Shigeki Kamigaichi

Growth and Development, and Auxin Polar Transport in Higher Plants under Microgravity Conditions in Space: BRIC-AUX on STS-95 Space Experiment

The principal objectives of the space experiment, BRIC-AUX on STS 95, were the integrated analysis of the growth and development of etiolated pea and maize seedlings in space and a study of the effects of microgravity conditions in space on auxin polar transport in these segments. Microgravity significantly affected the growth and development of etiolated pea and maize seedlings. Epicotyls of etiolated pea seedlings were the most oriented toward about 40 to 60 degrees from the vertical. Mesocotyls of etiolated maize seedlings were curved at random during space flight but coleoptiles were almost straight. Finally the growth inhibition of these seedlings in space was also observed. Roots of some pea seedlings grew toward to the aerial space of Plant Growth Chamber. Extensibilities of cell walls of the third internode of etiolated pea epicotyls and the top region of etiolated maize coleoptiles, which were germinated and grown under microgravity conditions in space, were significantly low as compared with those grown on the ground of the earth. Activities of auxin polar transport in the second internode segments of etiolated pea seedlings and coleoptile segments of etiolated maize seedlings were significantly inhibited and promoted, respectively, under microgravity conditions in space. These results strongly suggest that auxin polar transport as well as the growth and development of plants is controlled under gravity on the earth.

Morphogenesis of rice and Arabidopsis seedlings in space.

Oryza sativa L.) and Arabidopsis (A. thaliana L.) were cultivated for 68.5 hr in the RICE experiment on board during Space Shuttle STS-95 mission, and changes in their growth and morphology were analyzed. Microgravity in space stimulated elongation growth of both rice coleoptiles and Arabidopsis hypocotyls by making their cell walls extensible. In space, rice coleoptiles showed an inclination toward the caryopsis in the basal region and also a spontaneous curvature in the same direction in the elongating region. These inclinations and curvatures were more prominent in the Koshihikari cultivar compared to a dwarf cultivar, Tan-ginbozu. Rice roots elongated in various directions including into the air on orbit, but two thirds of the roots formed a constant angle with the axis of the caryopsis. In space, Arabidopsis hypocotyls also elongated in a variety of directions and about 10% of the hypocotyls grew into the agar medium. No clear curvatures were observed in the elongating region of Arabidopsis hypocotyls. Such a morphology of both types of seedlings was fundamentally similar to that observed on a 3-D clinostat. Thus, it was confirmed by the RICE experiment that rice and Arabidopsis seedlings perform an automorphogenesis under not only simulated but also true microgravity conditions.

Control of gravimorphogenesis by auxin : accumulation pattern of CS-IAA1 mRNA in cucumber seedlings grown in space and on the ground

Abstract. Cucumber (Cucumis sativus L.) seedlings grown in microgravity developed a peg on each side of the transition zone between hypocotyl and root, whereas seedlings grown in a horizontal position on the ground developed a peg on the concave side of the gravitropically bending transition zone. The morphological features of the space-grown seedlings were similar to those of seedlings grown in a vertical position on the ground with their radicles pointing down: both became two-pegged seedlings. Morphogenesis of cucumber seedlings is thus inhibited by gravity. Analysis by in-situ hybridization of an auxin-inducible gene, CS-IAA1, showed that its mRNA accumulated to a much greater extent on the lower side of the transition zone in the horizontally placed seedlings on the ground just prior to and during the initiation period of peg formation. On the other hand, when seedlings were grown in microgravity or in a vertical position on the ground, accumulation of CS-IAA1 mRNA occurred all around the transition zone. Accumulation of CS-IAA1 mRNA in horizontally grown seedlings appreciably decreased on the upper side of the transition zone and increased on the lower side upon gravistimulation, compared with the two-pegged seedlings. Application of IAA to seedlings in a horizontal position caused the development of a peg on each side of the transition zone, or a collar-like protuberance, depending on the concentration used. These results suggest that upon gravistimulation the auxin concentration on the upper side of the horizontally placed transition zone is reduced to a level below the threshold value necessary for peg formation. Space-grown seedlings of cucumber might develop two pegs symmetrically because the auxin level in the entire transition zone is maintained above the threshold. This spaceflight experiment verified for the first time that auxin does not redistribute in microgravity.

Morphogenesis in cucumber seedlings is negatively controlled by gravity.

Abstract. Seedlings of most cucurbitaceous plants develop a peg (protuberance caused by cell outgrowth) on the transition zone between the hypocotyl and root. The peg is necessary for removing the seed coat after germination. In our spaceflight experiments on the STS-95 space shuttle, Discovery, we found that cucumber (Cucumis sativus L.) seedlings grown under microgravity conditions developed two pegs symmetrically at the transition zone. Thus, cucumber seedlings potentially develop two pegs and do not require gravity for peg formation itself, but on the ground the development of one peg is suppressed in response to gravity. This may be considered as negative control of morphogenesis by gravity.

A Spaceflight Experiment for the Study of Gravimorphogenesis and Hydrotropism in Cucumber Seedlings

peg, on the transition zone between hypocotyl and root. Our spaceflight experiment verified that the lateral positioning of a peg in cucumber seedlings is modified by gravity. It has been suggested that auxin plays an important role in the gravity-controlled positioning of a peg on the ground. Furthermore, cucumber seedlings grown in microgravity developed a number of the lateral roots that grew towards the water-containing substrate in the culture vessel, whereas on the ground they oriented perpendicular to the primary root growing down. The response of the lateral roots in microgravity was successfully mimicked by clinorotation of cucumber seedlings on the three dimensional clinostat. However, this bending response of the lateral roots was observed only in an aeroponic culture of the seedlings but not in solid medium. We considered the response of the lateral roots in microgravity and on clinostat as positive hydrotropism that could easily be interfered by gravitropism on the ground. This system with cucumber seedlings is thus a useful model of spaceflight experiment for the study of the gravimorphogenesis, root hydrotropism and their interaction.

NASDA aquatic animal experiment facilities for space shuttle and ISS

National Space Development Agency of Japan (NASDA) has developed aquatic animal experiment facilities for NASA Space Shuttle use. Vestibular Function Experiment Unit (VFEU) was firstly designed and developed for physiological research using carp in Spacelab-J (SL-J, STS-47) mission. It was modified as Aquatic Animal Experiment Unit (AAEU) to accommodate small aquatic animals, such as medaka and newt, for second International Microgravity Laboratory (IML-2, STS-65) mission. Then, VFEU was improved to accommodate marine fish and to perform neurobiological experiment for Neurolab (STS-90) and STS-95 missions. We have also developed and used water purification system which was adapted to each facility. Based on these experiences of Space Shuttle missions, we are studying to develop advanced aquatic animal experiment facility for both Space Shuttle and International Space Station (ISS).

NASDA next generation Aquatic Habitat for Space Shuttle and ISS.

The National Space Development Agency of Japan (NASDA) has more than 20 years of experience developing aquatic animal experiment facilities. We are now studying the next-generation aquatic animal experiment facility or the Aquatic Habitat (AQH) for both Space Shuttle and International Space Station use. A prototype breeding system was designed and tested. Medaka adult fish were able to mate and spawn in this closed circulatory breeding system, and the larvae grew to adult fish and spawned on the 45th day after hatching. The water quality-control system using nitrifying bacteria worked well throughout the medaka breeding test. For amphibians, we also conducted the African clawed toad (Xenopus laevis) breeding test with the same specimen chambers, although a part of circulation loop was opened to air. Xenopus larvae grew and completed metamorphosis successfully in the small specimen chamber. The first metamorphic climax started on the 30th day and was completed on the 38th day.