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At least 109 records · Page 6

Generation and multiplication of plantlets from callus derived from Haplopappus gracilus (Nutt.) Gray and their karyotype analysis

Unopened flower heads of Haplopappus gracilis (2n = 4) provided primary explants for callus production and subsequent induction of organized growth. Callus was initiated from small (3-5 mm in length) floral buds with benzylaminopurine (BAP) (44.4 micromoles; 10 mg/l) and naphthalene acetic acid (NAA) (0.54 micromole; 0.1 mg/l). Lowering the BAP level to 4.44 micromoles (1 mg/l) but maintaining the NAA level, gave rise to organized but highly compressed shoot growing points from an otherwise undifferentiated callus mass. Shoots selected from such cultures were maintainable and could be proliferated by growing 1-1.5-cm stem tip cuttings on Murashige and Skoog basal medium (solidified with agar) containing 0.444 micromole (0.1 mg/l) BAP and 0.054 micromole (0.01 mg/l) NAA. The stem tip multiplication rates obtainable by these means permit reliable strategies for shoot multiplication or production of rooted plantlets. Prolonged subculture and maintenance of shoots on growth regulator-free medium leads to in vitro flowering and greatly reduces rooting capacity. Karyotype analysis of chromosomes from root tip cells at metaphase and chromosome measurements show that karyologically uniform plantlets (based on chromosome number and morphology) can be obtained.

NASA Discipline Plant Biology↗

Green and non-green callus induction from excised rice (Oryza sativa) embryos: effects of exogenous plant growth regulators

Calli were induced either from excised rice embryos or from whole seeds in the presence of 1 to 5 mg l-1 NAA. After 12 days of culture, calli were induced only from excised rice embryos. We found that excised embryos accumulated NAA up to 6 times higher concentration than did whole seeds. In the presence of 1 to 5 mg l-1 NAA and 2 to 10 mg l-1 kinetin, chlorophyllous calli were induced from excised rice embryos. Chlorophyll contents in the callus tissue increased with increasing kinetin concentration while percent callus induction decreased. The total chlorophyll content was linearly correlated with the ratio of kinetin to NAA in the medium.

NASA Discipline Number 40-50↗

Osteoblastic phenotype of rat marrow stromal cells cultured in the presence of dexamethasone, beta-glycerolphosphate, and L-ascorbic acid

We investigated the effects of the time course of addition of osteogenic supplements dexamethasone, beta-glycerolphosphate, and L-ascorbic acid to rat marrow stromal cells, and the exposure time on the proliferation and differentiation of the cells. It was the goal of these experiments to determine the time point for supplement addition to optimize marrow stromal cell proliferation and osteoblastic differentiation. To determine this, two studies were performed; one study was based on the age of the cells from harvest, and the other study was based on the duration of exposure to supplemented medium. Cells were seen to proliferate rapidly at early time points in the presence and absence of osteogenic supplements as determined by 3H-thymidine incorporation into the DNA of replicating cells. These results were supported by cell counts ascertained through total DNA analysis. Alkaline phosphatase (ALP) activity and osteocalcin production at 21 days were highest for both experimental designs when the cells were exposed to supplemented medium immediately upon harvest. The ALP levels at 21 days were six times greater for cells maintained in supplements throughout than for control cells cultured in the absence of supplements for both studies, reaching an absolute value of 75 x 10(-7) micromole/min/cell. Osteocalcin production reached 20 x 10(-6) ng/cell at 21 days in both studies for cells maintained in supplemented medium throughout the study, whereas the control cells produced an insignificant amount of osteocalcin. These results suggest that the addition of osteogenic supplements to marrow-derived cells early in the culture period did not inhibit proliferation and greatly enhanced the osteoblastic phenotype of cells in a rat model.

Non-NASA Center↗

Modeling of two-phase flow in membranes and porous media in microgravity as applied to plant irrigation in space

In traditional applications in soil physics it is convention to scale porous media properties, such as hydraulic conductivity, soil water diffusivity, and capillary head, with the gravitational acceleration. In addition, the Richards equation for water flux in partially saturated porous media also contains a gravity term. With the plans to develop plant habitats in space, such as in the International Space Station, it becomes necessary to evaluate these properties and this equation under conditions of microgravitational acceleration. This article develops models for microgravity steady state two-phase flow, as found in irrigation systems, that addresses critical design issues. Conventional dimensionless groups in two-phase mathematical models are scaled with gravity, which must be assigned a value of zero for microgravity modeling. The use of these conventional solutions in microgravity, therefore, is not possible. This article therefore introduces new dimensionless groups for two-phase models. The microgravity models introduced here determined that in addition to porous media properties, important design factors for microgravity systems include applied water potential and the ratio of inner to outer radii for cylindrical and spherical porous media systems.

Plant Roots/growth & development↗

Growth and maintenance of an embryogenic cell culture of daylily (Hemerocallis) on hormone-free medium

Callus cultures of the diploid daylily (Hemerocallis) clone Autumn Blaze' were initiated and maintained in hormone-containing nutrient medium. At various times (from 6 weeks to 1 year) after being initiated, hormone-derived cultures were evaluated for their ability to be maintained and to multiply on hormone-free medium at low pH (between pH 4 and 4.5). Cultures had to be exposed to hormone-containing medium for at least 12 weeks before they could be maintained on hormone-free medium at low pH. The transition to maintainability on low pH hormone-free medium included the production of many aberrant embryonal forms ( neomorphs'). However, all hormone-derived cultures tested consisted entirely of preglobular stage proembryos (PGSPs) after 12-24 weeks on low pH hormone-free medium. PGSP cultures have been maintained and multiplied as such for over 1 year on low pH hormone-free medium. PGSPs continue their development into various somatic embryo stages when cultured on hormone-free medium buffered at pH 5.8. The production of well-formed somatic embryos was greatly enhanced when PGSPs were plated on activated charcoal impregnated filter papers that were placed on top of the agar surface. The gross morphology and histology of the PGSPs and stages of somatic embryo development are presented. The work shows that the ability of hormone-free medium at low pH to permit PGSP multiplication without development into later stages of embryo development is not restricted to carrot.

Non-NASA Center↗

Simulation of gaseous diffusion in partially saturated porous media under variable gravity with lattice Boltzmann methods

Liquid distributions in unsaturated porous media under different gravitational accelerations and corresponding macroscopic gaseous diffusion coefficients were investigated to enhance understanding of plant growth conditions in microgravity. We used a single-component, multiphase lattice Boltzmann code to simulate liquid configurations in two-dimensional porous media at varying water contents for different gravity conditions and measured gas diffusion through the media using a multicomponent lattice Boltzmann code. The relative diffusion coefficients (D rel) for simulations with and without gravity as functions of air-filled porosity were in good agreement with measured data and established models. We found significant differences in liquid configuration in porous media, leading to reductions in D rel of up to 25% under zero gravity. The study highlights potential applications of the lattice Boltzmann method for rapid and cost-effective evaluation of alternative plant growth media designs under variable gravity.

NASA Center JSC↗

Phylogenetic and physiological diversity of microorganisms isolated from a deep greenland glacier ice core

We studied a sample from the GISP 2 (Greenland Ice Sheet Project) ice core to determine the diversity and survival of microorganisms trapped in the ice at least 120,000 years ago. Previously, we examined the phylogenetic relationships among 16S ribosomal DNA (rDNA) sequences in a clone library obtained by PCR amplification from genomic DNA extracted from anaerobic enrichments. Here we report the isolation of nearly 800 aerobic organisms that were grouped by morphology and amplified rDNA restriction analysis patterns to select isolates for further study. The phylogenetic analyses of 56 representative rDNA sequences showed that the isolates belonged to four major phylogenetic groups: the high-G+C gram-positives, low-G+C gram-positives, Proteobacteria, and the Cytophaga-Flavobacterium-Bacteroides group. The most abundant and diverse isolates were within the high-G+C gram-positive cluster that had not been represented in the clone library. The Jukes-Cantor evolutionary distance matrix results suggested that at least 7 isolates represent new species within characterized genera and that 49 are different strains of known species. The isolates were further categorized based on the isolation conditions, temperature range for growth, enzyme activity, antibiotic resistance, presence of plasmids, and strain-specific genomic variations. A significant observation with implications for the development of novel and more effective cultivation methods was that preliminary incubation in anaerobic and aerobic liquid prior to plating on agar media greatly increased the recovery of CFU from the ice core sample.

Bacteria/classification/genetics/isolation & purif↗

Factors enhancing Agrobacterium tumefaciens-mediated gene transfer in peanut (Arachis hypogaea L.)

Parameters enhancing Agrobacterium-mediated transfer of foreign genes to peanut (Arachis hypogaea L.) cells were investigated. An intron-containing beta-glucuronidase uidA (gusA) gene under the transcriptional control of CaMV 35S promoter served as a reporter. Transformation frequency was evaluated by scoring the number of sectors expressing GUS activity on leaf and epicotyl explants. The 'Valencia Select' market type cv. New Mexico was more amenable to Agrobacterium transformation than the 'runner' market type cultivars tested (Florunner, Georgia Runner, Sunrunner, or South Runner). The disarmed Agrobacterium tumefaciens strain EHA101 was superior in facilitating the transfer of uidA gene to peanut cells compared to the disarmed strain C58. Rinsing of explants in half-strength Murashige-Skoog (MS) media prior to infection by Agrobacterium significantly increased the transformation efficiency. The use of cocultivation media containing high auxin [1.0 or 2.5 mg/l (4.53 micromolar or 11.31 micromolar) 2,4-D] and low cytokinin [0.25 or 0.5 mg/l (1.0 micromolar or 2.0 micromolar) BA] promoted higher transformation than either hormone-free or thidiazuron-containing medium. The polarity of the epicotyl during cocultivation was important; explants incubated in an inverted (vertically) manner followed by a vertically upright position resulted in improved transformation and shoot regeneration frequencies. Preculture of explants in MS basal medium or with 2.5 mg thidiazuron per l prior to infection drastically decreased the number of transformed zones. The optimized protocol was used to obtain transient transformation frequencies ranging from 12% to 36% for leaf explants, 15% to 42% for epicotyls. Initial evidence of transformation was obtained by polymerase chain reaction and subsequently confirmed by Southern analysis of regenerated plants.

NASA Discipline Life Support Systems↗

A comparison of soil moisture sensors for space flight applications

Plants will be an important part of future long-term space missions. Automated plant growth systems require accurate and reliable methods of monitoring soil moisture levels. There are a number of different methods to accomplish this task. This study evaluated sensors using the capacitance method (ECH2O), the heat-pulse method (TMAS), and tensiometers, compared to soil water loss measured gravimetrically in a side-by-side test. The experiment monitored evaporative losses from substrate compartments filled with 1- to 2-mm baked calcinated clay media. The ECH2O data correlated well with the gravimetric measurements, but over a limited range of soil moisture. The averaged TMAS sensor data overstated soil moisture content levels. The tensiometer data appeared to track evaporative losses in the 0.5- to 2.5-kPa range of matric potential that corresponds to the water content needed to grow plants. This small range is characteristic of large particle media, and thus high-resolution tensiometers are required to distinguish changing moisture contents in this range.

NASA Center KSC↗

Discontinuous pore fluid distribution under microgravity--KC-135 flight investigations

Designing a reliable plant growth system for crop production in space requires the understanding of pore fluid distribution in porous media under microgravity. The objective of this experimental investigation, which was conducted aboard NASA KC-135 reduced gravity flight, is to study possible particle separation and the distribution of discontinuous wetting fluid in porous media under microgravity. KC-135 aircraft provided gravity conditions of 1, 1.8, and 10(-2) g. Glass beads of a known size distribution were used as porous media; and Hexadecane, a petroleum compound immiscible with and lighter than water, was used as wetting fluid at residual saturation. Nitrogen freezer was used to solidify the discontinuous Hexadecane ganglia in glass beads to preserve the ganglia size changes during different gravity conditions, so that the blob-size distributions (BSDs) could be measured after flight. It was concluded from this study that microgravity has little effect on the size distribution of pore fluid blobs corresponding to residual saturation of wetting fluids in porous media. The blobs showed no noticeable breakup or coalescence during microgravity. However, based on the increase in bulk volume of samples due to particle separation under microgravity, groups of particles, within which pore fluid blobs were encapsulated, appeared to have rearranged themselves under microgravity.

manned↗

Porous media matric potential and water content measurements during parabolic flight

Control of water and air in the root zone of plants remains a challenge in the microgravity environment of space. Due to limited flight opportunities, research aimed at resolving microgravity porous media fluid dynamics must often be conducted on Earth. The NASA KC-135 reduced gravity flight program offers an opportunity for Earth-based researchers to study physical processes in a variable gravity environment. The objectives of this study were to obtain measurements of water content and matric potential during the parabolic profile flown by the KC-135 aircraft. The flight profile provided 20-25 s of microgravity at the top of the parabola, while pulling 1.8 g at the bottom. The soil moisture sensors (Temperature and Moisture Acquisition System: Orbital Technologies, Madison, WI) used a heat-pulse method to indirectly estimate water content from heat dissipation. Tensiometers were constructed using a stainless steel porous cup with a pressure transducer and were used to measure the matric potential of the medium. The two types of sensors were placed at different depths in a substrate compartment filled with 1-2 mm Turface (calcined clay). The ability of the heat-pulse sensors to monitor overall changes in water content in the substrate compartment decreased with water content. Differences in measured water content data recorded at 0, 1, and 1.8 g were not significant. Tensiometer readings tracked pressure differences due to the hydrostatic force changes with variable gravity. The readings may have been affected by changes in cabin air pressure that occurred during each parabola. Tensiometer porous membrane conductivity (function of pore size) and fluid volume both influence response time. Porous media sample height and water content influence time-to-equilibrium, where shorter samples and higher water content achieve faster equilibrium. Further testing is needed to develop these sensors for space flight applications.

Parabolic Flight↗

Spirosymplokos deltaeiberi nov. gen., nov. sp.: variable-diameter composite spirochete from microbial mats

Large (up to 100 micrometers long), loosely coiled, free-living spirochetes with variable diameters (from 0.4 to 3 micrometers in the same cell) were seen at least 40 times between August 1990 and January 1993. These spirochetes were observed in mud water and enrichment media from highly specific habitats in intertidal evaporite flats at three disjunct localities, one in Spain and two in Mexico. All three are sites of commercial saltworks. Associated with Microcoleus chthonoplastes the large spirochetes from Spain display phototaxis and a composite organization. Shorter and smaller-diameter spirochetes are seen inside both healthy and spent periplasm of larger ones. Small spirochetes attached to large ones have been observed live. From two to twelve spirochete protoplasmic cylinders were seen inside a single common outer membrane. A distinctive granulated cytoplasm in which the granules are of similar diameter (20-32 nanometers) to that of the flagella (26 nanometers) was present. Granule diameters were measured in thin section and in negatively-stained whole-mount preparations. Based on their ultrastructure, large size, variable diameter, number of flagella (3 to 6), and phototactic behavior these unique spirochetes are formally named Spirosymplokos deltaeiberi. Under anoxic (or low oxygen) conditions they formed blooms in mixed culture in media selective for spirochetes. Cellobiose was the major carbon source in 80% seawater, the antibiotic rifampicin was added, mat from the original field site was present and tubes were incubated in the light at from 18-31 degrees C. Within 1-2 weeks populations of the large spirochete developed at 25 degrees C but they could not be transferred to fresh medium.

Non-NASA Center↗

Tip-growing cells of the moss Ceratodon purpureus Are gravitropic in high-density media

Gravity sensing in plants and algae is hypothesized to rely upon either the mass of the entire cell or that of sedimenting organelles (statoliths). Protonemata of the moss Ceratodon purpureus show upward gravitropism and contain amyloplasts that sediment. If moss sensing were whole-cell based, then media denser than the cell should prevent gravitropism or reverse its direction. Cells that were inverted or reoriented to the horizontal displayed distinct negative gravitropism in solutions of iodixanol with densities of 1.052 to 1.320 as well as in bovine serum albumin solutions with densities of 1.037 to 1.184 g cm(-3). Studies using tagged molecules of different sizes and calculations of diffusion times suggest that both types of media penetrate through the apical cell wall. Estimates of the density of the apical cell range from 1.004 to 1.085. Because protonemata grow upward when the cells have a density that is lower than the surrounding medium, gravitropic sensing probably utilizes an intracellular mass in moss protonemata. These data provide additional support for the idea that sedimenting amyloplasts function as statoliths in gravitropism.

NASA Discipline Plant Biology↗

Measurement of hydraulic characteristics of porous media used to grow plants in microgravity

Understanding the effect of gravity on hydraulic properties of plant growth medium is essential for growing plants in space. The suitability of existing models to simulate hydraulic properties of porous medium is uncertain due to limited understanding of fundamental mechanisms controlling water and air transport in microgravity. The objective of this research was to characterize saturated and unsaturated hydraulic conductivity (K) of two particle-size distributions of baked ceramic aggregate using direct measurement techniques compatible with microgravity. Steady state (Method A) and instantaneous profile measurement (Method B) methods for K were used in a single experimental unit with horizontal flow through thin sections of porous medium providing an earth-based analog to microgravity. Comparison between methods was conducted using a crossover experimental design compatible with limited resources of space flight. Satiated (natural saturation) K ranged from 0.09 to 0.12 cm s-1 and 0.5 to >1 cm s-1 for 0.25- to 1- and 1- to 2-mm media, respectively. The K at the interaggregate/intraaggregate transition was approximately 10(-4) cm s-1 for both particle-size distributions. Significant differences in log(10)K due to method and porous medium were less than one order of magnitude and were attributed to variability in air entrapment. The van Genuchten/Mualem parametric models provided an adequate prediction of K of the interaggregate pore space, using residual water content for that pore space. The instantaneous profile method covers the range of water contents relevant to plant growth using fewer resources than Method A, all advantages for space flight where mass, volume, and astronaut time are limited.

NASA Center JSC↗

Health significance and occurrence of injured bacteria in drinking water

Enteropathogenic and indicator bacteria become injured in drinking water with exposure to sublethal levels of various biological, chemical and physical factors. One manifestation of this injury is the inability to grow and form colonies on selective media containing surfactants. The resulting underestimation of indicator bacteria can lead to a false estimation of water potability. m-T7 medium was developed specifically for the recovery of injured coliforms (both "total" and fecal) in drinking water. The m-T7 method was used to survey operating drinking water treatment and distribution systems for the presence of injured coliforms that were undetected with currently used media. The mean recovery with m-Endo LES medium was less than 1/100 ml while it ranged between 6 and 68/100ml with m-T7 agar. The majority of samples giving positive results with m-T7 medium yielded no detectable coliforms with m-Endo LES agar. Over 95% of the coliform bacteria in these samples were injured. Laboratory experiments were also done to ascribe the virulence of injured waterborne pathogens. Enteropathogens including Salmonella typhimurium, Yersinia enterocolitica and Shigella spp. required up to 20 times the chlorine levels to produce the same injury in enterotoxigenic Escherichia coli (ETEC) and nonpathogenic coliforms. Similar results were seen with Y. enterocolitica exposed to copper. The recovery of ETEC was followed by delayed enterotoxin production, both in vitro and in the gut of experimental animals. This indicates that injured waterborne enteropathogenic bacteria can be virulent.

NASA Discipline Number 13-40↗

Cytoplasmic membrane changes during adaptation of the fresh water cyanobacterium Synechococcus 6311 to salinity

In this investigation, changes were characterized in cell structure and cytoplasmic membrane organization that occur when the freshwater cyanobacterium Synechococcus 6311 is transferred from 'low salt' (0.03 molar NaCl) to 'high salt' (0.5 molar NaCl) media (i.e. sea water concentration). Cells were examined at several time points after the imposition of the salt stress and compared to control cells, in thin sections and freeze fracture electron microscopy, and by flow cytometry. One minute after exposure to high salt, i.e. 'salt shock', virtually all intracellular granules disappeared, the density of the cytoplasm decreased, and the appearance of DNA material was changed. Glycogen and other granules, however, reappeared by 4 hours after salt exposure. The organization of the cytoplasmic membrane undergoes major reorganization following salt shock. Freeze-fracture electron microscopy showed that small intramembrane particles (diameter 7.5 and 8.5 nanometers) are reduced in number by two- to fivefold, whereas large particles, (diameters 14.5 and 17.5 nanometers) increase two- to fourfold in frequency, compared to control cells grown in low salt medium. The changes in particle size distribution suggest synthesis of new membrane proteins, in agreement with the known increases in respiration, cytochrome oxidase, and sodium proton exchange activity of the cytoplasmic membrane.

NASA Discipline Number 61-10↗

Delayed fertilization of anuran amphibian (Xenopus) eggs leads to reduced numbers of primordial germ cells

Several media were tested for the extent to which they promoted high fertilization efficiencies in ovulated, stripped Xenopus eggs. One medium was selected for maintaining eggs in a 'delayed fertilization' (DelF) condition. DelF eggs displayed several unusual characteristics, including shift of the center of gravity, prominent sperm entrance site, and occasional polyspermy. The frequency of normal pattern formation varied according to the length of time eggs were maintained in the DelF condition. Various developmental abnormalities were observed during gastrulation, neurulation, and organogenesis. Most abnormalities appeared, however, to be related to morphogenesis of the endoderm. Primordial germ cell (PGC) development was examined in DelF eggs which displayed normal external morphological features at the swimming tadpole stage. PGC counts were usually normal in short-duration (eg, 5 hr) DelF eggs, but frequently substantially reduced or completely diminished in longer-duration (eg, 25h) tadpoles. Six spawnings were compared and shown to exhibit considerable variability in fertility, morphogenesis, and PGC development. Yolk platelet shifts and developmental parameters were examined in two additional spawnings. The subcortical cytoplasm in which the germ plasm is normally localized appeared to be disrupted in longer duration DelF eggs. That observation may account for low PGC counts in DelF tadpoles.

NASA Discipline Number 40-20↗

Using mini-rockwool blocks as growing media for limited-cluster tomato production

Rockwool is an excellent growing medium for the hydroponic production of tomato; however, the standard size rockwool blocks [4 x 4 x 2.5 inches (10 x 10 x 6.3 cm) or 3 x 3 x 2.5 inches (7.5 x 7.5 x 6.3 cm)] are expensive. The following experiments were conducted with less expensive minirock wool blocks (MRBs), on rayon polyester material (RPM) as a bench top liner, to reduce the production cost of tomatoes (Lycopersicon esculentum) grown in a limited-cluster, ebb and flood hydroponic cultivation system. Fruit yield for single-cluster plants growing in MRBs [2 x 2 x 1.6 inches (5 x 5 x 4 cm) and 1.6 x 1.6 x 1.6 inches (4 x 4 x 4 cm)] was not significantly different from plants grown in larger sized blocks (3 x 3 x 2.5 inches). When the bench top was lined with RPM, roots penetrated the RPM, and an extensive root mat developed between the RPM and the bench top. The fruit yield from plants on RPM was significantly increased compared to plants without RPM due to increases in fruit size and fruit number. RPM also significantly reduced the incidence of blossom-end rot. In a second experiment, single- and double-cluster plants were grown on RPM. Fruit yield for double-cluster plants was 40% greater than for single-cluster plants due to an increase in fruit number, although the fruit were smaller in size. As in the first experiment, fruit yield for all plants grown in MRBs was not significantly different from plants grown in the larger sized blocks. MRBs and a RPM bench liner are an effective combination in the production of limited-cluster hydroponic tomatoes.

Non-NASA Center↗