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PubMed2005

Sites and regulation of auxin biosynthesis in Arabidopsis roots.

Auxin has been shown to be important for many aspects of root development, including initiation and emergence of lateral roots, patterning of the root apical meristem, gravitropism, and root elongation. Auxin biosynthesis occurs in both aerial portions of the plant and in roots; thus, the auxin required for root development could come from either source, or both. To monitor putative internal sites of auxin synthesis in the root, a method for measuring indole-3-acetic acid (IAA) biosynthesis with tissue resolution was developed. We monitored IAA synthesis in 0.5- to 2-mm sections of Arabidopsis thaliana roots and were able to identify an important auxin source in the meristematic region of the primary root tip as well as in the tips of emerged lateral roots. Lower but significant synthesis capacity was observed in tissues upward from the tip, showing that the root contains multiple auxin sources. Root-localized IAA synthesis was diminished in a cyp79B2 cyp79B3 double knockout, suggesting an important role for Trp-dependent IAA synthesis pathways in the root. We present a model for how the primary root is supplied with auxin during early seedling development.

Arabidopsis↗

PubMed1997

Cytoplasmic free calcium distributions during the development of root hairs of Arabidopsis thaliana.

In this study, confocal ratio analysis was used to image the relationship between cytoplasmic free calcium concentration ([Ca2+]c) and the development of root hairs of Arabidopsis thaliana. Although a localized change in [Ca2+]c that preceded or predicted the site of root hair initiation could not be detected, once initiated the majority of emerging root hairs showed an elevated [Ca2+]c (> 1 microM) in their apical cytoplasm, compared with 100-200 nM in the rest of the cell. These emerging root hairs then moved into a 3-5 h phase of sustained elongation during which they showed variable growth rates. Root hairs that were rapidly elongating exhibited a highly localized, elevated [Ca2+]c at the tip. Non-growing root hairs did not exhibit the [Ca2+]c gradient. The rhd-2 mutant, which is defective in sustained root hair growth, showed an altered [Ca2+]c distribution compared with wild-type. These results implicate [Ca2+]c in regulating the tip growth process. Treatment of elongating wild-type root hairs with the Ca2+ channel blocker verapamil (50 microM) caused dissipation of the elevated [Ca2+]c at the tip and cessation of growth, suggesting a requirement for Ca2+ channel activity at the root hair tip to maintain growth. Manganese treatment also preferentially quenched Indo-1 fluorescence in the apical cytoplasm of the root hair. As manganese is thought to enter cells through Ca(2+)-permeable channels, this result also suggests increased Ca2+ channel activity at the tip of the growing hair. Taken together, these data suggest that although Ca2+ does not trigger the initiation of root hairs, Ca2+ influx at the tip of the root hair leads to an elevated [Ca2+]c that may be required to sustain root hair elongation.

Arabidopsis↗

PubMed1985

Expression of plant genes during the development of pea root nodules.

The expression of plant genes involved in the pea-Rhizobium symbiosis was studied by analysing mRNA from root nodules. The RNA was translated in vitro and the translation products were separated by two-dimensional gel electrophoresis. The results show differential expression of nodulin genes during root nodule development. One gene encoding N-40' is expressed at a significant level 5 days before the leghemoglobin genes. Most other nodulin genes are expressed more of less concomitantly with the leghemoglobin genes whereas the N-21 mRNA is only present late during the development. In the development of ineffective root nodules induced by infection with different nod+fix- mutants of R. leguminosarum all nodulin genes are expressed except for the N-21 gene. The results suggest that neither bacteroid development, heme excretion nor nitrogen fixation are essential for the induction of nodulin gene expression in the host plant. Further, it appears that the amount of leghemoglobin in ineffective nodules is regulated at a post-transcriptional level.

Electrophoresis, Gel, Two-Dimensional↗

PubMed2003

An investigation of root-fractured permanent incisor teeth in children.

- The aim of this retrospective study was to determine the type of healing which occurred in root-fractured permanent incisor teeth in children. The objectives were to determine whether gender, age, stage of root development or location of the fracture affected the healing type. The method involved careful scrutiny of clinical records and radiographs of children who attended a unit of paediatric dentistry in a dental hospital. Relevant information was entered onto a data collection sheet. The results were tabulated and analysed by the chi2-tests using the SPSS statistical package. The results are based on 34 root-fractured teeth in 33 children aged 8-15 years. Root development was incomplete in 27 of the root-fractured teeth and complete in seven teeth. A good healing outcome was seen in 27 (79.4%) of the teeth and poor healing in 7 (20.6%). The only factor which was found to be statistically significantly related to healing was the stage of root development. It can be concluded that root-fractured teeth with immature roots have a better chance of showing good healing than teeth with mature roots.

Adolescent↗

PubMed2004

Green fluorescent protein fusions to Arabidopsis fimbrin 1 for spatio-temporal imaging of F-actin dynamics in roots.

The visualization of green fluorescent protein (GFP) fusions with microtubule or actin filament (F-actin) binding proteins has provided new insights into the function of the cytoskeleton during plant development. For studies on actin, GFP fusions to talin have been the most generally used reporters. Although GFP-Talin has allowed in vivo F-actin imaging in a variety of plant cells, its utility in monitoring F-actin in stably transformed plants is limited particularly in developing roots where interesting actin dependent cell processes are occurring. In this study, we created a variety of GFP fusions to Arabidopsis Fimbrin 1 (AtFim1) to explore their utility for in vivo F-actin imaging in root cells and to better understand the actin binding properties of AtFim1 in living plant cells. Translational fusions of GFP to full-length AtFim1 or to some truncated variants of AtFim1 showed filamentous labeling in transient expression assays. One truncated fimbrin-GFP fusion was capable of labeling distinct filaments in stably transformed Arabidopsis roots. The filaments decorated by this construct were highly dynamic in growing root hairs and elongating root cells and were sensitive to actin disrupting drugs. Therefore, the fimbrin-GFP reporters we describe in this study provide additional tools for studying the actin cytoskeleton during root cell development. Moreover, the localization of AtFim1-GFP offers insights into the regulation of actin organization in developing roots by this class of actin cross-linking proteins.

Actins↗

PubMed2003

The Sym35 gene required for root nodule development in pea is an ortholog of Nin from Lotus japonicus.

Comparative phenotypic analysis of pea (Pisum sativum) sym35 mutants and Lotus japonicus nin mutants suggested a similar function for the PsSym35 and LjNin genes in early stages of root nodule formation. Both the pea and L. japonicus mutants are non-nodulating but normal in their arbuscular mycorrhizal association. Both are characterized by excessive root hair curling in response to the bacterial microsymbiont, lack of infection thread initiation, and absence of cortical cell divisions. To investigate the molecular basis for the similarity, we cloned and sequenced the PsNin gene, taking advantage of sequence information from the previously cloned LjNin gene. An RFLP analysis on recombinant inbred lines mapped PsNin to the same chromosome arm as the PsSym35 locus and direct evidence demonstrating that PsNin is the PsSym35 gene was subsequently obtained by cosegregation analysis and sequencing of three independent Pssym35 mutant alleles. L. japonicus and pea root nodules develop through different organogenic pathways, so it was of interest to compare the expression of the two orthologous genes during nodule formation. Overall, a similar developmental regulation of the PsNin and LjNin genes was shown by the transcriptional activation in root nodules of L. japonicus and pea. In the indeterminate pea nodules, PsNin is highly expressed in the meristematic cells of zone I and in the cells of infection zone II, corroborating expression of LjNin in determinate nodule primordia. At the protein level, seven domains, including the putative DNA binding/dimerization RWP-RK motif and the PB1 heterodimerization domain, are conserved between the LjNIN and PsNIN proteins.

Amino Acid Sequence↗

PubMed1995

Replantation of 400 avulsed permanent incisors. 4. Factors related to periodontal ligament healing.

400 avulsed and replanted permanent teeth were examined for periodontal ligament (PDL) healing, using standardized radiographic and clinical examination procedures (i.e. percussion test and mobility test). The effect of various clinical factors was examined, such as age and sex of the patient, type of tooth replanted, presence of crown fracture or bone fracture, stage of root development (including apical diameter and length of the pulp), type and length of extra-alveolar storage, clinical contamination of the root surface, type of root surface cleansing procedure, type and length of splinting period and antibiotic therapy. Surface resorption was generally diagnosed after 12 months; while inflammatory resorption and replacement resorption (ankylosis) were usually observed after 1 month and 1-2 months respectively. Most resorptive processes were diagnosed within the first 2-3 years. However, although rarely, even after 5 and 10 years new resorptive processes could be diagnosed. A univariate statistical analysis of 272 teeth revealed 9 factors significantly related to PDL healing. A subsequent multivariate analysis revealed that the following 4 factors had the strongest impact upon PDL healing, in descending order of significance: Stage of root development; length of the dry extra-alveolar storage period; immediate replantation and length of the wet period (saliva or saline storage). Nonphysiological storage, such as homemade saline and sterilizing solutions (chloramine and alcohol) always led to root resorption. Storage in tap water for more than 20 minutes usually led to root resorption. The common denominator for all these factors related to PDL healing appears to be survival of the PDL cells along the root surface. Based on these findings, immediate replantation is recommended irrespective of stage of root development.

Analysis of Variance↗

PubMed2006

RPA, a class II ARFGAP protein, activates ARF1 and U5 and plays a role in root hair development in Arabidopsis.

The polar growth of plant cells depends on the secretion of a large amount of membrane and cell wall materials at the growing tip to sustain rapid growth. Small GTP-binding proteins, such as Rho-related GTPases from plants and ADP-ribosylation factors (ARFs), have been shown to play important roles in polar growth via regulating intracellular membrane trafficking. To investigate the role of membrane trafficking in plant development, a Dissociation insertion line that disrupted a putative ARF GTPase-activating protein (ARFGAP) gene, AT2G35210, was identified in Arabidopsis (Arabidopsis thaliana). Phenotypic analysis showed that the mutant seedlings developed isotropically expanded, short, and branched root hairs. Pollen germination in vitro indicated that the pollen tube growth rate was slightly affected in the mutant. AT2G35210 is specifically expressed in roots, pollen grains, and pollen tubes; therefore, it is designated as ROOT AND POLLEN ARFGAP (RPA). RPA encodes a protein with an N-terminal ARFGAP domain. Subcellular localization experiments showed that RPA is localized at the Golgi complexes via its 79 C-terminal amino acids. We further showed that RPA possesses ARF GTPase-activating activity and specifically activates Arabidopsis ARF1 and ARF1-like protein U5 in vitro. Furthermore, RPA complemented Saccharomyces cerevisiae glo3Delta gcs1Delta double mutant, which suggested that RPA functions as an ARFGAP during vesicle transport between the Golgi and the endoplasmic reticulum. Together, we demonstrated that RPA plays a role in root hair and pollen tube growth, most likely through the regulation of Arabidopsis ARF1 and ARF1-like protein U5 activity.

ADP-Ribosylation Factor 1↗

PubMed1991

Discovery of a rhizobial RNA that is essential for symbiotic root nodule development.

All of the Azorhizobium, Bradyrhizobium, and Rhizobium genes known to be involved in the development of nitrogen-fixing legume root nodules are genes that code for proteins. Here we report the first exception to this rule: the sra gene; it was discovered during the genetic analysis of a Bradyrhizobium japonicum Tn5 mutant (strain 259) which had a severe deficiency in colonizing soybean nodules. A DNA region as small as 0.56 kb cloned from the parental wild type restored a wild-type phenotype in strain 259 by genetic complementation. The sra gene was located on this fragment, sequenced, and shown to be transcribed into a 213-nucleotide RNA. Results obtained with critical point mutations in the sra gene proved that the transcript was not translated into protein; rather, it appeared to function as an RNA molecule with a certain stem-and-loop secondary structure. We also detected an sra homolog in Rhizobium meliloti which, when cloned and transferred to B. japonicum mutant 259, fully restored symbiotic effectiveness in that strain. We propose several alternative functions for the sra gene product, of which that as a regulatory RNA for gene expression may be the most probable one.

Amino Acid Sequence↗

PubMed1981

Relationship between cell damage in the periodontal ligament after replantation and subsequent development of root resorption. A time-related study in monkeys.

The etiology of root resorption subsequent to replantation of incisors was examined in green Vervet monkeys. Cell damage to the root surface due to the extraction procedures was registered histologically 1 week after replantation. The topographical distribution of cell damage was then related to the development of root resorption in similar replanted teeth with longer observation periods. Histometric analysis showed that surface-, inflammatory- and replacement resorption was significantly related to certain topographical locations on the root surface. These surfaces represented the "corner" surfaces of the root, where the maximum damage presumably would occur during the extraction procedure. A positive and highly significant correlation between cell damage in the cementoblast layer and the presence of surface-, inflammatory- and replacement resorption was found: replacement resorption was associated with the greatest loss of vital cementoblasts per unit root surface length. Furthermore, the distance from a potential resorption site to the nearest location on the root surface with a normal number of cementoblasts was found to be related to the type of root resorption. Thus, replacement resorption was found to be significantly related to the greatest distance from a site with normal numbers of cementoblasts compared to sites with no resorption or inflammatory resorption. Based on these findings, as well as previous experiments, a theory is presented for the etiology and pathogenesis of external root resorption after immediate replantation of mature teeth.

Animals↗

PubMed2003

The role of nutrient availability in regulating root architecture.

The ability of plants to respond appropriately to nutrient availability is of fundamental importance for their adaptation to the environment. Nutrients such as nitrate, phosphate, sulfate and iron act as signals that can be perceived. These signals trigger molecular mechanisms that modify cell division and cell differentiation processes within the root and have a profound impact on root system architecture. Important developmental processes, such as root-hair formation, primary root growth and lateral root formation, are particularly sensitive to changes in the internal and external concentration of nutrients. The responses of root architecture to nutrients can be modified by plant growth regulators, such as auxins, cytokinins and ethylene, suggesting that the nutritional control of root development may be mediated by changes in hormone synthesis, transport or sensitivity. Recent information points to the existence of nutrient-specific signal transduction pathways that interpret the external and internal concentrations of nutrients to modify root development. Progress in this field has led to the cloning of regulatory genes that play pivotal roles in nutrient-induced changes to root development.

Cell Differentiation↗

PubMed1985

Epithelial root-sheath changes during molar formation in the mouse.

The ultrastructure of the epithelial root-sheath was examined in the first mandibular molar teeth of 11, 16 and 21-day-old mice. The changing morphology of the sheath was related to root maturity. An initial, predominantly bicellular layer progressively shortened in an apical direction, the outer layer more so than the inner. The changing morphology appeared to involve maintenance of the inner layer largely at the expense of the outer. Some inner-layer cells persisted at spaced intervals adjacent to forming acellular cementum. Some epithelial rests, distant from the root surface, appeared to be formed early in root development at the time of initial root-sheath severance from the cervical loop of the enamel organ. Epithelial rests located along the acellular cementum appeared to arise from inner, rather than outer, epithelial root-sheath cells and at later stages in root development.

Animals↗

PubMed2026

Evolution and Expression Divergence of Legume PAL Genes Suggest Associations with Drought Response and Root Nodule Development.

Comparative genomic analyses provide insight into the mechanisms underlying gene-family evolution and crop adaptation. Here, we used the legume phenylalanine ammonia-lyase (PAL) gene family as a model and integrated pan-genomic, phylogenetic, molecular evolutionary, duplication-mode, and transcriptomic analyses, while developing GFtool for gene family identification. Across 45 genomes, we identified 302 PAL genes and classified them into five Groups. Groups 1-3 represented ancient lineages shared with outgroups, whereas Groups 4 and 5 were legume-specific. Molecular-clock analyses placed the divergence of Group 2 near the Paleocene-Eocene transition, while Groups 4 and 5 diversified from the middle Eocene to the early Oligocene. WGD/segmental duplication broadly contributed to PAL copy-number expansion, whereas tandem duplication was enriched in Group 5 of Papilionoideae. Group 2 genes showed drought-induced expression, whereas Group 5 genes were associated with early root nodule development. GFtool provides a scalable framework for gene-family studies.

Fabaceae↗

PubMed2004

The roles of latex and the vascular bundle in morphine biosynthesis in the opium poppy, Papaver somniferum.

The opium poppy, Papaver somniferum, is one of mankind's oldest medicinal plants. Opium poppy today is the commercial source of the narcotic analgesics morphine and codeine. Along with these two morphinans, opium poppy produces approximately eighty alkaloids belonging to various tetrahydrobenzylisoquinoline-derived classes. It has been known for over a century that morphinan alkaloids accumulate in the latex of opium poppy. With identification of many of the enzymes of alkaloid biosynthesis in this plant, biochemical data suggested involvement of multiple cell types in alkaloid biosynthesis in poppy. Herein the immunolocalization of five enzymes of alkaloid formation in opium poppy is reported: (R,S)-3'-hydroxy-N-methylcoclaurine 4'-O-methyltransferase central to the biosynthesis of tetrahydroisoquinoline-derived alkaloids, the berberine bridge enzyme of the sanguinarine pathway, (R,S)-reticuline 7-O-methyltransferase specific to laudanosine formation, and salutaridinol 7-O-acetyltransferase and codeinone reductase, which lead to morphine. In capsule and stem, both O-methyltransferases and the O-acetyltransferase are found predominantly in parenchyma cells within the vascular bundle, and codeinone reductase is localized to laticifers, the site of morphinan alkaloid accumulation. In developing root tip, both O-methyltransferases and the O-acetyltransferase are found in the pericycle of the stele, and the berberine bridge enzyme is localized to parenchyma cells of the root cortex. Laticifers are not found in developing root tip, and, likewise, codeinone reductase was not detected. These results provide cell-specific localization that gives a coherent picture of the spatial distribution of alkaloid biosynthesis in opium poppy.

Alkaloids↗

PubMed2004

Root length in transplanted premolars.

The objectives of the present prospective longitudinal study were to establish the survival and success rates for premolars with partly developed roots transplanted according to a set protocol, and to analyze the effect of varying pre-surgery root development on final root length. Standardized periapical radiographs of 132 consecutively transplanted premolars were obtained at predetermined intervals over a 4-year follow-up period. Survival and success rates were calculated on the basis of presence of teeth and recording of defined radiological variables, respectively. Final root length was compared to normative metric data for root morphology. From the total sample, 54 patients with 68 transplanted premolars suitable for analysis of root growth relative to initial root length were identified, and divided into subsamples according to root length pre-transplantation. Survival rates were 100% after 1, 2, and 3 years, and 98.6% 4 years post-transplantation. The success rates were 92.9%, 92.8%, 89%, and 90.5% in subsamples collected 1, 2, 3, and 4 years after transplantation, respectively. The transplants obtained on average 78% of normative root length, and by dichotomizing the transplanted teeth in subsamples with 'short' and 'long' roots pre-surgery, similar final root lengths were observed even if 'short' roots had the greatest increase. Very early transplantation (roots < 7 mm), however, tended to result in short final root length. It is concluded that the success and survival rates were comparable to results obtained in similar studies. Initial root length was a significant predictor of root length increase. Transplanting teeth with short roots is to risk short final root length.

Bicuspid↗

PubMed1998

Developmental appearance and distribution of bone sialoprotein and osteopontin in human and rat cementum.

BACKGROUND: Bone sialoprotein (BSP) and osteopontin (OPN), two major noncollagenous proteins (NCPs) in collagen-based mineralized tissues, have been implicated in mineral deposition and cell- and matrix-matrix interactions during root development. However, their role in cementogenesis is still a subject of debate. Since distribution of proteins is indicative of function, we have analyzed their temporo-spatial appearance in relation to that of cementum collagen. METHODS: Human premolars and rat molars at various stages of root development characterized by differing rates of formation were fixed in aldehyde and embedded in epoxy and LR White resin. Sections were processed for ultrastructural analysis and postembedding colloidal gold (immuno)cytochemistry. RESULTS: Incubations with antibodies against BSP and OPN and with lectins recognizing prominent sugars in these proteins generally revealed similar labeling patterns in both human and rat teeth, with gold particles accumulating mainly in the interfibrillar spaces. The lectin Helix pomatia, specific for N-acetyl-D-galactosamine, was distinctive in that it consistently reacted with human cementum, but only sporadically labeled rat cementum. Regardless of both the species and the stage of root development, mineralization initiated in mantle predentin in association with distinct foci immunoreactive for BSP and OPN. In human teeth, the deposition of cementum collagen began before the start of dentin mineralization and thus prior to any detectable labeling for BSP and OPN. However, at early stages of root formation in the rat, cementum collagen appeared after BSP and OPN accumulated on the root surface, whereas at advanced stages the deposition of cementum collagen, BSP and OPN coincided. CONCLUSIONS: The temporo-spatial differences in the appearance of BSP and OPN relative to cementum collagen correlate well with known differences in the speed of root elongation and explain the variable appearance of the dentino-cemental junction. The data reveal no causal relationship between BSP and OPN and the differentiation of cementoprogenitor cells and indicate that the distribution of collagen fibrils ultimately determines the amount and pattern of accumulation of these NCPs. There also is no consistent planar accumulation of BSP and OPN between dentin and cementum such as the cement lines found between "old" and "new" bone. It is concluded that the interlacement of collagen fibrils at the dentino-cemental junction, across which mineralization spreads, represents the primary attachment mechanism between cementum and dentin.

Adolescent↗

PubMed1998

Saline stress alters the temporal patterns of xylem differentiation and alternative oxidase expression in developing soybean roots

We conducted a coordinated biochemical and morphometric analysis of the effect of saline conditions on the differentiation zone of developing soybean (Glycine max L.) roots. Between d 3 and d 14 for seedlings grown in control or NaCl-supplemented medium, we studied (a) the temporal evolution of the respiratory alternative oxidase (AOX) capacity in correlation with the expression and localization of AOX protein analyzed by tissue-print immunoblotting; (b) the temporal evolution and tissue localization of a peroxidase activity involved in lignification; and (c) the structural changes, visualized by light microscopy and quantified by image digitization. The results revealed that saline stress retards primary xylem differentiation. There is a corresponding delay in the temporal pattern of AOX expression, which is consistent with the xylem-specific localization of AOX protein and the idea that this enzyme is linked to xylem development. An NaCl-induced acceleration of the development of secondary xylem was also observed. However, the temporal pattern of a peroxidase activity localized in the primary and secondary xylem was unaltered by NaCl treatment. Thus, the NaCl-stressed root was specifically affected in the temporal patterns of AOX expression and xylem development.

Journal Article↗

PubMed1981

Growth and development of cultured carrot cells and embryos under spaceflight conditions.

Morphogenetically competent proembryonic cells and well-developed somatic embryos of carrot at two levels of organization were exposed for 18.5 days to a hypogravity environment aboard the Soviet Biosatellite Cosmos 1129. It was confirmed that cultured totipotent cells of carrot can give rise to embryos with well-developed roots and minimally developed shoots. It was also shown that the space hypogravity environment could support the further growth of already-organized, later somatic embryonic stages and give rise to fully developed embryo-plantlets with roots and shoots.

Cells, Cultured↗