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The development and dynamics of the root apical meristem.

During the past ten years our concepts of the root apex and how it functions have expanded considerably. Through the use of several unique root tissue systems investigators have characterized the populations of cells which comprise the apex and have refined our views of the ways in which these populations interact to regulate aspects of root development. The aim of this paper is to present a summary of current thoughts on root development with emphasis on the activities of the cells comprising the root apex.

Allium↗

Expression of bone morphogenetic proteins and Msx genes during root formation.

Like crown development, root formation is also regulated by interactions between epithelial and mesenchymml tissues. Bone morphogenetic proteins (BMPs), together with the transcription factors Msx1 and Msx2, play important roles in these interactions during early tooth morphogenesis. To investigate the involvement of this signaling pathway in root development, we analyzed the expression patterns of Bmp2, Bmp3, Bmp4, and Bmp7 as well as Msx1 and Msx2 in the roots of mouse molars. Bmp4 was expressed in the apical mesenchyme and Msx2 in the root sheath. However, Bmps were not detected in the root sheath epithelium, and Msx transcripts were absent from the underlying mesenchyme. These findings indicate that this Bmp signaling pathway, required for tooth initiation, does not regulate root development, but we suggest that root shape may be regulated by a mechanism similar to that regulating crown shape in cap-stage tooth germs. Msx2 expression continued in the epithelial cell rests of Malassez, and the nearby cementoblasts intensely expressed Bmp3, which may regulate some functions of the fragmented epithelium.

Animals↗

Developing dorsal root ganglion neurons require trophic support from their central processes: evidence for a role of retrogradely transported nerve growth factor from the central nervous system to the periphery.

Injury to the peripheral processes produces a profound cell loss (40-50%) in the dorsal root ganglion of newborn rats. Although division of central processes produces little or no cellular change in sensory ganglion of adult animals, no information has been available on the effect of dorsal root section in developing dorsal root ganglion. We show that 6 days after dorsal rhizotomy on newborn rats, there is a 50% decrease in neuronal number in L5 dorsal root ganglion. A combined central and peripheral lesion of the sensory process results in a greater decrease in neuronal number (70%). Both of these effects can be prevented by the concomitant treatment with nerve growth factor. We also demonstrate that 125I-Ia-labeled nerve growth factor is retrogradely transported with high selectivity from the spinal cord to the dorsal root ganglion via the dorsal roots. The results indicate that trophic support for developing sensory neurons is provided through the central processes. This is presumably due to the uptake and retrograde transport of a trophic factor by the terminals of the central processes. The data suggest that nerve growth factor may be the trophic factor.

Animals↗

The circadian clock proteins PRR modulate root hair development via the RHD6/RSL module in Arabidopsis.

Root hairs, derived from trichoblasts, are critical for plant growth and environmental adaptation. Although environmental cues are known to influence root hair development, how endogenous timing systems such as the circadian clock integrate into the core transcriptional network governing root hair formation remains unclear. Here, we show that the circadian clock-associated protein PSEUDO-RESPONSE REGULATOR5 (PRR5) physically interacts with ROOT HAIR DEFECTIVE6 (RHD6) and RHD6 LIKE1 (RSL1), two basic helix-loop-helix transcription factors essential for root hair initiation. Genetic analyses suggest that PRR proteins contribute to root hair development under long-day conditions in Arabidopsis thaliana. Simultaneous disruption of PRR5, PRR7, and PRR9 results in defective root hairs, whereas PRR5 overexpression markedly increases root hair density and length. Transcriptomic and RT-qPCR analyses reveal that PRRs enhance the expression of RHD6, RSL1, and multiple downstream root hair-responsive genes, while modulating their temporal expression patterns. Furthermore, PRR5-mediated root hair promotion requires RHD6/RSL1, and PRR proteins enhance RHD6-dependent activation of the RSL4 promoter. PRRs also contribute to root hair development under phosphate-deficient and salt-stress conditions. Together, these findings establish a molecular framework in which PRR proteins regulate the RHD6/RSL network to coordinate root hair development and environmental responses.

Arabidopsis↗

Stimulation of carotenoid metabolism in arbuscular mycorrhizal roots.

Development of arbuscular mycorrhizal roots is correlated with accumulation of various isoprenoids, i.e. acyclic C(14) polyene 'mycorradicin' and C(13) cyclohexenone derivatives. We present data indicating a strong stimulation of carotenoid metabolism in such roots. Carotenoid profiling revealed mycorrhiza-specific accumulation of zeta-carotene in Zea mays and Medicago truncatula. Precursor accumulation after inhibition of phytoene desaturase (Pds) activity by norflurazon indicated an increased phytoene biosynthetic capacity in mycorrhizal roots of all species analyzed. Nicotiana tabacum plants transformed with a PDS promoter- GUS construct showed a cell-specific induction of PDS promoter activity in root cells containing arbuscules. Mycorradicin biosynthesis and, partially, mycorrhization were impaired in maize mutants deficient in carotenoid biosynthesis. These data indicate that (1) mycorradicin is probably synthesized via a C(40) precursor carotenoid, (2) carotenoid biosynthesis is induced in mycorrhizal roots, (3) induction occurs, at least partially, at the transcriptional level, and (4) that this may play a functional role during mycorrhization.

Carotenoids↗

Ethylene is a positive regulator of root hair development in Arabidopsis thaliana.

Evidence is provided that ethylene is a positive regulator of hair cell development in the root epidermis of Arabidopsis thaliana. Treatment of seedlings with increasing concentrations of the ethylene precursor, 1-aminocyclopropane-1-carboxylic acid (ACC) results in progressively more root hair cells developing in positions normally occupied by non-hair cells. Consistent with these findings are observations that treatments that block either ethylene synthesis or its perception reduce the number of root hairs. A model is proposed in which either ethylene or ACC is a signal involved in specifying the pattern of cell differentiation in the Arabidopsis root epidermis.

Arabidopsis↗

Inhibition of growth and development of root border cells in wheat by Al.

The production and development of border cells vary with genotype, and they are released in wheat at an earlier stage of root development than other species studied so far. No significant difference was observed in the maximum number of border cells between Al-tolerant (Atlas 66) and Al-sensitive (Scout 66) cultivars in the absence of Al treatment. Al seriously inhibited the production and release of border cells, resulting in clumping of border cells in Scout 66, but less clustering in Atlas 66. The number of border cells released from roots treated with Al is significantly less than that from roots grown without Al treatment. Al treatment induced the death of detached border cells in vitro and they were killed by a 20-h treatment with 25 micro m Al. No significant difference in survival percentage of detached border cells was observed between Atlas 66 and Scout 66, regardless of the presence or absence of Al. The removal of border cells from root tips of both Atlas 66 and Scout 66 enhanced the Al-induced inhibition of root elongation concomitant with increased Al accumulation in the root. These results suggest that border cells adhered to the root tips play a potential role in the protection of root from Al injury in wheat.

Journal Article↗

Radial hydraulic conductivity along developing onion roots.

Although most studies have shown that water uptake varies along the length of a developing root, there is no consistent correlation of this pattern with root anatomy. In the present study, water movement into three zones of onion roots was measured by a series of mini-potometers. Uptake was least in the youngest zone (mean hydraulic conductivity, Lpr = 1.5 x 10(-7) +/- 0.34 x 10(-7) m MPa-1 s-1; +/- SE, n = 10 roots) in which the endodermis had developed only Casparian bands and the exodermis was immature. Uptake was significantly greater in the middle zone (Lpr = 2.4 x 10(-7) +/- 0.43 x 10(-7) m MPa-1 s-1; +/- SE, n = 10 roots) which had a mature exodermis with both Casparian bands and suberin lamellae, and continued at this level in the oldest zone in which the endodermis had also developed suberin lamellae (Lpr = 2.8 x 10(-7) +/- 0.30 x 10(-7) m MPa-1 s-1; +/- SE, n = 10 roots). Measurements of the hydraulic conductivities of individual cells (Lp) in the outer cortex using a cell pressure probe indicated that this parameter was uniform in all three zones tested (Lp = 1.3 x 10(-6) +/- 0.01 x 10(-6) m MPa-1 s-1; +/- SE, n = 60 cells). Lp of the youngest zone was lowered by mercuric chloride treatment, indicating the involvement of mercury-sensitive water channels (aquaporins). Water flow in the older two root zones measured by mini-potometers was also inhibited by mercuric chloride, despite the demonstrated impermeability of their exodermal layers to this substance. Thus, water channels in the epidermis and/or exodermis of the older regions were especially significant for water flow. The results of this and previous studies are discussed in terms of two models. The first, which describes maize root with an immature exodermis, is the 'uniform resistance model' where hydraulic resistances are evenly distributed across the root cylinder. The second, which describes the onion root with a mature exodermis, is the 'non-uniform resistance model' where resistances can be variable and are concentrated in a certain layer(s) on the radial path.

Aquaporins↗

Genetic Control of Root Hair Development in Arabidopsis thaliana.

Visual examination of roots from 12,000 mutagenized Arabidopsis seedlings has led to the identification of more than 40 mutants impaired in root hair morphogenesis. Mutants from four phenotypic classes have been characterized in detail, and genetic tests show that these result from single nuclear recessive mutations in four different genes designated RHD1, RHD2, RHD3, and RHD4. The phenotypic analysis of the mutants and homozygous double mutants has led to a proposed model for root hair development and the stages at which the genes are normally required. The RHD1 gene product appears to be necessary for proper initiation of root hairs, whereas the RHD2, RHD3, and RHD4 gene products are required for normal hair elongation. These results demonstrate that root hair development in Arabidopsis is amenable to genetic dissection and should prove to be a useful model system to study the molecular mechanisms governing cell differentiation in plants.

Journal Article↗

The xipotl mutant of Arabidopsis reveals a critical role for phospholipid metabolism in root system development and epidermal cell integrity.

Phosphocholine (PCho) is an essential metabolite for plant development because it is the precursor for the biosynthesis of phosphatidylcholine, which is the major lipid component in plant cell membranes. The main step in PCho biosynthesis in Arabidopsis thaliana is the triple, sequential N-methylation of phosphoethanolamine, catalyzed by S-adenosyl-l-methionine:phosphoethanolamine N-methyltransferase (PEAMT). In screenings performed to isolate Arabidopsis mutants with altered root system architecture, a T-DNA mutagenized line showing remarkable alterations in root development was isolated. At the seedling stage, the mutant phenotype is characterized by a short primary root, a high number of lateral roots, and short epidermal cells with aberrant morphology. Genetic and biochemical characterization of this mutant showed that the T-DNA was inserted at the At3g18000 locus (XIPOTL1), which encodes PEAMT (XIPOTL1). Further analyses revealed that inhibition of PCho biosynthesis in xpl1 mutants not only alters several root developmental traits but also induces cell death in root epidermal cells. Epidermal cell death could be reversed by phosphatidic acid treatment. Taken together, our results suggest that molecules produced downstream of the PCho biosynthesis pathway play key roles in root development and act as signals for cell integrity.

Arabidopsis↗

Distribution and synthesis of type I and type III collagens in developing mouse molar tooth root.

The distribution and synthesis of type I and type III collagens in the mouse molar tooth root have been investigated by correlating light and electron immunohistochemical data. Purified rabbit antibodies were raised against mouse type I and type III collagens and indirect immunoperoxidase procedures were used. In these conditions, predentin, pre-bone, and pre-acellular cementum were intensely immunostained for type I collagen. Both optic and ultrastructural data confirmed the presence of type I collagen at the epithelio-mesenchymal junction, but Hertwig's basement membranes remained unlabelled. The odontoblasts including the short polarized ones, osteoblasts, some cells of pulp mesenchyme and the perifollicular cells possessed type I collagen immunoreactivity in the rough endoplasmic reticulum (RER), Golgi complex and the secretory vesicles. Type III collagen immunoreactivity was strong in the perifollicular mesenchyme, light in the pulp mesenchyme and absent from the epithelio-mesenchymal junction, the predentin, pre-bone and pre-acellular cementum. Intracellular immunolabelling was detected at the ultrastructural level in the perifollicular cells by a faint homogeneous peroxidase deposit in the RER cisternae. Finally, these results, compared with previous biochemical and morphological data, represent the first dynamic aspect of collagens distribution and synthesis in the mouse molar root development. In terms of cell differentiation, our data also suggest that type III collagen synthesis does not occur during the odontoblast process of differentiation.

Animals↗

Ethylene and phosphorus availability have interacting yet distinct effects on root hair development.

The hypothesis that ethylene participates in the regulation of root hair development by phosphorus availability in Arabidopsis thaliana was tested by chemically manipulating ethylene synthesis and response and with ethylene-insensitive mutants. Low phosphorus-induced root hair development could be mimicked by adding the ethylene precursor, 1-aminocyclopropane-1-carboxylate (ACC), to high phosphorus media, and inhibited by adding ethylene inhibitors to low phosphorus media. Ethylene-insensitive mutants showed a reduced response to low phosphorus, indicating ethylene involvement in root hair responses to phosphorus deficiency. To dissect the nature of this involvement, the morphological and anatomical changes associated with increased root hair density were investigated. Growth in low phosphorus resulted in smaller, more numerous cortical cells, resulting in a larger number of root hair-bearing epidermal cell files. Cortical cell number was not affected by ethylene inhibitors, ACC, or mutations reducing ethylene sensitivity in roots grown with low phosphorus, indicating that ethylene does not participate in this response. The exception was the eir1 mutation, which strongly reduced this change in radial anatomy, supporting a role for polar auxin transport in this process. Trichoblast cell length was reduced by low phosphorus availability in all genotypes, but even more so for ein2-1 and ein4. The proportion of epidermal cells forming hairs and root hair length were reduced in ethylene-insensitive mutants, especially in the presence of low phosphorus. These results demonstrate multiple effects of low phosphorus from the earliest stages of root hair development, and cross-talk between ethylene and phosphorus in the control of a subset of the low phosphorus effects, concentrating on those later in development.

Amino Acids, Cyclic↗

Expression of a Soybean Hydroxyproline-Rich Glycoprotein Gene Is Correlated with Maturation of Roots

A novel extensin gene has been identified in soybean (Glycine max L.) that encodes a hydroxyproline-rich glycoprotein (SbHRGP3) with two different domains. In this study expression of SbHRGP3 was investigated during soybean root development. SbHRGP was expressed in roots of mature plants, as well as seedlings, and showed a distinct pattern of expression during root development. The expression of SbHRGP3 increased gradually during root development of seedlings and reached a maximum while the secondary roots were maturing. The maximum expression level was contributed mainly by the secondary roots rather than by the primary root. Furthermore, expression of SbHRGP3 was preferentially detected in the regions undergoing maturation of the primary and secondary roots. These results imply that the expression of SbHRGP3 is regulated in an organ- and development-specific manner and that in soybean SbHRGP3 expression may be required for root maturation, presumably for the cessation of root elongation. Wounding and sucrose in combination enhanced expression of SbHRGP3 in roots, whereas both wounding and sucrose were required for the expression of SbHRGP3 in leaves.

Journal Article↗

Manipulation of root hair development and sorgoleone production in sorghum seedlings.

Sorghum (Sorghum bicolor) roots exude a potent bioherbicide sorgoleone. Previous work indicates that sorgoleone is produced in living root hairs. We have developed a mist system that resulted in abundant production of root hairs exuding sorgoleone and a mat system that significantly inhibited root hair development and consequently sorgoleone production. Applying Ag+ (an ethylene action inhibitor) at 1.2 mM to the seedlings grown in the mist system also inhibited root hair formation and elongation. Hypoxic conditions in the mist system did not result in the inhibition of root hair growth as compared to the standard air atmosphere (20.8% O2). Applying ethephon (an ethylene-releasing agent) at 0.031 mM to the roots of seedlings grown in the mat system with water running at 1 ml/min reversed the inhibition of root hair development by water movement. These results indicate that either water movement or ethylene can be utilized to manipulate root hair development and sorgoleone production in sorghum seedlings. It is hypothesized that water movement reduced the local ethylene concentration on the root surface and consequently inhibited root hair development of sorghum seedlings grown in the mat system.

Benzoquinones↗

Through form to function: root hair development and nutrient uptake.

Root hairs project from the surface of the root to aid nutrient and water uptake and to anchor the plant in the soil. Their formation involves the precise control of cell fate and localized cell growth. We are now beginning to unravel the complexities of the molecular interactions that underlie this developmental regulation. In addition, after years of speculation, nutrient transport by root hairs has been demonstrated clearly at the physiological and molecular level, with evidence for root hairs being intense sites of H(+)-ATPase activity and involved in the uptake of Ca(2+), K(+), NH(4)(+), NO(3)(-), Mn(2+), Zn(2+), Cl(-) and H(2)PO(4)(-).

Nitrogen↗

Control of root growth and development by cyclin expression.

Root development is plastic, with post-embryonic organogenesis being mediated by meristems. Although cell division is intrinsic to meristem initiation, maintenance and proliferative growth, the role of the cell cycle in regulating growth and development is unclear. To address this question, we examined the expression of cdc2 and cye genes, which encode the catalytic and regulatory subunits, respectively, of cyclin-dependent protein kinases that control progression through the cell cycle. Unlike cdc2, which is expressed not only in apical meristems but also before lateral root initiation in quiescent, pericycle cells arrested in the G2 phase of the cell cycle, cyc1At transcripts accumulate specifically in dividing cells immediately before cytokinesis. Ectopic expression of cyc1At under the control of the cdc2aAt promoter in Arabidopsis plants markedly accelerates growth without altering the pattern of lateral root development or inducing neoplasia. Thus cyclin expression is a limiting factor for growth, which in turn drives indeterminate development of the root system.

Arabidopsis↗

Dorsal root ganglia development in chicks following partial ablation of the neural crest.

To assess the effects of reduced competition for peripheral targets on developing brachial dorsal root ganglia (DRG), chick embryos were subjected to partial ablations of the brachial neural crest at stages 13 or 14 (Hamburger, V., and H.L. Hamilton (1951) J. Morphol. 88: 49-92), using an ophthalmological cauterization unit. In the initial studies reported here, ganglia developing from the remaining crest material were examined for ganglionic volume and neuronal size, neuronal number, and degenerative activity at stage 35. Results showed that the lesion procedure resulted in the reduction or absence of one or two ganglia on each side at the level of DRG 15 to DRG 17. Hypertrophies occurred in other ganglia remaining at these and at more rostral levels and ranged up to 220%. These hypertrophies were most pronounced, however, not in the ganglia adjacent to those lesioned but rather in more remote ganglia, including those at cervical levels. Accompanying these ganglionic changes were significant alterations in all three neuronal parameters examined. The findings clearly demonstrate a responsiveness of chick brachial DRG to reduced competition resulting from neural crest ablations and that such responsiveness occurs along several axial segments.

Animals↗

Histopathological evaluation of subcutaneous tissue reaction in mice to a calcium hydroxide paste developed for root canal fillings.

We aimed to evaluate the subcutaneous tissue reaction to a new calcium hydroxide paste for root canal fillings developed by Neo Dental Chemical Products Co., Tokyo, Japan. We injected calcium hydroxide paste and a water-based control subcutaneously into the dorsal area of 6-week-old anaesthetized male ddY mice. The tissue surrounding the injection sites was removed from mice in each group, 2 days, 1 week, 3 weeks and 12 weeks after the injection, and histopathologically examined. We found necrotic and degenerative changes, as well as foreign body reaction or proliferation of granulation tissue in specimens from both groups until 3 weeks after the injection. Some lymphocyte infiltration was observed in the 12-week specimens of the control group only. We conclude that calcium hydroxide paste is safe to use as a root canal filling material.

Animals↗