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At least 19 records
All sourcesNASA NTRSDOE OSTI
NASA NTRS2001

Auxins and tropisms

Differential growth of plants in response to the changes in the light and gravity vectors requires a complex signal transduction cascade. Although many of the details of the mechanisms by which these differential growth responses are induced are as yet unknown, auxin has been implicated in both gravitropism and phototropism. Specifically, the redistribution of auxin across gravity or light-stimulated tissues has been detected and shown to be required for this process. The approaches by which auxin has been implicated in tropisms include isolation of mutants altered in auxin transport or response with altered gravitropic or phototropic response, identification of auxin gradients with radiolabeled auxin and auxin-inducible gene reporter systems, and by use of inhibitors of auxin transport that block gravitropism and phototropism. Proteins that transport auxin have been identified and the mechanisms which determine auxin transport polarity have been explored. In addition, recent evidence that reversible protein phosphorylation controls this process is summarized. Finally, the data in support of several hypotheses for mechanisms by which auxin transport could be differentially regulated during gravitropism are examined. Although many details of the mechanisms by which plants respond to gravity and light are not yet clear, numerous recent studies demonstrate the role of auxin in these processes.

NASA Discipline Plant Biology↗
NASA NTRS1999

Differentiation and Tropisms in Space-Grown Moss

This grant supported a Space Shuttle experiment on the effects of microgravity on moss cells. Moss provides a rich system for gravitational and spaceflight research. The early phase of the moss life cycle consists of chains of cells that only grow only at their tips. In the moss Ceratodon purpureus these filaments (protonemata) grow away from gravity in the dark, in a process called gravitropism. The tipmost cells, the apical cells, contain heavy starch-filled bodies called amyloplasts that probably function in g-sensing and that sediment within the apical cell. The SPM-A (Space Moss aka SPAM) experiment flew in November - December, 1997 on STS-87 as part of the Collaborative US Ukrainian Experiment (CLTE). The experiment was accommodated in hardware purpose-built by NASA KSC and Bionetics and included Petri Dish Fixation Units (PDFU) and BRIC-LEDs. Together, this hardware allowed for the culture of the moss on agar in commercial petri dishes, for unilateral illumination with red light of varying intensity, and for chemical fixation in situ. The key findings of the spaceflight were quite unexpected. Neither the orientation of tip-growth nor the distribution of amyloplasts was random in microgravity.

Sack, Fred D.↗
NASA NTRS2000

How can plants tell which way is up?

Many people think of plants as essentially sessile organisms that do not actively respond to their environment. What could be further from the truth! In fact, plants are capable of a variety of movements, including the dramatic nastic responses (such as Venus fly trap closure) and the less sensational tropisms. These latter movements are directed growth responses to some type of external stimulus such as gravity (gravitropism, formerly known as geotropism) or light (phototropism). This paper describes some interesting exercises that are derived from recent work, including research that has led to experiments performed on two Space Shuttle missions in 1997 (Kiss et al. 1998). The study of tropisms can be a useful way to introduce students to plant biology in high school and introductory college courses. In our experience, students are fascinated by plant movements when they are presented in lectures and find laboratory experiences on this topic quite engaging. Laboratory work on plant tropisms can also be used to introduce important concepts in science such as hypothesis testing, quantitative analysis, and the use of statistics. The laboratory exercises described in this paper involve the higher plant Arabidopsis thaliana, which has become an important organism in molecular biology research and is the focus of an international plant genome project. Based on the material presented here, a number of plant gravitropism laboratory exercises with Arabidopsis that are simple in terms of equipment/materials and procedures can be developed. These exercises are robust in that they work well even in the hands of introductory students, and they can be expanded according to the individual instructor's needs. This paper describes two exercises that have been performed by beginning college students, and these exercises can easily be performed in biology classes in most high school settings.

Non-NASA Center↗
NASA NTRS2002

The kinetics of root gravitropism: dual motors and sensors

The Cholodny-Went theory of tropisms has served as a framework for investigation of root gravitropism for nearly three quarters of a century. Recent investigations using modern techniques have generated findings consistent with the classical theory, including confirmation of asymmetrical distribution of polar auxin transport carriers, molecular evidence for auxin asymmetry following gravistimulation, and generation of auxin response mutants with predictable lesions in gravitropism. Other results indicate that the classical model is inadequate to account for key features of root gravitropism. Initiation of curvature, for example, occurs outside the region of most rapid elongation and is driven by differential acceleration rather than differential inhibition of elongation. The evidence indicates that there are two motors driving root gravitropism, one of which appears not to be auxin regulated. We have recently developed technology that is capable of maintaining a constant angle of gravistimulation at any selected target region of a root while continuously monitoring growth and curvature kinetics. This review elaborates on the advantages of this new technology for analyzing gravitropism and describes applications of the technology that reveal (1) the existence of at least two phases to gravitropic motor output, even under conditions of constant stimulus input and (2) the existence of gravity sensing outside of the root cap. We propose a revised model of root gravitropism including dual sensors and dual motors interacting to accomplish root gravitropism, with only one of the systems linked to the classical Cholodny-Went theory.

Review↗
DOE OSTI2024

In vivo human T cell engineering with enveloped delivery vehicles

Viruses and virally derived particles have the intrinsic capacity to deliver molecules to cells, but the difficulty of readily altering cell-type selectivity has hindered their use for therapeutic delivery. Here, we show that cell surface marker recognition by antibody fragments displayed on membrane-derived particles encapsulating CRISPR–Cas9 protein and guide RNA can deliver genome editing tools to specific cells. Compared to conventional vectors like adeno-associated virus that rely on evolved capsid tropisms to deliver virally encoded cargo, these Cas9-packaging enveloped delivery vehicles (Cas9-EDVs) leverage predictable antibody–antigen interactions to transiently deliver genome editing machinery selectively to cells of interest. Antibody-targeted Cas9-EDVs preferentially confer genome editing in cognate target cells over bystander cells in mixed populations, both ex vivo and in vivo. By using multiplexed targeting molecules to direct delivery to human T cells, Cas9-EDVs enable the generation of genome-edited chimeric antigen receptor T cells in humanized mice, establishing a programmable delivery modality with the potential for widespread therapeutic utility.

42 ENGINEERING↗
DOE OSTI2025

Viral delivery of recombinases activates heritable genetic switches in plants

Viral vectors provide an increasingly versatile platform for transformation-free reagent delivery to plants. RNA viral vectors can be used to induce gene silencing, overexpress proteins, or introduce gene editing reagents; however, they are often constrained by carrying capacity or restricted tropism in germline cells. Site-specific recombinases that catalyze precise genetic rearrangements are powerful tools for genome engineering that vary in size and, potentially, efficacy in plants. In this work, we show that viral vectors based on tobacco rattle virus (TRV) deliver and stably express four recombinases ranging in size from ~0.6kb to ~1.5kb and achieve simultaneous marker removal and reporter activation through targeted excision in transgenic Nicotiana benthamiana lines. TRV vectors with Cre, FLP, CinH, and Integrase13 efficiently mediated recombination in infected somatic tissue and led to heritable modifications at high frequency. Here, an excision-activated Ruby reporter enabled simple and high-resolution tracing of infected cell lineages without the need for molecular genotyping. Together, our experiments broaden the scope of viral recombinase delivery and offer insights into infection dynamics that may be useful in developing future viral vectors.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2025

Data for "Viral Delivery of Recombinases Activates Heritable Genetic Switches in Plants"

Viral vectors provide an increasingly versatile platform for transformation-free reagent delivery to plants. RNA viral vectors can be used to induce gene silencing, overexpress proteins, or introduce gene editing reagents; however, they are often constrained by carrying capacity or restricted tropism in germline cells. Site-specific recombinases that catalyze precise genetic rearrangements are powerful tools for genome engineering that vary in size and, potentially, efficacy in plants. In this work, we show that viral vectors based on tobacco rattle virus (TRV) deliver and stably express four recombinases ranging in size from ∼0.6 to ∼1.5 kb and achieve simultaneous marker removal and reporter activation through targeted excision in transgenic Nicotiana benthamiana lines. TRV vectors with Cre, FLP, CinH, and Integrase13 efficiently mediated recombination in infected somatic tissue and led to heritable modifications at high frequency. An excision-activated Ruby reporter enabled simple and high-resolution tracing of infected cell lineages without the need for molecular genotyping. Together, our experiments broaden the scope of viral recombinase delivery and offer insights into infection dynamics that may be useful in developing future viral vectors.

gene editing↗
DOE OSTI2024

Structure and antigenicity of the divergent human astrovirus VA1 capsid spike

Human astrovirus (HAstV) is a known cause of viral gastroenteritis in children worldwide, but HAstV can cause also severe and systemic infections in immunocompromised patients. There are three clades of HAstV: classical, MLB, and VA/HMO. While all three clades are found in gastrointestinal samples, HAstV-VA/HMO is the main clade associated with meningitis and encephalitis in immunocompromised patients. To understand how the HAstV-VA/HMO can infect the central nervous system, we investigated its sequence-divergent capsid spike, which functions in cell attachment and may influence viral tropism. Here we report the high-resolution crystal structures of the HAstV-VA1 capsid spike from strains isolated from patients with gastrointestinal and neuronal disease. The HAstV-VA1 spike forms a dimer and shares a core beta-barrel structure with other astrovirus capsid spikes but is otherwise strikingly different, suggesting that HAstV-VA1 may utilize a different cell receptor, and an infection competition assay supports this hypothesis. Furthermore, by mapping the capsid protease cleavage site onto the structure, the maturation and assembly of the HAstV-VA1 capsid is revealed. Finally, comparison of gastrointestinal and neuronal HAstV-VA1 sequences, structures, and antigenicity suggests that neuronal HAstV-VA1 strains may have acquired immune escape mutations. Overall, our studies on the HAstV-VA1 capsid spike lay a foundation to further investigate the biology of HAstV-VA/HMO and to develop vaccines and therapeutics targeting it.

60 APPLIED LIFE SCIENCES↗
NASA NTRS1971

Simulated weightlessness studies by compensation

Plants behavioral reactions to continuous gravitational field directional reorientation by clinostat, discussing gravity compensation effects on tropism and forces required for geotropic response

Gordon, S. A.↗
NASA NTRS1972

Hair radioactivity as a measure of exposure to radioisotopes

Since many radioisotopes accumulate in hair, this tropism was investigated by comparing the radioactivity of shaved with plucked hair collected from rats at various time intervals up to 24 hrs after intravenous injection of the ecologically important radioisotopes, iodine-131, manganese-54, strontium-85, and zinc-65. The plucked hair includes the hair follicles where biochemical transformations are taking place. The data indicate a slight surge of each radioisotpe into the hair immediately after injection, a variation of content of each radionuclide in the hair, and a greater accumulation of radioactivity in plucked than in shaved hair. These results have application not only to hair as a measure of exposure to radioisotopes, but also to tissue damage and repair at the hair follicle.

Strain, W. H.↗
NASA NTRS1973

Effect of free fall on higher plants.

The influence of exposure to the free-fall state on the orientation, morphogenesis, physiology, and radiation response of higher plants is briefly summarized. It is proposed that the duration of the space-flight experiments has been to brief to permit meaningful effects of free fall on general biochemistry, growth, and development to appear. However, two types of significant effect did occur. The first is on differential growth - i.e., tropism and epinasty - resulting from the absence of a normal geostimulus. For these phenomena it is suggested that ground-based experiments with the clinostat would suffice to mimic the effect of the free-fall state. The second is an apparent interaction between the radiation response and some flight condition, yielding an enhanced microspore abortion, a disturbed spindle function, and a stunting of stamen hairs. It is suggested that this apparent interaction may be derived from a shift in the rhythm of the cell cycle, induced by the free fall.

Gordon, S. A.↗
NASA NTRS1985

Report of the Insect Development Group

Drosophila metanogaster was chosen as the insect species of choice, in regard to gravity response experiments involving normal reproduction and develop different strains. The specific gravity responses which might be affected by microgravity and are exhibited in normal reproduction and development include normal flight for courtship, mating and oviposition, tropisms for pupating or emergency of the adult, and crawling for gettering food by the larval instars at the organismic level. At the suborganismic elevel, it is believed that maturation of developing eggs in the virgin female and embryonic development of the developing egg could be affected by microgravity and warrant study.

Rockstein, M.↗
NASA NTRS1994

Autonomous mobile robot teams

This paper describes autonomous mobile robot teams performing tasks in unstructured environments. The behavior and the intelligence of the group is distributed, and the system does not include a central command base or leader. The novel concept of the Tropism-Based Cognitive Architecture is introduced, which is used by the robots in order to produce behavior transforming their sensory information to proper action. The results of a number of simulation experiments are presented. These experiments include worlds where the robot teams must locate, decompose, and gather objects, and defend themselves against hostile predators, while navigating around stationary and mobile obstacles.

Agah, Arvin↗
NASA NTRS1995

Plant Roots: The Hidden Half: Calcium and Gravitropism - Chapter 16

Environmental signals such as light and gravity control many aspects of plant growth and development. In higher plants, the directional growth of an organ in response to stimuli such as gravity and light is considered a tropic movement. Such movement could be either positive or negative with respect to a specific stimulus. In general, stems show a positive response to light and negative response to gravity. In contrast, most roots show a positive response to gravity and a negative response to light. Investigations on plant tropism date back a century when Darwin studied the phototropic responses of maize seedlings (Darwin). Although the precise mechanism of signal perception and transduction in roots is not understood, Darwin recognized over 100 years ago that the root cap is the probable site of signal perception. He discovered that the removal of the root cap eliminates the ability of roots to respond to gravity. Other investigators have since confirmed Darwin's observation (Konings; Evans et al.). In recent years, especially with the advent of the U.S. Space Program, there has been a renewed interest in understanding how plants respond to extracellular signals such as gravity (Halstead and Dutcher). Studies on the mechanisms involved in perception and transduction of gravity signal by roots would ultimately help us to better understand gravitropism and also to grow plants under microgravity conditions as in space. In this chapter, we restrict ourselves to the role of calcium in transduction of the gravity signal. In doing so, emphasis is given to the role of calcium-modulated proteins and their role in signal transduction in gravitropism. Detailed reviews on various other aspects of gravitropism (Scott, Torrey, Wilkins, Fim and Digby, Feldman, Pickard, Moore and Evans, Halstead and Dutcher, Poovaiah et al.) and on the role of calcium as a messenger in signal transduction in general have been published (Helper and Wayne, Poovaiah and Reddy, Roberts and Hartnon, Bowler and Chua, Gilroy and Trewavas). Plant roots have been widely used to study the transduction of gravity and light signals (Poovaiah et al., Roux and Serlin). Most roots show positive gravitropic response in either dark or light. However, roots of some varieties of plants (e.g., Zea mays L., cv Merit, and Zea rwvs L., cv Golden Cross Bantam 70) show positive gravitropic response only in light (Feldman, Miyazaki et al.). Investigations from various laboratories indicate that calcium acts as a messenger in transducing gravity and light signals in plant roots(Pickard, Evans et al., Pooviah et al.).

Poovaiah, B. W.↗
NASA NTRS2003

Root phototropism: how light and gravity interact in shaping plant form

The interactions among tropisms can be critical in determining the final growth form of plants and plant organs. We have studied tropistic responses in roots as an example of these type of interactions. While gravitropism is the predominant tropistic response in roots, phototropism also plays a role in the oriented growth in this organ in flowering plants. In blue or white light, roots exhibit negative phototropism, but red light induces positive phototropism. In the flowering plant Arabidopsis, the photosensitive pigments phytochrome A (phyA) and phytochrome B (phyB) mediate this positive red-light-based photoresponse in roots since single mutants (and the double phyAB mutant) were severely impaired in this response. While blue-light-based negative phototropism is primarily mediated by the phototropin family of photoreceptors, the phyA and phyAB mutants (but not phyB) were inhibited in this response relative to the WT. The differences observed in phototropic responses were not due to growth limitations since the growth rates among all the mutants tested were not significantly different from that of the WT. Thus, our study shows that the blue-light and red-light systems interact in plants and that phytochrome plays a key role in integrating multiple environmental stimuli.

Non-NASA Center↗
NASA NTRS2003

Phytochromes A and B mediate red-light-induced positive phototropism in roots

The interaction of tropisms is important in determining the final growth form of the plant body. In roots, gravitropism is the predominant tropistic response, but phototropism also plays a role in the oriented growth of roots in flowering plants. In blue or white light, roots exhibit negative phototropism that is mediated by the phototropin family of photoreceptors. In contrast, red light induces a positive phototropism in Arabidopsis roots. Because this red-light-induced response is weak relative to both gravitropism and negative phototropism, we used a novel device to study phototropism without the complications of a counteracting gravitational stimulus. This device is based on a computer-controlled system using real-time image analysis of root growth and a feedback-regulated rotatable stage. Our data show that this system is useful to study root phototropism in response to red light, because in wild-type roots, the maximal curvature detected with this apparatus is 30 degrees to 40 degrees, compared with 5 degrees to 10 degrees without the feedback system. In positive root phototropism, sensing of red light occurs in the root itself and is not dependent on shoot-derived signals resulting from light perception. Phytochrome (Phy)A and phyB were severely impaired in red-light-induced phototropism, whereas the phyD and phyE mutants were normal in this response. Thus, PHYA and PHYB play a key role in mediating red-light-dependent positive phototropism in roots. Although phytochrome has been shown to mediate phototropism in some lower plant groups, this is one of the few reports indicating a phytochrome-dependent phototropism in flowering plants.

NASA Discipline Plant Biology↗
NASA NTRS2002

Transcription profiling of the early gravitropic response in Arabidopsis using high-density oligonucleotide probe microarrays

Studies of plant tropisms, the directed growth toward or away from external stimuli such as light and gravity, began more than a century ago. Yet biochemical, physiological, and especially molecular mechanisms of plant tropic responses remain for the most part unclear. We examined expression of 8,300 genes during early stages of the gravitropic response using high-density oligonucleotide probe microarrays. Approximately 1.7% of the genes represented on the array exhibited significant expression changes within the first 30 min of gravity stimulation. Among gravity-induced genes were a number of genes previously implicated to be involved in gravitropism. However, a much larger number of the identified genes have not been previously associated with gravitropism. Because reorientation of plants may also expose plants to mechanical perturbations, we also compared the effects of a gentle mechanical perturbation on mRNA levels during the gravity response. It was found that approximately 39% of apparently gravity-regulated genes were also regulated by the mechanical perturbation caused by plant reorientation. Our study revealed the induction of complex gene expression patterns as a consequence of gravitropic reorientation and points to an interplay between the gravitropic and mechanical responses and to the extreme sensitivity of plants to even very gentle mechanical perturbations.

NASA Discipline Plant Biology↗