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Transformation and gene editing in the bioenergy grass Miscanthus
Miscanthus, a C4 member of Poaceae, is a promising perennial crop for bioenergy, renewable bioproducts, and carbon sequestration. Species of interest include nothospecies M. x giganteus and its parental species M. sacchariflorus and M. sinensis. Use of biotechnology-based procedures to genetically improve Miscanthus, to date, have only included plant transformation procedures for introduction of exogenous genes into the host genome at random, non-targeted sites. We developed gene editing procedures for Miscanthus using CRISPR/Cas9 that enabled the mutation of a specific (targeted) endogenous gene to knock out its function. Classified as paleo-allopolyploids (duplicated ancient Sorghum-like DNA plus chromosome fusion event), design of guide RNAs (gRNAs) for Miscanthus needed to target both homeologs and their alleles to account for functional redundancy. Prior research in Zea mays demonstrated that editing the lemon white1 (lw1) gene, involved in chlorophyll and carotenoid biosynthesis, via CRISPR/Cas9 yielded pale green/yellow, striped or white leaf phenotypes making lw1 a promising target for visual confirmation of editing in other species. Using sequence information from both Miscanthus and sorghum, orthologs of maize lw1 were identified; a multi-step screening approach was used to select three gRNAs that could target homeologs of lw1. Embryogenic calli of M. sacchariflorus, M. sinensis and M. x giganteus were transformed via particle bombardment (biolistics) or Agrobacterium tumefaciens introducing the Cas9 gene and three gRNAs to edit lw1. Leaves on edited Miscanthus plants displayed the same phenotypes noted in maize. Sanger sequencing confirmed editing; deletions in lw1 ranged from 1 to 26 bp in length, and one deletion (433 bp) encompassed two target sites. Confocal microscopy verified lack of autofluorescence (chlorophyll) in edited leaves/sectors. We developed procedures for gene editing via CRISPR/Cas9 in Miscanthus and, to the best of our knowledge, are the first to do so. This included five genotypes representing three Miscanthus species. Designed gRNAs targeted all copies of lw1 (homeologous copies and their alleles); results also confirmed lw1 made a good editing target in species other than Z. mays. The ability to target specific loci to enable endogenous gene editing presents a new avenue for genetic improvement of this important biomass crop.
Data for Transformation and Gene Editing in the Bioenergy Grass Miscanthus
Miscanthus, a C4 member of the family Poaceae, is a promising perennial crop for bioenergy, renewable bioproducts, and carbon sequestration. Species of interest include nothospecies Miscanthus x giganteus and its parental species M. sacchariflorus and M. sinensis . Use of biotechnology-based procedures to genetically improve miscanthus, to date, have only included plant transformation procedures for introduction of exogenous genes into the host genome at random, non-targeted sites.
Surface Meteorological Station - PNNL Short Tower, Grass Valley - Raw Data
In support of the Wind Forecasting Improvement Project, Pacific Northwest National Laboratory (PNNL) deployed surface meteorological stations in Oregon.
Surface Meteorological Station - PNNL Short Tower, Grass Valley - Reviewed Data
In support of the Wind Forecasting Improvement Project, Pacific Northwest National Laboratory (PNNL) deployed surface meteorological stations in Oregon.
Data from: Emerging wild virus of native grass bioenergy feedstock is well established in the Midwestern USA and associated with premature stand senescence
This dataset includes values for the prevalence of switchgrass mosaic virus (Genus Marafivirus, Family Tymoviridae) detected with molecular diagnostics (RT-PCR) in individual Panicum virgatum (switchgrass) plants and in Graminella leafhoppers that feed on them. Surveys were conducted in 15 sites in August 2012. Stands surveyed had been established for some time and represent a range of landscape contexts. Measures of stand height and percent senescence were also collected. Land cover composition surrounding each site was calculated from the USDA-NASS Cropland Data Layer and estimates of drought impact were derived from the US Drought Monitor.
Soil and root bacterial communities of Switchgrass and other biofuel grasses in the Michigan and Wisconsin (2018)
Meta-amplicon sequencing of the soil fungal community of Switchgrass (var. Cave-n-Rock) of the Great Lakes Bioenergy Research Center Marginal Lands Experiment. Samples were collected in 2018.
Thermal absorption in seeded grass Semiannual report
Thermal absorption of particle clouds in seeded gas
Interactions of technology and society: Impacts of improved airtransport. A study of airports at the grass roots
The feasibility of applying a particular conception of technology and social change to specific examples of technological development was investigated. The social and economic effects of improved airport capabilities on rural communities were examined. Factors which led to the successful implementation of a plan to construct sixty small airports in Ohio are explored and implications derived for forming public policies, evaluating air transportation development, and assessing technology.
Development of the University of Massachusetts Remote Sensing Program: A grass-roots approach
The early stages of development of a remote sensing center at a state land grant university are considered. Several critical strategies adopted early in the planning process which may be useful to other programs or institutions are described.
Light interception and leaf area estimates from measurements of grass canopy reflectance
Grassland is a major component of the earth's available land. The vast area and remoteness of this ecosystem makes it difficult to assess its condition and monitor productivity by traditional methods. Remote sensing potentially offers a rapid nondestructive approach for monitoring such ecosystems. A study was carried out in a tallgrass prairie site near Manhattan, Kansas, during the 1983 and 1984 seasons to investigate the feasibility of estimating light interception and green leaf area index (LAI) from measurements of canopy multispectral reflectance. Greenness (Gn) index was found to be strongly correlated with intercepted photosynthetically active radiation (PAR). Two methods, a direct regression (RGR) and an indirect approach (IND), were used to estimate LAI from Gn index. The LAI values estimated by RGR method were consistently lower than the measured ones; however, good agreement was obtained between the LAI values estimated by IND method and the measured LAI. This suggests that Gn transformation of canopy spectral reflectance is more closely related to the fraction of intercepted PAR by green foliage than the quantity of green LAI.
Quantification of biomass of the marsh grass Spartina alterniflora Loisel using Landsat Thematic Mapper imagery
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Use of vegetation indices to estimate intercepted solar radiation and net carbon dioxide exchange of a grass canopy
Results are presented from field experiments relating spectral reflectance to intercepted photosynthetically active radiation (PAR) and net CO2 exchange in a natural canopy composed of the marsh cordgrass (Spartina alterniflora). Reflectance measurements made by a hand-held radiometer with Landsat TM spectral wavebands are used to compute remote sensing indices such as the normalized difference vegetation index. Consideration is given to the impact of standing dead canopy material on the relationship between intercepted PAR and spectral vegetation indices and the impact of changes in photosynthetic efficiency on the relationship between vegetation indices and CO2 exchange rates. The results suggest that quantitative remote assessment of photosynthesis and net gas exchange in natural vegetation is feasible, especially if the analysis incorporates information on biological responses to environmental variables.
How to lay out and build an airplane landing field : notes on shape and size of plot, runway details, type and arrangement of buildings, drainage of field, best kind of grass and proper marking to aid pilots
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Towards a High Temporal Frequency Grass Canopy Thermal IR Model for Background Signatures
In this paper, we present our first results towards understanding high temporal frequency thermal infrared response from a dense plant canopy and compare the application of our model, driven both by slowly varying, time-averaged meteorological conditions and by high frequency measurements of local and within canopy profiles of relative humidity and wind speed, to high frequency thermal infrared observations. Previously, we have employed three-dimensional ray tracing to compute the intercepted and scattered radiation fluxes and for final scene rendering. For the turbulent fluxes, we employed simple resistance models for latent and sensible heat with one-dimensional profiles of relative humidity and wind speed. Our modeling approach has proven successful in capturing the directional and diurnal variation in background thermal infrared signatures. We hypothesize that at these scales, where the model is typically driven by time-averaged, local meteorological conditions, the primary source of thermal variance arises from the spatial distribution of sunlit and shaded foliage elements within the canopy and the associated radiative interactions. In recent experiments, we have begun to focus on the high temporal frequency response of plant canopies in the thermal infrared at 1 second to 5 minute intervals. At these scales, we hypothesize turbulent mixing plays a more dominant role. Our results indicate that in the high frequency domain, the vertical profile of temperature change is tightly coupled to the within canopy wind speed In the results reported here, the canopy cools from the top down with increased wind velocities and heats from the bottom up at low wind velocities. .
Comparison of Surface IR Radiometer and Near Surface Air Temperatures over a Grass Covered Field
In this paper, we shall not concern ourselves with any of the issues mentioned above, but instead will concentrate on two other fundamental measurements issues.
Coherent Microwave Scattering Model of Marsh Grass
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