Effects of gravitropic stress on the development of the primary root of lentil seedlings grown in space

Root growth and cell differentiation were analysed in lentil seedlings grown (1) in microgravity (F microg), (2) on the 1 x g centrifuge (F1 x g), (3) in microgravity and placed on the 1 x g centrifuge for 4 h [F(microg + 1 x g)], (4) on the 1 x g centrifuge and placed in microgravity for 4 h [F(1 x g + microg)]. In microgravity, there were strong oscillations of the root tip, even when the seedlings were grown first on the 1 x g centrifuge [F(1 x g + microg)]. In the [F(microg + 1 x g)] sample, the roots grown in microgravity were oblique with respect to the 1 x g acceleration when the seedlings were placed on the centrifuge. They were therefore gravistimulated. However, root length was similar in the 4 samples after 29 h of growth and growth rate of the root was the same between 25 h and 29 h although it appeared to be slightly greater in the [F(microg + 1 x g)] sample. Cell elongation was analysed as a function of the distance from the root cap junction. Cell length was similar in the seedlings grown in microgravity or on the 1 x g centrifuge. The transfer from the 1 x g centrifuge to microgravity [F(1 x g + microg)] did not modify cell elongation in the roots. Cell length in the roots which were grown in microgravity and gravistimulated [F(microg + 1 x g)] was different from that observed in microgravity but this was only due to gravistimulation. Thus, gravity does not have an effect on cell elongation when the roots are strictly oriented in the vertical position but it does as soon as the root tip deviates from this orientation.     

Random root movements in weightlessness

The dynamics of root growth was studied in weightlessness. In the absence of the gravitropic reference direction during weightlessness, root movements could be controlled by spontaneous growth processes, without any corrective growth induced by the gravitropic system. If truly random of nature, the bending behavior should follow socalled 'random walk' mathematics during weightlessness. Predictions from this hypothesis were critically tested.
In a Spacelab ESA-experiment, denoted RANDOM and carried out during the IML-2 Shuttle flight in July 1994, the growth of garden cress ( Lepidium sativum ) roots was followed by time lapse photography at 1-h intervals.
The growth pattern was recorded for about 20 h. Root growth was significantly smaller in weightlessness as compared to gravity (control) conditions.
It was found that the roots performed spontaneous movements in weightlessness. The average direction of deviation of the plants consistently stayed equal to zero, despite these spontaneous movements. The average squared deviation increased linearly with time as predicted theoretically (but only for 8–10 h).
Autocorrelation calculations showed that bendings of the roots, as determined from the 1-h photographs, were uncorrelated after about a 2-h interval.
It is concluded that random processes play an important role in root growth. Predictions from a random walk hypothesis as to the growth dynamics could explain parts of the growth patterns recorded. This test of the hypothesis required microgravity conditions as provided for in a space experiment.     

Graviperception of lentil seedling roots grown in space (Spacelab Dl Mission)

The growth and graviresponsiveness of roots were investigated in lentil seedlings (Lens culinaris L. cv. Verte du Puy) grown (1) in microgravity, (2) on a 1 g centrifuge in space, (3) in microgravity and then placed on the 1 g centrifuge for 3 h, (4) on the ground. Dry seeds were hydrated in space (except for the ground control) and incubated for 25 h at 22°C in darkness. At the end of the experiment, the seedlings were photographed and fixed in glutaraldehyde in a Biorack glove box. Root length was similar for seedlings grown in space and for the ground and the 1 g centrifuge controls. The direction of root growth in the microgravity sample deviated strongly from the initial orientation of the roots of the dry seeds. This deviation could be due to spontaneous curvatures similar to those observed on clinostats. When lentil seedlings were first grown in microgravity for 25 h and then placed on the 1 g centrifuge for 3 h, their roots bent strongly under the effect of the centrifugal acceleration. The amplitude of root curvature on the centrifuge was not significantly different from that observed on ground controls growing in the vertical position and placed in the horizontal position for 3 h. The gravisensitivity of statocytes differentiated in microgravity was similar to that of statocytes differentiated on earth. There were no qualitative differences in the ultrastructural features of the gravisensing cells in microgravity and in the 1 g centrifuge and ground controls. However, the distribution of statoliths in the gravisensing cells was different in microgravity: most of them were observed in the proximal part of these cells. Thus, these organelles were not distributed at random, which is in contradiction with results obtained with clinostats. The distal complex of endoplasmic reticulum in the statocytes was not in contact with the amyloplasts. Contact and pressure of amyloplasts on the tubules were not prerequisites for gravisensing. The results obtained are not in agreement with the hypothesis that the distal endoplasmic reticulum would be the transducer of the action of the statoliths.     

Root elongation against a constant force: experiment with a computerized feedback-controlled device

Axial force was applied to the root tip of corn (Zea mays L. cv. Merit) seedlings using a computerized, feedback-controlled mechanical device. The system's feedback capability allowed continuous control of a constant tip load, and the attached displacement transducer provided the time course of root elongation. Loads up to 7.5 g decreased the root elongation rate by 0.13 mm h-1 g-1, but loads 7.5 to 17.5 g decreased the growth rate by only 0.04 mm h-1 g-1. Loads higher than 18 g stopped root elongation completely. Measurement of the cross-sectional areas of the root tips indicated that the 18 g load had applied about 0.98 MPa of axial pressure to the root, thereby exceeding the root's ability to respond with increased turgor pressure. Recorded time-lapse images of loaded roots showed that radial thickening (swelling) occurred behind the root cap, whose cross-sectional area increased with tip load.     

Sensitivity to gravistimulus of lentil seedling roots grown in space during the IML 1 Mission of Spacelab

The gravitropic curvature of seedlings of lentil ( Lens culinaris L. cv. Verte du Puy) grown in microgravity and stimulated on the 1 g centrifuge for 5 to 60 min was followed by time lapse photography in near weightlessness in the frame of the IML 1 Mission of Spacelab. In microgravity, the root tip could overshoot the direction of the 1 g acceleration after bending, whereas roots stimulated on the ground did not reach the direction of the gravity vector. On earth, there is, therefore, a regulation (inhibition of root curvature), which is gravity dependent. In space, the initial rate of curvature as well as the amplitude of curvature varied as a function of the quantity of stimulation (Q, in g min). For a given quantity of stimulation, the rate of curvature remained constant for 80 min. The bending has thus a certain inertia, which is linked to the mechanism of differential growth. The presentation time (Tp) of the lentil root was calculated by extrapolation to zero curvature of the regression line representing either the initial rate of curvature or the amplitude of curvature at 2 h after the end of the stimulation. Tp was estimated to 27 and 26 s. respectively. These results confirm the values of Tp obtained by clinostats, and they also lead to a reconsideration of the causes of the kinetics of root curvature.     

The behaviour of normal and agravitropic transgenic roots of rapeseed (Brassica napus L.) under microgravity conditions

In the TRANSFORM experiment for IML-2 on the Space Shuttle Columbia, normal (wild type = WT) and genetically transformed agravitropic rapeseed roots were tested under microgravity conditions. The aim of the experiment was to determine if the wild-type roots behaved differently (growth, morphology, gravitropical sensitivity) from the transgenic roots. The appearance of the organelles and distribution of statoliths (i.e. amyloplasts with starch grains) in the gravitropic reactive cells (statocytes) under weightlessness was compared for the two types of roots. Attempts have also been made to regenerate new plants from the root material tested in space. Both the WT and the transgenic root types showed the expected increase in length during 36 h of photorecording. Contrary to the results of the ground controls, no significant difference in elongation rates was found between the WT and transgenic roots grown in orbit. However, there are indications that the total growth both in the WT and the transgenic roots was higher in the ground control than for roots in orbit. After a 60 min 1 x g stimulation of the roots on board the Shuttle, no detectable curvatures were obtained in either the transgenic or the WT roots. However, it cannot be excluded that a minute curvature development occurs in the root tips but was not detected due to technical reasons. The ultrastructure was well preserved in both the WT and the transgenic roots, despite the fact that the tissue was kept in the prefixative for over 3 weeks. No marked differences in ultrastructure were observed between the transformed root statocyte cells and the equivalent cells in the wild type. There were no obvious differences in root morphology during the orbital period. Light micrographs and morphometrical analysis indicate that the amyloplasts of both the wild type and transformed root statocytes are randomly distributed over the cells kept under micro-g conditions for 37 h after a 14 h stimulation on the 1 x g centrifuge. The main scientific conclusion from the TRANSFORM experiment is that the difference in growth found in the ground control between the WT and the transgenic root types seems to be eliminated under weightlessness. Explanations for this behaviour cannot be found in the root ultrastructure or in root morphology.     

Plastids: dynamic components of plant cell development

The gravitropic bending of maize roots, as a response to reorientation of the root within a gravitational field, was examined for sensitivity to exogenous applications of the cytoskeletal inhibitor, cytochalasin D. Agar blocks were impregnated with this inhibitor, and were applied either to the root cap or to the zone of root cell elongation. Root growth was normal with either treatment, if the roots were not repositioned with respect to the gravitational vector. When untreated roots were placed in a horizontal position with respect to gravity, a 40 degree bending response was observed within one hour. This bending also occurred when cytochalasin D was applied at high concentrations to the zone of root cell elongation. However, when cytochalasin D above 40 micrograms/ml was applied to the root cap, roots lost the ability of directional reorientation within the gravitational field, causing a random bending.     

Negative phototropism of rice root and its influencing factors

Some characteristics of the rice (Oryza sativa L.) root were found in the experiment of unilaterally irradiating the roots which were planted in water: (i) All the seminal roots, adventitious roots and their branched roots bent away from light, and their curvatures ranged from 25℃ to 60℃. The curvature of adventitious root of the higher node was often larger than that of the lower node, and even larger than that of the seminal root. (ii) The negative phototropic bending of the rice root was mainly due to the larger growth increment of root-tip cells of the irradiated side compared with that of the shaded side, (iii) Root cap was the site of light perception. If root cap was shaded while the root was irradiated the root showed no negative phototropism, and the root lost the characteristic of negative phototropism when root cap was divested. Rice root could resume the characteristic of negative phototropism when the new root cap grew up, if the original cells of root cap were well protected while root ca     

Changes in plant growth processes under microgravity conditions simulated by a three-dimensional clinostat

We developed a three-dimensional (3-D) clinostat to simulate a microgravity environment and studied the changes in plant growth processes under this condition. The rate of germination of cress (Lepidium sativum), maize (Zea mays), rice (Oryza sativa), pea (Pisum sativum), or azuki bean (Vigna angularis) was not affected on the clinostat. The clinostat rotation did not influence the growth rate of their roots or shoots, except for a slight promotion of growth in azuki roots and epicotyls. On the contrary, the direction of growth of plant organs clearly changed on the 3-D clinostat. On the surface of the earth, roots grow downward while shoots upward in parallel to the gravity vector. On the 3-D clinostat, roots of cress elongated along the direction of the tip of root primordia after having changed the direction continuously. Rice roots also grew parallel to the direction of the tip of root primordia. On the other hand, roots of maize, pea, and azuki bean grew in a random fashion. The direction of growth of shoots was more controlled even on the 3-D clinostat. In a front view of embryos, shoots grew mostly along the direction of the tip of primordia. In a side view, rice coleoptiles showed an adaxial (toward the caryopsis) while coleoptiles of maize and epicotyls of pea and azuki bean an abaxial curvature. The curvature of shoots became larger with their growth. Such an autotropism may have an important role in regulation of life cycle of higher plants under a microgravity environment.     

Effects of fire,mowing and nitrogen addition on root characteristics in tall-grass prairie

Abstract. Root harvests and root windows were used to study the influence of fire, mowing and nitrogen additions on root lengths, biomass, and nitrogen content in tall-grass prairie. Four years of nitrogen additions (10 g m² yr^?1) increased below-ground mass by 15 % and nitrogen concentration in that mass by 77 %. In general, live roots and rhizomes exhibited greater increases in nitrogen concentrations than detrital roots and rhizomes. After four years of treatment, live roots and rhizomes immobilized an additional 1.5 to 5 g/m² of nitrogen, depending upon specific treatment, while dead roots and rhizomes immobilized an additional 3 to 3.5 g/m². Average root growth parameters, as measured with root windows, were positively correlated with above-ground peak foliage biomass; however, the only significant correlation was between average new root growth and above-ground peak foliage biomass (r = 0.73, p ≤ 0.04). Root growth and decay, as measured by annual mean values for eight root windows over a four year interval, were insensitive to climatic and treatment effects.     

Automorphogenesis of maize roots under simulated microgravity conditions

Plant seedlings show exaggerated growth responses on a three-dimensional clinostat. Such an automorphogenesis appears to be one of major factors which govern the life cycle of higher plants under a microgravity environment. On the three-dimensional clinostat, maize roots exhibited curvatures in three different portions; 1) the basal region just protruding from the coleorhiza, 2) the region between the mature and the elongation zone, and 3) the elongation zone, several mm from the tip. Even non-clinostatted control roots showed some degree of curvature. The curvature occurred at random without any dorsiventrality. There was no difference in the osmotic concentration of the cell sap between the convex and the concave halves of any region. However, the convex, rapidly expanding side exhibited a higher extensibility of the cell wall in some regions, which appears to be a cause of the curvature. In order to understand the role of gravity in regulation of plant growth and development, we should clarify a series of events by which an automorphogenesis is induced under simulated microgravity conditions.     

Root growth and statocyte polarity in lentil seedling roots grown in microgravity or on a slowly rotating clinostat

The ability of clinostats to simulate microgravity was evaluated by comparing lentil ( Lens culinnrias L. cv. Verte du Puy) seedlings grown in space (Spacelab D1 Mission) with seedlings grown on a slowly rotating elinostat. Seeds were germinated and incubated for 25.5 h at 22°C (1) in microgravity, (2) on a 1g-centrifuge in space. (3) on a slowly rotating elinostat and (4) on the ground. Morphological (root length and orientation) and ultrastructural (distribution of amyloplasts, location of the nucleus in statocytes) parameters were studied. For clinostat experiments, two different configurations were employed: the longitudinal axis of the root was parallel (horizontal elinorotation) or perpendicular (vertical elinorotation) to the axis of rotation. the same configurations were used for the lg-controls. Root length and orientation were similar for roots grown on the clinostat and in microgravity. The amyloplasts were identically distributed in statocytes of horizontally clinorolated roots and in statocytes differentiated in microgravity. However, the location of the nucleus was similar in vertically rotated roots and microgravity samples. Since the involvement of the nucleus in graviperception is not known, it can be concluded that horizontal clinorotation simulates microgravity better than vertical elinorotation.     

Cell cycle and cell differentiation in lentil roots grown on a slowly rotating clinostat

Root growth was studied for seedlings of lentil ( Lens culinaris L. cv. Verte du Puy) grown for 27 h either on a slowly rotating clinostat (0.9 rev. min⁻¹) of vertically (controls). Horizontal clinorotation was employed, so that the longitudinal axis of the root was parallel to the axis of the rotation. Morphological (root length and orientation) and cellular (cell proliferation and cell elongation) parameters were studied. The cell cycle was also analysed by flow cytometry. Root length deviation of the roots from the initial orientation was observed on the clinostat; this deviation could be due to spontaneous oscillation. Cell elongation of the clinostat-rotated roots occurred closer to the tip than in the vertical roots, but the mitotic index was not modified. Clinorotation did not change the frequencies of the G1, S and G2 phases of the cell cycle. These results were compared to those obtained during the D1 mission on Spacelab, 1985. The effects of microgravity on root orientation and mitotic index were not simulated by clinorotation.     

Negative phototropism of rice root and its influencing factors

Some characteristics of the rice (Oryza sativa L.) root were found in the experiment of unilaterally irradiating the roots which were planted in water: (i) All the seminal roots, adventitious roots and their branched roots bent away from light, and their curvatures ranged from 25° to 60°. The curvature of adventitious root of the higher node was often larger than that of the lower node, and even larger than that of the seminal root, (ii) The negative phototropic bending of the rice root was mainly due to the larger growth increment of root-tip cells of the irradiated side compared with that of the shaded side, (iii) Root cap was the site of light perception. If root cap was shaded while the root was irradiated the root showed no negative phototropism, and the root lost the characteristic of negative phototropism when root cap was divested. Rice root could resume the characteristic of negative phototropism when the new root cap grew up, if the original cells of root cap were well protected while root cap was divested, (iv) The growth increment and curvature of rice root were both influenced by light intensity. Within the range of 0–100 μmol · m² -s⁻¹, the increasing of light intensity resulted in the decreasing of the growth increment and the increasing of the curvature of rice root, (v) The growth increment and the curvature reached the maximum at 30°C with the temperature treatment of 10–40°C. (vi) Blue-violet light could prominently induce the negative phototropism of rice root, while red light had no such effect. (vii) The auxin (IAA) in the solution, as a very prominent influencing factor, inhibited the growth, the negative phototropism and the gravitropism of rice root when the concentration of IAA increased. The response of negative phototropism of rice root disappeared when the concentration of IAA was above 10 mg · L⁻¹     

Root cap removal increases root penetration resistance in maize (Zea mays L)

The root cap assists the passage of the root through soil by means of its slimy mucilage secretion and by the sloughing of its outer cells. The root penetration resistance of decapped primary roots of maize (Zea mays L. cv. Mephisto) was compared with that of intact roots in loose (dry bulk density 1.0 g cm-3; penetration resistance 0.06 MPa) and compact soil (1.4 g cm-3; penetration resistance 1.0 MPa), to evaluate the contribution of the cap to decreasing the impedance to root growth. Root elongation rate and diameter were the same for decapped and intact roots when the plants were grown in loose soil. In compacted soil, however, the elongation rate of decapped roots was only about half that of intact roots, whilst the diameter was 30% larger. Root penetration resistances of intact and decapped seminal axis were 0.31 and 0.52 MPa, respectively, when the roots were grown in compacted soil. These results indicated that the presence of a root cap alleviates much of the mechanical impedance to root penetration, and enables roots to grow faster in compacted soils.     

D‐Root: a system for cultivating plants with the roots in darkness or under different light conditions

In nature roots grow in the dark and away from light (negative phototropism). However, most current research in root biology has been carried out with the root system grown in the presence of light. Here, we have engineered a device, called Dark‐Root (D‐Root), to grow plants in vitro with the aerial part exposed to the normal light/dark photoperiod while the roots are in the dark or exposed to specific wavelengths or light intensities. D‐Root provides an efficient system for cultivating a large number of seedlings and easily characterizing root architecture in the dark. At the morphological level, root illumination shortens root length and promotes early emergence of lateral roots, therefore inducing expansion of the root system. Surprisingly, root illumination also affects shoot development, including flowering time. Our analyses also show that root illumination alters the proper response to hormones or abiotic stress (e.g. salt or osmotic stress) and nutrient starvation, enhancing inhibition of root growth. In conclusion, D‐Root provides a growing system closer to the natural one for assaying Arabidopsis plants, and therefore its use will contribute to a better understanding of the mechanisms involved in root development, hormonal signaling and stress responses.     

Root productivity in a lowland tropical rain forest in Mexico

Fine root productivity was estimated in a lowland tropical rain forest at Los Tuxtlas (SE Mexico) and examined in relation to climatic factors. Two root diameter classes were defined (class I,<1 mm; class II, 1–3 mm). Total root productivity was estimated to 1.95 t ha^–1 year^–1, a value which is lower than those reported from other rain forest sites. Significant differences in root dry weight were found among months and between diameter classes throughout the year. Class I monthly means formed two groups: one corresponding to the months of highest precipitation, and the other to the relatively dry season. Class II monthly means also formed two groups, although these were unrelated to the regional precipitation pattern. A multiplicative regression model of productivity on precipitation was significant for both root classes when rainfall data of the previous month were used, while a linear regression model was significant only for class I roots when temperature data of two months before were used; these results suggest a delay in the effect of climatic conditions on root productivity. While the seasonal pattern of root productivity is clearly related to the annual rainfall distribution, the low total annual productivity may be related to the very high soil fertility at Los Tuxtlas.     

Nastic curvatures of wheat coleoptiles that develop in true microgravity

Dark-grown wheat coleoptiles developed strong curvatures within 5 h of being transferred in orbit from a 1 g centrifuge to microgravity during an experiment flown on the IML-1 shuttle mission. The curving tendency was strongest in seedlings that were immature, with coleoptiles shorter than 10 mm at the time of transfer. The curvature direction was non-random, and directed away from the caryopsis (the coleptile face adjacent to the caryopsis becoming convex). The curvatures were most marked in the basal third of the coleoptiles, contrasting with phototropic responses, which occur in the apical third. We interpret these curvatures as being nastic, and related to the curvatures commonly reported to occur during clinostat rotation treatments.     

Patterns of variability in the diameter of lateral roots in the banana root system

The relative importance of root system structure, plant carbon status and soil environment in the determination of lateral root diameter remains unclear, and was investigated in this study. Banana (Musa acuminata) plants were grown at various moderate levels of soil compaction in two distinct experiments, in a field experiment (FE) and in a glasshouse experiment (GE). Radiant flux density was 5 times lower in GE. The distribution of root diameter was measured for several root branching orders. Root diameters ranged between 0.09 and 0.52 mm for secondary roots and between 0.06 and 0.27 mm for tertiary roots. A relationship was found between the diameter of the parent bearing root and the median diameter of its laterals, which appears to be valid for a wide range of species. Mean lateral root diameter increased with distance to the base of the root and decreased with branching density [number of lateral roots per unit length of bearing root (cm(-1))]. Typical symptoms of low light availability were observed in GE. In this case, lateral root diameter variability was reduced. Although primary root growth was affected by soil compaction, no effects on lateral root diameter were observed.     

Linking above- and belowground phenology of hybrid walnut growing along a climatic gradient in temperate agroforestry systems

Background and aims

Plant phenology is a sensitive indicator of plant response to climate change. Observations of phenological events belowground for most ecosystems are difficult to obtain and very little is known about the relationship between tree shoot and root phenology. We examined the influence of environmental factors on fine root production and mortality in relation with shoot phenology in hybrid walnut trees (Juglans sp.) growing in three different climates (oceanic, continental and Mediterranean) along a latitudinal gradient in France.

Methods

Eight rhizotrons were installed at each site for 21 months to monitor tree root dynamics. Root elongation rate (RER), root initiation quantity (RIQ) and root mortality quantity (RMQ) were recorded frequently using a scanner and time-lapse camera. Leaf phenology and stem radial growth were also measured. Fine roots were classified by topological order and 0–1 mm, 1–2 mm and 2–5 mm diameter classes and fine root longevity and risk of mortality were calculated during different periods over the year.

Results

Root growth was not synchronous with leaf phenology in any climate or either year, but was synchronous with stem growth during the late growing season. A distinct bimodal pattern of root growth was observed during the aerial growing season. Mean RER was driven by soil temperature measured in the month preceding root growth in the oceanic climate site only. However, mean RER was significantly correlated with mean soil water potential measured in the month preceding root growth at both Mediterranean (positive relationship) and oceanic (negative relationship) sites. Mean RIQ was significantly higher at both continental and Mediterranean sites compared to the oceanic site. Soil temperature was a driver of mean RIQ during the late growing season at continental and Mediterranean sites only. Mean RMQ increased significantly with decreasing soil water potential during the late aerial growing season at the continental site only. Mean root longevity at the continental site was significantly greater than for roots at the oceanic and Mediterranean sites. Roots in the 0–1 mm and 1–2 mm diameter classes lived for significantly shorter periods compared to those in the 2–5 mm diameter class. First order roots (i.e. the primary or parents roots) lived longer than lateral branch roots at the Mediterranean site only and first order roots in the 0–1 mm diameter class had 44.5% less risk of mortality than that of lateral roots for the same class of diameter.

Conclusions

We conclude that factors driving root RER were not the same between climates. Soil temperature was the best predictor of root initiation at continental and Mediterranean sites only, but drivers of root mortality remained largely undetermined.