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Non-linear signaling for pattern formation?

Developmental pathways are often simplified as linear cascades of gene activation caused by initial signaling events. Recent data, however, support a more complex view in which non-linear responses to signals are important for pattern formation. First, a positive feedback loop may regulate cellular patterning in the Arabidopsis epidermis. Second, an asymmetric auxin distribution that organizes pattern in the distal root tip may be established by positive feedback.

Arabidopsis↗

Changes in hormonal balance and meristematic activity in primary root tips on the slowly rotating clinostat and their effect on the development of the rapeseed root system.

The morphometry of the root system, the meristematic activity and the level of indole-3-acetic acid (IAA), abscisic acid (ABA) and zeatin in the primary root tips of rapeseed seedlings were analyzed as functions of time on a slowly rotating clinostat (1 rpm) or in the vertical controls (1 rpm). The fresh weight of the root system was 30% higher throughout the growth period (25 days) in clinorotated seedlings. Morphometric analysis showed that the increase in biomass on the clinostat was due to greater primary root growth, earlier initiation and greater elongation of the secondary roots, which could be observed even in 5-day-old seedlings. However, after 15 days, the growth of the primary root slowed on the clinostat, whereas secondary roots still grew faster in clinorotated plants than in the controls. At this time, the secondary roots began to be initiated closer to the root tip on the clinostat than in the control. Analysis of the meristematic activity and determination of the levels in IAA, ABA and zeatin in the primary root tips demonstrated that after 5 days on the clinostat, the increased length of the primary root could be the consequence of higher meristematic activity and coincided with an increase in both IAA and ABA concentrations. After 15 days on the clinostat, a marked increase in IAA, ABA and zeatin, which probably reached supraoptimal levels, seems to cause a progressive disturbance of the meristematic cells, during a decrease of primary root growth between 15 and 25 days. These modifications in the hormonal balance and the perturbation of the meristematic activity on the clinostat were followed by a loss of apical dominance, which was responsible for the early initiation of secondary roots, the greater elongation of the root system and the emergence of the lateral roots near the tip of the primary root.

Abscisic Acid↗

Aluminium toxicity in rye (Secale cereale): root growth and dynamics of cytoplasmic Ca2+ in intact root tips.

Aluminium (Al) toxicity in rye (Secale cereale L.), an Al-resistant crop, was examined by measuring root elongation and cytoplasmic free activity of calcium ([Ca2+]cyt) in intact root apical cells. Measurement of [Ca2+]cyt, was achieved by loading a Ca2+-sensitive fluorescent probe. Fluo-3/AM ester, into root apical cells followed by detection of intracellular fluorescence using a confocal laser scanning microscope. After 20 min of exposure to 50 microM Al (pH 4-2) a slight increase in [Ca2+]cyt of root apical cells was observed, while the response of [Ca2+]cyt to 100 microM Al (pH 4.2) was faster and larger ([Ca2+]cyt increased by 46% in 10 min). Increases in [Ca2+]cyt were correlated with inhibition of root growth, generally measurable after 2 h. Addition of 400 microM malic acid (pH 4.2) largely ameliorated the effect of 100 microM Al on [Ca2+]cyt in root apical cells and protected root growth from Al toxicity. These results suggest that an increase in [Ca2+]cyt in root apical cells in rye is an early effect of Al toxicity and is followed by the secondary effect on root elongation.

Aluminum↗

A morphometric analysis of the redistribution of organelles in columella cells in primary roots of normal seedlings and agravitropic mutants of Hordeum vulgare.

The redistribution of organelles in columella cells of horizontally-oriented roots of Hordeum vulgare was quantified in order to determine what structural changes in graviperceptive (i.e., columella) cells are associated with the onset of the root gravicurvature. The sedimentation of amyloplasts is the only major change in cellular structure that correlates positively with the onset of root gravicurvature, which begins within 15 min after re-orientation. There is no consistent contact between sedimented amyloplasts and any other organelles. Nuclei are restricted to the proximal ends of columella cells in vertically-oriented roots, and remain there throughout gravicurvature after roots are oriented horizontally. Root gravicurvature does not involve significant changes in (1) the volume of columella cells, (2) the relative or absolute volumes of organelles in columella cells, or (3) the distribution of endoplasmic reticulum (ER). The size, number and sedimentation rates of amyloplasts in columella cells of non-graviresponsive roots of mutant seedlings are not significantly different from those of graviresponsive roots of normal seedlings. Similarly, there is no significant difference in (1) cellular volume, (2) distribution or surface area of ER, (3) patterns or rates of organelle redistribution in horizontally-oriented roots, (4) relative or absolute volumes of organelles in columella cells of graviresponsive and non-graviresponsive roots. These results suggest that the lack of graviresponsiveness by roots of mutant seedlings is probably not due to either (1) structural differences in columella cells, or (2) differences in patterns or rates of organelle redistribution as compared to that characteristic of graviresponsive roots. Thus, the basis of non-graviresponsiveness in this mutant is probably different from other agravitropic mutants so far studied.

Endoplasmic Reticulum↗

A morphometric analysis of the redistribution of organelles in columella cells of horizontally-oriented roots of Zea mays.

In order to determine what structural changes in graviperceptive cells are associated with onset of root gravicurvature, the redistribution of organelles in columella cells of horizontally-oriented, graviresponding roots of Zea mays has been quantified. Root gravicurvature began by 15 min after reorientation, and did not involve significant changes in the (i) volume of individual columella cells or amyloplasts, (ii) relative volume of any cellular organelle, (iii) number of amyloplasts per columella cell, or (iv) surface area of cellular location of endoplasmic reticulum. Sedimentation of amyloplasts began within 1 to 2 min after reorientation, and was characterized by an intensely staining area of cytoplasm adjacent to the sedimenting amyloplasts. By 5 min after reorientation, amyloplasts were located in the lower distal corner of columella cells, and, by 15 min after reorientation, overlaid the entire length of the lower cell wall. No consistent contact between amyloplasts and any cellular structure was detected at any stage of gravicurvature. Centrally-located nuclei initially migrated upward in columella cells of horizontally-oriented roots, after which they moved to the proximal ends of the cells by 15 min after reorientation. No significant pattern of redistribution of vacuoles, mitochondria, dictyosomes, or hyaloplasm was detected that correlated with the onset of gravicurvature. These results indicate that amyloplasts and nuclei are the only organelles whose movements correlate positively with the onset of gravicurvature by primary roots of this cultivar of Zea mays.

Cell Nucleus↗

Calcium and protein phosphorylation in the transduction of gravity signal in corn roots.

The involvement of calcium and protein phosphorylation in the transduction of gravity signal was studied using corn roots of a light-insensitive variety (Zea mays L., cv. Patriot). The gravitropic response was calcium-dependent. Horizontal placement of roots preloaded with 32P for three minutes resulted in changes in protein phosphorylation of polypeptides of 32 and 35 kD. Calcium depletion resulted in decreased phosphorylation of these phosphoproteins and replenishment of calcium restored the phosphorylation.

Calcium↗

Microgravity and clinorotation cause redistribution of free calcium in sweet clover columella cells.

In higher plants, calcium redistribution is believed to be crucial for the root to respond to a change in the direction of the gravity vector. To test the effects of clinorotation and microgravity on calcium localization in higher plant roots, sweet clover (Melilotus alba L.) seedlings were germinated and grown for two days on a slow rotating clinostat or in microgravity on the US Space Shuttle flight STS-60. Subsequently, the tissue was treated with a fixative containing antimonate (a calcium precipitating agent) during clinorotation or in microgravity and processed for electron microscopy. In root columella cells of clinorotated plants, antimonate precipitates were localized adjacent to the cell wall in a unilateral manner. Columella cells exposed to microgravity were characterized by precipitates mostly located adjacent to the proximal and lateral cell wall. In all treatments some punctate precipitates were associated with vacuoles, amyloplasts, mitochondria, and euchromatin of the nucleus. A quantitative study revealed a decreased number of precipitates associated with the nucleus and the amyloplasts in columella cells exposed to microgravity as compared to ground controls. These data suggest that roots perceive a change in the gravitational field, as produced by clinorotation or space flights, and respond respectively differently by a redistribution of free calcium.

Antimony↗

Responses of eucalypt species to aluminum: the possible involvement of low molecular weight organic acids in the Al tolerance mechanism.

Aluminum (Al) tolerance mechanisms in crop plants have been extensively researched, but our understanding of the physiological mechanisms underlying Al tolerance in trees is still limited. To investigate Al tolerance in eucalypts, seedlings of six species (Eucalyptus globulus Labill., Eucalyptus urophylla S.T. Blake, Eucalyptus dunnii Maiden, Eucalyptus saligna Sm., Eucalyptus cloeziana F. J. Muell. and Eucalyptus grandis w. Hill ex Maiden) and seedlings of six clones of Eucalyptus species were grown for 10 days in nutrient solutions containing Al concentrations varying from 0 to 2.5 microM (0 to 648 microM Al3+ activities). Root elongation of most species was inhibited only by high Al3+ activities. Low to intermediate Al3+ activities were beneficial to root elongation of all species and clones. Among the species tested, E. globulus and E. urophylla were more tolerant to Al toxicity, whereas E. grandis and E. cloeziana were more susceptible to Al-induced damage. Although E. globulus seedlings were tolerant to Al toxicity, they were highly sensitive to lanthanum (La), indicating that the tolerance mechanism is specific for Al. Fine roots accumulated more Al and their elongation was inhibited more than that of thick roots. In E. globulus, accumulation of Al in root tips increased linearly with increasing Al concentration in the nutrient solution. The majority of Al taken up was retained in the root system, and the small amounts of Al translocated to the shoot system were found mainly in older leaves. No more than 60% of the Al in the thick root tip was in an exchangeable form in the apoplast that could be removed by sequential citrate rinses. Gas chromatography/mass spectrometry and ion chromatography analyses indicated that root exposure to Al led to a greater than 200% increase in malic acid concentration in the root tips of all eucalypt species. The increase in malate concentration in response to Al treatment correlated with the degree of Al tolerance of the species. A small increase in citric acid concentration was also observed in all species, but there were no consistent changes in the concentrations of other organic acids in response to Al treatment. In all eucalypt species, Al treatment induced the secretion of citric and malic acid in root exudates, but no trend with respect to Al tolerance was observed. Thus, although malate and citrate exudation by roots may partially account for the overall high Al tolerance of these eucalypt species, it appears that tolerance is mainly derived from the internal detoxification of Al by complexation with malic acid.

Aluminum↗

Light- and gravity-sensing guidance systems in plants.

The present state of our knowledge of phototropic and geotropic responses in plant organs is summarized, and an attempt made to identify aspects of these responses that might be studied advantageously in a 0 g environment. The paper aims to provide a background for non-specialists and to initiate critical discussion of the best ways to exploit the facilities provided by the Spacelab project in the study of plant guidance systems.

Avena↗

The perception of gravity by a plant.

Ever since geotropism was first studied in plants, attempts have been made to create model systems which might simulate the perception by a plant of a gravitational change. The most resilient of these models, the so-called statolith theory, has now enjoyed a run of over 75 years and demonstrates its viability by reappearing in many different forms. It has shown its value by anticipating the now well understood graviperception mechanism in the Chara rhizoid and this will be described. However, it is unlikely that many features of this relatively simple system can be translated to the higher plant. We now know precisely, at least in many primary roots, the distribution and approximate numbers of the cells that perceive gravity. There is no reason to assume that an identical system operates in shoots, since it is now clear that the fundamental hormonal bases of these two systems are different. We also know much about the ultrastructure of many geoperceptive cells, but apart from speculative models a satisfactory explanation of this very rapid and flexible system eludes us. A possible model system is proposed and ways of testing it in zero gravity are suggested.

Chlorophyta↗

Hydrotropism and its interaction with gravitropism in maize roots.

We have partially characterized root hydrotropism and its interaction with gravitropism in maize (Zea mays L.). Roots of Golden Cross Bantam 70, which require light for orthogravitropism, showed positive hydrotropism; bending upward when placed horizontally below a hydrostimulant (moist cheesecloth) in 85% relative humidity (RH) and in total darkness. However, the light-exposed roots of Golden Cross Bantam 70 or roots of a normal maize cultivar, Burpee Snow Cross, showed positive gravitropism under the same conditions; bending downward when placed horizontally below the hydrostimulant in 85% RH. Light-exposed roots of Golden Cross Bantam 70 placed at 70 degrees below the horizontal plane responded positively hydrotropically, but gravitropism overcame the hydrotropism when the roots were placed at 45 degrees below the horizontal. Roots placed vertically with the tip down in 85% RH bent to the side toward the hydrostimulant in both cultivars, and light conditions did not affect the response. Such vertical roots did not respond when the humidity was maintained near saturation. These results suggest that hydrotropic and gravitropic responses interact with one another depending on the intensity of one or both factors. Removal of the approximately 1.5 millimeter root tip blocked both hydrotropic and gravitropic responses in the two cultivars. However, removal of visible root tip mucilage did not affect hydrotropism or gravitropism in either cultivar.

Darkness↗

Root gravitropism.

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Abscisic Acid↗

Gravitropism of roots: an evaluation of progress during the last three decades.

The response of plant roots to gravity has fascinated many botanists since the early days of plant physiology and much research has been devoted to the elucidation of the sequence of events between the physical reception of gravity and the visible growth response. In the last few decades the ideas on the graviresponse of roots have changed profoundly and much progress has been made in understanding parts of the process. One of the reasons for writing this review was my curiosity to know what has happened since the time I myself was involved in the study of root geotropism, as it was called, about 30 years ago. Some excellent reviews have appeared since then, e.g. Audus (1975), Jackson & Barlow (1981) and Moore & Evans (1986), which were more restricted in scope and, moreover, there have been several fascinating developments. The aim of this review is to discuss briefly all aspects of the graviresponse of roots and the progress made in understanding during the last three decades. Some data on other plant organs are included where appropriate.

Calcium↗

A gene expression map of the Arabidopsis root.

A global map of gene expression within an organ can identify genes with coordinated expression in localized domains, thereby relating gene activity to cell fate and tissue specialization. Here, we present localization of expression of more than 22,000 genes in the Arabidopsis root. Gene expression was mapped to 15 different zones of the root that correspond to cell types and tissues at progressive developmental stages. Patterns of gene expression traverse traditional anatomical boundaries and show cassettes of hormonal response. Chromosomal clustering defined some coregulated genes. This expression map correlates groups of genes to specific cell fates and should serve to guide reverse genetics.

Arabidopsis↗

Geoperception in primary and lateral roots of Phaseolus vulgaris (Fabaceae). II. Intracellular distribution of organelles in columella cells.

A morphometric analysis of the ultrastructures of columella cells in primary and lateral roots of Phaseolus vulgaris was performed to determine the precise location of cellular components in these cells. Roots were fixed in situ to preserve the in vivo ultrastructure of the cells. All cellular components in columella cells of both types of roots were distributed asymmetrically. The nucleus and vacuome were located primarily in the middle third of both types of columella cells. Dictyosomes, mitochondria, and amyloplasts were most abundant in the lower third of the columella cells in both types of roots. The distribution of amyloplasts was the most asymmetrical of all cellular components examined, with the lower third of the columella cells containing approximately 90% of the relative volume of amyloplasts in both types of roots. The distribution of cellular components in columella cells of primary roots was not significantly different from that of columella cells of lateral roots. These results indicate that differences in georesponsiveness of primary and lateral roots of P. vulgaris are probably due to factors other than the ultrastructures of their individual columella cells.

Cytoplasm↗