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The excitability of plant cells: with a special emphasis on characean internodal cells

This review describes the basic principles of electrophysiology using the generation of an action potential in characean internodal cells as a pedagogical tool. Electrophysiology has proven to be a powerful tool in understanding animal physiology and development, yet it has been virtually neglected in the study of plant physiology and development. This review is, in essence, a written account of my personal journey over the past five years to understand the basic principles of electrophysiology so that I can apply them to the study of plant physiology and development. My formal background is in classical botany and cell biology. I have learned electrophysiology by reading many books on physics written for the lay person and by talking informally with many patient biophysicists. I have written this review for the botanist who is unfamiliar with the basics of membrane biology but would like to know that she or he can become familiar with the latest information without much effort. I also wrote it for the neurophysiologist who is proficient in membrane biology but knows little about plant biology (but may want to teach one lecture on "plant action potentials"). And lastly, I wrote this for people interested in the history of science and how the studies of electrical and chemical communication in physiology and development progressed in the botanical and zoological disciplines.

NASA Discipline Plant Biology↗

Plant biology in space: proceedings of the International Workshop, Bad Honnef, Germany, June 24-27, 1996

Papers presented at the International Workshop on Plant Biology in Space include reviews, reports, and perspectives related to plant gravitational biology. Presentations focused on nine subject areas: gravitropism in unicellular plants, gravitropism in fungi, cell development, gravity perception in multicellular plants, gravity responses in multicellular plants, plant reproduction, evaluation of a clinostat for weightlessness simulation, biological life support systems, and future research.

NASA Discipline Plant Biology↗

Plants and gravity. Special issue

This issue of the Journal of Plant Growth Regulation explores the effects of gravity on plant growth and development from several perspectives. Most of the review papers consider plants and gravity from the viewpoint of ground-based laboratory research, and several papers consider gravitropism, the directed growth in response to gravity, in some detail. However, another approach to study the effects of gravity on plant is to effectively remove the force due to gravity. A very dramatic way to accomplish this goal is through the free-fall conditions achieved by spacecraft in low Earth orbit, so some of the authors have reviewed recent advances in spaceflight research with plant systems.

Non-NASA Center↗

Mechanical forces in plant growth and development

Plant cells perceive forces that arise from the environment and from the biophysics of plant growth. These forces provide meaningful cues that can affect the development of the plant. Seedlings of Arabidopsis thaliana were used to examine the cytoplasmic tensile character of cells that have been implicated in the gravitropic response. Laser-trapping technology revealed that the starch-containing statoliths of the central columella cells in root caps are held loosely within the cytoplasm. In contrast, the peripheral cells have starch granules that are relatively resistant to movement. The role of the actin cytoskeleton in affecting the tensile character of these cells is discussed. To explore the role that biophysical forces might play in generating developmental cues, we have developed an experimental model system in which protoplasts, embedded in a synthetic agarose matrix, are subjected to stretching or compression. We have found that protoplasts subjected to these forces from five minutes to two hours will subsequently elongate either at right angles or parallel to the tensive or compressive force vector. Moreover, the cortical microtubules are found to be organized either at right angles or parallel to the tensive or compressive force vector. We discuss these results in terms of an interplay of information between the extracellular matrix and the underlying cytoskeleton.

Non-NASA Center↗

The mechanosensory calcium-selective ion channel: key component of a plasmalemmal control centre?

Mechanosensory calcium-selective ion channels probably serve to detect not only mechanical stress but also electrical, thermal, and diverse chemical stimuli. Because all stimuli result in a common output, most notably a shift in second messenger calcium concentration, the channels are presumed to serve as signal integrators. Further, insofar as second messenger calcium in turn gives rise to mechanical, electrical, and diverse chemical changes, the channels are postulated to initiate regulatory feedbacks. It is proposed that the channels and the feedback loops play a wide range of roles in regulating normal plant function, as well as in mediating disturbance of normal function by environmental stressors and various pathogens. In developing evidence for the physiological performance of the channel, a model for a cluster of regulatory plasmalemmal proteins and cytoskeletal elements grouped around a set of wall-to-membrane and transmembrane linkers has proved useful. An illustration of how the model might operate is presented. It is founded on the demonstration that several xenobiotics interfere both with normal channel behaviour and with gravitropic reception. Accordingly, the first part of the illustration deals with how the channels and the control system within which they putatively operate might initiate gravitropism. Assuming that gravitropism is an asymmetric expression of growth, the activities of the channels and the plasmalemmal control system are extrapolated to account for regulation of both rate and allometry of cell expansion. Finally, it is discussed how light, hormones, redox agents and herbicides could in principle affect growth via the putative plasmalemmal control cluster or centre.

NASA Program Space Biology↗

The cytoskeleton and gravitropism in higher plants

The cellular and molecular mechanisms underlying the gravitropic response of plants have continued to elude plant biologists despite more than a century of research. Lately there has been increased attention on the role of the cytoskeleton in plant gravitropism, but several controversies and major gaps in our understanding of cytoskeletal involvement in gravitropism remain. A major question in the study of plant gravitropism is how the cytoskeleton mediates early sensing and signal transduction events in plants. Much has been made of the actin cytoskeleton as the cellular structure that sedimenting amyloplasts impinge upon to trigger the downstream signaling events leading to the bending response. There is also strong molecular and biochemical evidence that the transport of auxin, an important player in gravitropism, is regulated by actin. Organizational changes in microtubules during the growth response phase of gravitropism have also been well documented, but the significance of such reorientations in controlling differential cellular growth is unclear. Studies employing pharmacological approaches to dissect cytoskeletal involvement in gravitropism have led to conflicting results and therefore need to be interpreted with caution. Despite the current controversies, the revolutionary advances in molecular, biochemical, and cell biological techniques have opened up several possibilities for further research into this difficult area. The myriad proteins associated with the plant cytoskeleton that are being rapidly characterized provide a rich assortment of candidate regulators that could be targets of the gravity signal transduction chain. Cytoskeletal and ion imaging in real time combined with mutant analysis promises to provide a fresh start into this controversial area of research.

Review↗

Mature phenotype in Hemerocallis plantlets fortuitously generated in vitro

Daylily plantlets generated on semi-solid media from morphogenetically competent cells or morphogenetically competent cells regenerated from protoplasts can give rise in aseptic culture to plantlets with a mature phenotype. The individual leaves of these plantlets open to the extreme base so that no encircling leaf sheath is present. This permits the overlapping bases and leaves to assume an open fan-like arrangement. The occurrence of fans correlates with exceptionally tightly sealed culture vessels and experiments to date suggest a gaseous component is associated with this change of growth form. It has not been possible to fix the mature growth mode, however, and new leaf growth assumes the more normal juvenile phenotype when the gaseous environment is altered by admitting or exposure to room air.

NASA Discipline Plant Biology↗

Lateral root initiation in Marsilea quadrifolia. I. Origin and histogensis of lateral roots

In Marsilea quadrifolia, lateral roots arise from modified single cells of the endodermis located opposite the protoxylem poles within the meristematic region of the parent root. The initial cell divides in four specific planes to establish a five-celled lateral root primordium, with a tetrahedral apical cell in the centre and the oldest merophytes and the root cap along the sides. The cells of the merophyte divide in a precise pattern to give rise to the cells of the cortex, endodermis, pericycle, and vascular tissues of the emerging lateral root. Although the construction of the parent root is more complicated than that of lateral roots, patterns of cell division and tissue formation are similar in both types of roots, with the various tissues being arranged in similar positions in relation to the central axis. Vascular connection between the lateral root primordium and the parent root is derived from the pericycle cells lying between the former and the protoxylem members of the latter. It is proposed that the central axis of the root is not only a geometric centre, but also a physiological centre which determines the fate of the different cell types.

Non-NASA Center↗

The Acid Growth Theory of auxin-induced cell elongation is alive and well

Plant cells elongate irreversibly only when load-bearing bonds in the walls are cleaved. Auxin causes the elongation of stem and coleoptile cells by promoting wall loosening via cleavage of these bonds. This process may be coupled with the intercalation of new cell wall polymers. Because the primary site of auxin action appears to be the plasma membrane or some intracellular site, and wall loosening is extracellular, there must be communication between the protoplast and the wall. Some "wall-loosening factor" must be exported from auxin-impacted cells, which sets into motion the wall loosening events. About 20 years ago, it was suggested that the wall-loosening factor is hydrogen ions. This idea and subsequent supporting data gave rise to the Acid Growth Theory, which states that when exposed to auxin, susceptible cells excrete protons into the wall (apoplast) at an enhanced rate, resulting in a decrease in apoplastic pH. The lowered wall pH then activates wall-loosening processes, the precise nature of which is unknown. Because exogenous acid causes a transient (1-4 h) increase in growth rate, auxin must also mediate events in addition to wall acidification for growth to continue for an extended period of time. These events may include osmoregulation, cell wall synthesis, and maintenance of the capacity of walls to undergo acid-induced wall loosening. At present, we do not know if these phenomena are tightly coupled to wall acidification or if they are the products of multiple independent signal transduction pathways.

NASA Discipline Number 40-50↗

Wall extensibility: its nature, measurement and relationship to plant cell growth

Expansive growth of plant cells is controlled principally by processes that loosen the wall and enable it to expand irreversibly. The central role of wall relaxation for cell expansion is reviewed. The most common methods for assessing the extension properties of plant cell walls ( wall extensibility') are described, categorized and assessed critically. What emerges are three fundamentally different approaches which test growing cells for their ability (a) to enlarge at different values of turgor, (b) to induce wall relaxation, and (c) to deform elastically or plastically in response to an applied tensile force. Analogous methods with isolated walls are similarly reviewed. The results of these different assays are related to the nature of plant cell growth and pertinent biophysical theory. I argue that the extensibilities' measured by these assays are fundamentally different from one another and that some are more pertinent to growth than others.

Review, Academic↗

Recovery of gravitropism after basipetal centrifugation in protonemata of the moss Ceratodon purpureus

Apical cells of 5-day-old dark-grown protonemata of the moss Ceratodon purpureus (Hedw.) Brid. are negatively gravitropic and appear to utilize amyloplasts as statoliths. These cells exhibit a characteristic plastid zonation (five zones) with one zone (No. 3) specialized for the lateral sedimentation of amyloplasts. Basipetal centrifugation displaces all amyloplasts in the apical cell to the end wall. In basipetally centrifuged protonemata observed using infrared videomicroscopy, tip extension occurred with or without amyloplasts present in the apical dome. The initial return of upward curvature was always correlated with the return and sedimentation of amyloplasts in zone 3. Subsequent vigorous upward curvature was correlated with distinct amyloplast zonation and further sedimentation in zone 3. Initial downward ("wrong way") curvature, which often preceded upward curvature, correlated with the presence of amyloplasts in the apical dome (zone 1). These data support the hypotheses that nonsedimenting amyloplasts in zone 1 are necessary for initial downward curvature and that amyloplast sedimentation in zone 3 is necessary for upward curvature.

NASA Program Space Biology↗

If walls could talk

The plant cell wall is very complex, both in structure and function. The wall components and the mechanical properties of the wall have been implicated in conveying information that is important for morphogenesis. Proteoglycans, fragments of polysaccharides and the structural integrity of the wall may relay signals that influence cellular differentiation and growth control. Furthering our knowledge of cell wall structure and function is likely to have a profound impact on our understanding of how plant cells communicate with the extracellular environment.

Review↗

Mechanisms of the early phases of plant gravitropism

Gravitropism is directed growth of a plant or plant organ in response to gravity and can be divided into the following temporal sequence: perception, transduction, and response. This article is a review of the research on the early events of gravitropism (i.e., phenomena associated with the perception and transduction phases). The two major hypotheses for graviperception are the protoplast-pressure and starch-statolith models. While most researchers support the concept of statoliths, there are suggestions that plants have multiple mechanisms of perception. Evidence supports the hypothesis that the actin cytoskeleton is involved in graviperception/transduction, but the details of these mechanisms remain elusive. A number of recent developments, such as increased use of the molecular genetic approach, magnetophoresis, and laser ablation, have facilitated research in graviperception and have allowed for refinement of the current models. In addition, the entire continuum of acceleration forces from hypo- to hyper-gravity have been useful in studying perception mechanisms. Future interdisciplinary molecular approaches and the availability of sophisticated laboratories on the International Space Station should help to develop new insights into mechanisms of gravitropism in plants.

NASA Discipline Plant Biology↗

Gravity and light control of the developmental polarity of regenerating protoplasts isolated from prothallial cells of the fern Ceratopteris richardii

A procedure has been developed for isolating protoplasts from prothalli of Ceratopteris richardii which can be cultured and are capable of regeneration. Protoplasts were isolated from 2-week-old gametophytes in a medium containing wall-digesting enzymes in 0.5 M sucrose, followed by purification of the released protoplasts by floating them up into a 0.5 M sorbitol layer. Regeneration occurred over a period of 10-24 days, and, under optimal osmotic conditions, followed the developmental pattern seen during spore germination, in that the first division gave rise to a primary rhizoid. Thus, prothallial protoplasts are comparable to germinating spores as suitable models for studies of developmental polarity in single cells. As in germinating spores, the polarity of development in regenerating protoplasts is influenced by the vectors of gravity and unilateral light. However, the relative influence of light in fixing this polarity is greater in regenerating protoplasts, while in germinating spores, the influence of gravity is greater.

NASA Discipline Plant Biology↗

Development of the gametophyte of the fern Schizaea pusilla

Schizaea pusilla is a pteridophyte with several unique developmental characteristics. In contrast to most other fern species, S. pusilla gametophytes remain filamentous throughout their development, and the gametophytes are associated with an endophytic fungus which appears to be mycorrhizal. In terms of tropistic responses, apical filament cells of young gametophytes are negatively phototropic compared with germ filaments of other ferns which exhibit positive phototropism. Cryofixation (propane jet freezing and high-pressure freezing) in conjunction with freeze substitution electron microscopy was used to study young gametophytes. The results demonstrate that apical filament cells have a distinctive structural polarity and that rhizoids also can be successfully frozen by these methods. The cytoskeleton and endomembrane system were particularly well preserved in cryofixed cells. In addition, Schizaea gametophytes were used as a test system to evaluate potential artifacts of propane jet freezing and high pressure freezing. There was little apparent difference in ultrastructure between cells cryofixed by either freezing method. These gametophytes will be useful in determining the effectiveness of cryofixation techniques and as a model system in tip growth studies.

Non-NASA Center↗