Mechanics of root elongation and the effects of 3,5-diiodo-4-hydroxybenzoic acid (DIHB)

Summary This paper reports the results of two series of experiments. In the first the effects of DIHB on the rate of root elongation were compared on unstressed roots and on roots stressed by mechanical impedance and by inadequate levels of aeration. Barley plants were grown in beds of small glass spheres through which nutrient solution was circulated. Mechanical impedance of 25 kPa was applied by subjecting the beds to a confining pressure. Inadequate aeration was obtained by reducing the oxygen concentration in the nutrient solution to 5%. The second series examined possible effects of DIHB on the elastic modulus of root tips of wheat and pea. Elastic modulus gives an indication of the behaviour of roots in structured soil where penetration of peds can be limited by the buckling of root tips. The elastic modulus was measured in experiments of the static cantilever type on roots previously immersed in solutions of polyethylene glycol of different osmotic potential. Elastic modulus measurements can also detect any changes in turgor pressure and wilting characteristics of roots and can therefore help to identify the mechanisms of action of DIHB. DIHB caused increases in root elongation relative to controls in all cases: 26±5.7% in unstressed roots, 14±6.4% in mechanically impeded roots and 54±9.8% in roots growing in 5% oxygen. DIHB had no effect on the elastic modulus, osmotic or turgor pressure of the roots. It is concluded that DIHB acts by modifying the cell wall extensibility factor.     

Root elongation, water stress, and mechanical impedance: a review of limiting stresses and beneficial root tip traits

Root elongation in drying soil is generally limited by a combination of mechanical impedance and water stress. Relationships between root elongation rate, water stress (matric potential), and mechanical impedance (penetration resistance) are reviewed, detailing the interactions between these closely related stresses. Root elongation is typically halved in repacked soils with penetrometer resistances >0.8-2?MPa, in the absence of water stress. Root elongation is halved by matric potentials drier than about -0.5?MPa in the absence of mechanical impedance. The likelihood of each stress limiting root elongation is discussed in relation to the soil strength characteristics of arable soils. A survey of 19 soils, with textures ranging from loamy sand to silty clay loam, found that ～10% of penetration resistances were >2?MPa at a matric potential of -10?kPa, rising to nearly 50% >2?MPa at - 200?kPa. This suggests that mechanical impedance is often a major limitation to root elongation in these soils even under moderately wet conditions, and is important to consider in breeding programmes for drought-resistant crops. Root tip traits that may improve root penetration are considered with respect to overcoming the external (soil) and internal (cell wall) pressures resisting elongation. The potential role of root hairs in mechanically anchoring root tips is considered theoretically, and is judged particularly relevant to roots growing in biopores or from a loose seed bed into a compacted layer of soil.     

The effect of mechanical impedance on root growth in pea (Pisum sativum). II. Cell expansion and wall rheology during recovery

The aim of the present work was to determine the factors limiting growth in mechanically impeded roots. Pea roots were grown in compressed and uncompressed sand cores, and then removed and transferred to hydroponics. Root elongation was slowed in impeded sand cores and did not recover to the unimpeded rates until 60 h after transfer to the hydroponics system. Root diameter was greater in impeded roots, and only after 36 h in hydroponics was new root tissue produced of the same diameter as the unimpeded controls. The turgor pressure of the growing cells was measured with a turgor probe and was the same in both treatments. The slower elongation rate of the previously impeded roots was, therefore, the result of axial tightening of the cell walls. Cell length profiles suggested that axial cell wall tightening persisted in the unrestricted hydroponics system. Production of new cells in unrestricted conditions was required before root elongation returned to the unimpeded state. Osmotic potential was decreased by approximately 0.2 MPa in previously impeded roots compared with the unimpeded ones. This corresponds to a decrease in water potential of 0.2 MPa. These data are discussed in relation to regulation of cell extension, solute unloading and the penetration of compacted soils by roots.     

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.     

Penetrometer resistance,root penetration resistance and root elongation rate in two sandy loam soils

Root penetration resistance and elongation of maize seedling roots were measured directly in undisturbed cores of two sandy loam soils. Root elongation rate was negatively correlated with root penetration resistance, and was reduced to about 50 to 60% of that of unimpeded controls by a resistance of between 0.26 and 0.47 MPa. Resistance to a 30° semiangle, 1 mm diameter penetrometer was between about 4.5 and 7.5 times greater than the measured root penetration resistance. However, resistance to a 5° semiangle, 1 mm diameter probe was approximately the same as the resistnace to root penetration after subtracting the frictional component of resistance. The diameter of roots grown in the undisturbed cores was greater than that of roots grown in loose soil, probably as a direct result of the larger mechanical impedance in the cores.     

The effects of 3,5-diiodo-4-hydroxybenzoic acid on the oxidation of IAA and auxin-induced ethylene production by cress root segments

Summary In previous research here, 3,5-diiodo-4-hydroxybenzoic acid (DIHB) was shown to promote the elongation of roots of cress (Lepidium sativum) seedlings growing in light, and to inhibit the auxin-induced production of ethylene in this tissue. Although DIHB is a cofactor for the oxidation of indole-3-acetic acid (IAA) by horse-radish peroxidase, it inhibits the decarboxylation of [1-¹⁴C]IAA by segments excised from cress roots. The inhibition by DIHB of ethylene production by this tissue does not, therefore, arise from a reduction of IAA levels. These findings are discussed in relation to the effects of DIHB on cress root growth.Abbreviations IAA indole-3-acetic acid - DIHB 3,5-diiodo-4-hydroxybenzoic acid - DCP 2,4-dichlorophenol - 2,4-D 2,4-dichlorophenoxyacetic acid This study forms part of a research project to be submitted by M.L.R. for PhD degree and supported by a grant from Consejo Nacional de Ciencia y Tecnología (México).     

Root elongation of seedling peas through layered soil of different penetration resistances

Field soils contain localized zones of larger penetration resistance within peds and compacted layers, while cracks and biopores offer low resistance pathways to roots. Root responses to such localized conditions have not been investigated in detail. This study examined what happens to the root elongation rate when roots grew through a layer of hard soil into a layer of looser soil for a 4 day period. The experiment was performed twice; firstly with the shoot in continuous darkness, and secondly with it exposed to a day-night cycle to prevent etiolation of the shoot. Pea seedlings were grown in columns of a sandy loam soil which was packed to bulk densities of 0.85, 1.1, 1.3 or 1.4 Mg/m³ in the top layer and 0.85 Mg/m³ in the bottom layer. The root elongation rate in the top layer of 1.4 Mg/m³ soil (penetrometer resistance=1.8 MPa) was only 55% of the elongation rate in the top layer of 0.85 Mg/m³ soil (penetrometer resistance=0.06 MPa). The elongation rate of roots that had grown through the top layer of 1.4 Mg/m³ soil into the bottom layer of loose soil was reduced by some residual effect of the mechanical impedance. The root elongation rate in the bottom layer of loose soil decreased as the penetrometer resistance of the top layer of soil increased. The daily elongation rate of the roots in the bottom layer that had grown through the 1.4 Mg/m³ soil averaged only about 65% of the elongation rate of the roots that had grown through the 0.85 Mg/m³ soil. This residual effect of mechanical impedance on root elongation persisted for at least 2 days and was more severe in the day-night cycle experiment than in the dark experiment. These results have important implications for modelling root elongation in any soil in which the soil strength changes with distance or with time.     

Effect of post-planting loosening of unstable soil on the anatomy of corn root

Summary Corn plants were grown in pots to study the effect of post-planting loosening of an unstable sandy loam soil on the anatomy of roots. Roots from compact soil had more elaborate production of sclerified cells in the cortical and vascular tissue which may be considered as traits developed in such roots to resist external forces to prevent the deformation of interior cells. It appears that the compact layer that developes on unstable soils after irrigation, hinders the normal development of root structure. Its loosening decreases soil resistance and permits normal development of root anatomy.     

A biophysical analysis of root growth under mechanical stress

The factors controlling root growth in hard soils are reviewed alongside summarised results from our recent studies. The turgor in cells in the elongation zone of roots pushes the apex forward, resisted by the external pressure of the soil and the tension in the cell walls. The external pressure of the soil consists of the pressure required to deform the soil, plus a component of frictional resistance between the root and soil. This frictional component is probably small due to the continuous sloughing of root cap cells forming a low-friction lining surrounding the root. Mechanically impeded roots are not only thicker, but are differently shaped, continuing to increase in diameter for a greater distance behind the root tip than in unimpeded roots. The osmotic potential decreases in mechanically impeded roots, possibly due to accumulation of solutes as a result of the slower root extension rate. This more negative osmotic potential is not always translated into increased turgor pressure, and the reasons for this require further investigation. The persistent effect of mechanical impedance on root growth is associated with both a stiffening of cell walls in the axial direction, and with a slowing of the rate of cell production.     

Growth dynamics of mechanically impeded lupin roots: does altered morphology induce hypoxia?

BACKGROUND AND AIMS: Root axes elongate slowly and swell radially under mechanical impedance. However, temporal and spatial changes to impeded root apices have only been described qualitatively. This paper aims (a) to quantify morphological changes to root apices and (b) assess whether these changes pre-dispose young root tissues to hypoxia. METHODS: Lupin (Lupinus angustifolius) seedlings were grown into coarse sand that was pressurized through a diaphragm to generate mechanical impedance on growing root axes. In situ observations yielded growth rates and root response to hypoxia. Roots were then removed to assess morphology, cell lengths and local growth velocities. Oxygen uptake into excised segments was measured. KEY RESULTS: An applied pressure of 15 kPa slowed root extension by 75% after 10-20 h while the same axes thickened by about 50%. The most terminal 2-3 mm of axes did not respond morphologically to impedance, in spite of the slower flux of cells out of this region. The basal boundary of root extension encroached to within 4 mm of the apex (cf. 10 mm in unimpeded roots), while radial swelling extended 10 mm behind the apex in impeded roots. Oxygen demand by segments of these short, thick, impeded roots was significantly different from segments of unimpeded roots when the zones of elongation in each treatment were compared. Specifically, impeded roots consumed O2 faster and O2 consumption was more likely to be O2-limited over a substantial proportion of the elongation zone, making these roots more susceptible to O2 deficit. Impeded roots used more O2 per unit growth (measured as either unit of elongation or unit of volumetric expansion) than unimpeded roots. Extension of impeded roots in situ was O2-limited at sub-atmospheric O2 levels (21% O2), while unimpeded roots were only limited below 11% O2. CONCLUSIONS: The shift in the zone of extension towards the apex in impeded roots coincided with greater vulnerability to hypoxia even after soil was removed. Roots still encased in impeded soil are likely to suffer from marked O2 deficits.     

Ethylene as a possible mediator of light-induced inhibition of root growth

Eliasson, L. and Bollmark, M. 1988. Ethylene as a possible mediator of light-induced inhibition of root growth. - Physiol. Plant. 72: 605–609.
Pea seedlings ( Pisum sativum L. cv. Weibull's Marma) were used to investigate the possible role of ethylene in light-induced inhibition of root elongation. Illumination of the roots with white light inhibited root elongation by 40–50% and increased ethylene production by the roots about 4-fold. Our main approach was to use exogenous 1-aminocyclopropane-1-carboxylic acid (ACC), supplied in the growth solution, to monitor ethylene production of the roots independent of light treatment. Ethylene production of excised root tips increased with increasing ACC concentrations. The rate of ethylene production in dark-grown roots treated with 0.1 μ M ACC was similar to that caused by illumination. Low ACC concentrations (0.01–0.1 μ M ) decreased the rate of root elongation, especially in seedlings grown in the dark, and 0.1 μ M ACC inhibited elongation to about the same extent as light. In light the roots curved and grew partly plagiogravitropically. This effect was also simulated by the 0.1 μ M ACC treatment. At 1 μ M and higher concentrations, ACC inhibited root growth almost completely and caused conspicuous curvatures of the root tips both in light and darkness. Inhibitors of ethylene synthesis and action partially counteracted the inhibition of root elongation caused by light. These observations suggest that the increase in ethylene production caused by light is at least partly responsible for the decreased growth of light-exposed roots.     

Pre-germination effects on mechanically impeded root growth of barley (Hordeum vulgare L)

The effects of conditions pre-dating germination on growth rate of impeded barley cv. Harrington roots were measured using an agar-capillary tube technique. Seedling root tips were directed into glass capillary tubes twothirds filled with agar at eight concentrations ranging from 1.6 to 9.6%, equivalent to penetrometer resistances of 25 to 1240 kPa. The rate of unrestricted root elongation (growth in air) of seed stored for 13 months (old seed), and of seed grown for a second generation without subsequent storage (new seed) was compared with growth in agar over a 24-hour interval. Root elongation rate of old and new seed was identical in the absence of resistance. At low to intermediate agar concentrations, elongation was significantly slower in roots from old, compared with new seed. At high agar concentrations root growth of old and new seed was the same. In both old and new seed, root growth through agar was greater in seed that germinated after 24, compared with 48 h. Differences in impeded root growth between old and new seed were lost in progeny of the test seed. Environmental factors that pre-date germination are an important influence on the ability of seedling roots to elongate through soil.LRS Contribution no. 3879349LRS Contribution no. 3879349     

Root elongation is restricted by axial but not by radial pressures: so what happens in field soil?

Root distribution determines largely the zone of soil that roots have access to for water and nutrient uptake, and is of great importance especially if water and fertilizer input is restricted. Mechanical impedance is the major limitation to root elongation in many field soils. Until now, experiments have focused largely on the axial resistance to root growth. In a fascinating study of the radial forces exerted by the roots of chickpea, root extension, diameter change, and the radial forces that axially unimpeded roots exert are reported: Kolb et al. (this volume) record radial stresses of about 0.3?MPa that are broadly consistent with cell turgor pressures, but, interestingly, find no restriction to axial elongation. This result is in marked contrast to large decreases in elongation of pea radicles resulting from much smaller axial pressures reported elsewhere in the literature (e.g., an 85?% decrease in root elongation in response to axial pressures of?<?0.1?MPa). The situation is different also from that in homogeneous soil, where root penetration resistance pressures of 0.4-1.0?MPa are typically required to halt root elongation. Soil structure and strength properties will determine the balance of axial and radial pressures on an individual root tip, and hence the root elongation response. It appears that a degree of radial confinement may help roots to extend axially into hard soil. This result also complements recent findings that in strong field soils the availability of soil macropores has a large influence on regulating the root-elongation rates of seedlings.     

Contribution of root cap mucilage and presence of an intact root cap in maize (Zea mays) to the reduction of soil mechanical impedance

BACKGROUND AND AIMS: The impedance to root growth imposed by soil can be decreased by both mucilage secretion and the sloughing of border cells from the root cap. The aim of this study is to quantify the contribution of these two factors for maize root growth in compact soil. METHODS: These effects were evaluated by assessing growth after removing both mucilage (treatment I -- intact) and the root cap (treatment D -- decapped) from the root tip, and then by adding back 2 micro L of mucilage to both intact (treatment IM -- intact plus mucilage) and decapped (treatment DM -- decapped plus mucilage) roots. Roots were grown in either loose (0.9 Mg m(-3)) or compact (1.5 Mg m(-3)) loamy sand soils. Also examined were the effects of decapping on root penetration resistance at three soil bulk densities (1.3, 1.4 and 1.5 Mg m(-3)). KEY RESULTS: In treatment I, mucilage was visible 12 h after transplanting to the compact soil. The decapping and mucilage treatments affected neither the root elongation nor the root widening rates when the plants were grown in loose soil for 12 h. Root growth pressures of seminal axes in D, DM, I and IM treatments were 0.328, 0.288, 0.272 and 0.222 MPa, respectively, when the roots were grown in compact soil (1.5 Mg m(-3) density; 1.59 MPa penetrometer resistance). CONCLUSIONS: The contributions of mucilage and presence of the intact root cap without mucilage to the lubricating effect of root cap (percentage decrease in root penetration resistance caused by decapping) were 43 % and 58 %, respectively. The lubricating effect of the root cap was about 30 % and unaffected by the degree of soil compaction (for penetrometer resistances of 0.52, 1.20 and 1.59 MPa).     

Soil types with different texture affects development of Rhizoctonia root rot of wheat seedlings

The effect of different soil textures, sandy (97.5% sand, 1.6% silt, 0.9% clay), loamy sand (77% sand, 11% silt, 12% clay) and a sandy clay loam (69% sand, 7% silt, 24% clay), on root rot of wheat caused by Rhizoctonia solani Kühn Anastomosis Group (AG) 8 was studied under glasshouse conditions. The reduction in root and shoot biomass following inoculation with AG-8 was greater in sand than in loamy sand or sandy clay loam. Dry root weight of wheat in the sand, loamy sand and sandy clay loam soils infested with AG-8 was 91%, 55% and 28% less than in control uninfested soils. There was greater moisture retention in the loamy sand and sandy clay loam soils as compared to the sand in the upper 10–20 cm. Root penetration resistance was greater in loamy sand and sandy clay loam than in sand. Root growth in the uninfested soil column was faster in the sand than in the loamy sand and sandy clay loam soils, the roots in the sandy soil being thinner than in the other two soils. Radial spread of the pathogen in these soils in seedling trays was twice as fast in the sand in comparison to the loamy sand which in turn was more than twice that in the sandy clay loam soil. There was no evidence that differences among soils in pathogenicity or soil spread of the pathogen was related to their nutrient status. This behaviour may be related to the severity of the disease in fields with sandy soils as compared to those with loam or clay soils. This revised version was published online in June 2006 with corrections to the Cover Date.     

Penetration of very strong soils by seedling roots of different plant species

The abilities of seedling roots of twenty-two plant species to penetrate a strong growth medium were compared under controlled conditions. Seedlings were grown for 10 days in compression chambers filled with siliceous sandy soil at 0.2 kg kg^–1 water content and mean penetrometer resistance of 4.2 MPa. Root elongation and thickening were measured after growth. The results show that soil strength reduced the elongation of roots of all plant species by over 90% and caused the diameters of the roots to increase compared with control plants grown in vermiculite (0 MPa resistance).Differences in both root elongation and root diameter were observed among plant species. Generally, the roots of dicotyledons (with large diameters) penetrated the strong medium more than graminaceous monocotyledons (with smaller diameters). There was a significant positive correlation (r=0.78, p<0.05) between root diameter and elongation over all the species in the stressed plants. The species were ranked according to the relative root elongation and relative root thickening. Based on this ranking, lupin (Lupinus angustifolius), medic (Medicago scutelata) and faba bean (Vicia faba) were the species with the greatest thickening and elongation while wheat (Triticum aestivum), rhodesgrass (Chloris gayana) and barley (Hordeum vulgare) had the least. The weight of the seeds did not seem to influence either the thickening or elongation of the roots.     

Ethylene Evolution from Maize (Zea mays L.) Seedling Roots and Shoots in Response to Mechanical Impedance

The effect of mechanical impedance on ethylene evolution and growth of preemergent maize (Zea mays L.) seedlings was investigated by pressurizing the growth medium in triaxial cells in a controlled environment. Pressure increased the bulk density of the medium and thus the resistance to growth. The elongation of maize primary roots and preemergent shoots was severely hindered by applied pressures as low as 10 kilopascals. Following a steep decline in elongation at low pressures, both shoots and roots responded to additional pressure in a linear manner, but shoots were more severely affected than roots at higher pressures. Radial expansion was promoted in both organs by mechanical impedance. Primary roots typically became thinner during the experimental period when grown unimpeded. In contrast, pressures as low as 25 kilopascals caused a 25% increase in root tip diameter. Shoots showed a slight enhancement of radial expansion; however, in contrast to roots, the shoots increased in diameter even when growing unimpeded. Such morphological changes were not evident until at least 3 hours after initiation of treatment. All levels of applied pressure promoted ethylene evolution as early as 1 hour after application of pressure. After 1 hour, ethylene evolution rates had increased 10, 32, 70, and 255% at 25, 50, 75, and 100 kilopascals respectively, and continued to increase linearly for at least 10 hours. When intact corn seedlings were subjected to a series of hourly cycles of pressure, followed by relaxation, ethylene production rates increased or decreased rapidly, illustrating tight coupling between mechanical impedance and tissue response. Seedlings exposed to 1 microliter of ethylene per liter showed symptoms similar to those shown by plants grown under mechanical impedance. Root diameter increased 5 times as much as the shoot diameter. Pretreatment with 10 micromolar aminoethoxyvinyl glycine plus 1 micromolar silver thiosulfate maintained ethylene production rates of impeded seedlings at basal levels and restored shoot and root extension to 84 and 90% of unimpeded values, respectively. Our results support the hypothesis that ethylene plays a pivotal role in the regulation of plant tissue response to mechanical impedance.     

Numbers and locations of native bacteria on field-grown wheat roots quantified by fluorescence in situ hybridization (FISH)

Native bacteria, Pseudomonas and filamentous bacteria were quantified and localized on wheat roots grown in the field using fluorescence in situ hybridization (FISH). Seminal roots were sampled through the season from unploughed soil in a conservation farming system. Such soils are spatially heterogeneous, and many roots grow slowly through hard soil with cracks and pores containing dead roots remnant from previous crops. Root and rhizosphere morphology, and contact with soil particles were preserved, and autofluorescence was avoided by observing sections in the far-red with Cy5 and Cy5.5 fluorochromes. Spatial analyses showed that bacteria were embedded in a stable matrix (biofilm) within 11 microm of the root surface (range 2-30 microm) and were clustered on 40% of roots. Half the clusters co-located with axial grooves between epidermal cells, soil particles, cap cells or root hairs; the other half were not associated with visible features. Across all wheat roots, although variable, bacteria averaged 15.4 x 10(5) cells per mm(3) rhizosphere, and of these, Pseudomonas and filaments comprised 10% and 4%, respectively, with minor effects of sample time, and no effect of plant age. Root caps were most heavily colonized by bacteria along roots, and elongation zones least heavily colonized. Pseudomonas varied little with root development and were 17% of bacteria on the elongation zone. Filamentous bacteria were not found on the elongation zone. The most significant factor to rhizosphere populations along a wheat root, however, was contact with dead root remnants, where Pseudomonas were reduced but filaments increased to 57% of bacteria (P < 0.001). This corresponded with analyses of root remnants showing they were heavily colonized by bacteria, with 48% filaments (P < 0.001) and 1.4%Pseudomonas (P = 0.014). Efforts to manage rhizosphere bacteria for sustainable agricultural systems should continue to focus on root cap and mucilage chemistry, and remnant roots as sources of beneficial bacteria.     

Effect of auxin and ethylene on elongation of intact primary roots of maize (Zeamays L.)

We tested that the hypothesis that root elongation might be controlled by altering the level of ethylene in intact primary roots of maize(Zea mays L.). We measured root elongation in a short period using a computerized root auxanometer. Compounds which regulate ethylene production were applied to intact primary roots in different time periods. Root elongation was stimulated by the treatment with ethylene antagonists such as Co²⁺, aminoethoxyvinylglycine (AVG) and L-canaline. This result suggested that root elongation was closely related to ethylene level of intact primary roots. Furthermore, IAA- and 1-aminocyclopropane-1-carboxylic acid (ACC)-induced inhibition of root elongation was reversed by treatment with Co²⁺. The application of ACC to roots which have been exposed to IAA and Co²⁺ have no significant effect on root elongation. However, the inhibition of root elongation by ACC in roots previously treated with IAA and AVG became manifest when the applied IAA concentrations were lower. These results were consistent with the hypothesis that the level of ethylene in intact roots functions to moderate root elongation, and suggested that auxin-induced inhibition of root elongation results from auxin induced promotion of ethylene production.