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All sourcesNASA NTRSDOE OSTI
NASA NTRS2009

Microwave Sterilization and Depyrogenation System

A fully functional, microgravity-compatible microwave sterilization and depyrogenation system (MSDS) prototype was developed that is capable of producing medical-grade water (MGW) without expendable supplies, using NASA potable water that currently is available aboard the International Space Station (ISS) and will be available for Lunar and planetary missions in the future. The microwave- based, continuous MSDS efficiently couples microwaves to a single-phase, pressurized, flowing water stream that is rapidly heated above 150 C. Under these conditions, water is rapidly sterilized. Endotoxins, significant biological toxins that originate from the cell walls of gram-negative bacteria and which represent another defining MGW requirement, are also deactivated (i.e., depyrogenated) albeit more slowly, with such deactivation representing a more difficult challenge than sterilization. Several innovations culminated in the successful MSDS prototype design. The most significant is the antenna-directed microwave heating of a water stream flowing through a microwave sterilization chamber (MSC). Novel antenna designs were developed to increase microwave transmission efficiency. These improvements resulted in greater than 95-percent absorption of incident microwaves. In addition, incorporation of recuperative heat exchangers (RHxs) in the design reduced the microwave power required to heat a water stream flowing at 15 mL/min to 170 C to only 50 W. Further improvements in energy efficiency involved the employment of a second antenna to redirect reflected microwaves back into the MSC, eliminating the need for a water load and simplifying MSDS design. A quick connect (QC) is another innovation that can be sterilized and depyrogenated at temperature, and then cooled using a unique flow design, allowing collection of MGW at atmospheric pressure and 80 C. The final innovation was the use of in-line mixers incorporated in the flow path to disrupt laminar flow and increase contact time at a given flow rate. These technologies can be employed in small-scale systems for efficient production of MGW in the laboratory or in a range of larger systems that meet various industrial requirements. The microwave antennas can also be adapted to selectively sterilize vulnerable connections to ultra-pure water production facilities or biologically vulnerable systems where microorganisms may intrude.

Akse, James R.↗
NASA NTRS1992

Short-term reversibility of ultrastructural changes in pulmonary capillaries caused by stress failure

We previously showed that when the pulmonary capillaries in anesthetized rabbits are exposed to a transmural pressure (Ptm) of approximately 40 mmHg, stress failure of the walls occurs with disruption of the capillary endothelium, alveolar epithelium, or sometimes all layers. The present study was designed to determine whether some of the ultrastructural changes are rapidly reversible when the capillary pressure is reduced. To test this, the Ptm was raised to 52.5 cmH2O for 1 min of blood perfusion and then reduced to 12.5 cmH2O for 3 min of saline-dextran perfusion, followed by intravascular fixation at the same pressure. In another group of animals, the pressure was elevated for 1 min of blood and 3 min of saline-dextran before being reduced. The results were compared with previous studies in which the capillary pressures were maintained elevated at 52.5 cmH2O during the entire procedure. Control studies were also done at sustained low pressures. The results showed that the number of endothelial and epithelial breaks per millimeter and the total fraction area of the breaks were reduced when the pressure was lowered. For example, the number of endothelial breaks per millimeter decreased from 7.1 +/- 2.1 to 2.4 +/- 0.7, and the number of epithelial breaks per millimeter fell from 11.4 +/- 3.7 to 3.4 +/- 0.7. There was evidence that the breaks that closed were those that were initially small and were associated with an intact basement membrane. The results suggest that cells can move along their underlying matrix by rapid disengagement and reattachment of cell adhesion molecules, causing breaks to open or close within minutes.(ABSTRACT TRUNCATED AT 250 WORDS).

Non-NASA Center↗
NASA NTRS1995

Neurochemical phenotype of corticocortical connections in the macaque monkey: quantitative analysis of a subset of neurofilament protein-immunoreactive projection neurons in frontal, parietal, temporal, and cingulate cortices

The neurochemical characteristics of the neuronal subsets that furnish different types of corticocortical connections have been only partially determined. In recent years, several cytoskeletal proteins have emerged as reliable markers to distinguish subsets of pyramidal neurons in the cerebral cortex of primates. In particular, previous studies using an antibody to nonphosphorylated neurofilament protein (SMI-32) have revealed a consistent degree of regional and laminar specificity in the distribution of a subpopulation of pyramidal cells in the primate cerebral cortex. The density of neurofilament protein-immunoreactive neurons was shown to vary across corticocortical pathways in macaque monkeys. In the present study, we have used the antibody SMI-32 to examine further and to quantify the distribution of a subset of corticocortically projecting neurons in a series of long ipsilateral corticocortical pathways in comparison to short corticocortical, commissural, and limbic connections. The results demonstrate that the long association pathways interconnecting the frontal, parietal, and temporal neocortex have a high representation of neurofilament protein-enriched pyramidal neurons (45-90%), whereas short corticocortical, callosal, and limbic pathways are characterized by much lower numbers of such neurons (4-35%). These data suggest that different types of corticocortical connections have differential representation of highly specific neuronal subsets that share common neurochemical characteristics, thereby determining regional and laminar cortical patterns of morphological and molecular heterogeneity. These differences in neuronal neurochemical phenotype among corticocortical circuits may have considerable influence on cortical processing and may be directly related to the type of integrative function subserved by each cortical pathway. Finally, it is worth noting that neurofilament protein-immunoreactive neurons are dramatically affected in the course of Alzheimer's disease. The present results support the hypothesis that neurofilament protein may be crucially linked to the development of selective neuronal vulnerability and subsequent disruption of corticocortical pathways that lead to the severe impairment of cognitive function commonly observed in age-related dementing disorders.

Non-NASA Center↗
NASA NTRS2019

Effects of Altered Gravity on the Central Nervous System of Drosophila melanogaster

A comprehensive understanding of the effects of spaceflight and altered gravity on human physiology is necessary for continued human space exploration and long-term space habitation. Spaceflight includes multiple factors such as microgravity, hypergravity, ionizing radiation, physiological stress, and disrupted circadian rhythms and these have been shown to contribute to pathophysiological responses that target immunity, bone and muscle integrity, cardiovascular and nervous systems. In terrestrial conditions, some of these factors can lead to cancer and neuroimmunological disorders. In this study, we used a well-established spaceflight model organism, Drosophila melanogaster, to assess spaceflight-associated changes in the nervous system. We hypothesize that exposure to altered gravity triggers the oxidative stress response, leading to impairments in the nervous system. To test this hypothesis, we used two experimental paradigms: 1) hypergravity, using the ground-based chronic acceleration model, and 2) spaceflight conditions, which includes exposure to microgravity and in-flight space 1g controls. In our ground studies, acute hypergravity resulted in an induction of oxidative stress-related genes with an increase in reactive oxygen species (ROS) in fly brains. Additionally, we observed a depressed locomotor phenotype in these flies (p<0.05). These flies also show a decreased dopaminergic neuron counts in the fly brain upon exposure to acute hypergravity (p<0.05). Thus, the data suggest that altered gravity has a profound effect on the fly nervous system. Similarly, we observe behavioral impairments (p<0.001) and synaptic deficits, including decreased synaptic connections (p<0.05), in 3rd instar larvae which were developed in space. Furthermore, space-grown adults show a decrease in neuronal (p<0.05) and dendritic field (p<0.01) in adult brains coupled with an increased number of apoptotic cells (p<0.001), suggesting increased neuronal loss under spaceflight conditions. In summary, we observe that altered gravity leads to gross neurological deficits. To better understand the long-term effects of spaceflight on the nervous system, longitudinal and multigenerational changes were also identified. This study will help elucidate the different approaches to prevent nervous system dysfunction in astronauts during spaceflight, while also contributing to a better understanding of the pathways that are related to some CNS disorders on Earth.

nervous system↗
NASA NTRS2008

Intelligent Memory Module Overcomes Harsh Environments

Solar cells, integrated circuits, and sensors are essential to manned and unmanned space flight and exploration, but such systems are highly susceptible to damage from radiation. Especially problematic, the Van Allen radiation belts encircle Earth in concentric radioactive tori at distances from about 6,300 to 38,000 km, though the inner radiation belt can dip as low as 700 km, posing a severe hazard to craft and humans leaving Earth s atmosphere. To avoid this radiation, the International Space Station and space shuttles orbit at altitudes between 275 and 460 km, below the belts range, and Apollo astronauts skirted the edge of the belts to minimize exposure, passing swiftly through thinner sections of the belts and thereby avoiding significant side effects. This radiation can, however, prove detrimental to improperly protected electronics on satellites that spend the majority of their service life in the harsh environment of the belts. Compact, high-performance electronics that can withstand extreme environmental and radiation stress are thus critical to future space missions. Increasing miniaturization of electronics addresses the need for lighter weight in launch payloads, as launch costs put weight at a premium. Likewise, improved memory technologies have reduced size, cost, mass, power demand, and system complexity, and improved high-bandwidth communication to meet the data volume needs of the next-generation high-resolution sensors. This very miniaturization, however, has exacerbated system susceptibility to radiation, as the charge of ions may meet or exceed that of circuitry, overwhelming the circuit and disrupting operation of a satellite. The Hubble Space Telescope, for example, must turn off its sensors when passing through intense radiation to maintain reliable operation. To address the need for improved data quality, additional capacity for raw and processed data, ever-increasing resolution, and radiation tolerance, NASA spurred the development of the Radiation Tolerant Intelligent Memory Stack (RTIMS).

Source record↗
NASA NTRS1997

The Solar-Stellar Connection (NAG5-6124: SOHO Guest Investigator Program)

The following is a final report from the SOHO Guest Investigator program to use the SUMER far-UV spectrometer to obtain imaging spectroscopy in support of the goals of the so-called "solar-stellar connection." In particular, a major emphasis was utilization of the long-slit time-resolved maps of the solar surface in bright far-UV emission lines to deduce how particular aspects of the temporally and spatially averaged line profiles trace back to individual structural features of the magnetically disturbed outer atmosphere; to help interpret the unresolved line profiles from high quality stellar observations (say, with the Space Telescope Imaging Spectrograph). The researchers served two tours of duty in the SOHO Operations Center as SUMER planners, during which time we conducted an extensive series of observing programs. These can be divided into three general categories: surface mapping, translimb spectroscopy, and active region diagnostics. We have analyzed some of the large volumes of data to the point where we have presented them in poster papers, and in invited papers at national and international meetings. Listed below are the titles of the preliminary publications we have written, including brief abstracts to indicate the main results. (1) Chromospheric structure and Dynamics-- Observations -- The chromosphere is a highly structured dynamic 'layer' of the solar outer atmosphere. Here, not only are the effects of mechanical heating first evident (moving upward in altitude from the deep photosphere), but also the amount of nonradiative energy deposited is far greater than in the albeit much hotter overlying transition region and corona. Further, the chromosphere is by far the thickest zone of the solar atmosphere with respect to the pressure scale height. A major goal of stellar astrophysics is to understand how the chromosphere is heated and why it adopts its peculiar structure. A cursory examination of solar filtergrams and high-resolution movies demonstrates that much of the chromospheric "action" must be occurring on fine spatial scales and short times; particularly in the cell interior transient brightenings, but also in the longer-lived network fragments. That regime of investigation is far removed from what one usually associates with "synoptic" measurements. Nevertheless, synoptic observations of chromospheric indices, filtergrams, and globally-averaged profile parameters not only can provide important insight concerning the crucial role of the cycle-variable part of the solar magnetic field; but they also can forge a key link with analogous measurements of the stars, where often the phenomena can be significantly exaggerated from the solar case, but high spatial resolution reconnaissance is not even a remote possibility. In addition to discussing the synoptic aspects of chromospheric structure and dynamics, I summarized new insights into the general problem of the solar chromosphere that have been obtained recently with the SUMER far-ultraviolet spectrometer on SOHO. (2) CO and the Temperature Structure of the Solar Atmosphere -- The surface layers of the Sun provide a crucial boundary condition for many of the processes that occur in the deep interior. The stratification of the outer solar atmosphere once was thought to be well understood. However, studies of thermally sensitive molecular absorptions in the infrared revealed puzzling anomalies. Strong lines of the CO fundamental vibration-rotation bands near 5 microns showed very cool temperatures at the extreme limb, and remarkable off-limb emissions extending well into the supposedly hot chromosphere. The conflicting pictures of the photosphere/chromosphere interface, from the widely separated wavelength regimes, has raised suspicions that those "layers" of the atmosphere are much more inhomogeneous than previously suspected. One proposal is that the low chromosphere is dominated by cool gas, the "COmosphere," which is threaded by a network of persistent small-scale hot magnetic filaments and occasionally disrupted by localized acoustic disturbances. The COmosphere is capped by the merged fields of the network elements in the chromospheric "canopy." (3) Translimb Spectroscopy with SOHO/SUMER -- We have used SUMER to obtain deep exposures of the 1300-1400 A spectrum, at the extreme limb and off-limb. Previous "translimb" studies in the thermal infrared had revealed remarkable extensions of cold material (T is approximately equal to 3000 K)-traced by carbon monoxide emission lines-into the heart of the hot chromosphere. A main objective of our program was to search for corresponding far-UV signatures of the "thermally-bifurcated" low chromosphere; for example, radiatively fluoresced emissions of the CO A-X 4th-positive system (collisional excitation would be negligible in cold gas). We conducted two separate observing programs with SUMER. Both made use of the 1 inch-diameter circular aperture, translated across the limb in the minimum motor step increments of 0."375, along the central meridian in the Northern polar coronal hole. The first program executed for nine hours beginning 19UT 25 Oct 1996. The full wavelength range was 1340-1400 A. It was recorded in two overlapping segments, placing key regions of the spectrum alternately on the KBr and bare parts of the detector, to help isolate 2nd-order features. Each segment was integrated for 500 s, and 32 pairs were obtained to span a 12" swath centered on the optical limb. The second program was conducted 00-09UT 01 Dec 1996. It consisted of a single wavelength setting (1300-1340 A) with exposure time 500 s, but twice the spatial coverage of the earlier series: 64 steps, for a total displacement of 24". The strong chromospheric resonance lines of atomic oxygen (1302-1305 A) and ionized carbon (1334-1335 A) were observed on the bare part of the MCP camera.

Ayres, Thomas R.↗
NASA NTRS2003

Gap junctional communication modulates gene transcription by altering the recruitment of Sp1 and Sp3 to connexin-response elements in osteoblast promoters

Loss-of-function mutations of gap junction proteins, connexins, represent a mechanism of disease in a variety of tissues. We have shown that recessive (gene deletion) or dominant (connexin45 overexpression) disruption of connexin43 function results in osteoblast dysfunction and abnormal expression of osteoblast genes, including down-regulation of osteocalcin transcription. To elucidate the molecular mechanisms of gap junction-sensitive transcriptional regulation, we systematically analyzed the rat osteocalcin promoter for sensitivity to gap junctional intercellular communication. We identified an Sp1/Sp3 containing complex that assembles on a minimal element in the -70 to -57 region of the osteocalcin promoter in a gap junction-dependent manner. This CT-rich connexin-response element is necessary and sufficient to confer gap junction sensitivity to the osteocalcin proximal promoter. Repression of osteocalcin transcription occurs as a result of displacement of the stimulatory Sp1 by the inhibitory Sp3 on the promoter when gap junctional communication is perturbed. Modulation of Sp1/Sp3 recruitment also occurs on the collagen Ialpha1 promoter and translates into gap junction-sensitive transcriptional control of collagen Ialpha1 gene expression. Thus, regulation of Sp1/Sp3 recruitment to the promoter may represent a potential general mechanism for transcriptional control of target genes by signals passing through gap junctions.

Non-NASA Center↗
NASA NTRS2016

Proteomic Assessment of Fluid Shifts and Association with Visual Impairment and Intracranial Pressure in Twin Astronauts

BACKGROUND: Astronauts participating in long duration space missions are at an increased risk of physiological disruptions. The development of visual impairment and intracranial pressure (VIIP) syndrome is one of the leading health concerns for crew members on long-duration space missions; microgravity-induced fluid shifts and chronic elevated cabin CO2 may be contributing factors. By studying physiological and molecular changes in one identical twin during his 1-year ISS mission and his ground-based co-twin, this work extends a current NASA-funded investigation to assess space flight induced "Fluid Shifts" in association with the development of VIIP. This twin study uniquely integrates physiological and -omic signatures to further our understanding of the molecular mechanisms underlying space flight-induced VIIP. We are: (i) conducting longitudinal proteomic assessments of plasma to identify fluid regulation-related molecular pathways altered by long-term space flight; and (ii) integrating physiological and proteomic data with genomic data to understand the genomic mechanism by which these proteomic signatures are regulated. PURPOSE: We are exploring proteomic signatures and genomic mechanisms underlying space flight-induced VIIP symptoms with the future goal of developing early biomarkers to detect and monitor the progression of VIIP. This study is first to employ a male monozygous twin pair to systematically determine the impact of fluid distribution in microgravity, integrating a comprehensive set of structural and functional measures with proteomic, metabolomic and genomic data. This project has a broader impact on Earth-based clinical areas, such as traumatic brain injury-induced elevations of intracranial pressure, hydrocephalus, and glaucoma. HYPOTHESIS: We predict that the space-flown twin will experience a space flight-induced alteration in proteins and peptides related to fluid balance, fluid control and brain injury as compared to his pre-flight protein/peptide signatures. Conversely, the trajectory of these protein signatures will remain relatively constant in his ground based co-twin. METHODS: We are using proteomic and standard immunoelectrophoresis techniques to delineate the change in protein signatures throughout the course of a long duration space flight in relation to the development of VIIP. We are also applying a novel cell-based metaboloic organ system assay ("Organs on a Plate") to address how these circulating biomarkers affect physiological processes at the cellular and organ level which could result in VIIP symptoms. These molecular data will be correlated with physiological measures (eg. extra and intracellular fluid volume, vascular filling/flow patterns, MRI, and Optic Coherence Tomography. DISCUSSION: Pre- and in-flight data collection is in progress for the space-flown twin, and similar data have been obtained from the ground-based twin. Biosamples will be batch processed when received from ISS after the conclusion of the 1-year mission. Omic and Physiological measures from the twin astronauts will be compared to similar data being collected on twin subjects who participated in simulated microgravity study. bed rest study.

Rana, Brinda K.↗
NASA NTRSDate not supplied

A Pair of Trans-Golgi Network/Early Endosome-Localized Proteins Facilitate Cytoskeletal-Mediated Root Skewing in Arabidopsis Thaliana

Roots treated with the actin-disrupting compound Latrunculin B (LatB) show stronger gravitropic responses on Earth and dampened straightening responses on a 2-D clinostat. In related experiments using the Biological Research in Canisters (BRIC) hardware, it was found that knockouts to vegetative actin isoforms in Arabidopsis thaliana had more robust root skewing in microgravity and waved more strongly than wild type on hard agar surfaces. These results indicate that the actin cytoskeleton mediates directional root growth in space and on the ground. To gain new insights into the role of actin in directional root growth, we identified mutants that showed differential growth responses to low doses of LatB. In-depth studies of one mutant led to the identification of a trans-Golgi Network (TGN)/Early Endosome (EE)- localized protein that associates with actin. This protein called hypersensitive to LatB 1 (HLB1) colocalized with the ADP-ribosylation-factor guanine nucleotide exchange factor, MIN7/BEN1 (HOPM INTERACTOR7/BREFELDIN A-VISUALIZED ENDOCYTIC TRAFFICKING DEFECTIVE1), at the TGN/EE. HLB1 and MIN7/BEN1 were found to regulate exocytosis and endocytosis, respectively, suggesting that both proteins are involved in actin-mediated membrane traffic. Microtubules, another component of the cytoskeleton, is known to be involved in root skewing. Both hlb1 and min7/ben1 mutants exhibited dampened root skewing on the microtubule stabilizing compound taxol. Taken together, our results support the conclusion that cytoskeletal-membrane interactions contribute to directional root skewing in A. thaliana and is facilitated in part by the TGN/EE-localized HLB1 and MIN7/BEN1 proteins.

Plant Space Biology↗
NASA NTRS2000

Effects of hindlimb unloading on neuromuscular development of neonatal rats

We hypothesized that hindlimb suspension unloading of 8-day-old neonatal rats would disrupt the normal development of muscle fiber types and the motor innervation of the antigravity (weightbearing) soleus muscles but not extensor digitorum longus (EDL) muscles. Five rats were suspended 4.5 h and returned 1.5 h to the dam for nursing on a 24 h cycle for 9 days. To control for isolation from the dam, the remaining five littermates were removed on the same schedule but not suspended. Another litter of 10 rats housed in the same room provided a vivarium control. Fibers were typed by myofibrillar ATPase histochemistry and immunostaining for embryonic, slow, fast IIA and fast IIB isomyosins. The percentage of multiple innervation and the complexity of singly-innervated motor terminal endings were assessed in silver/cholinesterase stained sections. Unique to the soleus, unloading accelerated production of fast IIA myosin, delayed expression of slow myosin and retarded increases in standardized muscle weight and fiber size. Loss of multiple innervation was not delayed. However, fewer than normal motor nerve endings achieved complexity. Suspended rats continued unloaded hindlimb movements. These findings suggest that motor neurons resolve multiple innervation through nerve impulse activity, whereas the postsynaptic element (muscle fiber) controls endplate size, which regulates motor terminal arborization. Unexpectedly, in the EDL of unloaded rats, transition from embryonic to fast myosin expression was retarded. Suspension-related foot drop, which stretches and chronically loads EDL, may have prevented fast fiber differentiation. These results demonstrate that neuromuscular development of both weightbearing and non-weightbearing muscles in rats is dependent upon and modulated by hindlimb loading.

Non-NASA Center↗
NASA NTRSDate not supplied

A Dominant Arabidopsis Thaliana ACTIN7 Mutant for Studies of Cytoskeletal-Mediated Root Gravity and Spaceflight Stress Responses

A forward genetic screen for Arabidopsis thaliana mutants that exhibited differential sensitivity to the actin-disrupting compound, latrunculin B (LatB), was conducted to uncover new players involved in actin-mediated root gravity responses. This to the isolation of a mutant that exhibited robust primary root growth at 100 nM LatB, which is a concentration that severely inhibits wild-type root elongation. Phenotypic analysis revealed that hypocotyl elongation in the dark and root hair tip growth in the mutant could tolerate LatB concentrations that impaired these processes in wild type. A cross between the mutant and wild type resulted in progeny resistance to LatB, which indicated that the mutant is dominant (hereafter referred to as LBR1 for LatB Resistant1). Filamentous-actin (F-actin) organization in LBR1 primary roots remained intact at 100 nM LatB, while that of wild type exhibited fragmented F-actin. Next generation sequencing revealed that LBR1 had a single nucleotide polymorphism (SNP) in the AT5G09810 gene that changed cytosine to a thiamine at the first exon. AT5G09810 encodes ACTIN7 (ACT7), which is one of three vegetative actin isoforms in A. thaliana. The SNP in the ACT7 gene led to a change in a single amino acid from proline at position 32 to a serine. Transgenic complementation of LBR1 plants with wild-type ACT7 under the control of the ACT7 promoter (pACT7:ACT7) and wild-type plants with LBR1, which contained the proline to serine mutation, also under the ACT7 promoter (pACT7:LBR1) confirmed that LBR1 is ACT7. The pACT7:LBR1 construct was also able to confer LatB resistance to the act7-5 and act2-3 vegetative ACT mutants. Moreover, LBR1 exhibited partial tolerance to salt and low phosphate, and enhanced root skewing on a clinostat, suggesting that site-directed engineering of vegetative ACT presents a strategy for generating stress-tolerant plants for spaceflight applications and studies of actin-mediated gravity responses.

Plant Space Biology↗
NASA NTRSDate not supplied

A Dominant Arabidopsis Thaliana ACTIN7 Mutant for Studies of Cytoskeletal-Mediated Root Gravity and Spaceflight Stress Responses

A forward genetic screen for Arabidopsis thaliana mutants that exhibited differential sensitivity to the actin-disrupting compound, latrunculin B (LatB), was conducted to uncover new players involved in actin-mediated root gravity responses. This led to the isolation of a mutant that exhibited robust primary root growth at 100 nM LatB, which is a concentration that severely inhibits wild-type root elongation. Phenotypic analysis revealed that hypocotyl elongation in the dark and root hair tip growth in the mutant could tolerate LatB concentrations that impaired these processes in wild type. A cross between the mutant and wild type resulted in progeny resistance to LatB, which indicated that the mutant is dominant (hereafter referred to as LBR1 for LatB Resistant1). Filamentous-actin (F-actin) organization in LBR1 primary roots remained intact at 100 nM LatB, while that of wild type exhibited fragmented F-actin. Next generation sequencing revealed that LBR1 had a single nucleotide polymorphism (SNP) in the AT5G09810 gene that changed cytosine to a thiamine at the first exon. AT5G09810 encodes ACTIN7 (ACT7), which is one of three vegetative actin isoforms in A. thaliana. The SNP in the ACT7 gene led to a change in a single amino acid from proline at position 32 to a serine. Transgenic complementation of LBR1 plants with wild-type ACT7 under the control of the ACT7 promoter (pACT7:ACT7) and wild-type plants with LBR1, which contained the proline to serine mutation, also under the ACT7 promoter (pACT7:LBR1) confirmed that LBR1 is ACT7. The pACT7:LBR1 construct was also able to confer LatB resistance to the act7-5 and act2-3 vegetative ACT mutants. Moreover, LBR1 exhibited partial tolerance to salt and low phosphate, and enhanced root skewing on a clinostat, suggesting that site-directed engineering of vegetative ACT presents a strategy for generating stress-tolerant plants for spaceflight applications and studies of actin-mediated gravity responses.

Plant Space Biology↗