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At least 55 records · Page 3

The integral spliceosomal component CWC15 is required for development in Arabidopsis

Efficient mRNA splicing is a prerequisite for protein biosynthesis and the eukaryotic splicing machinery is evolutionarily conserved among species of various phyla. At its catalytic core resides the activated splicing complex Bact consisting of the three small nuclear ribonucleoprotein complexes (snRNPs) U2, U5 and U6 and the so-called NineTeen complex (NTC) which is important for spliceosomal activation. CWC15 is an integral part of the NTC in humans and it is associated with the NTC in other species. Here we show the ubiquitous expression and developmental importance of the Arabidopsis ortholog of yeast CWC15. CWC15 associates with core components of the Arabidopsis NTC and its loss leads to inefficient splicing. Consistent with the central role of CWC15 in RNA splicing, cwc15 mutants are embryo lethal and additionally display strong defects in the female haploid phase. Interestingly, the haploid male gametophyte or pollen in Arabidopsis, on the other hand, can cope without functional CWC15, suggesting that developing pollen might be more tolerant to CWC15-mediated defects in splicing than either embryo or female gametophyte.

59 BASIC BIOLOGICAL SCIENCES↗

Patch selection by bumble bees navigating discontinuous landscapes

Pollen and nectar resources are unevenly distributed over space and bees must make routing decisions when navigating patchy resources. Determining the patch selection process used by bees is crucial to understanding bee foraging over discontinuous landscapes. To elucidate this process, we developed four distinct probability models of bee movement where the size and the distance to the patch determined the attractiveness of a patch. A field experiment with a center patch and four peripheral patches of two distinct sizes and distances from the center was set up in two configurations. Empirical transition probabilities from the center to each peripheral patch were obtained at two sites and two years. The best model was identified by comparing observed and predicted transition probabilities, where predicted values were obtained by incorporating the spatial dimensions of the field experiment into each model’s mathematical expression. Bumble bees used both patch size and isolation distance when selecting a patch and could assess the total amount of resources available in a patch. Bumble bees prefer large, nearby patches. This information will facilitate the development of a predictive framework to the study of bee movement and of models that predict the movement of genetically engineered pollen in bee-pollinated crops.

54 ENVIRONMENTAL SCIENCES↗

Stress-regulated Arabidopsis GAT2 is a low affinity γ-aminobutyric acid transporter

Abstract The four-carbon non-proteinogenic amino acid γ-aminobutyric acid (GABA) accumulates to high levels in plants in response to various abiotic and biotic stress stimuli, and plays a role in C:N balance, signaling, and as a transport regulator. Expression in Xenopus oocytes and voltage-clamping allowed the characterization of Arabidopsis GAT2 (At5g41800) as a low affinity GABA transporter with a K0.5GABA ~8 mM. l-Alanine and butylamine represented additional substrates. GABA-induced currents were strongly dependent on the membrane potential, reaching the highest affinity and highest transport rates at strongly negative membrane potentials. Mutation of Ser17, previously reported to be phosphorylated in planta, did not result in altered affinity. In a short-term stress experiment, AtGAT2 mRNA levels were up-regulated at low water potential and under osmotic stress (polyethylene glycol and mannitol). Furthermore, AtGAT2 promoter activity was detected in vascular tissues, maturating pollen, and the phloem unloading region of young seeds. Even though this suggested a role for AtGAT2 in long-distance transport and loading of sink organs, under the conditions tested neither AtGAT2-overexpressing plants, atgat2 or atgat1 T-DNA insertion lines, nor atgat1 atgat2 doubleknockout mutants differed from wild-type plants in growth on GABA, amino acid levels, or resistance to salt and osmotic stress.

Plant Sciences↗

Photovoltaic Mini-Module Soiling Stations: Cooperative Research and Development (Final Report)

Photovoltaic (PV) panels can become soiled due to a variety of environmental factors. This soiling reduces the amount of light reaching the cells and thus their energy output. The ratio of actual output of the soiled device that of a clean device is known as the soiling ratio. The daily change in soiling ratio is known as the soiling rate. To study soiling for different glass coatings. Two soiling stations were fabricated. The stations were designed to monitor the short-circuit current of PV cells encapsulated with glass featuring different coatings and measure the daily soiling ratio with a pair of reference cells, one of which is automatically brushed daily. This work meets the need to understand how the glass coatings perform in additional environments. The stations are designed to quantify both the soiling ratio (the ratio between actual/expected photovoltaic panel power output) as well as differences in anti-soiling performance of different glass coatings. The design of the dirty/clean reference cell system is described by Toth et al. The soiling ratio is calculated by calculating the average irradiance measured within one hour of solar noon by each of the reference cells and then taking the ratio of these daily near-noon averages. The soiling ratio time series for the station deployed in Georgia is shown in Figure 1. This figure shows that the soiling ratio at the site has not yet fallen below 0.99, indicating that peak daily soiling losses have been below 1%. The site was chosen because it is thought to be affected by pollen soiling. With continued monitoring over the course of at least a full year, we expect to be able to observe and quantify pollen soiling events.

14 SOLAR ENERGY↗

VPS45 is required for both diffuse and tip growth of Arabidopsis thaliana cells

Introduction VPS45 belongs to the Sec1/Munc18 family of proteins, which interact with and regulate Qa-SNARE function during membrane fusion. We have shown previously that Arabidopsis thaliana VPS45 interacts with the SYP61/SYP41/VTI12 SNARE complex, which locates on the trans -Golgi network (TGN). It is required for SYP41 stability, and it functions in cargo trafficking to the vacuole and in cell expansion. It is also required for correct auxin distribution during gravitropism and lateral root growth. Results As vps45 knockout mutation is lethal in Arabidopsis, we identified a mutant, vps45-3 , with a point mutation in the VPS45 gene causing a serine 284-to-phenylalanine substitution. The VPS45-3 protein is stable and maintains interaction with SYP61 and SYP41. However, vps45-3 plants display severe growth defects with significantly reduced organ and cell size, similar to vps45 RNAi transgenic lines that have reduced VPS45 protein levels. Root hair and pollen tube elongation, both processes of tip growth, are highly compromised in vps45-3 . Mutant root hairs are shorter and thicker than those of wild-type plants, and are wavy. These root hairs have vacuolar defects, containing many small vacuoles, compared with WT root hairs with a single large vacuole occupying much of the cell volume. Pollen tubes were also significantly shorter in vps45-3 compared to WT. Discussion We thus show that VPS45 is essential for proper tip growth and propose that the observed vacuolar defects lead to loss of the turgor pressure needed for tip growth.

59 BASIC BIOLOGICAL SCIENCES↗

Chromoanagenesis from radiation-induced genome damage in Populus

Chromoanagenesis is a genomic catastrophe that results in chromosomal shattering and reassembly. These extreme single chromosome events were first identified in cancer, and have since been observed in other systems, but have so far only been formally documented in plants in the context of haploid induction crosses. The frequency, origins, consequences, and evolutionary impact of such major chromosomal remodeling in other situations remain obscure. Here, we demonstrate the occurrence of chromoanagenesis in poplar ( Populus sp . ) trees produced from gamma-irradiated pollen. Specifically, in this population of siblings carrying indel mutations, two individuals exhibited highly frequent copy number variation (CNV) clustered on a single chromosome, one of the hallmarks of chromoanagenesis. Using short-read sequencing, we confirmed the presence of clustered segmental rearrangement. Independently, we identified and validated novel DNA junctions and confirmed that they were clustered and corresponded to these rearrangements. Our reconstruction of the novel sequences suggests that the chromosomal segments have reorganized randomly to produce a novel rearranged chromosome but that two different mechanisms might be at play. Our results indicate that gamma irradiation can trigger chromoanagenesis, suggesting that this may also occur when natural or induced mutagens cause DNA breaks. We further demonstrate that such events can be tolerated in poplar, and even replicated clonally, providing an attractive system for more in-depth investigations of their consequences.

59 BASIC BIOLOGICAL SCIENCES↗

Wild bees and natural enemies prefer similar flower species and respond to similar plant traits

Designing wildflower habitats to support beneficial insects providing pollination and pest control services is important for supporting sustainable crop production. It is often desirable to support both groups of insects, making the selection of resource plants for insect conservation programs more challenging. Moreover, the process of selecting resource plants is complicated by the array of possible options in each region, and the need to provide resources over the entire growing season. Identifying traits shared by resource plants that are attractive to both bees and natural enemies can reduce the need to evaluate new plants in each region, by providing a guide for the types of plants expected to be rewarding to these insects. Using insect visitation data collected from replicated common garden plantings of native wildflower and shrub species from the Great Lakes region of the United States, we found a high degree of correlation between the abundance of bees and natural enemies visiting native plant species. These results were used to identify a set of 15 plant species that can provide resources for these insects throughout the summer. Across all tested species, pollen quantity per flower and the week of bloom were positively correlated with some, but not all, taxonomic groupings of beneficial insects. In contrast, floral area was consistently positively associated with visitation of both natural enemies and wild bees. This trait is easy to document and can allow for efficient local testing of potential resource plants, providing a faster path to implementing insect conservation in working landscapes.

54 ENVIRONMENTAL SCIENCES↗

Climate-eutrophication-anoxia interactions in Late Glacial Soppensee, Switzerland: Forcings, non-linear responses and recovery

Combined effects of climate warming and anthropogenic nutrient loadings lead to lake eutrophication and anoxia globally. Because of chemical feedbacks, lakes under multiple stressors often respond in non-linear ways. However, it remains unclear whether climate change alone can lead to non-linear lake responses in the absence of anthropogenic nutrient disturbances. Here, we investigate the interactions between climate variability, nutrient cycling and trophic state changes, mixing regimes, anoxia and related chemical feedback in a small kettle-hole lake in Switzerland during Late Glacial times (15.2–12.6 cal ka BP), a period known for high-amplitude climate change in pre-anthropogenic times. After its formation during Heinrich Stadial 1 (>15 cal ka BP), Soppensee was oligotrophic and well-mixed. Soppensee became eutrophic and developed anoxia at 14.25 cal ka BP. Phosphorus (P) was released from sediments through the reductive dissolution of Fe-oxyhydroxides, fuelling eutrophication. Eutrophication lagged the Bølling warming (14.65 cal ka BP) by 400 years, suggesting that rising temperatures were not the trigger for eutrophication. Instead, eutrophication responded non-linearly to forest closure (threshold at 76 % arboreal pollen AP), which shielded Soppensee from wind mixing, enhancing lake stratification, anoxia and P release, intensifying eutrophication. These conditions ended during the 200-years cold period of the Aegelsee Oscillation (GI-1d, ca. 14.0 cal ka BP) when the landscape regionally opened (AP<76 %); the lake became well-mixed, oxygenated and P was efficiently sequestered. Throughout the Allerød (13.9–12.8 cal ka BP), enhanced Fe input prompted diagenetic vivianite formation, sequestering P in sediments, naturally remediating lake eutrophication despite closed forests, warm temperatures, lake stratification and anoxia.

Environmental sciences↗

Chemical Imaging of Fine Mode Atmospheric Particles Collected from a Research Aircraft over Agricultural Fields

Vegetation and crops in agricultural areas are a significant source of both biogenic secondary organic aerosols and of directly emitted primary biological atmospheric particles (PBAPs) composed of plant fragments/debris, fungal spores, pollen, and bacteria among others. Seasonal crop harvesting significantly intensifies regional PBAP emissions and aerosolizes soil organic matter and dust particulates. In this paper, the atmospheric contributions and impact of PBAPs have been insufficiently investigated due to limited real-time observations and ambiguity in their detection by existing measurement techniques. We collected ambient particles on-board an instrumented Beechcraft BE76 Duchess research aircraft flying above actively harvested agriculture fields of corn and soybean crops. In this work, particle samples were analyzed using complementary chemical imaging techniques, including computer-controlled scanning electron microscopy coupled with elemental microanalysis to describe particle-type population and scanning transmission X-ray microscopy to provide quantitative metrics on particle internal heterogeneity. We show the particle-class contribution is consistent between corn and soybean harvesting across altitudes of 100–300 m AGL. The contribution of PBAPs in the size range of 0.15–1.25 μm is estimated to be 10–12% of ~39,000 analyzed particles. Determining viscosity with X-ray microanalysis has shown that the fine-mode organic particles collected are viscous/semisolid (10 2 –10 12 Pa s) while the majority of PBAP fragments are solid (>10 12 Pa s). The observation of submicrometer, solid carbonaceous fragments of biogenic origin has implications for the regional cloud condensation nuclei and ice nuclei budget. Therefore, seasonal harvesting of crops may play an important, yet unrecognized, role in regional cloud formation and climate.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

A neotropical perspective on the uniqueness of the Holocene among interglacials

Abstract Understanding how tropical systems have responded to large-scale climate change, such as glacial-interglacial oscillations, and how human impacts have altered those responses is key to current and future ecology. A sedimentary record recovered from Lake Junín, in the Peruvian Andes (4085 m elevation) spans the last 670,000 years and represents the longest continuous and empirically-dated record of tropical vegetation change to date. Spanning seven glacial-interglacial oscillations, fossil pollen and charcoal recovered from the core showed the general dominance of grasslands, although during the warmest times some Andean forest trees grew above their modern limits near the lake. Fire was very rare until the last 12,000 years, when humans were in the landscape. Here we show that, due to human activity, our present interglacial, the Holocene, has a distinctive vegetation composition and ecological trajectory compared with six previous interglacials. Our data reinforce the view that modern vegetation assemblages of high Andean grasslands and the presence of a defined tree line are aspects of a human-modified landscape.

Science & Technology - Other Topics↗

A 2-million-year-old ecosystem in Greenland uncovered by environmental DNA

Late Pliocene and Early Pleistocene epochs 3.6 to 0.8 million years ago had climates resembling those forecasted under future warming. Palaeoclimatic records show strong polar amplification with mean annual temperatures of 11–19 °C above contemporary values. The biological communities inhabiting the Arctic during this time remain poorly known because fossils are rare. Here we report an ancient environmental DNA (eDNA) record describing the rich plant and animal assemblages of the Kap København Formation in North Greenland, dated to around two million years ago. The record shows an open boreal forest ecosystem with mixed vegetation of poplar, birch and thuja trees, as well as a variety of Arctic and boreal shrubs and herbs, many of which had not previously been detected at the site from macrofossil and pollen records. The DNA record confirms the presence of hare and mitochondrial DNA from animals including mastodons, reindeer, rodents and geese, all ancestral to their present-day and late Pleistocene relatives. The presence of marine species including horseshoe crab and green algae support a warmer climate than today. The reconstructed ecosystem has no modern analogue. The survival of such ancient eDNA probably relates to its binding to mineral surfaces. Our findings open new areas of genetic research, demonstrating that it is possible to track the ecology and evolution of biological communities from two million years ago using ancient eDNA.

58 GEOSCIENCES↗

A native IgE in complex with profilin provides insights into allergen recognition and cross-reactivity

Allergies have become a rising health problem, where plentiful substances can trigger IgE-mediated allergies in humans. While profilins are considered minor allergens, these ubiquitous proteins are primary molecules involved in cross-reactivity and pollen-food allergy syndrome. Here we report the first crystal structures of murine Fab/IgE, with its chains naturally paired, in complex with the allergen profilin from Hevea brasiliensis (Hev b 8). The crystallographic models revealed that the IgE’s six complementarity-determining regions (CDRs) interact with the allergen, comprising a rigid paratope-epitope surface of 926 Å 2 , which includes an extensive network of interactions. Interestingly, we also observed previously unreported flexibility at Fab/IgE’s elbow angle, which did not influence the shape of the paratope. The Fab/IgE exhibits a high affinity for Hev b 8, even when using 1 M NaCl in BLI experiments. Finally, based on the encouraging cross-reactivity assays using two mutants of the maize profilin (Zea m 12), this antibody could be a promising tool in IgE engineering for diagnosis and research applications.

59 BASIC BIOLOGICAL SCIENCES↗

Spatially co-registered wide-field nonlinear optical imaging of living and complex biosystems in a total internal reflection geometry

Nonlinear optical microscopy that leverages an objective based total internal reflection (TIR) excitation scheme is an attractive means for rapid, wide-field imaging with enhanced surface sensitivity. Through select combinations of distinct modalities, one can, in principle, access complementary chemical and structural information for various chemical species near interfaces. Here, we report a successful implementation of such a wide-field nonlinear optical microscope system, which combines coherent anti-Stokes Raman scattering (CARS), two-photon fluorescence (TPF), second harmonic generation (SHG), and sum frequency generation (SFG) modalities on the same platform. The intense optical fields needed to drive these high order nonlinear optical processes are achieved through the use of femtosecond pulsed light in combination with the intrinsic field confinement induced by TIR over a large field of view. The performance of our multimodal microscope was first assessed through the experimental determination of its chemical fidelity, intensity and polarization dependences, and spatial resolution using a set of well-defined model systems. Subsequently, its unique capabilities were validated through imaging complex biological systems, including Hydrangea quercifolia pollen grains and Pantoea sp. YR343 bacterial cells. Specifically, the spatial distribution of different molecular groups in the former was visualized via vibrational contrast mechanisms of CARS, whereas co-registered TPF imaging allowed the identification of spatially localized intrinsic fluorophores. We further demonstrate the feasibility of our microscope for wide-field CARS imaging on live cells through independent characterization of cell viability using spatially co-registered TPF imaging. Overall, this approach to TIR enabled wide-field imaging is expected to provide new insights into bacterial strains and their interactions with other species in the rhizosphere in a time-resolved and chemically selective manner.

47 OTHER INSTRUMENTATION↗

Corona splashing triggered by a loose monolayer of particles

In nature, high-speed raindrops often impact and spread on particulate surfaces (e.g., soil, plant leaves with spores or pollen). We study the dynamics of droplet impact on a loosely packed monolayer of particles by combining experimental and mathematical approaches. We find that the presence of mobile particles lowers the critical impact velocity at which the droplet exhibits corona splashing, as the particle area fraction is systematically increased. We rationalize this experimental observation by considering the jamming of frictional particles at the spreading rim. Elucidating the splashing transition of the drop on a particulate bed can lead to a better understanding of soil loss and erosion from falling raindrops

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

SEED LIPID DROPLET PROTEIN1, SEED LIPID DROPLET PROTEIN2, and LIPID DROPLET PLASMA MEMBRANE ADAPTOR mediate lipid droplet–plasma membrane tethering

Membrane contact sites (MCSs) are interorganellar connections that allow for the direct exchange of molecules, such as lipids or Ca 2+ between organelles, but can also serve to tether organelles at specific locations within cells. Here, we identified and characterized three proteins of Arabidopsis thaliana that form a lipid droplet (LD)–plasma membrane (PM) tethering complex in plant cells, namely LD-localized SEED LD PROTEIN (SLDP) 1 and SLDP2 and PM-localized LD-PLASMA MEMBRANE ADAPTOR (LIPA). Using proteomics and different protein–protein interaction assays, we show that both SLDPs associate with LIPA. Disruption of either SLDP1 and SLDP2 expression, or that of LIPA, leads to an aberrant clustering of LDs in Arabidopsis seedlings. Ectopic co-expression of one of the SLDPs with LIPA is sufficient to reconstitute LD–PM tethering in Nicotiana tabacum pollen tubes, a cell type characterized by dynamically moving LDs in the cytosolic streaming. Furthermore, confocal laser scanning microscopy revealed both SLDP2.1 and LIPA to be enriched at LD–PM contact sites in seedlings. These and other results suggest that SLDP and LIPA interact to form a tethering complex that anchors a subset of LDs to the PM during post-germinative seedling growth in Arabidopsis.

59 BASIC BIOLOGICAL SCIENCES↗

A seed-like proteome in oil-rich tubers

There are numerous examples of plant organs or developmental stages that are desiccation-tolerant and can withstand extended periods of severe water loss. One prime example are seeds and pollen of many spermatophytes. However, in some plants, also vegetative organs can be desiccation-tolerant. One example are the tubers of yellow nutsedge ( Cyperus esculentus ), which also store large amounts of lipids similar to seeds. Interestingly, the closest known relative, purple nutsedge ( Cyperus rotundus ), generates tubers that do not accumulate oil and are not desiccation-tolerant. We generated nanoLC-MS/MS-based proteomes of yellow nutsedge in five replicates of four stages of tuber development and compared them to the proteomes of roots and leaves, yielding 2257 distinct protein groups. Our data reveal a striking upregulation of hallmark proteins of seeds in the tubers. A deeper comparison to the tuber proteome of the close relative purple nutsedge ( C. rotundus ) and a previously published proteome of Arabidopsis seeds and seedlings indicates that indeed a seed-like proteome was found in yellow but not purple nutsedge. This was further supported by an analysis of the proteome of a lipid droplet-enriched fraction of yellow nutsedge, which also displayed seed-like characteristics. One reason for the differences between the two nutsedge species might be the expression of certain transcription factors homologous to ABSCISIC ACID INSENSITIVE3, WRINKLED1, and LEAFY COTYLEDON1 that drive gene expression in Arabidopsis seed embryos.

59 BASIC BIOLOGICAL SCIENCES↗

PV O&M Cost Model [SWR-21-14]

This tool presents a method for calculating costs associated with the operation and maintenance (O&M) of photovoltaic (PV) systems. The tool compiles details regarding the cost and frequency of multiple O&M services to estimate annual O&M costs ($/year) for each year of an analysis period, the net present value ($) of life cycle costs accumulated over the analysis period, and the reserve account amount ($) that might be required to fund unexpected repairs. The reserve account includes parts inventory and is important for providing a source of funds to make repairs quickly and avoid lost production. This method allows a detailed selection of services to perform based on system size, market served (e.g., residential, commercial, or utility), type and configuration of system components (e.g., micro-, string, or central inverter), and site and environmental conditions (e.g., pollen, bird populations) which is an improvement over simple per unit valuations of O&M costs ($/kW/year). This model also distinguishes costs that vary from year to year and increase at different rates over time as modeled by heuristic failure distributions (e.g., Weibull or Lognormal distribution) based on actuarial data for many of the services.

Walker, Andy↗

Model of Operation-and-Maintenance Costs for Photovoltaic Systems

This article presents a method for calculating costs associated with operation and maintenance (O&M) of photovoltaic (PV) systems. It compiles details regarding the cost and frequency of multiple O&M services to estimate annual O&M costs ($\$$/year) for each year of an analysis period, the net present value ($\$$) of life cycle costs accumulated over the analysis period, and the reserve account amount ($\$$). Here we show that this method is an improvement over the previous averaged or levelized per-unit ($\$$/kW/year) valuations for estimating PV O&M costs, because it allows a detailed selection of services to perform based on system size, market served (e.g., residential, commercial, or utility), type and configuration of system components (e.g., micro-, string or central inverter) and site and environmental conditions (e.g., snow, pollen, bird populations). This model also distinguishes costs that vary from year to year and increase at different rates over time because of heuristic failure distributions (e.g. Weibull or Lognormal distribution) based on actuarial data for many of the services. This cost model was created by the PV O&M Working Group of researchers and industry, sponsored by DOE Solar Energy Technologies Office, and has been published in an on-line version hosted by SunSpec Alliance at apsuite.sunspec.org. A spreadsheet version is included with this paper as supplementary material.

14 SOLAR ENERGY↗