Vladimir Sychev

Gravity independence of seed-to-seed cycling in Brassica rapa.

Abstract. Growth of higher plants in the microgravity environment of orbital platforms has been problematic. Plants typically developed more slowly in space and often failed at the reproductive phase. Short-duration experiments on the Space Shuttle showed that early stages in the reproductive process could occur normally in microgravity, so we sought a long-duration opportunity to test gravitys role throughout the complete life cycle. During a 122-d opportunity on the Mir space station, full life cycles were completed in microgravity with Brassica rapa L. in a series of three experiments in the Svet greenhouse. Plant material was preserved in space by chemical fixation, freezing, and drying, and then compared to material preserved in the same way during a high-fidelity ground control. At sampling times 13 d after planting, plants on Mir were the same size and had the same number of flower buds as ground control plants. Following hand-pollination of the flowers by the astronaut, siliques formed. In microgravity, siliques ripened basipetally and contained smaller seeds with less than 20% of the cotyledon cells found in the seeds harvested from the ground control. Cytochemical localization of storage reserves in the mature embryos showed that starch was retained in the spaceflight material, whereas protein and lipid were the primary storage reserves in the ground control seeds. While these successful seed-to-seed cycles show that gravity is not absolutely required for any step in the plant life cycle, seed quality in Brassica is compromised by development in microgravity.

Mice in Bion-M 1 Space Mission: Training and Selection

After a 16-year hiatus, Russia has resumed its program of biomedical research in space, with the successful 30-day flight of the Bion-M 1 biosatellite (April 19–May 19, 2013). The principal species for biomedical research in this project was the mouse. This paper presents an overview of the scientific goals, the experimental design and the mouse training/selection program. The aim of mice experiments in the Bion-M 1 project was to elucidate cellular and molecular mechanisms, underlying the adaptation of key physiological systems to long-term exposure in microgravity. The studies with mice combined in vivo measurements, both in flight and post-flight (including continuous blood pressure measurement), with extensive in vitro studies carried out shortly after return of the mice and in the end of recovery study. Male C57/BL6 mice group housed in space habitats were flown aboard the Bion-M 1 biosatellite, or remained on ground in the control experiment that replicated environmental and housing conditions in the spacecraft. Vivarium control groups were used to account for housing effects and possible seasonal differences. Mice training included the co-adaptation in housing groups and mice adaptation to paste food diet. The measures taken to co-adapt aggressive male mice in housing groups and the peculiarities of “space” paste food are described. The training program for mice designated for in vivo studies was broader and included behavioral/functional test battery and continuous behavioral measurements in the home-cage. The results of the preliminary tests were used for the selection of homogenous groups. After the flight, mice were in good condition for biomedical studies and displayed signs of pronounced disadaptation to Earths gravity. The outcomes of the training program for the mice welfare are discussed. We conclude that our training program was effective and that male mice can be successfully employed in space biomedical research.

Genome-wide expression analysis of reactive oxygen species gene network in Mizuna plants grown in long-term spaceflight

BackgroundSpaceflight environment have been shown to generate reactive oxygen species (ROS) and induce oxidative stress in plants, but little is known about the gene expression of the ROS gene network in plants grown in long-term spaceflight. The molecular response and adaptation to the spaceflight environment of Mizuna plants harvested after 27 days of cultivation onboard the International Space Station (ISS) were measured using genome-wide mRNA expression analysis (mRNA-Seq).ResultsTotal reads of transcripts from the Mizuna grown in the ISS as well as on the ground by mRNA-Seq showed 8,258 and 14,170 transcripts up-regulated and down-regulated, respectively, in the space-grown Mizuna when compared with those from the ground-grown Mizuna. A total of 20 in 32 ROS oxidative marker genes were up-regulated, including high expression of four hallmarks, and preferentially expressed genes associated with ROS-scavenging including thioredoxin, glutaredoxin, and alternative oxidase genes. In the transcription factors of the ROS gene network, MEKK1-MKK4-MPK3, OXI1-MKK4-MPK3, and OXI1-MPK3 of MAP cascades, induction of WRKY22 by MEKK1-MKK4-MPK3 cascade, induction of WRKY25 and repression of Zat7 by Zat12 were suggested. RbohD and RbohF genes were up-regulated preferentially in NADPH oxidase genes, which produce ROS.ConclusionsThis large-scale transcriptome analysis revealed that the spaceflight environment induced oxidative stress and the ROS gene network activation in the space-grown Mizuna. Among transcripts altered in expression by space conditions, some were common genes response to abiotic and biotic stress. Furthermore, certain genes were exclusively up-regulated in Mizuna grown on the ISS. Surprisingly, Mizuna grew in space normally, as well as on the ground, demonstrating that plants can acclimate to long-term exposure in the spaceflight environment by reprogramming the expression of the ROS gene network.

Study of the effects of the outer space environment on dormant forms of microorganisms, fungi and plants in the 'Expose-R' experiment

Investigations of the effects of solar radiation combined with the spaceflight factors on biological objects were performed in the «EXPOSE-R» experimenton the outersurface ofISS. After more than 1 year of outerspaceexposure,thesporesofmicroorganismsandfungi,aswellastwospeciesofplantseedswereanalysed for viability and the set of biological properties. The experiment provided evidence that not only bacterial and fungalsporesbutalsodormantformsofplantshadthecapabilitytosurvivealong-termexposuretoouterspace. Received 11 August 2014, accepted 6 November 2014

Acute transcriptional up-regulation specific to osteoblasts/osteoclasts in medaka fish immediately after exposure to microgravity

Bone loss is a serious problem in spaceflight; however, the initial action of microgravity has not been identified. To examine this action, we performed live-imaging of animals during a space mission followed by transcriptome analysis using medaka transgenic lines expressing osteoblast and osteoclast-specific promoter-driven GFP and DsRed. In live-imaging for osteoblasts, the intensity of osterix- or osteocalcin-DsRed fluorescence in pharyngeal bones was significantly enhanced 1 day after launch; and this enhancement continued for 8 or 5 days. In osteoclasts, the signals of TRAP-GFP and MMP9-DsRed were highly increased at days 4 and 6 after launch in flight. HiSeq from pharyngeal bones of juvenile fish at day 2 after launch showed up-regulation of 2 osteoblast- and 3 osteoclast- related genes. Gene ontology analysis for the whole-body showed that transcription of genes in the category “nucleus” was significantly enhanced; particularly, transcription-regulators were more up-regulated at day 2 than at day 6. Lastly, we identified 5 genes, c-fos, jun-B-like, pai-1, ddit4 and tsc22d3, which were up-regulated commonly in the whole-body at days 2 and 6, and in the pharyngeal bone at day 2. Our results suggested that exposure to microgravity immediately induced dynamic alteration of gene expression levels in osteoblasts and osteoclasts.

The study of the genetic effects in generation of pea plants cultivated during the whole cycle of ontogenesis on the board of RS ISS

Results of studies on growth and development of offspring of two genetically marked dwarf pea lines planted during the whole ontogenesis cycle in the Lada space greenhouse on board of Russian Segment of International Space Station (RS ISS) are presented. The offspring of M1 and M2 plants grown from seeds formed during space flight was examined under conditions of Earth-based cultivation. It had been shown that growth and developmental characteristics, frequency of chromosome aberrations in primary root meristem and level of molecular polymorphism revealed in individual plants via RAPD method show no significant differences between offspring of “space-grown” and control seeds.

Studying the Phenomenon of Dormancy: Why it is Important for Space Exploration

Investigations to forward the use of animal and plant anabiosis, e.g. cryptobiosis and some other forms of dormancy, in space exploration highlight five notable programs on exobiology. The authors give an outline of each program and list the biological species from bacteria to vertebrates and higher plants that have a resting phase within the life cycle and have been selected for in-space studies. Biomedical support of humans in the absence of factors important to sustenance and development of every living thing is one of the indisputable aspects of space exploration. A critical aspect of the biomedical support framework is creation of the central ecological life support systems (CELSS) and, therefore, investigations in this area are no less important than designing space vehicles. Development of life support systems (LSS), including systems incorporating the biological cycle, has been pursued since the initial space flights of cosmonauts. The ground-based test experiment with CELSS performed in the USSR in the period from the early 1960s to 1980s demonstrated that, though simple in design, these systems were capable of regenerating atmosphere, water, and food and thus adequately provided the necessities of human subjects (Gitelson et al. 1975; Shepelev 1975; Meleshko & Shepelev 1996; Sychev et al. 2002, 2003). Implementation of CELSS for space crews requires prior all-around tests and studies in order to: ● Determine the biological impacts of the space flight factors on the life of individual organisms, as well as communities (populations and biocenoses) ● Develop technologies for cultivating highly productive populations of autotrophs and heterotrophs in the zero-gravity environment ● Design hardware to sustain the vital functions of autotrophs and heterotrophs as members of space crew CELSS ● Search for methods to preserve the gene pool aboard the space vehicle and on the planetary outposts ● Optimize CELSS with consideration for microgravity and constant radiation exposure According to the results of CELSS-related investigations in space flight, microgravity impedes tremendously both functioning of the biological systems components and their integration into a uniform system. There are some hardware and technologies that can make up for the lack of gravity; yet, some problems cannot be resolved technically. Modeling of even simplified ecosystems for remote planetary outposts, e.g. on Mars, instantly raises the issue of long-term transportation of the whole

A hazard analysis critical control point plan applied to the Lada vegetable production units (VPU) to ensure the safety of space grown vegetables

The Lada Vegetable Production Unit (VPU), flight hardware currently deployed on the Russian module of the International Space Station (ISS), is being used to validate the food safety of fresh “space-grown” crops. The final objective of this project is the development of a hazard analysis and critical control point (HACCP) plan for Lada grown crops to minimize potential microbial risks to the astronauts. Following FDA guidelines for the development of a HACCP plan, the identification of hazards and critical control points associated with the production of consumable crops grown in the Lada VPU and the establishment of preventative procedures to minimize risk were performed through the collection of baseline microbiological data and testing of pre and post harvest sanitization protocols. Microbiological data collected from both plant tissue and hardware (e.g., root modules) returned from a variety of crops grown on ISS and ground based experiments have been done to define normal microbial loads and understand the fate and survival of human associated pathogens in the Lada VPU. Protocols have been tested to determine the effectiveness of a sanitizer approved by the FDA and USDA (Pro-San) to achieve acceptable levels of microbes on VPU surfaces and vegetables. These data have been used to develop the HACCP plan outlined here for crops grown in a VPU designed for a space environment.

Oxidative Stress and Antioxidant Capacity in Barley Grown under Space Environment

The gene expression and enzyme activity of superoxide dismutase, catalase, and ascorbate peroxidase in the space-grown barley were not significantly different from those of the ground-grown barley. Cu2+ reducing and radical scavenging activities in an extract of the space-grown barley were lower than those of the ground-grown barley by 0.7 fold, suggesting that the space environment does not induce oxidative stress, and reduces antioxidant capacity in plants.

Cytoskeleton structure and total methylation of mouse cardiac and lung tissue during space flight

The purpose of this work was to evaluate the protein and mRNA expression levels of multiple cytoskeletal proteins in the cardiac and lung tissue of mice that were euthanized onboard the United States Orbital Segment of the International Space Station 37 days after the start of the SpaceX-4 mission (September 2014, USA). The results showed no changes in the cytoskeletal protein content in the cardiac and lung tissue of the mice, but there were significant changes in the mRNA expression levels of the associated genes, which may be due to an increase in total genome methylation. The mRNA expression levels of DNA methylases, the cytosine demethylases Tet1 and Tet3, histone acetylase and histone deacetylase did not change, and the mRNA expression level of cytosine demethylase Tet2 was significantly decreased.