Plant biology in space: recent accomplishments and recommendations for future research.

Gravity has shaped the evolution of life since its origin. However, experiments in the absence of this overriding force, necessary to precisely analyse its role, e.g. for growth, development, and orientation of plants and single cells, only became possible with the advent of spaceflight. Consequently, this research has been supported especially by space agencies around the world for decades, mainly for two reasons: first, to enable fundamental research on gravity perception and transduction during growth and development of plants; and second, to successfully grow plants under microgravity conditions with the goal of establishing a bioregenerative life support system providing oxygen and food for astronauts in long-term exploratory missions. For the second time, the International Space Life Sciences Working Group (ISLSWG), comprised of space agencies with substantial life sciences programmes in the world, organised a workshop on plant biology research in space. The present contribution summarises the outcome of this workshop. In the first part, an analysis is undertaken, if and how the recommendations of the first workshop held in Bad Honnef, Germany, in 1996 have been implemented. A chapter summarising major scientific breakthroughs obtained in the last 15 years from plant research in space concludes this first part. In the second part, recommendations for future research in plant biology in space are put together that have been elaborated in the various discussion sessions during the workshop, as well as provided in written statements from the session chairs. The present paper clearly shows that plant biology in space has contributed significantly to progress in plant gravity perception, transduction and responses - processes also relevant for general plant biology, including agricultural aspects. In addition, the interplay between light and gravity effects has increasingly received attention. It also became evident that plants will play a major role as components of bioregenerative life support and energy systems that are necessary to complement physico-chemical systems in upcoming long-term exploratory missions. In order to achieve major progress in the future, however, standardised experimental conditions and more advanced analytical tools, such as state-of-the-art onboard analysis, are required.

Gravitational biology within the German Space Program: goals, achievements, and perspectives.

Gravity plays an important role for the evolution, orientation and development of organisms. Most of us, however, tend to overlook its importance because--due to its constant presence from the beginning of evolution some 4 billion years ago--this environmental parameter is almost hardwired into our interpretation of reality. This negligence of gravity is the more surprising as we all have our strong fights with this factor, especially during the very early and again during the late phases of our lives. On the other hand, scientists have been fascinated to observe the effects of gravity especially on plants and microorganisms for more than a hundred years, since Darwin and Sachs demonstrated the role of the root cap for downward growing plants. Different experimental approaches are nowadays available in order to change the influence of gravity and to study the corresponding influences on the physiology of biological systems. With the advent of spaceflight, a long-term nearly nullification of gravity is possible. Utilisation of this so-called "microgravity" condition for research in life sciences thus became an important asset in the space programs of various space agencies around the world. The German Space Life Sciences Program is managed--like all other space programs and activities in Germany--by the German Aerospace Center (DLR) in its role as space agency for Germany. Within the current space program, approved by the German government in May 2001, the overall goal for its life sciences part was defined as to gain scientific knowledge and to disclose new application potential by research under space conditions, especially by utilising the microgravity environment of the International Space Station. Three main scientific fields have been identified in collaboration with the scientific community: integrative human physiology, biotechnological applications of the microgravity environment, and fundamental biology of gravity and radiation responses (i.e., gravitational and radiation biology). In the present contribution, specific goals as well as achievements and perspectives of research in gravitational biology are given. In addition, some information is provided on spaceflight opportunities available.

Life science experiments during the German-Russian MIR '97 mission.

Manned spaceflight has been an important element of the German space program over the last decades. This is demonstrated by the nationally managed space missions Spacelab D-l (1985), D-2 (1993), and MIR '92 as well as by the participation in the 1st Spacelab mission FSLP (1983), the NASA missions IML-1 (1992) and IML-2 (1994), as well as in the ESA missions EUROMIR '94 and '95. On February 12th, this year, the German cosmonaut Reinhold Ewald was launched together with his Russian colleagues Wasilij Zibliew and Alexander Lasudkin onboard of a Soyuz spacecraft for another stay of a German cosmonaut onboard of the Russian Space Station MIR. This mission--the so-called German/Russian MIR '97--was, of course, another cornerstone with regard to the cooperation between Russian and German space organizations. The cooperation in the area of manned missions began 1978 with the flight of the German cosmonaut Sigmund Jahn onboard of Salyut 6, at that time a cooperation between the Soviet Union and the German Democratic Republic in the frame of the Interkosmos Program. In March 1992, it was followed by the flight of Klaus Dietrich Flade with his stay onboard of MIR. After two further successful ESA missions, EUROMIR '94 and '95 with the two German cosmonauts Ulf Merbold and Thomas Reiter and with a marked contribution of German scientists, the decision was taken to perform another German/Russian MIR mission, the so-called MIR '97. In Germany, MIR'97 was managed and performed in a joint effort between several partners. DARA, the German Space Agency, was responsible for the overall program and project management, while DLR, the German Aerospace Research Establishment, was responsible for the cosmonaut training, for medical operations, for the mission control at GSOC in Oberpfaffenhofen as well as for user support.

DARA vestibular equipment onboard MIR.

In space, the weightless environment provides a different stimulus to the otolith organs of the vestibular system, and the resulting signals no longer correspond with the visual and other sensory signals sent to the brain. This signal conflict causes disorientation. To study this and also to understand the vestibular adaptation to weightlessness, DARA has developed scientific equipment for vestibular and visuo-oculomotoric investigations. Especially, two video-oculography systems (monocular--VOG--and binocular--BIVOG, respectively) as well as stimuli such as an optokinetic stimulation device have successfully been employed onboard MIR in the frame of national and European missions since 1992. The monocular VOG was used by Klaus Flade during the MIR '92 mission, by Victor Polyakov during his record 15 months stay onboard MIR in 1993/94 as well as by Ulf Merbold during EUROMIR '94. The binocular version was used by Thomas Reiter and Sergej Avdeyev during the 6 months EUROMIR '95 mission. PIs of the various experiments include H. Scherer and A. Clarke (FU Berlin), M. Dieterichs and S. Krafczyk (LMU Munchen) from Germany as well as C.H. Markham and S.G. Diamond from the United States. Video-Oculography (VOG) is a technique for examining the function of the human balance system located in the inner ear (vestibular system) and the visio-oculomotor interactions of the vestibular organ. The human eye movements are measured, recorded and evaluated by state-of-the-art video techniques. The method was first conceived and designed at the Vestibular Research Laboratory of the ENT Clinic in Steglitz, FU Berlin (A. Clarke, H. Scherer). Kayser-Threde developed, manufactured and tested the facilities for space application under contract to DARA. Evaluation software was first provided by the ENT Clinic, Berlin, later by our subcontractor Sensomotoric Instruments (SMI), Teltow. Optokinetic hardware to support visuo-oculomotoric investigations, has been shipped to MIR for EUROMIR '95 and has successfully been used in conjunction with VOG by ESA astronaut Thomas Reiter. Most recently, BIVOG aboard MIR will be reused in the frame of German/Russian joint experiment sessions employing two Russian cosmonauts from August 1997 to January 1998.

Light-stimulated respiration in the green algaDunaliella tertiolecta: Involvement of the ultraviolet/blue-light photoreceptor(s) and phytochrome?

Starch breakdown and respiratory O2 uptake in the green algaDunaliella tertiolecta (Butcher) are stimulated not only by blue, but also by red light. In the present study, attempts are described to identify the photoreceptor(s) involved. Fluence rate-response curves with different slopes in the ultraviolet (UV)/blue and in the red spectral region as well as differences in the kinetics and in the unfluence of dark pre-incubation on the stimulation of respiratory O2 uptake by blue and red light strongly indicate the action of two photoreceptors. Since the effect of red light shows some far-red reversibility, and since simultaneous irradiation with red and far-red light decreases the effectiveness of red light, the involvement of phytochrome - in addition to the UV/blue photoreceptor(s) - is suggested in the light-stimulated respiration inDunaliella.

[Enhancement of pyruvate kinase activity by blue light or glucose in a chlorophyll-free Chlorella-mutant]. / Über Aktivitätssteigerungen der Pyruvatkinase durch Blaulicht oder Glucose bei einer chlorophyllfreien Chlorella-Mutante.

Crude extracts of dark-kept resting cells of a chlorophyll-free, carotenoid-containing mutant of Chlorella vulgaris Beijerinck (211-11h/20) were found to convert 14.44±0.77 nmol PEP per min and mg protein into pyruvate by the action of pyruvate kinase (=PK; EC 2.7.1.40). When such cells were exposed to blue light (λ<550 nm, ∼300 µW cm(-2)) for 3 hrs the PK-activity/protein of their crude extracts rose to 21.47±1.30, i.e., it was enhanced by 43%. Poisoning with 10(-3) mol cycloheximide or with 150 µg actinomycin D/ml prevented the effect of blue light by 80-90% (Table 1). This result points to an induction of enzyme synthesis in blue light. Addition of 1% glucose in the dark resulted in an increase in PK-activity, too. Three hrs after application of glucose the PK-activity was 28.05±1.88 nmol/min and mg protein, which was 94% greater than in the control. The effect of glucose was also largely preventable by cycloheximide (10(-3) mol) or by actinomycin D (150 µg/ml) (Table 2). These results lead to the conclusion that blue light may induce the synthesis of PK by supplying free sugars at the site of enzyme synthesis. The assumption is supported by the observation that in hot water extracts of blue illuminated cells in which glucose oxidation had been poisoned by. 10(-2) mol monoiodoacetic acid there was 60% more glucose, glucose-6-phosphate, fructose-6-phosphate and sucrose detectable than in extracts of equally poisoned algae from darkness (Table 3). It is suggested that blue light activates a system for the transport of sugar out of the chloroplast, which results in the induction of respiratory enzyme synthesis and thus in enhanced respiration.