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At least 145 records · Page 8

Overexpression of phospholipid: diacylglycerol acyltransferase in Brassica napus results in changes in lipid metabolism and oil accumulation

The regulation of lipid metabolism in oil seeds is still not fully understood and increasing our knowledge in this regard is of great economic, as well as intellectual, importance. Oilseed rape (Brassica napus) is a major global oil crop where increases in triacylglycerol (TAG) accumulation have been achieved by overexpression of relevant biosynthetic enzymes. In this study, we expressed Arabidopsis phospholipid: diacylglycerol acyltransferase (PDAT1), one of the two major TAG-forming plant enzymes in B. napus DH12075 to evaluate its effect on lipid metabolism in developing seeds and to estimate its flux control coefficient. Despite several-fold increase in PDAT activity, seeds of three independently generated PDAT transgenic events showed a small but consistent decrease in seed oil content and had altered fatty acid composition of phosphoglycerides and TAG, towards less unsaturation. Mass spectrometry imaging of seed sections confirmed the shift in lipid compositions and indicated that PDAT overexpression altered the distinct heterogeneous distributions of phosphatidylcholine (PC) molecular species. Similar, but less pronounced, changes in TAG molecular species distributions were observed. Our data indicate that PDAT exerts a small, negative, flux control on TAG biosynthesis and could have under-appreciated effects in fine-tuning of B. napus seed lipid composition in a tissue-specific manner. This has important implications for efforts to increase oil accumulation in similar crops.

Biochemistry & Molecular Biology↗

Metabolic engineering of p –hydroxybenzoate in poplar lignin

Ester-linked p-hydroxybenzoate occurs naturally in poplar lignin as pendent groups that can be released by mild alkaline hydrolysis. These ‘clip-off’ phenolics can be separated from biomass and upgraded into diverse high-value bioproducts. We introduced a bacterial chorismate pyruvate lyase gene into transgenic poplar trees with the aim of producing more p-hydroxybenzoate from chorismate, itself a metabolic precursor to lignin. By driving heterologous expression specifically in the plastids of cells undergoing secondary wall formation, this strategy achieved a 50% increase in cell-wall-bound p-hydroxybenzoate in mature wood and nearly 10 times more in developing xylem relative to control trees. Comparable amounts also remained as soluble p-hydroxybenzoate-containing xylem metabolites, pointing to even greater engineering potential. Mass spectrometry imaging showed that the elevated p-hydroxybenzoylation was largely restricted to the cell walls of fibres. Finally, transgenic lines outperformed control trees in assays of saccharification potential. Here, this study highlights the biotech potential of cell-wall-bound phenolate esters and demonstrates the importance of substrate supply in lignin engineering.

4-hydroxybenzoic acid↗

Fungal organic acid uptake of mineral-derived K is dependent on distance from carbon hotspot

Fungal mineral weathering regulates the bioavailability of inorganic nutrients from mineral surfaces to organic matter and increase the bioavailable fraction of nutrients. Such weathering strategies are classified as biomechanical or biochemical. In the case of fungal uptake of mineral nutrients through biochemical weathering, it is widely hypothesized that uptake of inorganic nutrients occurs through organic acid chelation, but such processes have not been directly visualized. This is in part due to challenges in probing the complex and heterogeneous soil environment. Here, using an epoxy-based, mineral-doped soil micromodel platform, which emulates soil mineralogy and porosity, we visualize the molecular mechanisms of mineral weathering. Mass spectrometry imaging revealed differences in the distribution of fungal exudates, citric acid, and tartaric acid on the soil micromodels in presence of minerals. Citric acid was detected closer to the nutrient-rich inoculation point, whereas tartaric acid was highly abundant away from inoculation point. This suggested that the organic acid exuded by the fungi depended on the proximity from the carbon-rich organic substrate at the point of inoculation. Using a combination of X-ray fluorescence and X-ray near edge structure analysis, we identified citric acid- and tartaric acid-bound K within fungal hyphae networks grown in the presence of minerals. Combined, our results provide direct evidence that fungi uptake and transport mineral derived nutrient organic acid chelation. The results of this study provided unprecedented visualization of fungal uptake and transport of K + , while resolving the indirect weathering mechanism of fungal K uptake from mineral interfaces.

59 BASIC BIOLOGICAL SCIENCES↗

Visualizing Microbial Community Dynamics via a Controllable Soil Environment

Visualizing microbial community dynamics within soil environments remains challenging. Here, we developed a custom-built SoilBox system that facilitates probing various changes occurring in microbial community development and molecular processes when exposed to environmental perturbations (e.g., temperature changes, drought, nutrient availability). Using confocal microscopy along with mass spectrometry imaging, we demonstrate the power of this system by correlating microbial growth and spatial metabolic information from soil communities under different moisture regimes.

59 BASIC BIOLOGICAL SCIENCES↗

Protein N -Glycans in Healthy and Sclerotic Glomeruli in Diabetic Kidney Disease

Diabetes is expected to directly affect renal glycosylation; yet to date, there has not been a comprehensive evaluation of alterations in N-glycan composition in the glomeruli of patients with diabetic kidney disease (DKD). Here, we used untargeted mass spectrometry imaging to identify N-glycan structures in healthy and sclerotic glomeruli in formalin-fixed paraffin-embedded sections from needle biopsies of five patients with DKD and three healthy kidney samples. Regional proteomics was performed on glomeruli from additional biopsies from the same patients to compare the abundances of enzymes involved in glycosylation. Secondary analysis of single-nucleus RNA sequencing (snRNAseq) data were used to inform on transcript levels of glycosylation machinery in different cell types and states. We detected 120 N-glycans, and among them, we identified 12 of these protein post-translated modifications that were significantly increased in glomeruli. All glomeruli-specific N-glycans contained an N-acetyllactosamine epitope. Five N-glycan structures were highly discriminant between sclerotic and healthy glomeruli. Sclerotic glomeruli had an additional set of glycans lacking fucose linked to their core, and they did not show tetra-antennary structures that were common in healthy glomeruli. Orthogonal omics analyses revealed lower protein abundance and lower gene expression involved in synthesizing fucosylated and branched N-glycans in sclerotic podocytes. In snRNAseq and regional proteomics analyses, we observed that genes and/or proteins involved in sialylation and N-acetyllactosamine synthesis were also downregulated in DKD glomeruli, but this alteration remained undetectable by our spatial N-glycomics assay. Integrative spatial glycomics, proteomics, and transcriptomics revealed protein N-glycosylation characteristic of sclerotic glomeruli in DKD.

60 APPLIED LIFE SCIENCES↗

Spatial mapping of plant N-glycosylation cellular heterogeneity inside soybean root nodules provided insights into legume-rhizobia symbiosis

Although ubiquitously present, information on the function of complex N-glycan posttranslational modification in plants is very limited and it is often neglected. In this work, we adopted an enzyme-assisted matrix-assisted laser desorption/ionization mass spectrometry imaging strategy to visualize the distribution and identity of N-glycans in soybean root nodules at a cellular resolution. We additionally performed proteomics analysis to probe the potential correlation to proteome changes during symbiotic rhizobia-legume interactions. Our ion images reveal that intense N-glycosylation occurs in the sclerenchyma layer, and inside the infected cells within the infection zone, while morphological structures such as the cortex, uninfected cells, and cells that form the attachment with the root are less N-glycosylated. Notably, we observed different N-glycan profiles between soybean root nodules infected with wild type rhizobia and those infected with mutant rhizobia incapable of efficiently fixing atmospheric nitrogen. The majority of complex N-glycan structures, particularly those with characteristic Lewis-a epitopes, are overexpressed in the mutant nodules. Our proteomic results revealed that these glycans likely originated from proteins that maintain the redox balance crucial for proper nitrogen fixation, but also from enzymes involved in N-glycan and phenylpropanoid biosynthesis. These findings indicate possible involvement of Lewis-a glycans in these critical pathways during legume-rhizobia symbiosis.

59 BASIC BIOLOGICAL SCIENCES↗

High spectral resolution remote sensing of the land

Spectral remote sensing has been practiced on a large scale since the launch of Landsat 1 in 1972. The limited information contained in this spectrally undersampled data set has led to the development of sophisticated statistical-inferential methods for data analysis. The results are usually limited by the availability of ground truth information. Recent technological developments have made it feasible to create narrow-band, contiguous, spectral image data sets that make possible the identification of surface cover materials based on the complete reflectance spectrum for each picture element. This capability will revolutionize the use of remote sensing data and require new deterministic image processing techniques to extract the full information content from the data. Sensors, based on the concept of imaging spectrometry and the new technology of area array infrared detectors, have been constructed and are candidates for Shuttle and space platform flights.

Goetz, A. F. H.↗

Remote sensing of coastal wetlands

Various aircraft and satellite sensors for detecting and mapping wetlands properties are examined. The uses of color IR photography to map coastal vegetation, and of Landsat MSS and TM and SPOT data to quantify biomass and productivity for large wetland areas are discussed. For spectral estimation of biomass and productivity, the relation between radiance and biomass needs to be studied; the quantity and orientation of dead biomass and the amount of soil reflectance in comparison with vegetation reflectance in a given target area affect the spectral estimation of biomass. The radiometric evaluation of brackish wetland, and remote sensing in mangroves are described. The collection of images in narrow, contiguous spectral band using imaging spectrometry is considered.

Hardisky, M. A.↗

Spaceborne imaging radar on EOS

A multiparameter synthetic aperture imaging radar is planned as a facility instrument for the Earth Orbiting System (EOS). This sensor will operate at L, C, and X and at all possible polarizations (HH, VV, HV, VH), thus allowing the acquisition of detailed information about the surface physical and electrical properties. When combined with the visible and IR imaging spectrometry data and the surface topography, a full description of the surface structure, composition, thermal properties and physical properties could then be extracted.

Elachi, Charles↗

Concurrent Image Processing Executive (CIPE). Volume 1: Design overview

The design and implementation of a Concurrent Image Processing Executive (CIPE), which is intended to become the support system software for a prototype high performance science analysis workstation are described. The target machine for this software is a JPL/Caltech Mark 3fp Hypercube hosted by either a MASSCOMP 5600 or a Sun-3, Sun-4 workstation; however, the design will accommodate other concurrent machines of similar architecture, i.e., local memory, multiple-instruction-multiple-data (MIMD) machines. The CIPE system provides both a multimode user interface and an applications programmer interface, and has been designed around four loosely coupled modules: user interface, host-resident executive, hypercube-resident executive, and application functions. The loose coupling between modules allows modification of a particular module without significantly affecting the other modules in the system. In order to enhance hypercube memory utilization and to allow expansion of image processing capabilities, a specialized program management method, incremental loading, was devised. To minimize data transfer between host and hypercube, a data management method which distributes, redistributes, and tracks data set information was implemented. The data management also allows data sharing among application programs. The CIPE software architecture provides a flexible environment for scientific analysis of complex remote sensing image data, such as planetary data and imaging spectrometry, utilizing state-of-the-art concurrent computation capabilities.

Lee, Meemong↗

Wide-Angle, Reflective Strip-Imaging Camera

Proposed camera images thin, striplike portion of field of view of 180 degrees wide. Hemispherical concave reflector forms image onto optical fibers, which transfers it to strip of photodetectors or spectrograph. Advantages include little geometric distortion, achromatism, and ease of athermalization. Uses include surveillance of clouds, coarse mapping of terrain, measurements of bidirectional reflectance distribution functions of aerosols, imaging spectrometry, oceanography, and exploration of planets.

Vaughan, Arthur H.↗

HIRIS, the instrument and its science

The High Resolution Imaging Spectrometer (HIRIS) is a facility instrument slated for flight on the second EOS series AM platforms. HIRIS is designed to acquire 24 km wide, 30 m pixel images in 192 spectral bands simultaneously in the 0.4-2.45 micron wavelength region. With pointing mirrors it can sample any place on Earth, except the poles, every 2 days. HIRIS operates at the intermediate scale between the human and the global and therefore links studies of Earth surface processes to global monitoring carried out by lower resolution instruments. So far, over 50 science data products from HIRIS images have been identified in the fields of atmospheric gases, clouds, snow and ice, water, vegetation, and rocks and soils. The key attribute of imaging spectrometry that makes it possible to derive quantitative information from the data is the large number of contiguous, spectral bands. Therefore, spectrum-matching techniques can be applied. Such techniques are not possible with present-day, multispectral scanner data.

Goetz, Alexander F. H.↗

Use of AVIRIS data to the definition of optimised specifications for land applications with future spaceborne imaging spectrometers

Recent experience with airborne imaging spectrometers demonstrated the advantages of narrow band sensors over broad band scanners for characterizing the nature, extent, and physical status of typical land surfaces. Information on key spectral features associated with various land surfaces can be obtained from the data of such instruments, which can be used to simulate spaceborne imaging spectrometer data and to assess their information content if comprehensive underpinning is provided by ground data. The collection of such information was an issue of airborne imaging spectrometer campaigns like the NASA MAC-Europe 1991. Airborne and ground data obtained from different test sites in Europe are utilized for a comparative analysis of the spectral signatures of various land surfaces (vegetation, bare soils and rocks, and mixed soil/rock-vegetation) as seen from different imaging spectrometers like Airborne Visible/Infrared Imaging Spectrometer (AVIRIS), GERIS 63 band scanner, and CASI. The following items are discussed: (1) the significance of different spectral regions within the wavelength interval between 0.4 m and 2.5 m for the differentiation of different land units; (2) recommendations on the optimum band selection and band-widths to be used for the application of future satellite-based imaging spectrometers for land applications; (3) the boundaries for the detection of plant features in mixed-soil plant spectra and the influence of different soil properties on the mixture of the spectra; (4) recommendations on the optimum spatial resolution and recording dates for the discrimination of spectral features of various surface types; and (5) evaluation of different data compressing techniques for the optimum extraction of spectral information from imaging spectrometry data.

Bodechtel, J.↗

Using dark current data to estimate AVIRIS noise covariance and improve spectral analyses

Starting in 1994, all AVIRIS data distributions include a new product useful for quantification and modeling of the noise in the reported radiance data. The 'postcal' file contains approximately 100 lines of dark current data collected at the end of each data acquisition run. In essence this is a regular spectral-image cube, with 614 samples, 100 lines and 224 channels, collected with a closed shutter. Since there is no incident radiance signal, the recorded DN measure only the DC signal level and the noise in the system. Similar dark current measurements, made at the end of each line are used, with a 100 line moving average, to remove the DC signal offset. Therefore, the pixel-by-pixel fluctuations about the mean of this dark current image provide an excellent model for the additive noise that is present in AVIRIS reported radiance data. The 61,400 dark current spectra can be used to calculate the noise levels in each channel and the noise covariance matrix. Both of these noise parameters should be used to improve spectral processing techniques. Some processing techniques, such as spectral curve fitting, will benefit from a robust estimate of the channel-dependent noise levels. Other techniques, such as automated unmixing and classification, will be improved by the stable and scene-independence noise covariance estimate. Future imaging spectrometry systems should have a similar ability to record dark current data, permitting this noise characterization and modeling.

Boardman, Joseph W.↗

Using foreground/background analysis to determine leaf and canopy chemistry

Spectral Mixture Analysis (SMA) has become a well established procedure for analyzing imaging spectrometry data, however, the technique is relatively insensitive to minor sources of spectral variation (e.g., discriminating stressed from unstressed vegetation and variations in canopy chemistry). Other statistical approaches have been tried e.g., stepwise multiple linear regression analysis to predict canopy chemistry. Grossman et al. reported that SMLR is sensitive to measurement error and that the prediction of minor chemical components are not independent of patterns observed in more dominant spectral components like water. Further, they observed that the relationships were strongly dependent on the mode of expressing reflectance (R, -log R) and whether chemistry was expressed on a weight (g/g) or are basis (g/sq m). Thus, alternative multivariate techniques need to be examined. Smith et al. reported a revised SMA that they termed Foreground/Background Analysis (FBA) that permits directing the analysis along any axis of variance by identifying vectors through the n-dimensional spectral volume orthonormal to each other. Here, we report an application of the FBA technique for the detection of canopy chemistry using a modified form of the analysis.

Pinzon, J. E.↗

Extraction of ozone and chlorophyll-A distribution from AVIRIS data

The potential of airborne imaging spectrometry for assessing and monitoring natural resources is studied. Therefore, an AVIRIS scene of the NASA's MacEurope 1991 campaign - acquired in Central Switzerland - is used. The test site consists of an urban area, the Lake Zug with its surrounding fields, the Rigi mountain in the center of the test site, and the Lake of Four Cantons. The region is covered by the AVIRIS flight #910705, run 6 and 7 of the NASA ER-2 aircraft resulting in an average nominal pixel size of about 18 m. Simultaneous to the ER-2 overflight spectroradiometric measurements have been taken in various locations. Preselected reference targets were measured in the field with a GER Mark V spectroradiometer, and radiance measurements were taken to the lake using a Li-Cor LI 1800UW specroradiometer below and above the water surface. A comprehensive meteorological data set was obtained by joining the POLLUMET experiment which carried out measurements to investigate the summer smog in Switzerland on the same day. The quality assessment for the actual data set can be found in detail in Meyer et al. A parametric approach calculating the location of the airplane was used to simulate the observation geometry. This parametric preprocessing procedure, which takes care of effects of flight line and attitude variations as well as the pixel-by-pixel topographic corrections is described in Meyer.

Schaepman, M.↗

Predicting Photosynthetic Fluxes from Spectral Reflectance of Leaves and Canopies

The central hypothesis of this study has been that photosynthetic efficiency and capacity can be predicted from 'physiological reflectance indices' derived from spectral reflectance of leaves and canopies. I have approached this topic with a combination of laboratory and field experiments, and have also explored the potential of deriving a meaningful physiological index from imaging spectrometry (e.g. AVIRIS). A few highlights are presented below. The main emphasis has been on the 'Photochemical Reflectance Index' (PRI), derived from reflectance at 531 nm and 570 nm. Unlike most 'conventional' vegetation indices (e.g. NDVI), PRI changes rapidly both with illumination and physiological state, because it detects the interconversion of xanthophyll cycle pigments, which serve as photoregulatory pigments and control energy distribution for the photosynthetic system. This approach has differed dramatically from most remote sensing in that it has emphasized temporal variation in narrow-band spectral signatures, instead of spatial patterns of broadband indices. Our primary conclusion has been that PRI works well as an index of photosynthetic light-use efficiency at the leaf scale, much in the same way as the fluorescence index DeltaF/Fm. However, unlike DeltaF/Fm which must be measured at close scales, PRI can be sampled at a range of spatial scales, presenting the possibility of monitoring photosynthetic fluxes remotely.

Gamon, John A.↗

Surveying Dead Trees and CO2-Induced Stressed Trees Using AVIRIS in the Long Valley Caldera

Since 1980 the Long Valley Caldera in the eastern Sierra Nevada (California) has shown signs of renewed volcanic activity. Frequent earthquakes, a re-inflation of the caldera, hydrothermal activity and gas emissions are the outer symptoms of this renewed activity. In 1990 and 1991 several areas of dying trees were found around Mammoth Mountain. The cause of the die off of the trees was first sought in the persistent drought in the preceding years. However, the trees died regardless of age and species. Farrar et al. (1995) started a soil-gas survey in 1994 in the dead-tree areas and found carbon dioxide concentrations ranging from 30 to 96% at soil depths between 30 and 60 cm. CO2 concentrations in the atmosphere are usually around 0.03% and in the soil profile CO2 levels do commonly not exceed 4 to 5%. Although not much is known about the effect of high levels of carbon dioxide in the soil profile on roots, it is most likely that the trees are dying due to oxygen deprivation: the CO2 drives the oxygen out of the soil. So far, four sites of dead trees have been mapped around Mammoth Mountain. The two largest dying trees sites are located near Horseshoe Lake and near Mammoth Mountain Main Lodge covering approximately an area of 10 and 8 ha respectively. Analysis of the gas composition regarding the He-3/He-4 ratio and the percentage biogenic carbon reveals the source of the gas: the magma body beneath the Long Valley Caldera. Until recently it was not known that volcanoes release abundant carbon dioxide from their flanks as diffuse soil emanations. As a result of the magma gas emission around Mammoth Mountain there is an excellent sequence of dead trees, stressed trees, healthy trees and bare soil surfaces. This research site provides excellent opportunities to: (1) Study the capabilities of imaging spectrometry to map stressed (and dead) pine and fir species; (2) Study methods to separate the vivid vegetation, stressed vegetation and dead vegetation from the soil background of glacial deposits and crystalline rocks. The dead tree areas are located on the flanks of Mammoth Mountain (N:37 deg 37' 45" and W:119 deg 02' 05") at an elevation between 2600 and 3000 meters. The area is covered by an open type of Montane Forest. The dominant tree species are Lodgepole Pine (Pinus contorta), the Red Fir (Abies magnifica) and the Jeffrey Pine (Pinus jeffreyi). The soil surface near Horseshoe Lake is generally fairly bright. The surface is covered by glacial deposits (till) consisting mainly of weathered granitic rocks.

deJong, Steven M.↗