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The relationship between nodulin gene expression and the Rhizobium nod genes in Vicia sativa root nodule development.

The role of the Rhizobium nod genes in the induction of nodulin gene expression was examined by analyzing nodules formed on vetch roots by bacterial strains containing only the nod region. Introduction of an 11-kb cloned nod region of the R. leguminosarum sym plasmid pRL1JI into sym plasmid-cured rhizobia conferred on the recipient strains the ability to induce nodules in which all nodulin genes were expressed. This proves that from the sym plasmid only the nod region is involved in the induction of nodulin gene expression. A transconjugant of Agrobacterium carrying the same nod region induces nodules in which only early nodulin gene expression is detected. Thus, the nod region is essential for the induction of early nodulin gene expression. In this case, nodule cytology may indicate that a defense response of the plant interferes with the induction of late nodulin gene expression. Indirect evidence is presented that indeed the Rhizobium nod genes are also in some way involved in the induction of the expression of late noduling genes. The combination between histological data and pattern of nodulin gene expression furthermore reveals a correlation between nodule structure and nodulin gene expression. This correlation may aid in speculations about the functions of nodulins.

Blotting, Western↗

Remodelling processes following transplantation of dental germs in dogs.

Dental germs with different amounts of root development were transplanted from the left side to the right side, and vice versa, in young beagle dogs. The unerupted premolars had been taken out by osteotomy. After transplantation, the dental germs were covered with mucosa. The observation period lasted 7 postoperative weeks. Histological examination of the remodelling processes of the bone and tooth-tissue was carried out by polychromatic sequential labelling. All the transplanted dental germs remained vital, and the eruption continued to the normal occlusion level. From this, it follows that eruption of teeth does not depend on root development alone. In some cases, root length growth stopped immediately as a result of transplantation. Either a stumpy apex was formed, or resorption took place. In other cases root growth continued. There was a bend in the root at the point of transplantation, and the subsequently formed root section was shorter and thinner than in normal teeth.

Animals↗

Auxin distribution in Lotus japonicus during root nodule development.

For this work, Lotus japonicus transgenic plants were constructed expressing a fusion reporter gene consisting of the genes beta-glucuronidase (gus) and green fluorescent protein (gfp) under control of the soybean auxin-responsive promoter GH3. These plants expressed GUS and GFP in the vascular bundle of shoots, roots and leafs. Root sections showed that in mature parts of the roots GUS is mainly expressed in phloem and vascular parenchyma of the vascular cylinder. By detecting GUS activity, we describe the auxin distribution pattern in the root of the determinate nodulating legume L. japonicus during the development of nodulation and also after inoculation with purified Nod factors, N-naphthylphthalamic acid (NPA) and indoleacetic acid (IAA). Differently than white clover, which forms indeterminate nodules, L. japonicus presented a strong GUS activity at the dividing outer cortical cells during the first nodule cell divisions. This suggests different auxin distribution pattern between the determinate and indeterminate nodulating legumes that may be responsible of the differences in nodule development between these groups. By measuring of the GFP fluorescence expressed 21 days after treatment with Nod factors or bacteria we were able to quantify the differences in GH3 expression levels in single living roots. In order to correlate these data with auxin transport capacity we measured the auxin transport levels by a previously described radioactive method. At 48 h after inoculation with Nod factors, auxin transport showed to be increased in the middle root segment. The results obtained indicate that L. japonicus transformed lines expressing the GFP and GUS reporters under the control of the GH3 promoter are suitable for the study of auxin distribution in this legume.

Alphaproteobacteria↗

Localized changes in apoplastic and cytoplasmic pH are associated with root hair development in Arabidopsis thaliana.

Morphogenesis in plants is characterized by highly regulated cell enlargement. However, the mechanisms controlling and localizing regions of growth remain essentially unknown. Root hair formation involves the induction of a localized cell expansion in the lateral wall of a root epidermal cell. This expanded region then enters a second phase of localized growth called tip growth. Root hair formation therefore provides a model in which to study the cellular events involved in regulating localized growth in plants. Confocal ratio imaging of the pH of the cell wall revealed an acidification at the root hair initiation site. This acidification was present from the first morphological indications of localized growth, but not before, and was maintained to the point where the process of root hair initiation ceased and tip growth began. Preventing the wall acidification with pH buffers arrested the initiation process but growth resumed when the wall was returned to an acidic pH. Cytoplasmic pH was found to be elevated from approximately 7.3 to 7. 7 at the initiation site, and this elevation coincided with the acidification of the wall. Preventing the localized increase in cytoplasmic pH with 10 mM butyrate however did not inhibit either the wall acidification or the initiation process. In contrast, there was no detectable gradient in pH associated with the apex of tip growing root hairs, but both elevated apoplastic pH and butyrate treatment irreversibly inhibited the tip growth process. Thus the processes of tip growth and initiation of root hairs show differences in their pH requirements. These results highlight the role of localized control of apoplastic pH in the control of cell architecture and morphogenesis in plants.

Arabidopsis↗

Mineral trioxide aggregate pulpotomies: a case series outcomes assessment.

BACKGROUND: The greatest threats to developing teeth are dental caries and traumatic injury. A primary goal of all restorative treatment is to maintain pulp vitality so that normal root development or apexogenesis can occur. If pulpal exposure occurs, then a pulpotomy procedure aims to preserve pulp vitality to allow for normal root development. Historically, calcium hydroxide has been the material of choice for pulpotomy procedures. Recently, an alternative material called mineral trioxide aggregate (MTA) has demonstrated the ability to induce hard-tissue formation in pulpal tissue. The authors describe the clinical and radiographic outcome of a series of cases involving the use of MTA in pulpotomy procedures. METHODS: Twenty-three cases in 18 patients were treated with MTA pulpotomy procedures in an endodontic private practice. All of the patients had been referred to the practice for diagnosis and treatment of a symptomatic tooth. All of the authors provided treatment. Pulpal exposures were either due to caries or complicated enamel dentin fractures. RESULTS: Nineteen teeth in 14 patients were available for recall. The mean time of recall was 19.7 months. Of the 19 cases, 15 involved healed teeth, and three involved teeth that were healing. One of 19 cases involved a tooth with persistent disease. CONCLUSIONS: MTA may be useful as a substitute for calcium hydroxide in pulpotomy procedures. Further research, however, is required to clarify this conclusion. CLINICAL IMPLICATIONS: MTA conceivably could replace calcium hydroxide as the material of choice for pulpotomy procedures, if future research continues to show promising results.

Adolescent↗

Prognosis of root-fractured permanent incisors--prediction of healing modalities.

A population of 85 patients, comprising 95 root-fractured permanent incisors followed up to 11 years, was studied prospectively for the type of fracture healing that occurred (i.e. union of the fragments by interposition of hard tissue (HT) or connective tissue (CT), or nonunion characterized by interposition of granulation tissue between the fragments (GT)). Initial treatment was provided according to established treatment guidelines by the attending oral surgeon at the emergency room. Follow-up examination and treatment were provided by 2 of the authors. It appeared that GT could be diagnosed after approximately 3 weeks, while HT or CT could be diagnosed approximately 6 weeks after trauma. Many factors considered one at a time were found to have a significant or nearly significant effect on the type of fracture healing that occurred. However, a multivariate regression analysis revealed that the following factors were significantly related to fracture healing by HT: a large diameter of the apical foramen and severity of luxation of the coronal fragment (concussion/subluxation greater than lateral luxation greater than extrusion); fracture healing by CT: the presence of restorations in the injured teeth at the time of injury and the presence of marginal periodontal disease; and fracture nonhealing by GT: type of fixation (i.e. orthodontic band fixation versus acid etch or no fixation), antibiotic therapy, a constricted apical foramen, increased loosening of the coronal fragment, and stage of root development (GT never occurred in teeth with open apices). It was previously demonstrated following luxation injuries that type of luxation, stage of root development and type of fixation (orthodontic bands versus acid etch or no fixation) determined the prognosis of pulp survival. It therefore appears that the general factors that are able to predict the type of healing seen after root fracture are the same as those after luxation injuries, supporting the hypothesis that root fractures are another form of luxation injury, this time of only the coronal fragment.

Humans↗

Proliferative and degenerative events in the early development of chick dorsal root ganglia. I. Normal development.

Development of the chick dorsal root ganglia was examined in 4.5- to 9.5-day embryos. Tritiated thymidine (3H-TdR) and autoradiography was used to analyze proliferative activity and the Feulgen procedure to analyze degenerative activity in ganglia 12-17. Proliferative activity was found to be elevated through 4.5 days of incubation when as many as 14% of the ganglionic cells become labelled following a one-hour exposure to 3H-TdR. By 6.5 to 7.5 days proliferative activity decreases to 2-4% in the lateroventral (LV) regions and to approximately 1% in the mediodorsal (MD) regions of the ganglia. However, there appears to be increased proliferative activity by the end of the experimental period at 9.5 days. Birthdate studies demonstrate that large-scale neuronal production occurs between 4.5 and 6.5 days in the LV regions and between 4.5 and 7.5 days in the MD regions. After those times ganglionic proliferative activity must be largely nonneuronal in nature. This nonneuronal proliferation is greater in LV than in MD regions and in brachial than in nonbrachial ganglia. Degenerative activiy was found to be absent from the ganglia until after 4.5 days of incubation. It then increases rapidly, and by 5.5 days 5% of the LV cells in nonbrachial ganglia are degenerating. Degenerative activity then declines but is still present at 9.5 days. In contrast to results of an earlier study (Hamburger and Levi-Montalcini, '49), degenerative activity was also found in the LV region of brachial ganglia and the MD regions of brachial and nonbrachial ganglia. The pattern of LV degenerative activity in brachial ganglia is similar to that in nonbrachial ganglia, but the level of activity is lower. In the MD regions degenerative activity increases throughout the experimental period, and by 9.5 days as many as 4% of the MD cells are degenerating.

Age Factors↗

Radiographic and clinical manifestations of the impacted mandibular third molar.

Observations were made on 200 impacted mandibular third molars that were removed from 200 unselected patients under local anesthesia. Sixty percent of the teeth were found to be in apparent relationship with the inferior dental canal: 19 % in true relationship and 41 % in superimposition. The vertical impacted molars were mainly found to be in true relationship with the inferior dental canal. On comparison of the clinical postoperative diagnosis with the radiographic preoperative findings an error of only 0.8 % was shown. Most of the molars showed fully developed roots in the third decade of life. Most of the teeth in this series were vertically impacted having mostly fully developed roots. There was a significant correlation between the patients suffering from pain, trismus, and swelling. Swelling and trismus were the mostly affected postoperative sequelae.

Adolescent↗

[Biological effects of rare earth elements and their action mechanisms].

This paper reviewed the effects of rare earth elements on plant root development, biomass, quality, and resistance against stress. Applying appropriate amount of rare earth elements could not only promote seed germination and root development, increase plant biomass, but also improve harvest quality and plant resistance against stress. The plant uptake characteristics of rare earth elements, as well as their contents, distribution, existing forms, and cytolocalization in plants were discussed, with the focus on the effects of rare earth elements on photosynthesis and chlorophyll formation, nutrient uptake, relationships between rare earth elements and calcium, and action mechanisms of rare earth elements on cell membrane and enzymes. Rare earth elements could enhance chlorophyll content and improve photosynthetic rate, and thus, could increase plant biomass. When an appropriate amount of rare earth elements was applied, the uptake of nutrients by plant and their transformation and utilization were promoted. Rare earth elements could replace calcium ion to participate in enzymatic reactions, maintain the osmosis and stability of cell membrane, promote the protection function of cell membrane, and enhance the plant resistance capability against stress. In the end, further researches on rare earth elements were suggested.

Biomass↗

Autogenous free tooth transplantation in man by a 2-stage operation technique. A longitudinal intra-individual radiographic assessment.

Insufficient nutrition to the root surface of autotransplanted mature human teeth after transplantation could be an explanation for the high rates of complications such as root resorption and loss of the tooth transplant. Insufficient nutrition is suggested as being due to the distance between the recipient site tissue and the root surface, and the presence of a blood-clot between the tissues. Improved nutrition could be achieved by transplanting into contact with the vascularised connective tissue of a healing wound. To test this hypothesis, a series of human teeth with fully developed roots were autotransplanted to recipient sites prepared and left to heal for 14 days before transplantation. This study is a follow-up of 57 transplants with a minimum observation time of 1 year. Periodic radiographic examinations of the teeth were performed up to 5 years after transplantation. The results presented, showed a lower incidence of root resorption and extractions than previously reported for teeth with fully developed roots.

Adolescent↗

Effect of horizontal clinorotation on the root system development and on lipid breakdown in rapeseed (Brassica napus) seedlings.

Seedlings of Brassica napus were cultivated on a slowly rotating clinostat (1 rpm) or in the vertical control for 5 d. The root growth, the cotyledonary reserves and the transport of 14C-labeled sucrose from cotyledons to root system were studied in both cultural conditions. The biomass (fresh weight) of the root system was 35% higher in the horizontally clinorotated seedlings than in the controls. This increase was correlated with a greater degradation of reserve lipids and faster accumulation of sucrose in the cotyledons. The activity of isocitrate lyase, one of the two enzymes necessary to conversion of lipids into glucids, was also greater in the cotyledons of clinorotated seedlings. The labeling distribution of 14C in the cotyledons, the hypocotyl and the root system after 30, 60 and 120 min of application of 14C-labeled sucrose on the cotyledons showed higher translocation of the cotyledonary sucrose to the root system of clinorotated seedlings. In addition, we studied the effects of clinorotation on the biomass of the excised root system (of 10 d old seedlings) cultivated in a medium containing 1% sucrose. The horizontally clinorotated root system grew more than that of the controls. These results showed that the horizontal clinorotation acted on the root system growth and provoked a higher sucrose translocation from source to sink, i.e. from cotyledons to root system.

Biological Transport↗

Paracrine production of nerve growth factor during rat dorsal root ganglion development.

Nerve growth factor is a target derived growth factor. In the peripheral nervous system, it is produced by tissues innervated by the sympathetic nervous system and small sensory neurons. In the present study, we tested the hypothesis that an alternate source of nerve growth factor must be available to support dorsal root ganglion neurons before they make connection with the target. Using reverse transcriptase polymerase chain reaction (RT-PCR), we detected nerve growth factor mRNA at embryonic day 12 to 17, but not in adult dorsal root ganglia. In situ hybridization studies revealed positive staining in satellite/supportive cells juxtaposed to dorsal root ganglion neurons. Our study suggests that nerve growth factor from supporting cells may have a paracrine function during development of primary sensory neurons.

Animals↗

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.

Arabidopsis↗

Specific localization of the respiratory alternative oxidase in meristematic and xylematic tissues from developing soybean roots and hypocotyls.

We used tissue printing and specific immunostaining to examine the localization of the alternative oxidase (AOX) protein in correlation with measurements of AOX capacity. Selected root and hypocotyl regions were analyzed during the first 14 d of growth. It is shown that AOX protein is localized in the apical meristem and in developing xylem. The temporal pattern of expression is coincident with the evolution of AOX capacity. Data suggest that AOX expression is linked to xylem differentiation. Since heat is a major product of the alternative pathway, we speculate that thermogenesis is implicated in morphogenesis.

Gene Expression Regulation, Developmental↗

Carbon metabolism in developing soybean root nodules: the role of carbonic anhydrase.

A full-length cDNA clone encoding carbonic anhydrase (CA) was isolated from a soybean nodule cDNA library. In situ hybridization and immunolocalization were performed in order to assess the location of CA transcripts and protein in developing soybean nodules. CA transcripts and protein were present at high levels in all cell types of young nodules, whereas in mature nodules they were absent from the central tissue and were concentrated in cortical cells. The results suggested that, in the earlier stages of nodule development, CA might facilitate the recycling of CO2 while at later stages it may facilitate the diffusion of CO2 out of the nodule system. In parallel, sucrose metabolism was investigated by examination of the temporal and spatial transcript accumulation of sucrose synthase (SS) and phosphoenolpyruvate carboxylase (PEPC) genes, with in situ hybridization. In young nodules, high levels of SS gene transcripts were found in the central tissue as well as in the parenchymateous cells and the vascular bundles, while in mature nodules the levels of SS gene transcripts were much lower, with the majority of the transcripts located in the parenchyma and the pericycle cells of the vascular bundles. High levels of expression of PEPC gene transcripts were found in mature nodules, in almost all cell types, while in young nodules lower levels of transcripts were detected, with the majority of them located in parenchymateous cells as well as in the vascular bundles. These data suggest that breakdown of sucrose may take place in different sites during nodule development.

Amino Acid Sequence↗

Cellular organisation of the Arabidopsis thaliana root.

The anatomy of the developing root of Arabidopsis is described using conventional histological techniques, scanning and transmission electron microscopy. The root meristem is derived from cells of the hypophysis and adjacent cells of the embryo proper. The postembryonic organization of the root is apparent in the mature embryo and is maintained in the growing primary root after germination. Cell number and location is relatively invariant in the primary root, with 8 cortical and endodermal cell files but more variable numbers of pericycle and epidermal cells. The organisation of cells in lateral roots is similar to that of the primary root but with more variability in the numbers of cell files in each layer. [3H]thymidine labeling of actively growing roots indicates that a quiescent centre of four central cells (derived from the hypophysis) is located between the root cap columella and the stele. This plate of four cells is surrounded by three groups of cells in, proximal, distal and lateral positions. The labeling patterns of these cells suggest that they are the initials for the files of cells that comprise the root. They give rise to four sets of cell files: the stele, the cortex and endodermis, the epidermis and lateral root-cap and the columella. A model of meristem activity is proposed based on these data. This description of Arabidopsis root structure underpins future work on the developmental genetics of root morphogenesis.

Arabidopsis↗