Auxin action on proton influx in corn roots and its correlation with growth

At concentrations inhibitory to the elongation of corn (Zea mays L.) roots, the auxins, indole-3-acetic acid (IAA) and α-naphthaleneacetic acid (α-NAA), cause an increase in the pH of the bathing medium; this increase occurs with an average latent period shorter than the latent period for the inhibitory effect of these auxins on elongation. Indole-2-carboxylic acid, an inactive structural analogue of IAA, and β-naphthaleneacetic acid, an inactive analogue of α-NAA, affect neither growth nor the pH of the medium. Since acid pH is known to promote and basic pH to inhibit root elongation, the data are consistent with the hypothesis that hormone-induced modification of cell-wall pH plays a role in the control of elongation of roots, as has been proposed for elongation of stems and coleoptiles.     

Apical meristem organization and lack of establishment of the quiescent center in Cactaceae roots with determinate growth

Some species of Cactaceae from the Sonoran Desert are characterized by a determinate growth pattern of the primary root, which is important for rapid lateral-root formation and seedling establishment. An analysis of the determinate root growth can be helpful for understanding the mechanism of meristem maintenance in plants in general. Stenocereus gummosus (Engelm.) Gibson & Horak and Pachycereus pringlei (S. Watson) Britton & Rose are characterized by an open type of root apical meristem. Immunohistochemical analysis of 5-bromo-2-deoxyuridine incorporation into S. gummosus showed that the percentage of cells passing through the S-phase in a 24-h period is the same within the zone where a population of relatively slowly proliferating cells could be established and above this zone in the meristem. This indicated the absence of the quiescent center (QC) in S. gummosus. During the second and the third days of growth, in the distal meristem portion of P. pringlei roots, a compact group of cells that had a cell cycle longer than in the proximal meristem was found, indicating the presence of the QC. However, later in development, the QC could not be detected in this species. These data suggest that during post-germination the absence of the establishment of the QC within the apical meristem and limited proliferative activity of initial cells are the main components of a determinate developmental program and that establishment of the QC is required for maintenance of the meristem and indeterminate root growth in plants.Abbreviations QC quiescent center - RCP root cap-protoderm - BrdU 5-bromo-2-deoxyuridine - FITC fluorescein isothiocyanate - DAPI 4,6-diamidino-2-phenylindole     

Auxin binding in roots: A comparison between maize roots and coleoptiles

Auxin binding onto membrane fractions of primary roots of maize seedlings has been demonstrated using naphth-1yl-acetic acid (NAA) and indol-3yl-acetic acid (IAA) as ligands. This binding is compared with the already well characterized interaction between auxins and coleoptile membranes. The results indicate that while kinetic parameters are of the same order for root and coleoptile binding, a number of differences occur with respect to location in cells and relative affinity. The possible significance of the existence of such binding sites in root cells is discussed in relation to auxin action.Abbreviations 4-Cl-PA 4-chlorophenoxyacetic acid - EDTA ethylene diamine tetracetic acid - IAA indol-3yl-acetic acid - MCPA 2-methyl-4-chlorophenoxyacetic acid - NAA naphth-1yl-acetic acid - 2-NAA naphth-2yl-acetic acid - Tris 2-amino-2-(hydroxymethyl) propane-1,3 diol - TIBA 2,3,5 triiodobenzoic acid - NPA naphthylphthalamic acid - PCIB 4-chlorophenoxyisobutyric acid - PCPP 4-chlorophenoxyisopropionic acid - 2,4-D 2,4-dichlorophenoxyacetic acid     

Origin of the quiescent center in the root of Capsella bursa-pastoris (L.) Medik

The origin of the quiescent center in the embryonic radicle of Capsella bursa-pastoris was investigated by in-situ hybridization to cellular polyadenylic-acid-containing RNA using [³H]polyuridylic acid as a probe. In the globular embryo, autoradiographic silver grains were localized in all cells of the presumptive root apex except in the hypophysis. As the inner cell formed by a transverse division of the hypophysis cut off new cells toward the central procambial cylinder of the embryo, these cells remained characteristically unlabeled, in contrast to the labeled cells of the rest of the embryo. In the embryonic radicles of mature seeds and of seedlings, cells derived from the hypophysis appeared as a nonmeristematic, unlabeled, hemispherical group, bounded by the procambium to the inside and the root epidermis to the outside. When root tips excised from 2-d-old seedlings were incubated in [methyl-³H]thymidine, sectioned, and autoradiographed, cells derived from the inner cell of the hypophysis were found to be unlabeled, thus showing that they constitute the specific cells of the quiescent center. These results present evidence for the single-cell origin of the quiescent center in an angiosperm root and a role for the hypophysis in it.Abbreviations poly(A)⁺RNA polyadenylicacid-containing RNA - [³H]poly(U) [³H]polyuridylic acid - QC quiescent center This work was supported in part by National Science Foundation grants PCM-7902898 and DCB-8709092.     

Rapid-growth responses of corn root segments: Effect of auxin on elongation

The effect of indole-3-acetic acid (IAA) on the elongation rates of 2 mm corn (Zea mays L.) root segments induced by citrate-phosphate buffer (or unbuffered) solutions of pH 4.0 and 7.0 was studied. At pH 7.0, auxin initially reduced the elongation rate in both buffered and unbuffered solutions. Only in buffer at pH 7.0 was auxin at a concentration of 0.1 M found to promote the elongation rate though briefly. THis promoted rate represented only ca. 20% of the rate achieved with only buffer at pH 4.0. Auxin in pH 4.0 buffered and unbuffered solutions only served to reduce the elongation rates of root segments. Some comparative experiments were done using 2 mm corn coleoptile segments. Auxin (pH 6.8) promoted the elongation rate of coleoptile segments to a level equal or greater than the maximal H ion-induced rate. The two responses of root segments to auxin are compared to auxin action in coleoptile growth.     

Principal directions of growth and the generation of cell patterns in wild-type and gib-1 mutant roots of tomato (Lycopersicon esculentum Mill.) grown in vitro

The patterns of cell growth and division characteristic of the apex of tomato roots grown in vitro were simulated by computer using a growth tensor (GT). The GT was used to clarify the basis of the altered cell patterns found within apices of roots whose gibberellin levels had been depressed by mutation (at the GIB-1 locus) or through application of the gibberellin-biosynthesis inhibitor, 2S,3S paclobutrazol. At the pole of wild-type roots, where the cell files of the cortex converge, there are commonly only one or two tiers of cortical cells sandwiched between the pole of the stele and the cap initials. By contrast, root apices of the gib-1 mutant contain additional tiers in this region. The development of these additional tiers is suppressed when roots of the mutant are grown in the presence of gibberellic acid (GA₃), but could be induced in wild-type roots when they are grown in 2S,3S paclobutrazol. The wild-type cell pattern can be simulated using the GT and by the application of appropriate rules that govern cell growth and division. The induced variations in cell pattern are interpreted as being due to displacements, within the apex, of the principal directions of growth (PDGs), which are represented, in part, by the set of periclines and anticlines seen in the cell wall network; these, in turn, are utilized in the specification of the GT. During normal (wild-type) root growth, the PDGs maintain a stable pattern and the corresponding cell pattern is also stable. However, in order to interpret the cellular behaviour found in wild-type roots grown in 2S, 3S paclobutrazol, simulation using the GT shows that, if the pattern of PDGs is destabilized and displaced distally along the root axis, the cell pattern reorganizes into that typical of gib-1 mutant roots. Conversely, the cell pattern of gib-1 roots, which reverts to wild-type upon exposure to GA₃, can be simulated if the PDGs are displaced proximally to the inside of the apex whereupon the number of cortical tiers at the root pole decreases. These results suggest a link between endogenous gibberellin level and the specification of the PDGs in the growing tomato root apex. Furthermore, the evidence of cell patterns from gib-1 roots suggests that, in order to achieve stability of PDGs with concomitant stable cellular patterning, an optimal gibberellin level is necessary. In practice, this can be attained by culturing the mutant roots in medium containing 1 M GA₃.Abbreviations GA₃ gibberellic acid - GT growth tensor - NCS natural coordinate system - PDG principal direction of growth - QC quiescent centre - RERG relative elemental rate of growth We are grateful to the former Agricultural and Food Research Council for financial support under the International Scientific Interchange Scheme to enable J.N. to work at Long Ashton Research Station, and to K. Kurczyski (Silesian University, Katowice, Poland) for help in writing a computer program for cell proliferation. Preparation of the model for growth and division was supported in part by a grant from the Committee for Scientific Research, Poland.     

Acid growth effects in maize roots: Evidence for a link between auxin-economy and proton extrusion in the control of root growth

The role of proton excretion in the growth of apical segments of maize roots has been examined. Growth is stimulated by acidic buffers and inhibited by neutral buffers. Organic buffers such as 2[N-morpholino] ethane sulphonic acid (MES) — 2-amino-2-(hydroxymethyl)propane-1,3 diol (Tris) are more effective than phosphate buffers in inhibiting growth. Fusicoccin(FC)-induced growth is also inhibited by neutral buffers. The antiauxins 4-chlorophenoxyisobutyric acid (PCIB) and 2-(naphthylmethylthio) propionic acid (NMSP) promote growth and H⁺-excretion over short time periods; this growth is also inhibited by neutral buffers. We conclude that growth of maize roots requires proton extrusion and that regulation of root growth by indol-3yl-acetic acid (IAA) may be mediated by control of this proton extrusion.Abbreviations IAA indol-3yl-acetic acid - ABA abscisic acid - FC fusicoccin - PCIB 4-chlorophenoxy-isobutyric acid - MES 2(N-morpholino)ethane sulphonic acid - Tris 2-amino-2-(hydroxymethyl) propane-1,3-diol - NMSP 2-(naphthylmethylthio)propionic acid     

The quiescent center in roots of maize: initiation,maintenance and role in organization of the root apical meristem

Summary Using roots of maize, we tested the hypothesis that the origin and maintenance of the quiescent center (QC) are a consequence of polar auxin supply. Exposing roots to the polar auxin transport inhibitor 2,3,5-triiodobenzoic acid (TIBA), or to low temperature (4 °C, with subsequent return to 24 °C), enhances mitotic frequency within the QC. In both treatments, the QC most typically is activated at its distal face, and the protoderm/dermatogen undergoes several periclinal divisions. As a result, the root body penetrates and ruptures the root cap junction and the characteristic closed apical organization changes to open. A QC persists during these changes in apical organization, but it is diminished in size. The data from the TIBA-treated roots suggest a role for auxin in the origin and maintenance of the QC, and further, that alterations in QC dimensions are a consequence of polar auxin supply. We hypothesize that the root cap, and specifically the root cap initials, are important in regulating polar auxin movements towards the root apex, and hence are important in determining the status of the QC.Abbreviations QC quiescent center - TIBA 2,3,5-triiodobenzoic acid Dedicated to the memory of Professor John G. Torrey     

Latent nitrate reductase activity is associated with the plasma membrane of corn roots

Latent nitrate reductase activity (NRA) was detected in corn (Zea mays L., Golden Jubilee) root microsome fractions. Microsome-associated NRA was stimulated up to 20-fold by Triton X-100 (octylphenoxy polyethoxyethanol) whereas soluble NRA was only increased up to 1.2-fold. Microsome-associated NRA represented up to 19% of the total root NRA. Analysis of microsomal fractions by aqueous two-phase partitioning showed that the membrane-associated NRA was localized in the second upper phase (U₂). Analysis with marker enzymes indicated that the U₂ fraction was plasma membrane (PM). The PM-associated NRA was not removed by washing vesicles with up to 1.0 M NACl but was solubilized from the PM with 0.05% Triton X-100. In contrast, vanadate-sensitive ATPase activity was not solubilized from the PM by treatment with 0.1% Triton X-100. The results show that a protein capable of reducing nitrate is embedded in the hydrophobic region of the PM of corn roots.Abbreviations L₁ first lower phase - NR nitrate reductase - NRA nitrate-reductase activity - PM plasma membrane - T:p Triton X-100 (octylphenoxy polyethoxyethanol) to protein ratio - U₂ second upper phase     

Enzymes of asparagine synthesis in maize roots

An asparagine synthetase which is active with either glutamine or NH ₄ ⁺ has been found in maize (Zea mays L.) roots. Unlike the enzyme obtained from legume cotyledons, the maize-root enzyme is only slightly more efficient with glutamine (Km, 1.0 mM) than with NH ₄ ⁺ (Km, 2.0–3.0 mM). The activity of this enzyme is higher in the mature root than in the root-tip region, i.e. root cells develop a capacity to make asparagine from glutamine or NH ₄ ⁺ as they mature. -Cyanoalanine synthetase is also present in maize roots. The apparent Km for cysteine is 2.6 mM and for cyanide is 0.57 mM. The enzyme is more active in the root tip than in mature root tissue. Thus, if asparagine were made in the root tip, the cyanide pathway could represent the mechanism of synthesis. It is our contention, however, that this potential is not realized under normal conditions because ¹⁴C-experiments performed previously have indicated a limited availability of both CN and cysteine in the maize root.     

Gradients in maize roots: local elongation and pH

The elongation rate, the gradient of the local elongation rate and the surface pH of maize roots were measured over 12 h. A data bank was constituted by storing these values. By sorting these results on the basis of different elongation rates, different classes of root were obtained. Two classes were chosen: the low-growth roots and the high-growth roots. The mean growth of these two root classes was stable with time and differed significantly from one another. The surface pH of the elongation zone was the same for the roots of these two classes, but the roots selected for their higher growth rate had a larger acid efflux in this zone.     

The influence of mechanical resistance and phosphate supply on morphology and function of corn roots

Conclusions The influence of mechanical soil resistance on shoot growth can be explained qualitatively by its influence on root morphology. When relating ion uptake quantitatively to a certain root parameter (length, surface area, weight), availability of nutrients in the different soil layers must be taken into consideration.     

A negative image of the quiescent centre in regenerating root apices of Zea mays

Usually the presence of the quiescent centre in roots is demonstrated by the absence of labelled nuclei following treatment of the root with appropriate radioactive markers. By modification of the pulselabelling technique, a negative image of the quiescent center, showing more intense labelling from [³H]thymidine than the surrounding area, was obtained in regenerating root apices of Zea mays L.     

Spectral dependence of the light-induced geotropic response in Zea roots

The induction by light of geotropic responsiveness in the primary roots of Zea mays L. (cv. Golden Cross Bantam 70) was found to be governed by the all-or-none law. The response was induced by light energies above a threshold value, but the maximal curvature of geo-stimulated roots was constand irrespective of the light energy above that threshold. The action spectrum for this light effect showed a large peak at 650, a small peak at 410, and a shoulder at 663 nm. The effect of red light was not reversed by far-red light. Thus, the geotropic response in Zea roots may not be controlled by phytochrome.     

Modeling the formation of root apices

The formation of new root apices from small groups of cells with different cellular patterns has been simulated using an existing model based on growth tensors. To generate an apex, a steady growth field was used. The pattern of cells evolved to approach the steady state. Two extreme types of progressions have been obtained : one leading to an apex with a single or a few apical cells, and the other to an apex with a quiescent centre. The change of structure while applying a steady growth tensor indicates that development may involve a succession of discrete growth tensors.     

Suppression of gravitropic response of primary roots by submergence

Primary roots of six plant species were placed horizontally either in humid air or under water, and their growth and gravitropic responses were examined. In air, all the roots showed a normal gravitropic curvature. Under water without aeration, roots of rice (Oryza sativa L.), oat (Avena sativa L.), azuki bean (Vigna angularis Ohwi et Ohashi), and cress (Lepidium sativum L.) curved downward at almost same rate as in air, whereas the curvature of roots of maize (Zea mays L.) and pea (Pisum sativum L.) was strongly suppressed. Submergence did not cause a decrease in growth rate of these roots. When roots of maize and pea were placed horizontally under water without aeration and then rotated in three dimensions on a clinostat in air, they showed a significant curvature, suggesting that the step suppressed by submergence is not graviperception but the subsequent signal transmission or differential growth process. Constant bubbling of air through the water partly restored the gravitropic curvature of maize roots and completely restored that of pea roots. The curvature of pea roots was also partly restored by the addition of an inhibitor of ethylene biosynthesis, aminooxyacetic acid. In air, ethylene suppressed the gravitropic curvature of roots of maize and pea. Furthermore, the level of ethylene in the intercellular space of the roots was increased by submergence. These results suggest that the accumulation of ethylene in the tissue is at least partly involved in suppression of transmission of the gravity signal or of differential growth in maize and pea roots under conditions of submergence.Abbreviations AOA aminooxyacetic acid - 3-D three-dimensional Dedicated to Professor Andreas Sievers on the occasion of his retirementWe thank Professor H. Suge and Drs. H. Takahashi and H. Kataoka, Tohoku University and Dr. T. Suzuki, Yamagata University, for helpful suggestions. The present study was supported in part by a Grant for Basic Research in Space Station Utilization from the Institute of Space and Astronautical Science, Japan.     

Water stress and indol-3yl-acetic acid content of maize roots

Water-stress conditions were applied to the apical 12 mm of intact or excised roots ofZea mays L. (cv. LG 11) using mannitol solutions (0 to 0.66 M) and changes in weight, water content, growth and IAA level of these roots were investigated. With increasing stress a decrease in growth, correlated with an increased IAA level, was observed. The largest increase in IAA (about 2.7-fold) was found in the apical 5 mm of the root and was obtained under a stress corresponding to an osmotic potential of −1.39 MPa in the solution. This stress led to an isotonic state in the cells after 1 h. When the duration of water stress (−1.09 MPa) was increased to 2 or 3 h, no further increase in the IAA content was observed in the root segments. This indicated that there was no correlation between a hypothetical passive penetration of mannitol in the cells and IAA content. Indol-3yl-acetic acid rose to the same level in excised as in intact roots. In both cases, IAA accumulation was apparently independent of the hydrolysis of the conjugated form. The caryopsis and shoot seem not to be necessary to induce the increase of the IAA level in the roots during water stress (−1.09 MPa). Therefore, there seems to be a high rate of IAA biosynthesis in excised maize roots under water-stress conditions. Exodiffusion of IAA was observed during an immersion in either buffer or stress (−1.09 MPa) solution. In both cases, this IAA efflux into the medium represented about 50% of the endogenous level. Considering the present results, IAA appears to play an important part in the regulation of maize root metabolism and growth under water deficiency.     

Differential growth of georeacting maize roots

The growth rate of the two sides of 10-mm apical segments prepared from primary roots and of intact primary roots of maize has been analyzed in both vertical and horizontal positions, using a filming method allowing continuous growth recording. The data showed that the georeaction began by a decrease in the overall elongation rate of the roots. This inhibition is effective on the lower side of the bending zone, where the growth is practically stopped during the period of maximum rate of geocurvature. In contrast, the growth is slightly enhanced on the upper part of the elongating zone.     

Root growth regulation and gravitropism in maize roots does not require the epidermis

We have earlier published observations showing that endogenous alterations in growth rate during gravitropism in maize roots (Zea mays L.) are unaffected by the orientation of cuts which remove epidermal and cortical tissue in the growing zone (Björkman and Cleland, 1988, Planta 176, 513–518). We concluded that the epidermis and cortex are not essential for transporting a growth-regulating signal in gravitropism or straight growth, nor for regulating the rate of tissue expansion. This conclusion has been challenged by Yang et al. (1990, Planta 180, 530–536), who contend that a shallow girdle around the entire perimeter of the root blocks gravitropic curvature and that this inhibition is the result of a requirement for epidermal cells to transport the growth-regulating signal. In this paper we demonstrate that the entire epidermis can be removed without blocking gravitropic curvature and show that the position of narrow girdles does not affect the location of curvature. We therefore conclude that the epidermis is not required for transport of a growth-regulating substance from the root cap to the growing zone, nor does it regulate the growth rate of the elongating zone of roots.