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Analysis of GMO Plum Plant Culture in System Operations Failure

GMO plum trees are being evaluated at the Kennedy Space Center as a possible candidate for future space crops. Previously conducted horticultural testing compared the performance of several plum genotypes in controlled environment chambers, resulting in a down-selection to the NASA-11 genotype. Precursory studies determined the water use requirements to sustain the plants as well as the feasibility of grafting non-GMO plum scions onto GMO plum rootstocks of NASA-5, NASA-10, and NASA-11 genotypes. This study follows the growth and horticultural progress of plum trees and in-vitro cultures from August 2017 to November 2017, and provides supplemental support for future GMO plum studies. The presence of Hurricane Irma in early September 2017 resulted in the plants undergoing material deterioration from major changes to their overall horticultural progress.

life support

Microbiological Horticultural Internship Final Abstract

GMO dwarf plum (Prunus domestica) is being evaluated as a candidate food crop for long duration space flight missions. A project was undertaken to develop a protocol for transferring selected genetic lines of GMO plum (previously maintained in pots and propagated by cuttings at NASA's Kennedy Space Center in Florida) into in vitro tissue culture. In vitro culture may reduce the space, materials, and labor required to maintain the current lines of GMO plum and better preserve them for future study. Fresh plant material from three selected GMO plum lines (NASA-5, NASA-10, and NASA-11) and a non-modified control line (Control-5) were processed aseptically into in vitro culture on four separate occasions. The impact of multiple treatments on the successful growth of GMO plum tissue in vitro were tested: Parent explant tissue type (leaf petioles, stem nodes containing buds and internodes without buds), tissue sterilization method [soaking in 10 bleach only (5 min for petioles or 10 min for nodesinternodes), or soaking in 70 EtOH (30 sec) followed by 10 bleach (5 min for petioles and 10 min for nodesinternodes)], and media type [three Murashige and Skoog-based medias (SGM, SRM, and SRM+2,4-D) and one recipe containing woody plant media (WPM)]. 22.2 of the plates containing tissue sterilized with bleach alone developed microbial contamination after two weeks, while only 11.8 of plates containing tissue sterilized sequentially with EtOH and bleach developed contamination. Node bud tissue from all four genetic lines of plum produced leafy plantlets on SGM and SRM media after 4-6 weeks. The most numerous and well-developed plantlets were present on SGM. Upon reaching suitable size, plantlets were transferred to larger media containers for further growth. Some node bud growth occurred on SRM+2,4-D and WPM 2.5 weeks after plating, however as of yet no pieces on SRM+2,4-D have adequate development for transferring. Tissue pieces from NASA-5 plated on WPM are developing leaves and will be ready for transferring soon. Petioles and internode tissue lacking bud meristem failed to produce any plantlets on any plates, however petioles developed large masses of undifferentiated callus tissue on SRM+2,4-D media. These callused pieces were then transferred to SRM+TDZ media, which resulted in even larger callus growth but no differentiation. All four selected plum lines were successfully transitioned into in vitro culture. Nodes from NASA-5 and NASA-10 lines produced the most numerous and well-developed leafy plantlets in vitro, while those from NASA-11 and Control-5 were generally smaller, slower growing and less numerous. The best method overall was to use young stem node tissue with buds, surface sterilize the pieces sequentially with 70 EtOH and 10 bleach, and then plate them onto SGM media. Future areas of study will include introducing additional genetic lines of GMO plum into in vitro culture, attempting to induce shoot growth in petiole callus tissue, testing methods (such as cold storage) that extend the time interval between transferring explants into new media, and testing viability of plantlets transferred from in vitro culture back to traditional pot culture.

bioregenerative life support

Biological/Horticultural Internship Final Report

A study was conducted to determine water use requirements of genetically modified (GMO) dwarf plum. GMO plum and unmodified standard plum plants were grown in a controlled environment chamber under varying CO2 concentrations (400 ppm, 1500 ppm, and 5000 ppm). Pepper plants were also grown in the chamber for additional comparison. Leaf stomatal conductance, biomass accumulation, soil moisture and pot weights were measured; Stomatal conductance of GMO plum and pepper plants decreased at sustained elevated CO2 concentrations. The stomatal conductance rates of the standard plums, however, increased at sustained elevated CO2 concentrations. Further data analysis (statistical analysis, biomass, soil moisture and pot weight measurements) is ongoing and required to gain better understanding of the data. An additional proof-of-concept study was undertaken to determine the feasibility of grafting unmodified standard plum scions onto genetically modified rootstocks as a propagation method. Bud grafts were performed on three GMO plum rootstocks: NASA-5, NASA-10, and NASA-11. All of the standard plum buds grafted onto NASA-5 and NASA-10 rootstocks began growing, indicating that this grafting method is highly successful for the formation of a graft union and initial bud growth. However, bud growth during stem elongation was curtailed on several grafts due to a combination of nutritional deficiency and physical damage/obstruction of the grafted tissues. Bud growth on the NASA-5 rootstock occurred sooner than in grafts on the NASA-10 rootstock, while only one bud graft has shown growth on the NASA-11 rootstock thus far. These marked differences in the onset of bud growth suggest genotypic differences between the rootstocks may affect bud graft vigor. Mature standard plum scions grown on the NASA-5 rootstock appeared to retain most or all of the physical characteristics of the standard plum donor plant.

GMO

Interaction of a Model Peptide with a Water--Bilayer System

We discuss a molecular dynamics study of the alanine dipeptide at the interface between water and a glycerol-1-monooleate (GMO) bilayer. The dipeptide is interfacially active and incorporates into the bilayer without disrupting its structure. The interfacial region that is readily penetrated by the dipeptide spans the entire head group portion of the bilayer. The polar groups of the alanine dipeptide mostly remain well solvated by water and the oxygen atoms of GMO, and conformations of the dipeptide are characterized by (phi, psi) angles typical of alpha-helix and beta-sheet structures. When the molecule is deeper in the bilayer, the C(sub 7eq) state also becomes stable. The barrier to the isomerization reaction at the interface is lower than in bulk phases. After 7 ns of trajectories, the system is not fully equilibrated, due to slow collective motions involving GMO head groups. These result in decreased mobility and lower rates of isomerization of the dipeptide at the interface.

Pohorille, A.

Using the Flow-3D General Moving Object Model to Simulate Coupled Liquid Slosh - Container Dynamics on the SPHERES Slosh Experiment: Aboard the International Space Station

The SPHERES Slosh Experiment (SSE) is a free floating experimental platform developed for the acquisition of long duration liquid slosh data aboard the International Space Station (ISS). The data sets collected will be used to benchmark numerical models to aid in the design of rocket and spacecraft propulsion systems. Utilizing two SPHERES Satellites, the experiment will be moved through different maneuvers designed to induce liquid slosh in the experiment's internal tank. The SSE has a total of twenty-four thrusters to move the experiment. In order to design slosh generating maneuvers, a parametric study with three maneuvers types was conducted using the General Moving Object (GMO) model in Flow-30. The three types of maneuvers are a translation maneuver, a rotation maneuver and a combined rotation translation maneuver. The effectiveness of each maneuver to generate slosh is determined by the deviation of the experiment's trajectory as compared to a dry mass trajectory. To fully capture the effect of liquid re-distribution on experiment trajectory, each thruster is modeled as an independent force point in the Flow-3D simulation. This is accomplished by modifying the total number of independent forces in the GMO model from the standard five to twenty-four. Results demonstrate that the most effective slosh generating maneuvers for all motions occurs when SSE thrusters are producing the highest changes in SSE acceleration. The results also demonstrate that several centimeters of trajectory deviation between the dry and slosh cases occur during the maneuvers; while these deviations seem small, they are measureable by SSE instrumentation.

Schulman, Richard

Structure and functions of simple membrane-water interfaces

The structure and functions of the earliest ancestors of contemporary cells are focal points in studies of the origin of life. Probably the first cell-like structures were vesicles - closed, spheroidal structures with aqueous medium trapped inside. The membranous walls of vesicles were most likely bilayers composed of simple amphiphilic material available on early earth. The membrane studied was composed of glycerol 1-monooleate (GMO). Glycerol forms the polar head group and the oily tail contains 18 carbon atoms. All head groups have been found to be located in two narrow regions at the interfaces with water. The membrane interior, formed by the hydrophobic tails, is quite fluid with chain disorder increasing towards the center of the bilayer. These results are in agreement with x-ray and neutron scattering data from related bilayers. The width of the membrane is not constant, but fluctuates in time and space. Occasional thinning defects in the membrane, observed during the course of the simulations, may have a significant influence on rates of passive transport of small molecules across membranes. It has been found that water penetrates the head group region but not the oily interior of the membrane. Water molecules near the interface are oriented by dipoles of the head groups. The resulting electrostatic potential across the interface, determined in our simulations, has been found to be markedly larger than across the water-oil interface. This quantity has been implicated as the source of selectivity, with respect to the sign of the charge, as an ion approaches the interface and during transport of hydrophobic ions across membranes.

Pohorille, A.