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161 records · Page 9

Dryland feedbacks to future climate change: how species mortality and replacement will affect coupled biogeochemical cycles and energy balance (Final Technical Report)

The overall goal of the dryland climate manipulation experiment is to improve our understanding and ability to forecast dryland responses to increasing temperatures and altered precipitation regimes. Drylands cover over 40% of Earth’s terrestrial surface and are predicted to increase in size 11-23% by 2100. Yet our understanding of this vast biome, which exchanges enormous amounts of CO 2 and energy with the atmosphere, is exceedingly poor. This project aims to transform and improve that understanding in order to reduce uncertainty and increase the confidence with which we can make global predictions of future climate. We focus on both above- and belowground processes, and explore temperature controls over critical aspects of carbon and nutrient cycling for dryland plants, soil, and microbes. We have 60 climate manipulation plots where plants (from above the canopy) and soils are warmed using infrared lamps and where precipitation has been altered using targeted watering regimes. We have measured myriad aspects of plant physiology, morphology, carbon exchange, energy exchange, and phenology, as well as analogous climate-induced changes to biological soil crusts and soil microbial communities. Our goals are to investigate temperature responses and acclimation potential of dryland plant and soil processes. Concurrent soil incubation experiments complement the in-situ soil warming component and enable more controlled mechanistic investigations of temperature response on microbial function. Automated soil CO 2 efflux chambers within the plots provide phenomenal opportunity to improve our understanding of dryland CO 2 exchange with the atmosphere and are coupled with plant CO 2 exchange measurements. The project has been exceptionally successful, with 35 peer-reviewed publications (and more in review), including in high-profile journals such as Nature Climate Change and PNAS. We have given over fifty presentations on the work and it has been featured in many high-profile media outlets, such as Discover Magazine and on NPR.

54 ENVIRONMENTAL SCIENCES↗

Where Have All of the Electrons Gone? Understanding the Role of Alternative Electron Flow and O 2 Reduction in Balancing Cellular Redox (Final Technical Report)

An informed understanding of the fundamentals of light-driven electron transfer reactions in photoautotrophic organisms is necessary to enable improved photosynthetic efficiencies for the production of renewable fuels and novel biomaterials. To obtain high photosynthetic yields, light energy must be efficiently coupled to the fixation of CO 2 . Sub-optimal environmental conditions and metabolic routing (caused by blocks in biosynthetic routes) can severely impact the conversion of light energy to biomass and lead to reactive oxygen production, which in turn can cause cellular damage and productivity losses. Hence, plants, algae, and photosynthetic bacteria have evolved a network of alternative outlets to sustain the flow of photosynthetically derived-electrons. Our research was focused on the nature and integration of these outlets. The data obtained in this project will inform efforts to rationally engineer crop plants and algae to improve photosynthetic yields by enhancing electron transfer reactions to CO 2 reduction and targeted biomass accumulation. A major project research thrust was focused on identifying and quantifying the flow of photosynthetic reductant through electron transfer circuits that result in the light-dependent reduction of O 2 . New energy management strategies were identified that are used when normal carbon assimilation pathways are compromised due to nutrient deprivation, and/or by a reduction in starch synthesis/carbon storage, all conditions resulting in highly reduced intracellular redox pools. The green alga Chlamydomonas reinhardtii, and likely many other algae, have multiple O 2 -reducing pathways that play critical roles in maintaining cellular metabolic balance and scavenging electrons that could potentially cause cellular damage, which in some instances leads to reduced photosynthetic yields but increased fitness. We explored the activities of three essential outlets associated with Chlamydomonas reinhardtii photosynthetic electron transport: (1) reduction of O 2 to H 2 O through Flavodiiron proteins (FLVs) and (2) Plastid Terminal Oxidases (PTOX), and (3) the synthesis of starch. Real-time measurements of O 2 exchange demonstrated that FLVs immediately engage during dark to light transitions, allowing electron transport when the CBBC is not fully activated. Under these conditions, we quantified, for the first time, maximal FLV activity and its overall capacity to direct photosynthetic electrons towards O 2 reduction. However, when starch synthesis is compromised, a greater proportion of electrons is directed toward O 2 reduction through the FLVs, while PTOX, which is sensitive to the PQ pool redox state, is activated. This suggests, that starch synthesis has an important role in priming/regulating CBBC and electron transport. We also identified a biological ‘switch’ in the green alga Chlamydomonas reinhardtii that reversibly restricts photosynthetic electron transport (PET) at the cytochrome b 6 f complex when reductant and ATP generated by PET are in excess of the capacity of carbon metabolism to utilize these products; we specifically show a restriction at this switch when sta6 mutant cells, which cannot synthesize starch, are limited for nitrogen (growth inhibition) and subjected to a dark to light transition. This restriction causes diminished electron flow to PSI, which prevents PSI photodamage, and the plastid alternative oxidase (PTOX) becomes fully activated, serving as an electron valve that dissipates excitation energy absorbed by PSII, thereby lessening PSII photoinhibition. Furthermore, illumination of the cells following the dark acclimation gradually diminishes the restriction at the switch. Future engineering of these switches may allow more effective electron transfer to lipid (biofuel) pathways and diminish the number of electrons “wasted” in the reduction of O 2 to water. Lastly, we explored metabolic routing of electrons during algal fermentation and discovered multiple novel pathways that are activated when the preferred anoxic routes are blocked. These provide valuable products (e.g. lactate, glycerol) that can be used in broad portfolio of biotechnological applications.

59 BASIC BIOLOGICAL SCIENCES↗

Epigenetic Control of Drought Response in Sorghum (EPICON)

Genetic manipulation of crops to increase the presence of desirable traits has been critical to increasing agricultural productivity. These changes have primarily involved modification of the plant’s DNA sequence. However, there is increasing evidence that environmental responses are also mediated by epigenetics, which involves heritable changes without changes in DNA sequence. Epigenetic changes have been shown to play a major role in regulating plant responses to drought, an increasing problem worldwide due to climate change. In general, exposure of plants to water limitation triggers epigenetic changes, which include remodeling of chromatin, the network of DNA, RNA and various proteins making up chromosomes, and related changes in regulatory mechanisms. EPICON’s efforts focus on unraveling the role epigenetic signals play in acclimation to and recovery from drought through effects on individual transcription factors or transcriptional networks that direct entire metabolic pathways. To achieve this goal we will follow responses to water deprivation in sorghum, a widely cultivated cereal with recognized drought tolerance. In EPICON’s field trials, sorghum will be grown under controlled irrigation conditions. Leaf and root samples will be taken to perform molecular phenotyping to track changes in epigenetic, transcriptomic, metabolomic and proteomic footprints. Analysis of this data will provide a better understanding of the epigenetic processes related to drought tolerance, leading to our ultimate goal of identifying transcriptional regulators and pathways controlling drought resistance. The identified genetic targets and their regulatory pathways will be used in future efforts to improve growth of sorghum and other crops in the field and in marginal lands under water-limiting conditions.

59 BASIC BIOLOGICAL SCIENCES↗

Understanding spatial and temporal drivers of variation in tree hydraulic processes and their consequences for climate feedbacks (Final Technical Report)

This is the final technical report from the first phase of a project that changed institutions. The grant was titled “Understanding spatial and temporal drivers of variation in tree hydraulic processes and their consequences for climate feedbacks.” The overall objectives of this project were to (1) provide model‐compatible datasets of key plant hydraulic traits and status for model evaluation, parameterization and validation and (2) use these data to pinpoint ecosystem responses to a changing hydroclimate by addressing both long‐term climatic drying and episodic extreme droughts. We planned to address the objectives with three research activities to quantify plant responses to chronic water stress and episodic drought: (1) generate high frequency observations of soil and plant hydraulic data across different landscape positions at multiple sites, (2) quantify plant hydraulic trait plasticity in response to experimental soil moisture reduction in situ in two central hardwood forests, and (3) simulate the carbon consequences of incorporating plant hydrodynamics and plant acclimation to water stress in the DOE‐sponsored plant hydrodynamics model FATES‐HYDRO. As of the transfer of this project to another institution, we had made substantial progress on activities 1 and 2, and started activity 3.

54 ENVIRONMENTAL SCIENCES↗

SPRUCE Photosynthesis and Respiration of Picea mariana and Larix laricina in SPRUCE Experimental Plots, 2016-2017

This dataset contains empirical physiological, morphological, and chemical data collected on two dominant conifer species, Picea mariana and Larix laricina, between May 2016 and August 2017 at the SPRUCE (Spruce and Peatland Responses Under Changing Environments) experimental site within the Marcell Experimental Forest in northern Minnesota, USA. Analyses assessed the response of conifers to whole-ecosystem warming and elevated atmospheric carbon dioxide (CO2) concentrations. Measurements include light-saturated net photosynthesis and dark respiration at ambient atmospheric carbon dioxide (CO2) concentration (2016), CO2 response curves of light-saturated net photosynthesis (2017), leaf morphology (leaf mass per unit leaf area) and nitrogen content (on both mass and area basis). Leaf photosynthesis and respiration were measured with portable photosynthesis systems (LI6400XT, LI6800, LI-COR Biosciences, USA). In 2016, photosynthesis and respiration were measured at a common temperature of 25 ˚C to assess thermal acclimation, and at growth temperatures (for each of the five treatment temperatures) to assess field performance. In 2017, CO2 response curves of net photosynthesis (so-called A-Ci curves) were conducted at different leaf temperatures (15, 25, 32.5, 40 and 45 ˚C) for assessment of biochemical parameters and their temperature sensitivity. In addition, dark respiration measurements at different leaf temperatures (10, 15, 20, 25, 30, 35, 40, and 45 ºC) were also taken. This dataset contains one data file in comma-separate values (*.csv) and Microsoft Excel (*.xlsx) format. Additional metadata are provided: a data dictionary and a file-level metadata file in comma-separate values (.csv) format and a user guide in PDF (*.pdf) format.

54 ENVIRONMENTAL SCIENCES↗

The survivor strain: isolation and characterization of Phormidium yuhuli AB48, a filamentous phototactic cyanobacterium with biotechnological potential

Despite their recognized potential, current applications of cyanobacteria as microbial cell factories remain in early stages of development. This is partly due to the fact that engineered strains are often difficult to grow at scale. This technical challenge contrasts with the dense and highly productive cyanobacteria populations thriving in many natural environments. It has been proposed that the selection of strains pre-adapted for growth in industrial photobioreactors could enable more productive cultivation outcomes. Here, we described the initial morphological, physiological, and genomic characterization of Phormidium yuhuli AB48 isolated from an industrial photobioreactor environment. P. yuhuli AB48 is a filamentous phototactic cyanobacterium with a growth rate comparable to Synechocystis sp . PCC 6803. The isolate forms dense biofilms under high salinity and alkaline conditions and manifests a similar nutrient profile to Arthrospira platensis ( Spirulina ). We sequenced, assembled, and analyzed the P. yuhuli AB48 genome, the first closed circular isolate reference genome for a member of the Phormidium genus. We then used cultivation experiments in combination with proteomics and metabolomics to investigate growth characteristics and phenotypes related to industrial scale cultivation, including nitrogen and carbon utilization, salinity, and pH acclimation, as well as antibiotic resistance. These analyses provide insight into the biological mechanisms behind the desirable growth properties manifested by P. yuhuli AB48 and position it as a promising microbial cell factory for industrial-scale bioproduction[221, 1631].

59 BASIC BIOLOGICAL SCIENCES↗

Environmental Tuning of Homologs of the Orange Carotenoid Protein-Encoding Gene in the Cyanobacterium Fremyella diplosiphon

The orange carotenoid protein (OCP) family of proteins are light-activated proteins that function in dissipating excess energy absorbed by accessory light-harvesting complexes, i.e., phycobilisomes (PBSs), in cyanobacteria. Some cyanobacteria contain multiple homologs of the OCP-encoding gene ( ocp ). Fremyella diplosiphon , a cyanobacterium studied for light-dependent regulation of PBSs during complementary chromatic acclimation (CCA), contains several OCP homologs – two full-length OCPs, three Helical Carotenoid Proteins (HCPs) with homology to the N-terminus of OCP, and one C-terminal domain-like carotenoid protein (CCP) with homology to the C-terminus of OCP. We examined whether these homologs are distinctly regulated in response to different environmental factors, which could indicate distinct functions. We observed distinct patterns of expression for some OCP, HCP, and CCP encoding genes, and have evidence that light-dependent aspects of ocp homolog expression are regulated by photoreceptor RcaE which controls CCA. RcaE-dependent transcriptional regulator RcaC is also involved in the photoregulation of some hcp genes. Apart from light, additional environmental factors associated with cellular redox regulation impact the mRNA levels of ocp homologs, including salt, cold, and disruption of electron transport. Analyses of conserved sequences in the promoters of ocp homologs were conducted to gain additional insight into regulation of these genes. Several conserved regulatory elements were found across multiple ocp homolog promoters that potentially control differential transcriptional regulation in response to a range of environmental cues. The impact of distinct environmental cues on differential accumulation of ocp homolog transcripts indicates potential functional diversification of this gene family in cyanobacteria. These genes likely enable dynamic cellular protection in response to diverse environmental stress conditions in F . diplosiphon .

59 BASIC BIOLOGICAL SCIENCES↗

Genome-Wide Study of Hsp90 Gene Family in Cabbage (Brassica oleracea var. capitata L.) and Their Imperative Roles in Response to Cold Stress

Heat shock protein 90 (Hsp90) plays an important role in plant developmental regulation and defensive reactions. Several plant species have been examined for the Hsp90 family gene. However, the Hsp90 gene family in cabbage has not been well investigated to date. In this study, we have been discovered 12 BoHsp90 genes in cabbage ( Brassica oleracea var. capitata L.). These B. oleracea Hsp90 genes were classified into five groups based on phylogenetic analysis. Among the five groups, group one contains five Hsp90 genes, including BoHsp90-1 , BoHsp90-2 , BoHsp90-6 , BoHsp90-10 , and BoHsp90-12 . Group two contains three Hsp90 genes, including BoHsp90-3 , BoHsp90-4 , and BoHsp90 . Group three only includes one Hsp90 gene, including BoHsp90-9 . Group four were consisting of three Hsp90 genes including BoHsp90-5 , BoHsp90-7 , and BoHsp90-8 , and there is no Hsp90 gene from B. oleracea in the fifth group. Synteny analysis showed that a total of 12 BoHsp90 genes have a collinearity relationship with 5 Arabidopsis genes and 10 Brassica rapa genes. The promoter evaluation revealed that the promoters of B. oleracea Hsp90 genes included environmental stress-related and hormone-responsive cis-elements . RNA-seq data analysis indicates that tissue-specific expression of BoHsp90-9 and BoHsp90-5 were highly expressed in stems, leaves, silique, and flowers. Furthermore, the expression pattern of B. oleracea BoHsp90 exhibited that BoHsp90-2, BoHsp90-3, BoHsp90-7, BoHsp90-9, BoHsp90-10, and BoHsp90-11 were induced under cold stress, which indicates these Hsp90 genes perform a vital role in cold acclimation and supports in the continual of normal growth and development process. The cabbage Hsp90 gene family was found to be differentially expressed in response to cold stress, suggesting that these genes play an important role in cabbage growth and development under cold conditions.

Sajad, Shoukat↗

Homologs of Phycobilisome Abundance Regulator PsoR Are Widespread across Cyanobacteria

During chromatic acclimation (CA), cyanobacteria undergo shifts in their physiology and metabolism in response to changes in their light environment. Various forms of CA, which involves the tuning of light-harvesting accessory complexes known as phycobilisomes (PBS) in response to distinct wavelengths of light, have been recognized. Recently, a negative regulator of PBS abundance, PsoR, about which little was known, was identified. We used sequence analyses and bioinformatics to predict the role of PsoR in cyanobacteria and PBS regulation and to examine its presence in a diverse range of cyanobacteria. PsoR has sequence similarities to the β-CASP family of proteins involved in DNA and RNA processing. PsoR is a putative nuclease widespread across Cyanobacteria, of which over 700 homologs have been observed. Promoter analysis suggested that psoR is co-transcribed with upstream gene tcpA. Multiple transcription factors involved in global gene regulation and stress responses were predicted to bind to the psoR-tcpA promoter. The predicted protein–protein interactions with PsoR homologs included proteins involved in DNA and RNA metabolism, as well as a phycocyanin-associated protein predicted to interact with PsoR from Fremyella diplosiphon (FdPsoR). The widespread presence of PsoR homologs in Cyanobacteria and their ties to DNA- and RNA-metabolizing proteins indicated a potentially unique role for PsoR in CA and PBS abundance regulation.

Layer, Alicia↗

Novel Protocol for Acute In Situ Ecotoxicity Test Using Native Crustaceans Applied to Groundwater Ecosystems

Current standardized laboratory test protocols use model species that have limitations to accurately assess native species responses to stressors. We developed and tested a novel acute in situ protocol for testing field-collected organisms. We used Asellus aquaticus and NaCl as a reference toxicant to test for the effects of location (laboratory vs. in situ), medium (synthetic vs. field water), substrate (presence vs. absence), and protocol replicability. We further tested the protocol using groundwater-adapted isopods: Proasellus assaforensis for the effect of location, P. cavaticus of medium and P.lusitanicus of substrate. Our results showed that A.aquaticus’ lethality obtained with the novel acute in situ protocol did not significantly differ from those from laboratory testing. However, laboratory tested P.assaforensis showed a higher sensitivity, suggesting that its acclimation to laboratory conditions might have pernicious effects. A. aquaticus and P. cavaticus showed a higher mortality using synthetic medium in situ and under laboratory conditions, which overestimated the stressor’s effect. Besides, substrate use had no significant effect. The novel acute in situ protocol allows the use of native species under realistic scenarios. It is particularly well adapted for assessing the risk of groundwater ecosystems but it can be applied to a wide range of ecosystems.

Castaño-Sánchez, Andrea↗

Experimental trials of species-specific bat flight responses to an ultrasonic deterrent

Unintended consequences of increasing wind energy production include bat mortalities from wind turbine blade strikes. Ultrasonic deterrents (UDs) have been developed to reduce bat mortalities at wind turbines. Our goal was to experimentally assess the species-specific effectiveness of three emission treatments from the UD developed by NRG Systems. We conducted trials in a flight cage measuring approximately 60 m × 10 m × 4.4 m (length × width × height) from July 2020 to May 2021 in San Marcos, Texas, USA. A single UD was placed at either end of the flight cage, and we randomly selected one for each night of field trials. Trials focused on a red bat species group (Lasiurus borealis and Lasiurus blossevillii; n = 46) and four species: cave myotis (Myotis velifer; n = 57), Brazilian free-tailed bats (Tadarida brasiliensis; n = 73), evening bats (Nycteceius humeralis; n = 53), and tricolored bats (Perimyotis subflavus; n = 17). The trials occurred during three treatment emissions: low (emissions from subarrays at 20, 26, and 32 kHz), high (emissions from subarrays at 38, 44, and 50 kHz), and combined (all six emission frequencies). We placed one wild-captured bat into the flight cage for each trial, which consisted of an acclimation period, a control period with the UD powered off, and the three emission treatments (order randomly selected), each interspersed with a control period. We tracked bat flight using four thermal cameras placed outside the flight cage. We quantified the effectiveness of each treatment by comparing the distances each bat flew from the UD during each treatment vs. the control period using quantile regression. Additionally, we conducted an exploratory analysis of differences between sex and season and sex within season using analysis of variance. Broadly, UDs were effective at altering the bats’ flight paths as they flew farther from the UD during treatments than during controls; however, results varied by species, sex, season, and sex within season. For the red bat group, bats flew farther from the UD during all treatments than during the control period at all percentiles (p < 0.001), and treatments were comparable in effectiveness. For cave myotis, all percentile distances were farther from the UD during each of the treatments than during the control, except the 90th percentile distance during high, and low was most effective. For evening bats and Brazilian free-tailed bats, results were inconsistent, but high and low were most effective, respectively. For tricolored bats, combined and low were significant at the 10th–75th percentiles, high was significant at all percentiles, and combined was most effective. Results suggest UDs may be an effective means of reducing bat mortalities due to wind turbine blade strikes. We recommend that continued research on UDs focus on low emission treatments, which have decreased sound attenuation and demonstrated effectiveness across the bat species evaluated in this study.

17 WIND ENERGY↗

Adaptive Laboratory Evolution for algal strain improvement: methodologies and applications

Originally developed in bacterial and fungal models, Adaptive Laboratory Evolution (ALE) experimentally selects for targeted phenotypes by allowing cells containing beneficial random genetic mutations to replicate more quickly in the presence of environmental stress. When applied to microalgae, this technique has been used to increase growth rate, stress tolerance, substrate utilization, and product yield. This review discusses ALE experimental design parameters, and summarizes key results and lessons learned from recent microalgal ALE studies. By documenting the current state of the field, we provide insight on the ways microalgal ALE might be used to develop commercially viable biotechnology strains.

09 BIOMASS FUELS↗

Linking Community‐Climate Disequilibrium to Ecosystem Function

Turnover in species composition often lags behind the pace of climate change, resulting in mismatches between climate and communities. However, the impact of these community‐climate disequilibria on ecosystem functions is rarely considered, and current methods for measuring disequilibria assume that species ranges were, until recently, in equilibrium with climate. Here, in this work, we develop a simple theoretical model to address both of these problems by linking community‐climate disequilibrium with ecosystem functioning. We show how disequilibrium can impair functioning in the near‐term even when climate change is expected to enhance functioning in the long‐term. Responses are most likely to change over time in communities where turnover is slow, the impact of disequilibrium counteracts the direct effects of climate on ecosystem function, and pre‐existing disequilibrium is large. These findings emphasise the importance of precise and unbiased estimates of community‐climate disequilibria for improving ecological forecasts. By fitting our model to time series of both climate and ecosystem function from a metacommunity simulation, we show the potential for community‐climate disequilibrium to be inferred without direct knowledge about species' distributions or climatic tolerances. We end by outlining a research agenda to apply dynamic disequilibrium concepts and test novel hypotheses across diverse ecosystems.

climate change↗

Rising CO 2 and warming reduce global canopy demand for nitrogen

Nitrogen (N) limitation has been considered as a constraint on terrestrial carbon uptake in response to rising CO 2 and climate change. By extension, it has been suggested that declining carboxylation capacity (V cmax ) and leaf N content in enhanced-CO 2 experiments and satellite records signify increasing N limitation of primary production. We predicted Vcmax using the coordination hypothesis and estimated changes in leaf-level photosynthetic N for 1982–2016 assuming proportionality with leaf-level V cmax at 25°C. The whole-canopy photosynthetic N was derived using satellite-based leaf area index (LAI) data and an empirical extinction coefficient for V cmax , and converted to annual N demand using estimated leaf turnover times. The predicted spatial pattern of V cmax shares key features with an independent reconstruction from remotely sensed leaf chlorophyll content. Predicted leaf photosynthetic N declined by 0.27% yr -1 , while observed leaf (total) N declined by 0.2–0.25% yr -1 . Predicted global canopy N (and N demand) declined from 1996 onwards, despite increasing LAI. Leaf-level responses to rising CO 2 , and to a lesser extent temperature, may have reduced the canopy requirement for N by more than rising LAI has increased it. This finding provides an alternative explanation for declining leaf N that does not depend on increasing N limitation.

54 ENVIRONMENTAL SCIENCES↗

Data and code for: "Soil respiration response to simulated precipitation change depends on ecosystem type and study duration"

Data, code, and figures from our meta-analysis of the effect of precipitation change on soil respiration, with an emphasis on ecosystem variability and the effect of study duration. We combined the results of 81 separate studies to determine the effect of altered rainfall on the amount of carbon that leaves soil in the form of carbon dioxide, a major component of global carbon cycling. In addition we looked at how long the changes lasted, and how different soil properties and the intensity of precipitation changes affected the study results. We found that more precipitation resulted in greater amounts of carbon dioxide leaving the soil, and less precipitation resulted in less. However, the changes weakened over time in ecosystems that typically receive plenty of rainfall (e.g., forests), in contrast to ecosystems that typically receive little rainfall (e.g., deserts) where changes strengthened over time. Changes in the amount of carbon dioxide that left the soil were also affected by the amount of carbon in the soil, which impacts how much water soil can hold.

54 ENVIRONMENTAL SCIENCES↗

Next-generation sequencing dataset of genome-scale CRISPRi in Synechococcus sp. PCC 7002 across seven conditions

A 33,298-member sgRNA library developed for Synechococus sp. PCC 7002 was screened with two replicates across seven growth conditions and sequenced with Illumina NextSeq (paired end, 2x150 bp) for a total of ~700M reads. The original plasmid library and the library after transformation into a dCas9-containing and dCas9-absent strain were also sequenced as a reference for initial sgRNA abundance.

genome- wide screens environmental acclimation spe↗