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NASA NTRS2024

3D Printed Materials Characterization for Rapid Prototyping and Plant Growth

Through KSC IRTD funding in 2022, this project brought a list of 18, 3D printed filaments into formal characterization testing to provide a reference for their behaviors under relevant applications. The project format set up a series of tests to expose 3D printed specimens. A total of 1,989 individual 3D printed test specimens were sent across KSC to be scrutinized by three laboratories to fulfill a multidisciplinary assessment of each material TRL. Testing started with 18 materials. Initially, seed germination assays in the PPA, sample materials were enclosed in petri dishes with lettuce seeds on damp germination paper. No significant impacts on lettuce seed germination were observed in this testing. Next, sample coupons were printed and sent for materials testing to the KSC Analysis/Mechanical and Environmental Testing Laboratory, where they were subjected to 14- and 30-day soak periods in solutions used to provide nutrients to plants or to sanitize hardware before and after use. Following a long soak typical of a 30-day plant growout in Hoagland’s solution, 14 materials gained more than 10% of their own mass. This indicated an increased potential for leaching or providing conditions that are not food safe. Materials that exceeded 15% absorption by mass were eliminated from further testing. Based off this result, the team continued with a core list of nine filaments to fulfill Tensile, Flexural, Biofilm formation, and plant growth testing. Those materials were PLA (Raise3D), ABS (Raise3D), PETG (PolyethyleneTerephthalate Glycol) (Raise3D), ASA (Acrylonitrile Styrene Acrylate) (Raise3D), PC (Polycarbonate) (Raise3D), TPU (Thermoplastic polyurethane)-95 (Raise3D), PLA Copper (Gizmodorks), PP (Polypropylene) (Braskem), and HIPS (High Impact Polystyrene) (Gizmodorks). Testing also quantified the spectral impact of using different color 3D printed surfaces in a growth chamber. The material used for spectral testing was PLA. Printing employed a standard surface texture representative of all materials. It was shown through Tensile Testing (ASTM D638-22) that the breaking force of a 3D printed part greatly varied depending on layer orientation. This is common through all materials, and demonstrates that the strength of a 3D printed component can be maximized by layering the material normal to the primary force on the part. Four-point flexural testing (ASTM D790) provided quantities of interest, Flexural modulus, Flexural strength, Flexural stress, and strain at break within a 5% strain limit from each of nine materials. Biofilm formation testing was conducted in the Molecular and Microbiological Laboratory. Testing completed on specimens from each material showed equal formation on the surface. Additional plant growth testing was conducted in the PPA beyond the initial germination testing. The final assessment documents that three materials (PLA, ABS, and PC) have reached TRL 6 through extensive testing, and ultimate end-to-end applied use in experimental or testing conditions (flight and ground). TRL 5 materials (ASA, TPU-95, PLA Copper, PP, PETG, and HIPS) have all been successfully applied in Research and Development for crop growth applications and are ready to be applied in formal testing. TRL 4 materials Nylon910, PLA Carbon Fiber, PPA CF, PPA Glass Fiber (GF), NinjaFlex, and P-filament 721 are materials that were able to be printed and tested, but have yet to show data meeting applied requirements. TRL 3 NylonX, Flex TPE-185, and Nylon were unable to be reliably printed to fulfill testing. These results provide researchers with reference for materials to use during plant growth experimentation, and also set a standard for future characterization work applying 3D printing and materials to testing, research, and experimentation.

Gioia Massa↗

DOE OSTI2016

Fatty acid esters produced by Lasiodiplodia theobromae function as growth regulators in tobacco seedlings

The Botryosphaeriaceae are a family of trunk disease fungi that cause dieback and death of various plant hosts. This work sought to characterize fatty acid derivatives in a highly virulent member of this family, Lasiodiplodia theobromae. Nuclear magnetic resonance and gas chromatography-mass spectrometry of an isolated compound revealed (Z, Z)-9,12-ethyl octadecadienoate, (trivial name ethyl linoleate), as one of the most abundant fatty acid esters produced by L. theobromae. A variety of naturally produced esters of fatty acids were identified in Botryosphaeriaceae. In comparison, the production of fatty acid esters in the soil-borne tomato pathogen Fusarium oxysporum, and the non-phytopathogenic fungus Trichoderma asperellum was found to be limited. Ethyl linoleate, ethyl hexadecanoate (trivial name ethyl palmitate), and ethyl octadecanoate, (trivial name ethyl stearate), significantly inhibited tobacco seed germination and altered seedling leaf growth patterns and morphology at the highest concentration (0.2 mg/mL) tested, while ethyl linoleate and ethyl stearate significantly enhanced growth at low concentrations, with both still inducing growth at 98 ng/mL. This work provides new insights into the role of naturally esterified fatty acids from L. theobromae as plant growth regulators with similar activity to the well-known plant growth regulator gibberellic acid. - Highlights: • Lasiodiplodia theobromae produces a wide variety of fatty acid esters in natural substrates. • Ethyl stearate and ethyl linoleate inhibit tobacco germination at 0.2 mg/mL. • Ethyl stearate and ethyl linoleate induce tobacco germination at 98 ng/mL. • Tobacco growth increase in ethyl stearate and ethyl linoleate parallels gibberellic acid. • A role as plant growth regulators is proposed for fatty acid esters.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2011

Soil bioassays as tools for sludge compost quality assessment

Composting is a waste management technology that is becoming more widespread as a response to the increasing production of sewage sludge and the pressure for its reuse in soil. In this study, different bioassays (plant germination, earthworm survival, biomass and reproduction, and collembolan survival and reproduction) were assessed for their usefulness in the compost quality assessment. Compost samples, from two different composting plants, were taken along the composting process, which were characterized and submitted to bioassays (plant germination and collembolan and earthworm performance). Results from our study indicate that the noxious effects of some of the compost samples observed in bioassays are related to the low organic matter stability of composts and the enhanced release of decomposition endproducts, with the exception of earthworms, which are favored. Plant germination and collembolan reproduction inhibition was generally associated with uncomposted sludge, while earthworm total biomass and reproduction were enhanced by these materials. On the other hand, earthworm and collembolan survival were unaffected by the degree of composting of the wastes. However, this pattern was clear in one of the composting procedures assessed, but less in the other, where the release of decomposition endproducts was lower due to its higher stability, indicating the sensitivity and usefulness of bioassays for the quality assessment of composts.

09 BIOMASS FUELS↗

DOE OSTI2014

Optimization of Eisenia fetida stocking density for the bioconversion of rock phosphate enriched cow dung–waste paper mixtures

Highlights: • Vermidegradation of RP-enriched waste mixtures is dependent on E. fetida stocking density. • A stocking density of 12.5 g-worms kg{sup -1} resulted in highly humified vermicomposts. • P release from RP-enriched waste vermicomposts increases with E. fetida stocking density. • RP-enriched waste vermicomposts had no inhibitory effect on seed germination. - Abstract: Vermitechnology is gaining recognition as an environmental friendly waste management strategy. Its successful implementation requires that the key operational parameters like earthworm stocking density be established for each target waste/waste mixture. One target waste mixture in South Africa is waste paper mixed with cow dung and rock phosphate (RP) for P enrichment. This study sought to establish optimal Eisenia fetida stocking density for maximum P release and rapid bioconversion of RP enriched cow dung–paper waste mixtures. E. fetida stocking densities of 0, 7.5, 12.5, 17.5 and 22.5 g-worms kg{sup −1} dry weight of cow dung–waste paper mixtures were evaluated. The stocking density of 12.5 g-worms kg{sup −1} resulted in the highest earthworm growth rate and humification of the RP enriched waste mixture as reflected by a C:N ratio of <12 and a humic acid/fulvic acid ratio of >1.9 in final vermicomposts. A germination test revealed that the resultant vermicompost had no inhibitory effect on the germination of tomato, carrot, and radish. Extractable P increased with stocking density up to 22.5 g-worm kg{sup −1} feedstock suggesting that for maximum P release from RP enriched wastes a high stocking density should be considered.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

NASA NTRS1990

A direct screening procedure for gravitropism mutants in Arabidopsis thaliana (L.) Heynh

In order to isolate gravitropism mutants of Arabidopsis thaliana (L.) Heynh. var Estland for the genetic dissection of the gravitropism pathway, a direct screening procedure has been developed in which mutants are selected on the basis of their gravitropic response. Variability in hypocotyl curvature was dependent on the germination time of each seed stock, resulting in the incorrect identification of several lines as gravitropism mutants when a standard protocol for the potentiation of germination was used. When the protocol was adjusted to allow for differences in germination time, these lines were eliminated from the collection. Out of the 60,000 M2 seedlings screened, 0.3 to 0.4% exhibited altered gravitropism. In approximately 40% of these mutant lines, only gravitropism by the root or the hypocotyl was altered, while the response of the other organ was unaffected. These data support the hypothesis that root and hypocotyl gravitropism are genetically separable.

NASA Discipline Number 29-20↗

DOE OSTI2025

Evaluating the combined effects of light and water availability on the early growth and physiology of Tamarindus indica : Implications for restoration

Abstract Premise The tamarind tree (Tamarindus indica) is a species of significant cultural, economic, and ecological value, with a pantropical distribution. However, the tamarind is experiencing a decline in wild populations in its native range, but the reasons for its decline remain unknown. Methods We examined the critical early life‐history stages for tamarind establishment to understand how varying levels of light and water availability and watering frequency affect its regeneration. Through three greenhouse experiments, we assessed the impact of these resources on the germination, survival, growth, and physiological responses of tamarind seedlings and saplings. Results Water availability was critical for seed germination, but not light levels or pre‐germination treatments. Light was the primary limiting factor for seedling growth. Tamarinds in high light availability grew taller, had more biomass and larger diameter, but the effect of light was modulated by water availability, indicating that there was an interaction between both resources. Water and light affected specific leaf area and leaf dry matter content but not biomass allocation, root‐to‐shoot ratio, or stomatal conductance. Water availability influenced sapling growth, but watering frequency did not, indicating a resilience of tamarind saplings to changes in rainfall periodicity but a sensitivity to total rainfall amounts. Conclusions Our study underscores the importance of considering both light and water availability in tamarind restoration efforts and contribute to understanding plant responses and trade‐offs under different levels of critical resources. Our findings will inform conservation strategies to support the regeneration and long‐term survival ofTamarindus indicain its native habitats.

Plant Sciences↗

DOE OSTI2016

Phosphorylation of spore coat proteins by a family of atypical protein kinases

The modification of proteins by phosphorylation occurs in all life forms and is catalyzed by a large superfamily of enzymes known as protein kinases. We recently discovered a family of secretory pathway kinases that phosphorylate extracellular proteins. One member, family with sequence similarity 20C (Fam20C), is the physiological Golgi casein kinase. While examining distantly related protein sequences, we observed low levels of identity between the spore coat protein H (CotH), and the Fam20C-related secretory pathway kinases. CotH is a component of the spore in many bacterial and eukaryotic species, and is required for efficient germination of spores in Bacillus subtilis; however, the mechanism by which CotH affects germination is unclear. In this paper, we show that CotH is a protein kinase. The crystal structure of CotH reveals an atypical protein kinase-like fold with a unique mode of ATP binding. Examination of the genes neighboring cotH in B. subtilis led us to identify two spore coat proteins, CotB and CotG, as CotH substrates. Furthermore, we show that CotH-dependent phosphorylation of CotB and CotG is required for the efficient germination of B. subtilis spores. Finally and collectively, our results define a family of atypical protein kinases and reveal an unexpected role for protein phosphorylation in spore biology.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2021

Characterization of growth and development of sorghum genotypes with differential susceptibility to Striga hermonthica

Two sorghum varieties, Shanqui Red (SQR) and SRN39, have distinct levels of susceptibility to the parasitic weed Striga hermonthica, which have been attributed to different strigolactone composition within their root exudates. Root exudates of the Striga-susceptible variety Shanqui Red (SQR) contain primarily 5-deoxystrigol, which has a high efficiency for inducing Striga germination. SRN39 roots primarily exude orobanchol, leading to reduced Striga germination and making this variety resistant to Striga. The structural diversity in exuded strigolactones is determined by a polymorphism in the LOW GERMINATION STIMULANT 1 (LGS1) locus. Yet, the genetic diversity between SQR and SRN39 is broad and has not been addressed in terms of growth and development. Here, we demonstrate additional differences between SQR and SRN39 by phenotypic and molecular characterization. A suite of genes related to metabolism was differentially expressed between SQR and SRN39. Increased levels of gibberellin precursors in SRN39 were accompanied by slower growth rate and developmental delay and we observed an overall increased SRN39 biomass. The slow-down in growth and differences in transcriptome profiles of SRN39 were strongly associated with plant age. Additionally, enhanced lateral root growth was observed in SRN39 and three additional genotypes exuding primarily orobanchol. In summary, we demonstrate that the differences between SQR and SRN39 reach further than the changes in strigolactone profile in the root exudate and translate into alterations in growth and development.

09 BIOMASS FUELS↗

DOE OSTI2013

Development of Genomic and Genetic Tools for Foxtail Millet, and Use of These Tools in the Improvement of Biomass Production for Bioenergy Crops-Transformation Technologies (Final Report)

Foxtail millet (Setaria italica L.) is a warm-season, C4 annual crop commonly grown for grain and forage worldwide. It has a relatively short generation time, yet produces hundreds of seeds per inflorescence. The crop is inbred and it has a small-size genome (~500 Mb). These features make foxtail millet an attractive grass model, especially for bioenergy crops. While a number of genomic tools have been established for foxtail millet, including a fully sequenced genome and molecular markers, the objectives of this project were to develop a tissue culture system, determine the best explant(s) for tissue culture, optimize transient gene expression, and establish a stable transformation system for foxtail millet cultivar Yugu1. In optimizing a tissue culture medium for the induction of calli and somatic embryos from immature inflorescences and mature seed explants, Murashige and Skoog medium containing 2.5 mg l-1 2,4-dichlorophenoxyacetic acid and 0.6 mg l-1 6- benzylaminopurine was determined to be optimal for callus induction of foxtail millet. The efficiency of callus induction from explants of immature inflorescences was significantly higher at 76% compared to that of callus induction from mature seed explants at 68%. The calli induced from this medium were regenerated into plants at high frequency (~100%) using 0.2 mg l-1 kinetin in the regeneration media. For performing transient gene expression, immature embryos were first isolated from inflorescences. Transient expression of the GUS reporter gene in immature embryos was significantly increased after sonication, a vacuum treatment, centrifugation and the addition of L-cysteine and dithiothreitol, which led to the efficiency of transient expression at levels greater than 70% after Agrobacterium inoculation. Inoculation with Agrobacterium was also tested with germinated seeds. The radicals of germinated seeds were pierced with needles and dipped into Agrobacterium solution. This method achieved a 10% transient expression efficiency. Throughout these analyses, using plasmids with the hygromycin selectable marker, it was determined that 1.5 mg l-1 hygromycin was the optimal dose for genetic transformation of foxtail millet. In contrast, the nptII selectable marker appeared to yield many escapes. Three methods of transformation were employed in an attempt to produce stable transformants. An in planta transformation experiment, similar to the floral dip method used in Arabidopsis, which utilized a red fluorescent protein pporRFP from coral Porites porites and the hygromycin selectable marker, was tested using immature inflorescences. Although several plants were PCR positive using endpoint and Real-Time PCR and there was transient expression using pporRFP and GUS reporters, no plants were positive on Southern blot. Dipping in Agrobacterium may damage the anther or the pistil because seed production was significantly reduced. Agrobacterium transformation using embryogenic calli was also tested. Although hundreds of plants were regenerated from selection, none were positive using PCR. The third method was to wound germinated seeds with an Agrobacterium coated needle, but none of the plants were PCR positive. Although the Yugu1 genotype was recalcitrant to genetic transformation, several avenues of future research should be considered for foxtail millet. Calli from different foxtail millet genotypes should be screened and selected for regeneration potential, and some genotypes may be more amenable to transformation. Additional selectable markers should also be tested as hygromycin appears to be too stringent and there are too many escapes with nptII. This project has provided training for the following personnel: Dr. Xinlu Chen (postdoc), Xiaomei Liu (postdoc), Jayashree Desai (postdoc) and Kyle Berk (Undergraduate researcher). Conference presentations and peer-reviewed journal articles partly supported by this grant includes the following: 1. Baxter H., Equi R., Chen X, Berk K. and Zale J. Establishing Efficient in vitro Protocols For Foxtail Millet (Setaria italica L. cv. Yugi 1). Plant & Animal Genomes XVIII Conference XVIII, San Diego, California, January 2010 2. Chen X, Zale J and Chen F. The Regeneration and Transformation of Foxtail Millet (Setaria italica), A Model Biofuel Crop. Genomic Science Awardee Meeting IX and USDA-DOE Plant Feedstock Genomics for Bioenergy Awardee Meeting, Crystal City, Virginia, April 2011 3. Chen, F., Tholl, D., Bohlmann, J., and Pichersky, E. (2011) The family of terpene synthases in plants: A mid-size family of genes for specialized metabolism that is highly diversified throughout the kingdom. Plant J. 66: 212-229.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2021

Plant Growth Study: Growth of Three Plant Species in 100-OL-1 Operable Unit Soils

Soil contamination from historical application of lead arsenate pesticides persists in the 100-OL-1 Operable Unit (OU) of the U.S. Department of Energy’s Hanford Site in Benton County, Washington. A remedial investigation used a portable X-ray fluorescence analyzer to estimate and map the concentrations of metals in surface soil in the 133 decision units comprising the OU. There is variability in lead and arsenic concentrations among the non-contiguous decision units, and an incomplete understanding of the ecological and human risks from soil contamination. While general screening criteria for lead are reported in literature, limited information is available on the impacts of heavy metals on site-specific plant and animal life. To address this information gap, a study on plant growth in Hanford soils was conducted using native bluegrass (Poa secunda), invasive cheatgrass (Bromus tectorum), and buttercrunch lettuce (Lactuca sativa). The latter is included because it is a common species used in plant growth studies. Soil samples were collected from a single general location within the 100-OL-1 OU, prepared, and categorized as having low, medium, or high concentrations of lead and arsenic, with the high concentration soil having concentrations of up to 3,400 and 790 mg/kg lead and arsenic, respectively. Additionally, a synthetic soil was prepared and used as a control, in accordance with standard plant growth protocols. Fifty-four seeds of each plant species were planted in each of the four soil types following the procedure outlined by the Washington State Department of Ecology. Germination rates and biomass measurements were recorded for 20 days. Both bluegrass and lettuce germination rates appeared to be reduced at the highest lead and arsenic concentrations (Pb: 3400 mg/kg, As: 790 mg/kg), while cheatgrass germination rates were unaffected. Total biomass for all species appeared to be related to the relative concentrations of lead and arsenic in the soil. Results of this growth study agree reasonably well with previous Hanford Site plant growth studies and provide additional data for assessing ecological risk at the Hanford Site.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2022

Hanford Site Rare Plant Monitoring Report for Calendar Year 2022

Rare plants are monitored on the Hanford Site as distribution and abundance data of current and historic rare plant populations are necessary to make well-informed environmental and ecological resource management decisions. The Central Hanford Rare Plant Management Plan (DOE/RL-2021-35) guides rare plant monitoring methodology in the context of resource management at the Hanford Site. In 2022, a rare plant monitoring effort occurred on the portion of the Hanford Site (referred to herein as Central Hanford) managed by the U.S. Department of Energy, Richland Operations Office (DOE-RL; Figure 1). The goal of this effort was to revisit known occurrences of upland rare plant populations on Central Hanford to improve baseline information on locations of rare plant populations on the Hanford Site. Rare upland plants were thoroughly surveyed in 2021 but unusually low precipitation that year limited germination of many upland annual plants. Surveys in 2022 focused on rare upland annual species after the Hanford Site received higher than average spring precipitation that was expected to increase germination. Rainfall from April 2022 to June 2022 was 76% higher than average; however, these months were also colder than average. Winter rainfall from January 2022 to March 2022 was 84% lower than average. The high spring precipitation in 2022 appears to have triggered the germination of some populations of rare upland annuals but the dry winter and cold spring may have inhibited others. Population stability, viability, and threats were evaluated and will be used to inform future management of rare plant species.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2013

Technique for rapid establishment of American lotus in remediation efforts

A technique for increasing the establishment rate of American lotus (Nelumbo lutea) and simplifying planting was developed as part of a pond remediation project. Lotus propagation techniques typically require scarification of the seed, germination in heated water, and planting in nursery containers. Then mature (~ 1 yr) nursery-grown stock is transferred to planting site or scarified seed are broadcast applied. Mature plants should grow more quickly, but can be sensitive to handling, require more time to plant, and cost more. Scarified seeds are easier to plant and inexpensive, but have a lag time in growth, can fail to germinate, and can be difficult to site precisely. We developed an intermediate technique using small burlap bags that makes planting easier, provides greater germination success, and avoids lag time in growth. Data on survival and growth from experiments using mature stock, scarified seeds, and bag lotus demonstrate that bag lotus grow rapidly in a variety of conditions, have a high survival rate, can be processed and planted easily and quickly, and are very suitable for a variety of remediation projects

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS1984

Aromatic biosynthesis in pine tissues

Pinus elliotti is a woody plant species responsive to gravity and capable of synthesizing large quantities of lignin. Lignification begins very quickly after germination; lignin is detected in the vascular region within 4 days after germination and rapidly progresses up the hypocotyl. Young pine seedlings bend in response to geostimulation for about 10 days after germination, with the most rapid response time occurring in 4- to 5-day-old seedlings. Various chemicals were used to establish their effects on the geotropic response in this gymnosperm species. IAA completely arrests the geotropic response for 18 to 24 hr. Afterward the seedlings respond geostimulation as if they were not treated. The same pattern of response will occur with a second IAA treatment. If the synthetic auxin, 2-4,D, is used, the georesponse is permanently blocked. The method of application does not appear to be critical; addition of auxin to only one side of the seedling gave results similar to those obtained by treating the entire seedling.

Cowles, J. R.↗

NASA NTRS1991

Space Exposed Experiment Developed for Students (SEEDS) P-0004-2

This cooperative endeavor of NASA Headquarters, the NASA Langley Research Center, and the George W. Park Seed Company, resulted in the distribution, by the end of March, 1990, of approximately 132,000 space exposed experiment developed for students (SEEDS) kits to 64,000 teachers representing 40,000 classrooms and 3.3 million kindergarden through university students. Kits were sent to every state, as well as to 30 foreign countries. Preliminary radiation data indicates that layer A received 725 rads, while layer D received 350 rads. Germination rate was reported to be 73.8 percent for space exposed seeds and 70.3 percent for earth based control seeds. Tests conducted within the first six months after retrieval indicated space exposed seeds germinated in an average of 8.0 days, while earth based control seeds' average germination rate was 8.3 days. Some mutations (assumed to be radiation induced) reported by students and Park Seed include plants that added a leaf instead of the usual flower at the end of the flower front and fruit produced from a flower with a variegated calyx bore seeds producing albino plants, while fruit from a flower with a green calyx from the same plant bore seeds produced green plants.

Grigsby, Doris K.↗

NASA NTRS1994

Seed viability detection using computerized false-color radiographic image enhancement

Seed radiographs are divided into density zones which are related to seed germination. The seeds which germinate have densities relating to false-color red. In turn, a seed sorter may be designed which rejects those seeds not having sufficient red to activate a gate along a moving belt containing the seed source. This results in separating only seeds with the preselected densities representing biological viability lending to germination. These selected seeds demand a higher market value. Actual false-coloring isn't required for a computer to distinguish the significant gray-zone range. This range can be predetermined and screened without the necessity of red imaging. Applying false-color enhancement is a means of emphasizing differences in densities of gray within any subject from photographic, radiographic, or video imaging. Within the 0-255 range of gray levels, colors can be assigned to any single level or group of gray levels. Densitometric values then become easily recognized colors which relate to the image density. Choosing a color to identify any given density allows separation by morphology or composition (form or function). Additionally, relative areas of each color are readily available for determining distribution of that density by comparison with other densities within the image.

Vozzo, J. A.↗

NASA NTRS2002

Effects of Disinfectants in Water on Mir- and Earth-Grown Wheat

Iodine and silver fluoride are used to purify water onboard U. S. Shuttles and the Russian Space Station, Mir, respectively. In 1995, iodine-treated water, which ranged from 1.0-4.0 mg x kg(exp -1) with a mean of 2.9 mg x kg(exp -1), was applied to Super Dwarf wheat (Triticum aestivum L.) plants when Mir water (grey or tech grade) became scarce. The potential phytotoxicity of iodine on Super Dwarf wheat is an unknown. Since use of iodine-treated water was not part of the experiment, we sought to determine whether it accounted for the subsequent poor wheat seedling growth and floral development onboard the Mir. Super Dwarf wheat seeds were imbibed in iodine or silver fluoride concentrations of 0.0, 1.0, 2.0, 4.0, 8.0 or 16.0 mg x kg(exp -1) for 96 h at 4 C. Five seeds were then planted per 13.3 cm x 13.3 cm pots containing a granular clinoptilolite (Cp) zeolite (1 -2 mm dia.) and placed in Percival(TM) growth chambers programmed for 20/15 C and 18/6 h d/n regime. Plants were irrigated with distilled water, and Iodine- or silver fluoride-treated distilled water. In separate experiments, seeds were treated as above and germination and early seedling growth were determined by examining seedling responses to disinfectants in rolled paper towels. Silver fluoride had very little effect on wheat seed germination. By contrast, iodine reduced germination at all treatment levels. Seedlings exposed to 1.0, 2.0, and 4.0 mg x kg(exp -1) of iodine or silver fluoride levels exhibited a slight stimulation in shoot and root growth. Both disinfectants at 8 and 16 mg x kg(exp -1) showed significantly (p is less than or equal to 0.01) reduced seedling shoot and root lengths and fresh biomasses compared to the control and lower disinfectant levels. The number of spikelets per spike, florets per spikelet, seeds per spike and seed weight were also significantly reduced at the 8 and 16 mg x kg(exp -1) compared to the control and lower levels of disinfectant. Based on these ground-based post-flight analyses, the levels of iodine- and/or silver fluoride-treated water used on Mir-grown plants onboard the Mir did not cause the poor growth and development of the wheat plants.

Campbell, William .F.↗

NASA NTRS2000

Ceratopteris richardii: a productive model for revealing secrets of signaling and development

Ceratopteris richardii is an aquatic fern grown in tropical and subtropical regions of the world. It is proven to be a productive model system for studies in the genetics, biochemistry, and cell biology of basic biologic processes that occur in early gametophytic development. It provides several advantages to biologists, especially those interested in gravitational biology, polarity development, and in the genetics of sexual development. It is easy to culture, has a relatively short life cycle, and offers an array of attractive features that facilitate genetic studies. The germination and early development of large populations of genetically identical spores are easy to synchronize, and both the direction of polarity development and cell-level gravity responses can be measured and readily manipulated within the first 24 h of spore development. Although there is no reliable transformation system available yet in Ceratopteris, recent studies suggest that the technique of RNA interference can be used to block translation of specific genes in a related fern, Marsilea, and current studies will soon reveal the applicability of this approach, as well as of other transformation approaches, in Ceratopteris. A recently completed expressed sequence tag (EST) sequencing project makes available the partial sequence of more than 2000 cDNAs, representing a significant percentage of the genes being expressed during the first 24 h of spore germination, when many developmentally interesting processes are occurring. A microarray of these ESTs is being constructed, so especially for those scientists interested in basic cellular phenomena that occur early in spore germination, the availability of the ESTs and of the microarray will make Ceratopteris an even more attractive model system.

NASA Discipline Plant Biology↗

NASA NTRS1999

Early root cap development and graviresponse in white clover (Trifolium repens) grown in space and on a two-axis clinostat

White clover (Trifolium repens) was germinated and grown in microgravity aboard the Space Shuttle (STS-60, 1994; STS-63, 1995), on Earth in stationary racks and in a slow-rotating two-axis clinostat. The objective of this study was to determine if normal root cap development and early plant gravity responses were dependent on gravitational cues. Seedlings were germinated in space and chemically fixed in orbit after 21, 40, and 72 h. Seedlings 96 h old were returned viable to earth. Germination and total seedling length were not dependent on gravity treatment. In space-flown seedlings, the number of cell stories in the root cap and the geometry of central columella cells did not differ from those of the Earth-grown seedlings. The root cap structure of clinorotated plants appeared similar to that of seedlings from microgravity, with the exception of three-day rotated plants, which displayed significant cellular damage in the columella region. Nuclear polarity did not depend on gravity; however, the positions of amyloplasts in the central columella cells were dependent on both the gravity treatment and the age of the seedlings. Seedlings from space, returned viable to earth, responded to horizontal stimulation as did 1 g controls, but seedlings rotated on the clinostat for the same duration had a reduced curvature response. This study demonstrates that initial root cap development is insensitive to either chronic clinorotation or microgravity. Soon after differentiation, however, clinorotation leads to loss of normal root cap structure and plant graviresponse while microgravity does not.

manned↗