Differential growth resulting in the specification of different types of cellular architecture in root meristems

Symplastic growth of plant organs may be described by a continuous growth tensor field. In tensorial analysis of meristems, the trajectories of periclinal and anticlinal cell walls represent trajectories of the principal directions of growth (PDGs); this follows from the maintenance of mutual orthogonality between periclinal and anticlinal wall trajectories during growth. Periclinal and anticlinal cell divisions are also oriented in the principal planes of growth. The growth tensor for the root apex is specified in such a way that the principal directions of the tensor fit the pattern of periclinal and anticlinal walls in the apex, and that the grid formed by material particles aligned along PDG trajectories preserve this alignment during growth. Two growth tensors are formulated--one giving a maximum and the other giving a minimum of the volumetric relative elemental growth rate at the region of the initial cell(s). Temporal sequences of deformation of a grid formed by lines coinciding with the principal directions of growth are shown. The formation of cellular patterns in root apices is simulated. Two types of patterns are obtained: one with an apical cell and merophytes, and another with files of cells converging towards a quiescent centre.     

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.     

Clonal analysis of the Arabidopsis root confirms that position, not lineage, determines cell fate

 The cellular organization of the Arabidopsis thaliana (L.) Heynh. root meristem suggests that a regular pattern of cell divisions occurs in the root tip. Deviations from this pattern of division might be expected to disrupt the organization of cells and tissues in the root. A clonal analysis of the 3-d-old primary root meristem was carried out to determine if there is variability in division patterns, and if so to discover their effect on cellular organization in the root. Clones induced in the seedling meristem largely confirmed the predicted pattern of cell divisions. However, the cellular initials that normally give rise to the different cell files in the root were shown to exhibit some instability. For example, it was calculated that a lateral root cap/epidermal initial is displaced every 13 d. Furthermore, the existence of large marked clones that included more than two adjacent cell layers suggests that intrusive growth followed by cell division may occur at low frequency, perhaps in response to local cell deaths in the meristem. These findings support the view that even in plant organs with stereotypical cell division patterns, positional information is still the key determinant of cell fate. Received: 27 August 1999 / Accepted: 4 December 1999     

The simulation model of growth and cell divisions for the root apex with an apical cell in application to Azolla pinnata

In contrast to seed plants, the roots of most ferns have a single apical cell which is the ultimate source of all cells in the root. The apical cell has a tetrahedral shape and divides asymmetrically. The root cap derives from the distal division face, while merophytes derived from three proximal division faces contribute to the root proper. The merophytes are produced sequentially forming three sectors along a helix around the root axis. During development, they divide and differentiate in a predictable pattern. Such growth causes cell pattern of the root apex to be remarkably regular and self-perpetuating. The nature of this regularity remains unknown. This paper shows the 2D simulation model for growth of the root apex with the apical cell in application to Azolla pinnata. The field of growth rates of the organ, prescribed by the model, is of a tensor type (symplastic growth) and cells divide taking principal growth directions into account. The simulations show how the cell pattern in a longitudinal section of the apex develops in time. The virtual root apex grows realistically and its cell pattern is similar to that observed in anatomical sections. The simulations indicate that the cell pattern regularity results from cell divisions which are oriented with respect to principal growth directions. Such divisions are essential for maintenance of peri-anticlinal arrangement of cell walls and coordinated growth of merophytes during the development. The highly specific division program that takes place in merophytes prior to differentiation seems to be regulated at the cellular level.     

Cadmium stress suppresses lateral root formation by interfering with the root clock

A biological clock activated by oscillating signals, known as root clock, has been linked to lateral root (LR) formation and is essential for regular LR spacing along the primary root. However, it remains unclear how this internal mechanism is influenced by environmental factors known to affect the LR pattern. Here, we report that excessive cadmium (Cd) inhibits LR formation by disrupting the lateral root cap (LRC)‐programmed cell death (PCD)‐regulated root clock. Cd restricts the frequency of the oscillating signal rather than its amplitude. This could be attributed to the inhibition on meristematic activity by Cd, which resulted in decreased LRC cell number and LRC‐PCD frequency. Genetic evidence further showed that LRC cell number is positively correlated with root resistance to Cd. Our study reveals root cap dynamics as a novel mechanism mediating root responses to Cd, providing insight into the signalling pathways of the root clock responding to environmental cues.     

Developmental anatomy and auxin response of lateral root formation in Ceratopteris richardii

The homosporous fern Ceratopteris richardii exhibits a homorhizic root system where roots originate from the shoot system. These shoot-borne roots form lateral roots (LRs) that arise from the endodermis adjacent to the xylem poles, which is in contrast to flowering plants where LR formation arises from cell division in the pericycle. A detailed study of the fifth shoot-borne root showed that one lateral root mother cell (LRMC) develops in each two out of three successive merophytes. As a result, LRs emerge alternately in two ranks from opposite positions on a parent root. From LRMC initiation to LR emergence, three developmental stages were identified based on anatomical criteria. The addition of auxins (either indole-3-acetic acid or indole-3-butyric acid) to the growth media did not induce additional LR formation, but exogenous applications of both auxins inhibited parent root growth rate. Application of the polar auxin-transport inhibitor N-(1-naphthyl)phthalamic acid (NPA) also inhibited parent root growth without changing the LR initiation pattern. The results suggest that LR formation does not depend on root growth rate per se. The result that exogenous auxins do not promote LR formation in C. richardii is similar to reports for certain species of flowering plants, in which there is an acropetal LR population and the formation of the LRs is insensitive to the application of auxins. It also may indicate that different mechanisms control LR development in non-seed vascular plants compared with angiosperms, taking into consideration the long and independent evolutionary history of the two groups.     

Influence of a weak DC electric field on root meristem architecture

BACKGROUND AND AIMS: Electric fields are an important environmental factor that can influence the development of plants organs. Such a field can either inhibit or stimulate root growth, and may also affect the direction of growth. Many developmental processes directly or indirectly depend upon the activity of the root apical meristem (RAM). The aim of this work was to examine the effects of a weak electric field on the organization of the RAM. METHODS: Roots of Zea mays seedlings, grown in liquid medium, were exposed to DC electric fields of different strengths from 0.5 to 1.5 V cm(-1), with a frequency of 50 Hz, for 3 h. The roots were sampled for anatomical observation immediately after the treatment, and after 24 and 48 h of further undisturbed growth. KEY RESULTS: DC fields of 1 and 1.5 V cm(-1) resulted in noticeable changes in the cellular pattern of the RAM. The electric field activated the quiescent centre (QC): the cells of the QC penetrated the root cap junction, disturbing the organization of the closed meristem and changing it temporarily into the open type. CONCLUSIONS: Even a weak electric field disturbs the pattern of cell divisions in plant root meristem. This in turn changes the global organization of the RAM. A field of slightly higher strength also damages root cap initials, terminating their division.     

Repression of early lateral root initiation events by transient water deficit in barley and maize

The formation of lateral roots (LRs) is a key driver of root system architecture and developmental plasticity. The first stage of LR formation, which leads to the acquisition of founder cell identity in the pericycle, is the primary determinant of root branching patterns. The fact that initiation events occur asynchronously in a very small number of cells inside the parent root has been a major difficulty in the study of the molecular regulation of branching patterns. Inducible systems that trigger synchronous lateral formation at predictable sites have proven extremely valuable in Arabidopsis to decipher the first steps of LR formation. Here, we present a LR repression system for cereals that relies on a transient water-deficit treatment, which blocks LR initiation before the first formative divisions. Using a time-lapse approach, we analysed the dynamics of this repression along growing roots and were able to show that it targets a very narrow developmental window of the initiation process. Interestingly, the repression can be exploited to obtain negative control root samples where LR initiation is absent. This system could be instrumental in the analysis of the molecular basis of drought-responsive as well as intrinsic pathways of LR formation in cereals.     

Effect of mechanical stress on Zea root apex. I. Mechanical stress leads to the switch from closed to open meristem organization

The effect of mechanical stress on the root apical meristem (RAM) organization of Zea mays was investigated. In the experiment performed, root apices were grown through a narrowing of either circular (variant I) or elliptical (variant II) shape. This caused a mechanical impedance distributed circumferentially or from the opposite sides in variant I and II, respectively. The maximal force exerted by the growing root in response to the impedance reached the value of 0.15 N for variant I and 0.08 N for variant II. Significant morphological and anatomical changes were observed. The changes in morphology depended on the variant and concerned diminishing and/or deformation of the cross-section of the root apex, and buckling and swelling of the root. Anatomical changes, similar in both variants, concerned transformation of the meristem from closed to open, an increase in the number of the cell layers at the pole of the root proper, and atypical oblique divisions of the root cap cells. After leaving the narrowing, a return to both typical cellular organization and morphology of the apex was observed. The results are discussed in terms of three aspects: the morphological response, the RAM reorganization, and mechanical factors. Assuming that the orientation of division walls is affected by directional cues of a tensor nature, the changes mentioned may indicate that a pattern of such cues is modified when the root apex passes through the narrowing, but its primary mode is finally restored.     

Form matters: morphological aspects of lateral root development

Background

The crucial role of roots in plant nutrition, and consequently in plant productivity, is a strong motivation to study the growth and functioning of various aspects of the root system. Numerous studies on lateral roots, as a major determinant of the root system architecture, mostly focus on the physiological and molecular bases of developmental processes. Unfortunately, little attention is paid either to the morphological changes accompanying the formation of a lateral root or to morphological defects occurring in lateral root primordia. The latter are observed in some mutants and occasionally in wild-type plants, but may also result from application of external factors.

Scope and Conclusions

In this review various morphological aspects of lateral branching in roots are analysed. Morphological events occurring during the formation of a typical lateral root are described. This process involves dramatic changes in the geometry of the developing organ that at early stages are associated with oblique cell divisions, leading to breaking of the symmetry of the cell pattern. Several types of defects in the morphology of primordia are indicated and described. Computer simulations show that some of these defects may result from an unstable field of growth rates. Significant changes in both primary and lateral root morphology may also be a consequence of various mutations, some of which are auxin-related. Examples reported in the literature are considered. Finally, lateral root formation is discussed in terms of mechanics. In this approach the primordium is considered as a physical object undergoing deformation and is characterized by specific mechanical properties.     

Growth and cellular patterns in the petal epidermis of Antirrhinum majus: empirical studies

Background and Aims

Analysis of cellular patterns in plant organs provides information about the orientation of cell divisions and predominant growth directions. Such an approach was employed in the present study in order to characterize growth of the asymmetrical wild-type dorsal petal and the symmetrical dorsalized petal of the backpetals mutant in Antirrhinum majus. The aims were to determine how growth in an initially symmetrical petal primordium leads to the development of mature petals differing in their symmetry, and to determine how specific cellular patterns in the petal epidermis are formed.

Methods

Cellular patterns in the epidermis in both petal types over consecutive developmental stages were visualized and characterized quantitatively in terms of cell wall orientation and predominant types of four-cell packets. The data obtained were interpreted in terms of principal directions of growth (PDGs).

Key Results

Both petal types grew predominantly along the proximo-distal axis. Anticlinal cell walls in the epidermis exhibited a characteristic fountain-like pattern that was only slightly modified in time. New cell walls were mostly perpendicular to PDG trajectories, but this alignment could change with wall age.

Conclusions

The results indicate that the predominant orientation of cell division planes and the fountain-like cellular pattern observed in both petal types may be related to PDGs. The difference in symmetry between the two petal types arises because PDG trajectories in the field of growth rates (growth field) controlling petal growth undergo gradual redefinition. This redefinition probably takes place in both petal types but only in the wild-type does it eventually lead to asymmetry in the growth field. Two scenarios of how redefinition of PDGs may contribute to this asymmetry are considered.     

Cytokinin inhibits lateral root initiation but stimulates lateral root elongation in rice (Oryza sativa)

Research in lateral root (LR) development mainly focuses on the role of auxin. This article reports the effect of cytokinins (kinetin and trans-zeatin) on LR formation in rice (Oryza sativa L.). Our results showed that cytokinin has an inhibitory effect on LR initiation and stimulatory effect on LR elongation. Both KIN and ZEA at a concentration of 1 microM and above completely inhibited lateral root primordium (LRP) formation. The inhibitory effect of cytokinin on LR initiation required a continuous presence of KIN or ZEA in the growth solution. Cytokinin did not show any inhibitory effect on LR emergence from the seminal root once LRPs had been formed. The LRPs that developed in cytokinin-free solution can emerge normally in the solution containing inhibitory concentration (1 microM) of KIN and ZEA. The KIN and ZEA treatment dramatically stimulated LR elongation at all the concentrations tested. Maximum LR elongation was observed at a concentration of 0.01 microM KIN and 0.001 microM ZEA. The epidermal cell length increased significantly in LRs of cytokinin treated seedlings compared to those of untreated control. This result indicates that the stimulation of LR elongation by cytokinin is due to increased cell length. Exogenously applied auxin counteracted the effect of cytokinin on LR initiation and LR elongation, suggesting that cytokinin acts on LR elongation through an auxin dependent pathway.     

SCHIZORIZA controls an asymmetric cell division and restricts epidermal identity in the Arabidopsis root

The primary root of Arabidopsis has a simple cellular organisation. The fixed radial cell pattern results from stereotypical cell divisions that occur in the meristem. Here we describe the characterisation of schizoriza (scz), a mutant with defective radial patterning. In scz mutants, the subepidermal layer (ground tissue) develops root hairs. Root hairs normally only form on epidermal cells of wild-type plants. Moreover, extra periclinal divisions (new wall parallel to surface of the root) occur in the scz root resulting in the formation of supernumerary layers in the ground tissue. Both scarecrow (scr) and short root (shr) suppress the extra periclinal divisions characteristic of scz mutant roots. This results in the formation of a single layered ground tissue in the double mutants. Cells of this layer develop root hairs, indicating that mis-specification of the ground tissue in scz mutants is uncoupled to the cell division defect. This suggests that during the development of the ground tissue SCZ has two distinct roles: (1) it acts as a suppressor of epidermal fate in the ground tissue, and (2) it is required to repress periclinal divisions in the meristem. It may act in the same pathway as SCR and SHR.     

Tornado1 and tornado2 are required for the specification of radial and circumferential pattern in the Arabidopsis root

The cell layers of the Arabidopsis primary root are arranged in a simple radial pattern. The outermost layer is the lateral root cap and lies outside the epidermis that surrounds the ground tissue. The files of epidermal and lateral root cap cells converge on a ring of initials (lateral root cap/epidermis initial) from which the epidermal and lateral root cap tissues of the seedling are derived, once root growth is initiated after germination. Each initial gives rise to a clone of epidermal cells and a clone of lateral root cap cells. These initial divisions in the epidermal/lateral root cap initial are defective in tornado1 (trn1) and trn2 plants indicating a requirement for TRN1 and TRN2 for initial cell function. Furthermore, lateral root cap cells develop in the epidermal position in trn1 and trn2 roots indicating that TRN1 and TRN2 are required for the maintenance of the radial pattern of cell specification in the root. The death of these ectopic lateral root cap cells in the elongation zone (where lateral root cap cells normally die) results in the development of gaps in the epidermis. These observations indicate that TRN1 and TRN2 are required to maintain the distinction between the lateral root cap and epidermis and suggest that lateral root cap fate is the default state. It also suggests that TRN1 and TRN2 repress lateral root cap fate in cells in the epidermal location. Furthermore, the position-dependent pattern of root hair and non-root hair cell differentiation in the epidermis is defective in trn1 and trn2 mutants. Together these results indicate that TRN1 and TRN2 are required for the maintenance of both the radial pattern of tissue differentiation in the root and for the subsequent circumferential pattern within the epidermis.     

Abscisic acid promotes pre-emergence stages of lateral root development in Medicago truncatula

The plant root system is important for plant anchorage and nutrition. Among the different characteristics of the root system, root branching is a major factor of plasticity and adaptation to changing environments. Indeed, many biotic and abiotic stresses, such as drought or symbiotic interactions, influence root branching. Many studies concerning root development and root branching were performed on the model plant Arabidopsis thaliana, but this model plant has a very simplified root structure and is not able to establish any symbiotic interactions. We have recently described 7 stages for lateral root development in the model legume Medicago truncatula and found significant differences in the tissular contribution of root cell layers to the formation of new lateral roots (LR). We have also described 2 transgenic lines expressing the DR5:GUS and DR5:VENUS-N7 reporter genes that are useful to follow LR formation at early developmental stages. Here, we describe the use of these transgenic lines to monitor LR developmental responses of M. truncatula to the phytohormone abscisic acid (ABA) which is a major actor of stress and symbiotic interactions. We show that ABA promotes the formation of new lateral root primordia and their development, mostly at the late, pre-emergence stage.     

Lateral root initiation: one step at a time

Plant growth relies heavily on a root system that is hidden belowground, which develops post-embryonically through the formation of lateral roots. The de novo formation of lateral root organs requires tightly coordinated asymmetric cell division of a limited number of pericycle cells located at the xylem pole. This typically involves the formation of founder cells, followed by a number of cellular changes until the cells divide and give rise to two unequally sized daughter cells. Over the past few years, our knowledge of the regulatory mechanisms behind lateral root initiation has increased dramatically. Here, I will summarize these recent advances, focusing on the prominent role of auxin and cell cycle activity, and elaborating on the three key steps of pericycle cell priming, founder cell establishment and asymmetric cell division. Taken together, recent findings suggest a tentative model in which successive auxin response modules are crucial for lateral root initiation, and additional factors provide more layers of control.