Green fluorescent protein in transgenic plants. Brassica transformation
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NASA Discipline Life Sciences Technologies↗
SEARCH · Engineering Papers
Results for “Plant transformation”
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Spatiotemporal relationships between growth and microtubule orientation as revealed in living root cells of Arabidopsis thaliana transformed with green-fluorescent-protein gene construct GFP-MBD
Arabidopsis thaliana plants were transformed with GFP-MBD (J. Marc et al., Plant Cell 10: 1927-1939, 1998) under the control of a constitutive (35S) or copper-inducible promoter. GFP-specific fluorescence distributions, levels, and persistence were determined and found to vary with age, tissue type, transgenic line, and individual plant. With the exception of an increased frequency of abnormal roots of 35S GFP-MBD plants grown on kanamycin-containing media, expression of GFP-MBD does not appear to affect plant phenotype. The number of leaves, branches, bolts, and siliques as well as overall height, leaf size, and seed set are similar between wild-type and transgenic plants as is the rate of root growth. Thus, we conclude that the transgenic plants can serve as a living model system in which the dynamic behavior of microtubules can be visualized. Confocal microscopy was used to simultaneously monitor growth and microtubule behavior within individual cells as they passed through the elongation zone of the Arabidopsis root. Generally, microtubules reoriented from transverse to oblique or longitudinal orientations as growth declined. Microtubule reorientation initiated at the ends of the cell did not necessarily occur simultaneously in adjacent neighboring cells and did not involve complete disintegration and repolymerization of microtubule arrays. Although growth rates correlated with microtubule reorientation, the two processes were not tightly coupled in terms of their temporal relationships, suggesting that other factor(s) may be involved in regulating both events. Additionally, microtubule orientation was more defined in cells whose growth was accelerating and less stringent in cells whose growth was decelerating, indicating that microtubule-orienting factor(s) may be sensitive to growth acceleration, rather than growth per se.
NASA Discipline Plant Biology↗
Biological degradation and composition of inedible sweetpotato biomass
Many challenges are presented by biological degradation in a bioregenerative Controlled Ecological Life Support System (CELSS) as envisioned by the U.S. National Aeronautics and Space Administration (NASA). In the studies conducted with biodegradative microorganism indigenous to sweetpotato fields, it was determined that a particle size of 75 microns and incubation temperature of 30 degrees C were optimal for degradation. The composition of the inedible biomass and characterization of plant nutrient solution indicated the presence of potential energy sources to drive microbial transformations of plant waste. Selected indigenous soil isolates with ligno-cellulolytic or sulfate-reducing ability were utilized in biological studies and demonstrated diversity in ability to reduce sulfate in solution and to utilize alternative carbon sources: a lignin analog--4-hydroxy, 3-methoxy cinnamic acid, cellulose, arabinose, glucose, sucrose, mannitol, galactose, ascorbic acid.
Vegetables/chemistry/metabolism↗
Expression of pH-sensitive green fluorescent protein in Arabidopsis thaliana
This is the first report on using green fluorescent protein (GFP) as a pH reporter in plants. Proton fluxes and pH regulation play important roles in plant cellular activity and therefore, it would be extremely helpful to have a plant gene reporter system for rapid, non-invasive visualization of intracellular pH changes. In order to develop such a system, we constructed three vectors for transient and stable transformation of plant cells with a pH-sensitive derivative of green fluorescent protein. Using these vectors, transgenic Arabidopsis thaliana and tobacco plants were produced. Here the application of pH-sensitive GFP technology in plants is described and, for the first time, the visualization of pH gradients between different developmental compartments in intact whole-root tissues of A. thaliana is reported. The utility of pH-sensitive GFP in revealing rapid, environmentally induced changes in cytoplasmic pH in roots is also demonstrated.
NASA Discipline Plant Biology↗
The Ca(2+) status of the endoplasmic reticulum is altered by induction of calreticulin expression in transgenic plants
To investigate the endoplasmic reticulum (ER) Ca(2+) stores in plant cells, we generated tobacco (Nicotiana tabacum; NT1) suspension cells and Arabidopsis plants with altered levels of calreticulin (CRT), an ER-localized Ca(2+)-binding protein. NT1 cells and Arabidopsis plants were transformed with a maize (Zea mays) CRT gene in both sense and antisense orientations under the control of an Arabidopsis heat shock promoter. ER-enriched membrane fractions from NT1 cells were used to examine how altered expression of CRT affects Ca(2+) uptake and release. We found that a 2.5-fold increase in CRT led to a 2-fold increase in ATP-dependent (45)Ca(2+) accumulation in the ER-enriched fraction compared with heat-shocked wild-type controls. Furthermore, after treatment with the Ca(2+) ionophore ionomycin, ER microsomes from NT1 cells overproducing CRT showed a 2-fold increase in the amount of (45)Ca(2+) released, and a 2- to 3-fold increase in the amount of (45)Ca(2+) retained compared with wild type. These data indicate that altering the production of CRT affects the ER Ca(2+) pool. In addition, CRT transgenic Arabidopsis plants were used to determine if altered CRT levels had any physiological effects. We found that the level of CRT in heat shock-induced CRT transgenic plants correlated positively with the retention of chlorophyll when the plants were transferred from Ca(2+)-containing medium to Ca(2+)-depleted medium. Together these data are consistent with the hypothesis that increasing CRT in the ER increases the ER Ca(2+) stores and thereby enhances the survival of plants grown in low Ca(2+) medium.
NASA Discipline Plant Biology↗
The design of exact nonlinear model followers
A practical approach to the design of control systems for strongly nonlinear, multivariable, time-dependent plants is described. The structure of the control system is that of an exact model follower. The model dynamics are decoupled, linear, constant, and of the order of the plant. The plant state and controls are transformed so that the plant, when viewed through these transformations, looks like the simple model. Regulation of disturbances is accomplished by means of the transformed state and controls. Conditions for transformability into linear models, the appropriate models, and the construction of the transformations are discussed. The approach is illustrated on a trajectory autopilot for a helicopter.
Meyer, G.↗
Transgenic plants with increased calcium stores
The present invention provides transgenic plants over-expressing a transgene encoding a calcium-binding protein or peptide (CaBP). Preferably, the CaBP is a calcium storage protein and over-expression thereof does not have undue adverse effects on calcium homeostasis or biochemical pathways that are regulated by calcium. In preferred embodiments, the CaBP is calreticulin (CRT) or calsequestrin. In more preferred embodiments, the CaBP is the C-domain of CRT, a fragment of the C-domain, or multimers of the foregoing. In other preferred embodiments, the CaBP is localized to the endoplasmic reticulum by operatively associating the transgene encoding the CaBP with an endoplasmic reticulum localization peptide. Alternatively, the CaBP is targeted to any other sub-cellular compartment that permits the calcium to be stored in a form that is biologically available to the plant. Also provided are methods of producing plants with desirable phenotypic traits by transformation of the plant with a transgene encoding a CaBP. Such phenotypic traits include increased calcium storage, enhanced resistance to calcium-limiting conditions, enhanced growth and viability, increased disease and stress resistance, enhanced flower and fruit production, reduced senescence, and a decreased need for fertilizer production. Further provided are plants with enhanced nutritional value as human food or animal feed.
Wyatt, Sarah↗
Cereal transformation through particle bombardment
The review focuses on experiments that lead to stable transformation in cereals using microprojectile bombardment. The discussion of biological factors that affect transformation examines target tissues and vector systems for gene transfer. The vector systems include reporter genes, selectable markers, genes of agronomic interest, and vector constructions. Other topics include physical parameters that affect DNA delivery, selection of stably transformed cells and plant regeneration, and analysis of gene expression and transmission to the progeny.
NASA Discipline Plant Biology↗
Evaluation and interpretation of Thematic Mapper ratios in equations for estimating corn growth parameters
Reflectance and agronomic Thematic Mapper (TM) data were analyzed to determine possible data transformations for evaluating several plant parameters of corn. Three transformation forms were used: the ratio of two TM bands, logarithms of two-band ratios, and normalized differences of two bands. Normalized differences and logarithms of two-band ratios responsed similarly in the equations for estimating the plant growth parameters evaluated in this study. Two-term equations were required to obtain the maximum predictability of percent ground cover, canopy moisture content, and total wet phytomass. Standard error of estimate values were 15-26 percent lower for two-term estimates of these parameters than for one-term estimates. The terms log(TM4/TM2) and (TM4/TM5) produced the maximum predictability for leaf area and dry green leaf weight, respectively. The middle infrared bands TM5 and TM7 are essential for maximizing predictability for all measured plant parameters except leaf area index. The estimating models were evaluated over bare soil to discriminate between equations which are statistically similar. Qualitative interpretations of the resulting prediction equations are consistent with general agronomic and remote sensing theory.
Dardner, B. R.↗
Antisense and sense expression of cDNA coding for CYP73A15, a class II cinnamate 4-hydroxylase, leads to a delayed and reduced production of lignin in tobacco
A number of plant species contain the class II of genes encoding the cytochrome P450, CYP73, the cognate protein of which cinnamic acid 4-hydroxylase, is the second enzyme of the phenylpropanoid pathway. In order to begin to determine possible functionality, tobacco has been transformed with a truncated French bean class II cinnamate hydroxylase (CYP73A15) in the sense and antisense orientations. Signals for C4H protein could be detected in vascular tissue from wild-type plants using heterologous probes. The transformed plants showed a normal phenotype, even though detectable C4H protein was much reduced in tissue prints. Young propagated transformants displayed a range of reduced C4H activities, as well as either reduced or no phloroglucinol-stainable lignin. However, all mature tobacco plants showed the accumulation of lignin, even though its deposition was apparently delayed. This was not due to induction of tyrosine ammonia-lyase activity, which was not detected, but instead it is presumed due to sufficient C4H residual activity. Analysis of the lignin content of the plants showed reductions of up to 30% with a slightly reduced syringyl to guaiacyl ratio as compared to wild type. This reduction level was favourable in comparison with some other targets in the lignification pathway that have been manipulated including that of class I cinnamate 4-hydroxylase. It is proposed that the class II cinnamate 4-hydroxylase might also function in lignification in a number of species including French bean and tobacco, based on these data.
NASA Discipline Plant Biology↗
Relation of agronomic and multispectral reflectance characteristics of spring wheat canopies
The relationships between crop canopy variables such as leaf area index (LAI) and their multispectral reflectance properties were investigated along with the potential for estimating canopy variables from remotely sensed reflectance measurements. Reflectance spectra over the 0.4 to 2.5 micron wavelength range were acquired during each of the major development stages of spring wheat canopies at Williston, North Dakota, during three seasons. Treatments included planting date, N fertilization, cultivar, and soil moisture. Agronomic measurements included development stage, biomass, LAI, and percent soil cover. High correlations were found between reflectance and percent cover, LAI, and biomass. A near infrared wavelength band, 0.76 to 0.90 microns, was most important in explaining variation in LAI and percent cover, while a middle infrared band, 2.08 to 2.35 microns, explained the most variation in biomass and plant water content. Transformations, including the near infrared/red reflectance ratio and greenness index, were also highly correlated to canopy variables. The relationship of canopy variables to reflectance decreased as the crop began to ripen. the canopy variables could be accurately predicted using measurements from three to five wavelength bands. The wavelength bands proposed for the thematic mapper sensor were more strongly related to the canopy variables than the LANDSAT MSS bands.
Bauer, M. E.↗
Making Mars habitable
The possibility is considered that the atmosphere and climate of Mars could be altered to allow terrestrial life forms, and possibly human beings, to survive on the surface. Production of CFCs or other greenhouse gases on Mars would warm the surface enough for the regolith and polar caps to release their CO2 and raise atmospheric pressure to 100 mbar. If a large regolith and polar CO2 reservoirs exist, the pressure would continue to rise on its own. If these are absent, additional CO2 would have to be released from carbonate minerals. At this point, perhaps between 100 and 100,000 yrs, Mars might be suitable for plants. If there is a mechanism for sequestering the reduced carbon, these plants could slowly transform the CO2 to produce an O2-rich atmosphere in perhaps 100,000 yrs. If sufficient N2 could be released from putative soil deposits and the CO2 level could be kept low enough, then a human-breathable atmosphere would be produced.
Mckay, Christopher P.↗
Recycling plant, human and animal wastes to plant nutrients in a closed ecological system
The essential minerals for plant growth are nitrogen, phosphorous, potassium (macronutrients), calcium, magnesium, sulfur (secondary nutrients), iron, manganese, boron, copper, zinc, chlorine, sodium, and molybdenum (micronutrients). The first step in recycling wastes will undoubtedly be oxidation of carbon and hydrogen to CO2 and H2O. Transformation of minerals to plant nutrients depends upon the mode of oxidation to define the state of the nutrients. For the purpose of illustrating the type of processing required, ash and off-gas compositions of an incineration process were assumed and subsequent processing requirements were identified. Several processing schemes are described for separating out sodium chloride from the ash, leading to reformulation of a nutrient solution which should be acceptable to plants.
Meissner, H. P.↗
Modeling Demographic-Driven Vegetation Dynamics and Ecosystem Biogeochemical Cycling in NASA GISS’s Earth System Model (ModelE-BiomeE v.1.0)
We developed a demographic vegetation model, BiomeE, to improve the modeling of vegetation dynamics and ecosystem biogeochemical cycles in the NASA Goddard Institute of Space Studies' ModelE Earth system model. This model includes the processes of plant growth, mortality, reproduction, vegetation structural dynamics, and soil carbon and nitrogen storage and transformations. The model combines the plant physiological processes of ModelE's original vegetation model, Ent, with the plant demographic and ecosystem nitrogen processes that have been represented in the Geophysical Fluid Dynamics Laboratory's LM3-PPA. We used nine plant functional types to represent global natural vegetation functional diversity, including trees, shrubs, and grasses, and a new phenology model to simulate vegetation seasonal changes with temperature and precipitation fluctuations. Competition for light and soil resources is individual based, which makes the modeling of transient compositional dynamics and vegetation succession possible. Overall, the BiomeE model simulates, with fidelity comparable to other models, the dynamics of vegetation and soil biogeochemistry, including leaf area index, vegetation structure (e.g., height, tree density, size distribution, and crown organization), and ecosystem carbon and nitrogen storage and fluxes. This model allows ModelE to simulate transient and long-term biogeophysical and biogeochemical feedbacks between the climate system and land ecosystems. Furthermore, BiomeE also allows for the eco-evolutionary modeling of community assemblage in response to past and future climate changes with its individual-based competition and demographic processes.
Biogeochemical cycles↗
Comparison of N2O Emissions from Soils at Three Temperate Agricultural Sites
Nitrous oxide (N2O) flux simulations by four models were compared with year-round field measurements from five temperate agricultural sites in three countries. The field sites included an unfertilized, semi-arid rangeland with low N2O fluxes in eastern Colorado, USA; two fertilizer treatments (urea and nitrate) on a fertilized grass ley cut for silage in Scotland; and two fertilized, cultivated crop fields in Germany where N2O loss during the winter was quite high. The models used were daily trace gas versions of the CENTURY model, DNDC, ExpertN, and the NASA-Ames version of the CASA model. These models included similar components (soil physics, decomposition, plant growth, and nitrogen transformations), but in some cases used very different algorithms for these processes. All models generated similar results for the general cycling of nitrogen through the agro-ecosystems, but simulated nitrogen trace gas fluxes were quite different. In most cases the simulated N20 fluxes were within a factor of about 2 of the observed annual fluxes, but even when models produced similar N2O fluxes they often produced very different estimates of gaseous N loss as nitric oxide (NO), dinitrogen (N2), and ammonia (NH3). Accurate simulation of soil moisture appears to be a key requirement for reliable simulation of N2O emissions. All models simulated the general pattern of low background fluxes with high fluxes following fertilization at the Scottish sites, but they could not (or were not designed to) accurately capture the observed effects of different fertilizer types on N2O flux. None of the models were able to reliably generate large pulses of N2O during brief winter thaws that were observed at the two German sites. All models except DNDC simulated very low N2O fluxes for the dry site in Colorado. The US Trace Gas Network (TRAGNET) has provided a mechanism for this model and site intercomparison. Additional intercomparisons are needed with these and other models and additional data sets; these should include both tropical agro-ecosystems and new agricultural management techniques designed for sustainability.
Frolking, S. E.↗
Implicit model following and parameter identification of unstable aircraft
A transformation in the s-plane is described which has utility in implicit model-following optimal control design application and in estimation or parameter identification problems. The objective of the transformation is, for the control problem, to achieve an unstable closed-loop system, and, for the estimation problem, to alleviate algorithm convergence problems that may arise in identifying unstable systems. For the control problem, the transformation is a shift along the real (sigma) axis of the plant and model poles and zeros. This transformation is shown to be equivalent to a modified performance index but offers the advantage of compatibility with existing optimal control solution algorithms. For the estimation problem, the data are multiplied by an exponential function and the assumed measurement and process noise covariances are appropriately modified. Examples of both control and estimation applications are presented.
Lebacqz, J. V.↗
Dynamics of microorganism populations in recirculating nutrient solutions
This overview covers the basic microbial ecology of recirculating hydroponic solutions. Examples from NASA and Soviet Controlled Ecological Life Support Systems (CELSS) tests and the commercial hydroponic industry will be used. The sources of microorganisms in nutrient solutions include air, water, seeds, plant containers and plumbing, biological vectors, and personnel. Microbial fates include growth, death, and emigration. Important microbial habitats within nutrient delivery systems are root surfaces, hardware surfaces (biofilms), and solution suspension. Numbers of bacteria on root surfaces usually exceed those from the other habitats by several orders of magnitude. Gram negative bacteria dominate the microflora with fungal counts usually much lower. Trends typically show a decrease in counts with increasing time unless stressed plants increase root exudates. Important microbial activities include carbon mineralization and nitrogen transformations. Important detrimental interactions include competition with plants, and human and plant pathogenesis.
Strayer, R. F.↗
Dynamics of microorganism populations in recirculating nutrient solutions
This overview covers the basic microbial ecology of recirculating hydroponic solutions. Examples from NASA and Soviet CELSS tests and the commercial hydroponic industry will be used. The sources of microorganisms in nutrient solutions include air, water, seeds, plant containers and plumbing, biological vectors, and personnel. Microbial fates include growth, death, and emigration. Important microbial habitats within nutrient delivery systems are root surfaces, hardware surfaces (biofilms), and solution suspension. Numbers of bacteria on root surfaces usually exceed those from the other habitats by several orders of magnitude. Gram negative bacteria dominate the microflora with fungal counts usually much lower. Trends typically show a decrease in counts with increasing time unless stressed plants increase root exudates. Important microbial activities include carbon mineralization and nitrogen transformations. Important detrimental interactions include competition with plants, and human and plant pathogenesis.
Strayer, R. F.↗