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Sack, Fred D.

Publications and source records attributed to Sack, Fred D..

Gravitropic moss cells default to spiral growth on the clinostat and in microgravity during spaceflight

In addition to shoots and roots, the gravity (g)-vector orients the growth of specialized cells such as the apical cell of dark-grown moss protonemata. Each apical cell of the moss Ceratodon purpureus senses the g-vector and adjusts polar growth accordingly producing entire cultures of upright protonemata (negative gravitropism). The effect of withdrawing a constant gravity stimulus on moss growth was studied on two NASA Space Shuttle (STS) missions as well as during clinostat rotation on earth. Cultures grown in microgravity (spaceflight) on the STS-87 mission exhibited two successive phases of non-random growth and patterning, a radial outgrowth followed by the formation of net clockwise spiral growth. Also, cultures pre-aligned by unilateral light developed clockwise hooks during the subsequent dark period. The second spaceflight experiment flew on STS-107 which disintegrated during its descent on 1 February 2003. However, most of the moss experimental hardware was recovered on the ground, and most cultures, which had been chemically fixed during spaceflight, were retrieved. Almost all intact STS-107 cultures displayed strong spiral growth. Non-random culture growth including clockwise spiral growth was also observed after clinostat rotation. Together these data demonstrate the existence of default non-random growth patterns that develop at a population level in microgravity, a response that must normally be overridden and masked by a constant g-vector on earth.

Non-NASA Center↗

Development of Gravity Sensitive Plant Cells (Ceratodon) in Microgravity

Protonemata of the moss Ceratodon are tip-growing cells that grow up in the dark. This cell type is unique compared to cells in almost any other organism, since the growth of the plant cell itself is completely oriented by gravity. Thus, both the processes of gravity sensing and the gravity response occur in the same cell. Gravity sensing appears to rely upon amyloplasts (starch-filled plastids) that sediment. This sedimentation occurs in specific zones and plastid zonation is complex with respect to plastid morphology, distribution, and gravity. Microtubules restrict the extent of plastid sedimentation (i.e., they are load-bearing). Light also is important since apical cells have a phytochrome-based positive phototropism, light quality influences plastid zonation and sedimentation (photomorphogenesis), and red light suppresses gravitropism at higher but not lower light intensities. Many of these processes were examined in a 16 day spaceflight experiment, "SPM-A" space moss" or "SPAM)) on STS-87 that landed in December, 1997. The work described here involves the definition of a second flight experiment that builds upon the data and questions arising from STS-87. Effort was directed towards further definition of an experiment for the Shuttle (dubbed "SOS" for "Son of SPAM"). Our current target is STS 107 that is scheduled to fly in January 2001. This definition addressed two goals of the STS107 experiment. The goals of the current experiment were to determine whether the cytoskeleton plays a role in maintaining and generating an apical (non-random) plastid distribution in microgravity and to determine the development and extent of clockwise spiral tip-growth in microgravity.

Sack, Fred D.↗

Differentiation and Tropisms in Space-Grown Moss

This grant supported a Space Shuttle experiment on the effects of microgravity on moss cells. Moss provides a rich system for gravitational and spaceflight research. The early phase of the moss life cycle consists of chains of cells that only grow only at their tips. In the moss Ceratodon purpureus these filaments (protonemata) grow away from gravity in the dark, in a process called gravitropism. The tipmost cells, the apical cells, contain heavy starch-filled bodies called amyloplasts that probably function in g-sensing and that sediment within the apical cell. The SPM-A (Space Moss aka SPAM) experiment flew in November - December, 1997 on STS-87 as part of the Collaborative US Ukrainian Experiment (CLTE). The experiment was accommodated in hardware purpose-built by NASA KSC and Bionetics and included Petri Dish Fixation Units (PDFU) and BRIC-LEDs. Together, this hardware allowed for the culture of the moss on agar in commercial petri dishes, for unilateral illumination with red light of varying intensity, and for chemical fixation in situ. The key findings of the spaceflight were quite unexpected. Neither the orientation of tip-growth nor the distribution of amyloplasts was random in microgravity.

Sack, Fred D.↗

Re-Evaluation of the Role of Starch in Gravitropic Sensing

Plant organs grow toward or away from gravity as a way to orient those organs for optimizing growth. Starch has long been thought to be important in sensing the direction of the g-vector in gravitropism, but that hypothesis has also evoked controversy. We have previously shown that starch-deficient mutants of Arabidopsis (TC7) and Nicotiana (NS458) are impaired in their gravitropism. While this suggests that starch is not necessary for reduced gravitropism, it also indicates that the mass of the starch contributes to sensing when present and thus is necessary for full gravitropic sensitivity. The research supported by this grant focused on three related projects, (1) the effect of light on hypocotyl gravitropism in NS458, (2) the effects of root phototropism on measurements of gravitropic sensitivity, and (3) the effects of starch overproduction on sedimentation and gravitropism. Collectively, our results provide additional strong support for the importance of starch in gravitropic sensing. First, by accounting for negative phototropism in roots of two starchless mutants of Arabidopsis we have established that these mutants are much less sensitive to gravity than previously thought. This work also demonstrates the importance of designing experimental protocols that remove the influence of root phototropism on measuring root gravitropism. Second, light apparently promotes gravitropism in starch-deficient Nicotiana hypocotyls by increasing the trace amounts of starch in the plastids, by inducing limited plastid sedimentation and thus by presumably increasing the signal provided by plastid mass. And finally, we show that excess starch in Arabidopsis seedlings has little effect on gravitropic sensitivity implying that the sensing system is already saturated. However, in light-grown stems where this mutation results in starch accumulation and where the wild-type practically lacks starch in the sensing cells, the mutant is much more sensitive than the wild-type again showing that the loss of starch depresses gravity sensing.

Sack, Fred D.↗

Gravitropism and Autotropism in Cress Roots

The overall purpose of this experiment was to study how cress roots respond to a withdrawal of a gravity stimulus i.e. when and how much the roots straighten (autotropism) after curving (gravitropism). This question was studied both in extensive ground-based research and in microgravity on BioRack.

Sack, Fred D.↗

Cellular polarity and interactions in plant graviperception

Presented are results of studies on the mechanisms of gravitropic sensing in higher and lower plants. Gravitropic roots of the aquatic angiosperm, Limnobium, were found to have sedimented amyloplasts in their elongation zone but not in their rootcap; nuclei were found to sediment in the elongation zone as well. Another study attempted to understand how plastid sedimentation occurs in vertical Ceratodon cells and how this sedimentation is regulated. To determine whether the cytoskeleton restricts plastid sedimentation, the effects of amiprophos-methyl (APM) and cytochalasin (CD) on plastid position were qualified. Results suggest that microtubules restrict the sedimentation of plastids along the length of the cell and that microtubules are load-bearing for all the plastids in the apical cell, demonstrating the importance of the cytoskeleton in maintaining organelle position and cell organization against the force of gravity. Physcomitrella and Funaria were also studied. Results suggest that gravitropism may be relatively common in moss protonemata and reinforce the idea that amyloplast mass functions in gravitropic sensing.

Sack, Fred D.↗