mixed pixels, enabling differentiation between plants and leaves within one scene. We find that imager SIF correlates well with diurnal changes in leaf‐level physiology and gross primary productivity under well‐watered conditions. These relationships deteriorate throughout the dry down event. Our results demonstrate an advancement in SIF imaging with new possibilities in remotely sensing plant canopies from the leaf to the ecosystem. These data can be used to resolve outstanding questions regarding SIF's meaning and usefulness in terrestrial ecosystem monitoring.

60 APPLIED LIFE SCIENCES↗

Selective Detection of Intermediate-Amplitude Motion by Solid-State NMR

The coexistence of rigid and mobile molecules or molecular segments abounds in biomolecular assemblies. Examples include the carbohydrate-rich cell walls of plants and intrinsically disordered proteins that contain rigid β-sheet cores. In solid-state nuclear magnetic resonance (NMR) spectroscopy, dipolar polarization transfer experiments are well suited for detecting rigid components, whereas scalar-coupling experiments are well suited for detecting highly mobile components. However, few NMR methods are available to detect the segments that undergo intermediate-amplitude fast motion. Here, we introduce two NMR experiments, a two-dimensional T 2H -filtered CP-hCH correlation and a three-dimensional J-INADEQUATE CCH correlation, to observe this intermediate-amplitude motion. Both experiments involve 1 H detection under fast magic-angle spinning (MAS). By combining 1 H transverse relaxation (T 2H ) filters with dipolar polarization transfer, we suppress the signals of both highly rigid and highly mobile species, thus revealing the signals of intermediate mobile species. 1 H detection under fast MAS is crucial for distinguishing the different motional amplitudes. Here, we demonstrate these techniques on several plant cell wall samples and show that they allow the selective detection and resolution of certain hemicellulose and pectin signals, which are usually masked by the signals of the rigid cellulose and the highly dynamic pectins in purely dipolar and scalar NMR spectra.

59 BASIC BIOLOGICAL SCIENCES↗

ER-associated VAP27-1 and VAP27-3 proteins functionally link the lipid-binding ORP2A at the ER-chloroplast contact sites

Abstract The plant endoplasmic reticulum (ER) contacts heterotypic membranes at membrane contact sites (MCSs) through largely undefined mechanisms. For instance, despite the well-established and essential role of the plant ER-chloroplast interactions for lipid biosynthesis, and the reported existence of physical contacts between these organelles, almost nothing is known about the ER-chloroplast MCS identity. Here we show that the Arabidopsis ER membrane-associated VAP27 proteins and the lipid-binding protein ORP2A define a functional complex at the ER-chloroplast MCSs. Specifically, through in vivo and in vitro association assays, we found that VAP27 proteins interact with the outer envelope membrane (OEM) of chloroplasts, where they bind to ORP2A. Through lipidomic analyses, we established that VAP27 proteins and ORP2A directly interact with the chloroplast OEM monogalactosyldiacylglycerol (MGDG), and we demonstrated that the loss of the VAP27-ORP2A complex is accompanied by subtle changes in the acyl composition of MGDG and PG. We also found that ORP2A interacts with phytosterols and established that the loss of the VAP27-ORP2A complex alters sterol levels in chloroplasts. We propose that, by interacting directly with OEM lipids, the VAP27-ORP2A complex defines plant-unique MCSs that bridge ER and chloroplasts and are involved in chloroplast lipid homeostasis.

59 BASIC BIOLOGICAL SCIENCES↗

Root exudate composition reflects drought severity gradient in blue grama (Bouteloua gracilis)

Abstract Plant survival during environmental stress greatly affects ecosystem carbon (C) cycling, and plant–microbe interactions are central to plant stress survival. The release of C-rich root exudates is a key mechanism plants use to manage their microbiome, attracting beneficial microbes and/or suppressing harmful microbes to help plants withstand environmental stress. However, a critical knowledge gap is how plants alter root exudate concentration and composition under varying stress levels. In a greenhouse study, we imposed three drought treatments (control, mild, severe) on blue grama ( Bouteloua gracilis Kunth Lag. Ex Griffiths), and measured plant physiology and root exudate concentration and composition using GC–MS, NMR, and FTICR. With increasing drought severity, root exudate total C and organic C increased concurrently with declining predawn leaf water potential and photosynthesis. Root exudate composition mirrored the physiological gradient of drought severity treatments. Specific compounds that are known to alter plant drought responses and the rhizosphere microbiome mirrored the drought severity-induced root exudate compositional gradient. Despite reducing C uptake, these plants actively invested C to root exudates with increasing drought severity. Patterns of plant physiology and root exudate concentration and composition co-varied along a gradient of drought severity.

59 BASIC BIOLOGICAL SCIENCES↗

Trading water for carbon in the future: Effects of elevated CO 2 and warming on leaf hydraulic traits in a semiarid grassland

Abstract The effects of climate change on plants and ecosystems are mediated by plant hydraulic traits, including interspecific and intraspecific variability of trait phenotypes. Yet, integrative and realistic studies of hydraulic traits and climate change are rare. In a semiarid grassland, we assessed the response of several plant hydraulic traits to elevated CO 2 (+200 ppm) and warming (+1.5 to 3°C; day to night). For leaves of five dominant species (three graminoids and two forbs), and in replicated plots exposed to 7 years of elevated CO 2 , warming, or ambient climate, we measured: stomatal density and size, xylem vessel size, turgor loss point, and water potential (pre‐dawn). Interspecific differences in hydraulic traits were larger than intraspecific shifts induced by elevated CO 2 and/or warming. Effects of elevated CO 2 were greater than effects of warming, and interactions between treatments were weak or not detected. The forbs showed little phenotypic plasticity. The graminoids had leaf water potentials and turgor loss points that were 10% to 50% less negative under elevated CO 2 ; thus, climate change might cause these species to adjust their drought resistance strategy away from tolerance and toward avoidance. The C4 grass also reduced allocation of leaf area to stomata under elevated CO 2 , which helps explain observations of higher soil moisture. The shifts in hydraulic traits under elevated CO 2 were not, however, simply due to higher soil moisture. Integration of our results with others' indicates that common species in this grassland are more likely to adjust stomatal aperture in response to near‐term climate change, rather than anatomical traits; this contrasts with apparent effects of changing CO 2 on plant anatomy over evolutionary time. Future studies should assess how plant responses to drought may be constrained by the apparent shift from tolerance (via low turgor loss point) to avoidance (via stomatal regulation and/or access to deeper soil moisture).

54 ENVIRONMENTAL SCIENCES↗