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Hypergravity is a condition where the force of gravity exceeds that on the surface of the Earth and can be simulated by centrifugation. Previously, a significant increase in root growth phenotype was observed when wheat seeds were exposed to hypergravity (10 g for 12 h). In the present study, we investigated the molecular basis of this change through root transcriptome. The data revealed a total of 3765 up-regulated and 2102 down-regulated transcripts in response to hypergravity. GO enrichment analysis revealed hormonal responses, cell division, and cell-wall-related terms were significantly enriched in hypergravity. The increased isoform level expression of transcripts involved in auxin biosynthesis, transport, and signaling was observed. Further, enhanced expression of cell division transcripts and down-regulation of cell number regulator genes suggests rapid cell division. Overexpression of cellulose and hemicellulose biosynthesis transcripts suggests demand for cell-wall constituents. Collectively, this study identified candidate genes associated with hypergravity-induced enhanced root growth.

Hypergravity—an evolutionarily novel environment has been exploited to comprehend the response of living organisms including plants in the context of extra-terrestrial applications. Recently, researchers have shown that hypergravity induces desired phenotypic variability in seedlings. In the present study, we tested the utility of hypergravity as a novel tool in inducing reliable phenotype/s for potential terrestrial crop improvement applications. To investigate, bread wheat seeds (UAS-375 genotype) were subjected to hypergravity treatment (10×g for 12, and 24 h), and evaluated for seedling vigor and plant growth parameters in both laboratory and greenhouse conditions. It was also attempted to elucidate the associated biochemical and hormonal changes at different stages of vegetative growth. Resultant data revealed that hypergravity treatment (10×g for 12 h) significantly enhanced root length, root volume, and root biomass in response to hypergravity. The robust seedling growth phenotype may be attributed to increased alpha-amylase and TDH enzyme activities observed in seeds treated with hypergravity. Elevated total chlorophyll content and Rubisco (55 kDa) protein expression across different stages of vegetative growth in response to hypergravity may impart physiological benefits to wheat growth. Further, hypergravity elicited robust endogenous phytohormones dynamics in root signifying altered phenotype/s. Collectively, this study for the first time describes the utility of hypergravity as a novel tool in inducing reliable root phenotype that could be potentially exploited for improving wheat varieties for better water usage management.

While it has been shown that astronauts suffer immune disorders after spaceflight, the underlying causes are still poorly understood and there are many variables to consider when investigating the immune system in a complex environment. Additionally, there is growing evidence that suggests that not only is the immune system being altered, but the pathogens that infect the host are significantly influenced by spaceflight and ground-based spaceflight conditions. In this study, we demonstrate that Serratia marcescens (strain Db11) was significantly more lethal to Drosophila melanogaster after growth on the International Space Station than ground-based controls, but the increased virulence phenotype of S. marcescens did not persist after the bacterial cultures were passaged on the ground. Increased virulence was also observed in bacteria that were grown in simulated microgravity conditions on the ground using the rotating wall vessel. Increased virulence of the space-flown bacteria was similar in magnitude between wild-type flies and those that were mutants for the well-characterized immune pathways Imd and Toll, suggesting that changes to the host immune system after infection are likely not a major factor contributing towards increased susceptibility of ground-reared flies infected with space-flown bacteria. Characterization of the bacteria shows that at later timepoints spaceflight bacteria grew at a greater rate than ground controls in vitro, and in the host. These results suggest complex physiological changes occurring in pathogenic bacteria in space environments, and there may be novel mechanisms mediating these physiological effects that need to be characterized.

While it has been shown that astronauts suffer immune disorders after spaceflight, the underlying causes are still poorly understood and there are many variables to consider when investigating the immune system in a complex environment. Additionally, there is growing evidence that suggests that not only is the immune system being altered, but the pathogens that infect the host are significantly influenced by spaceflight and ground-based spaceflight conditions. In this study, we demonstrate that Serratia marcescens (strain Db11) was significantly more lethal to Drosophila melanogaster after growth on the International Space Station than ground-based controls, but the increased virulence phenotype of S. marcescens did not persist after the bacterial cultures were passaged on the ground. Increased virulence was also observed in bacteria that were grown in simulated microgravity conditions on the ground using the rotating wall vessel. Increased virulence of the space-flown bacteria was similar in magnitude between wild-type flies and those that were mutants for the well-characterized immune pathways Imd and Toll, suggesting that changes to the host immune system after infection are likely not a major factor contributing towards increased susceptibility of ground-reared flies infected with space-flown bacteria. Characterization of the bacteria shows that at later timepoints spaceflight bacteria grew at a greater rate than ground controls in vitro, and in the host. These results suggest complex physiological changes occurring in pathogenic bacteria in space environments, and there may be novel mechanisms mediating these physiological effects that need to be characterized.

Vascular patterning is a key, genetically responsive phylogenetic classifier of tissues in major organisms flown in space, such as the wings of Drosophila melanogaster (the fruit fly), mouse retina, and leaves of Arabidopsis thaliana. Phenotypes of increasingly abnormal ectopic wing venation in the highly stereotyped Drosophila wing generated by overexpressing the H-C2 construct of Notch antagonist Hairless (Johannes and Preiss, 2002) were mapped and quantified by NASA’s VESsel GENeration Analysis (VESGEN) software. By several confirming vascular parameters, the eight stereotyped wing veins remained quite constant in wild type compared to Class 5 H-C2, the most perturbed category of the H-C2 overexpression phenotypes. However, ectopic veins increased in number from 1 in the wild type, to 18 in Class 5 H-C2. We therefore demonstrate the feasibility of using VESGEN to quantify microscopic images of altered wing venation in Drosophila melanogaster. We further determined that several of the signal transduction pathways affecting wing vein patterning were altered by spaceflight, according to gene expression differences observed in our transcriptomic data from a previous shuttle flight experiment. Future studies will help characterize the extent to which these gene expression changes can cause even subtle developmental changes using model organisms, such as Drosophila. Therefore, we propose that the sensitive analyses provided by VESGEN software will not only serve as a useful tool to map the genetics of wing vein patterning for terrestrial applications, but also for future phenotypic studies with Drosophila for spaceflight missions.

The evolution of karyotypically stabilized short-lived (SL) and long-lived (LL) cytoraces in the laboratory have been established and validated through our previous lifespan studies. In the present investigation, we examined the possible reason(s) for the differential longevity among selected members of SL and LL cytoraces, employing the well known paraquat (PQ) resistance bioassay. Exposure of these races to varying concentrations of PQ revealed relatively higher resistance among LL cytoraces than SL cytoraces, as evident by the lower incidence of mortality. Biochemical analysis for endogenous markers of oxidative stress revealed that LL-2 cytorace exhibited lower reactive oxygen species (ROS) and lipid peroxidation (LPO) levels, higher activity levels of superoxide dismutase (SOD), and coupled with higher levels of reduced glutathione (GSH) compared with the levels found in SL-2 cytorace. These findings suggest that the higher susceptibility of SL cytoraces to PQ challenge may be, at least in part, related to the higher endogenous levels of oxidative stress markers. Although the precise mechanisms responsible for the longer longevity among LL cytoraces of the nasuta–albomicans complex of Drosophila merits further investigation, our data suggest that the relatively longer lifespan may be related to the status of endogenous markers that renders them more resistant towards oxidative-stress-mediated lethality, as evident in the PQ assay.

Environmental exposure to the oxidant-producing herbicide, paraquat (PQ) (1,1'-dimethyl-4,4'-bipyridinium dichloride) has long been implicated as a risk factor in Parkinson's disease (PD). PQ-induced oxidative stress has been exploited as a model to screen putative neuroprotective compounds employing Drosophila. In the present study, we investigated the prophylactic efficacy of Bacopa monnieri (BM) against PQ-induced oxidative stress, mitochondrial dysfunctions and lethality. Exposure of adult male flies (Oregon K) to PQ alone (40 mM in 5% sucrose) resulted in 50% mortality at 48 h. Prophylaxis (7 days) with BM extract (0.1%) offered significant protection (40%) against PQ-induced mortality. Further, oxidative impairments and mitochondrial dysfunctions were monitored among Drosophila exposed to PQ (20, 40 mM) for 24 h. Significant induction of oxidative stress was observed in terms of enhanced malondialdehyde and hydroperoxide levels, and elevated activities of antioxidant enzymes (catalase and SOD). Mitochondrial dysfunctions included of significant reduction in the activities of succinate dehydrogenase (23%), complex I-III (26%), and complex II-III (30%) enzymes. Interestingly, prophylaxis with BM extract prevented the oxidative stress induction by PQ and restored the activity of ETC complexes, suggesting clearly its specific effect on the mitochondria. While the precise mechanism of action of BM needs further investigations, it may be related to its ability to enhance antioxidant defences and thus mitigate PQ-induced oxidative stress in Drosophila.

Altered gravity can perturb normal development and induce corresponding changes in gene expression. Understanding this relationship between the physical environment and a biological response is important for NASA's space travel goals. We use RNA-Seq and qRT-PCR techniques to profile changes in early Drosophila melanogaster pupae exposed to chronic hypergravity (3g, or three times Earth's gravity). During the pupal stage, D. melanogaster rely upon gravitational cues for proper development. Assessing gene expression changes in the pupae under altered gravity conditions helps highlight gravity-dependent genetic pathways. A robust transcriptional response was observed in hypergravity-treated pupae compared to controls, with 1513 genes showing a significant (q