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AKT1 and TRH1 are required during root hair elongation in Arabidopsis.

TRH1 is a member of the AtKT/AtKUP/AtHAK family of potassium carriers that is required for root hair elongation and AKT1 is an inward rectifying potassium channel expressed in the root epidermis, endodermis and cortex of Arabidopsis thaliana. Plants homozygous for the trh1-1 mutation form short root hairs. The Trh1(-) phenotype cannot be suppressed by growing plants homozygous for the trh1-1 mutation in the presence of high external KCl concentration. This indicates an absolute requirement for TRH1 in root hair tip growth. Plants homozygous for the akt1-1 mutation develop longer root hairs than the wild type when grown in 0 mM external potassium, but develop shorter hairs than the wild type when grown in higher concentrations [>10 mM] of potassium. These data indicate that both TRH1 and AKT1 are active in the root hair over a wide range of external potassium concentrations, but suggest they have different functions in the growing hair cell.

Arabidopsis↗

Nerve growth factor receptor TrkA is down-regulated during postnatal development by a subset of dorsal root ganglion neurons.

Nerve growth factor (NGF), signaling through its receptor tyrosine kinase, TrkA, is required for the survival of all small and many intermediate-sized murine dorsal root ganglion (DRG) neurons during development, accounting for 80% of the total DRG population. Surprisingly, NGF/TrkA-dependent neurons include a large population that does not express TrkA in adult mice (Silos-Santiago et al., 1995). This finding suggests two hypotheses: Neurons lacking TrkA in the adult may express TrkA during development, or they may be maintained through a paracrine mechanism by TrkA-expressing neurons. To determine whether TrkA is expressed transiently by DRG neurons that lack the receptor in adulthood, we examined the distribution of TrkA protein during development. We show here that TrkA expression is strikingly developmentally regulated. Eighty percent of DRG neurons expressed TrkA during embryogenesis and early postnatal life, whereas only 43% expressed TrkA at postnatal day (P) 21. Because the period of TrkA down-regulation corresponds with a critical period during which nociceptive phenotype can be altered by NGF (see Lewin and Mendell [1993] Trends Neurosci. 16:353-359), we examined whether NGF modulates the down-regulation of TrkA. Surprisingly, neither NGF deprivation nor augmentation altered the extent of TrkA down-regulation. Our results demonstrate a novel form of regulation of neurotrophin receptor expression that occurs late in development. All DRG neurons that require NGF for survival express TrkA during embryogenesis, and many continue to express TrkA during a postnatal period when neuronal phenotype is regulated by NGF. The subsequent down-regulation of TrkA is likely to be importantly related to functional distinctions among nociceptive neurons in maturity.

Aging↗

Prognosis of luxated non-vital maxillary incisors treated with calcium hydroxide and filled with gutta-percha. A retrospective clinical study.

A material of 885 luxated, non-vital incisors was evaluated radiographically with respect to healing of periodontal tissues including inflammatory root resorption and occurrence of ankylosis and cervical root fractures. The results were assessed after completion of calcium hydroxide treatment and 4 years after filling of the root canal with gutta-percha. After treatment with calcium hydroxide, periapical healing occurred in 95% of the teeth. Four years after filling with gutta-percha, periapical healing was present in 91% of the teeth. In the remaining teeth, recurrent or persistent periapical radiolucency was more frequent in overfilled than adequately filled teeth (P = 0.0001). There was no difference between immature and mature teeth. Inflammatory root resorption healed in 192 of 197 teeth (97%); in 5 teeth it developed into ankylosis. Ankylosis occurred in 13 teeth, all of which were intruded into the alveolar bone at the time of injury. The frequency of cervical root fractures was markedly higher in immature than mature teeth (P greater than 0.0001). Among immature teeth, the frequency of fractures was dependent on the stage of root development (chi 2 = 31.6) and ranged from 77% in teeth with the least to 28% in teeth with the most developed roots. The frequency of fractures was also related to the defects after healing of inflammatory root resorption in the cervical area of the root, significant at P less than 0.0001.

Calcium Hydroxide↗

The alfalfa (Medicago sativa) TDY1 gene encodes a mitogen-activated protein kinase homolog.

Development of root nodules, specifically induction of cortical cell division for nodule initiation, requires expression of specific genes in the host and microsymbiont. A full-length cDNA clone and the corresponding genomic clone encoding a MAP (mitogen-activated protein) kinase homolog were isolated from alfalfa (Medicago sativa). The genomic clone, TDY1, encodes a 68.9-kDa protein with 47.7% identity to MMK4, a previously characterized MAP kinase homolog from alfalfa. TDY1 is unique among the known plant MAP kinases, primarily due to a 230 amino acid C-terminal domain. The putative activation motif, Thr-Asp-Tyr (TDY), also differs from the previously reported Thr-Glu-Tyr (TEY) motif in plant MAP kinases. TDY1 messages were found predominantly in root nodules, roots, and root tips. Transgenic alfalfa and Medicago truncatula containing a chimeric gene consisting of 1.8 kbp of 5' flanking sequence of the TDY1 gene fused to the beta-glucuronidase (GUS) coding sequence exhibited GUS expression primarily in the nodule parenchyma, meristem, and vascular bundles, root tips, and root vascular bundles. Stem internodes stained intensely in cortical parenchyma, cambial cells, and primary xylem. GUS activity was observed in leaf mesophyll surrounding areas of mechanical wounding and pathogen invasion. The promoter was also active in root tips and apical meristems of transgenic tobacco. Expression patterns suggest a possible role for TDY1 in initiation and development of nodules and roots, and in localized responses to wounding.

3' Untranslated Regions↗

Three-year root caries incidence and risk modeling in older adults in North Carolina.

OBJECTIVES: The purposes of this study were to describe the incidence of root caries and to identify its risk factors in a representative sample of older adults. METHODS: Root caries incidence was estimated and multivariate risk assessment models were developed to identify predictors for root caries in a three-year follow-up study of 234 black and 218 white noninstitutionalized adults aged 65 and older residing in North Carolina. RESULTS: During the observation period, 29 percent of blacks developed root caries, compared to 39 percent of whites (P < .05). The mean net DFS increment per person was 0.55 +/- 0.13 root surfaces for blacks vs 0.80 +/- 0.21 for whites (P > .32). Multivariate logistic regression analysis indicated that blacks wearing a partial denture, having some root fragments, having an average gingival recession > or = 2 mm, and being free of P. intermedia were at greater risk for developing new root caries. The model for whites showed that retired people with their most severe gingival recession > or = 4 mm, an average probing pocket depth > or = 2 mm, and taking antihistamines were more likely to develop new lesions. CONCLUSIONS: These findings suggest that older blacks had less risk of root caries than whites, and in both groups indicators of poor periodontal status increased the risk of root caries.

Aged↗

Analysis of bacterial communities in the rhizosphere of chrysanthemum via denaturing gradient gel electrophoresis of PCR-amplified 16S rRNA as well as DNA fragments coding for 16S rRNA.

The effect of developing chrysanthemum roots on the presence and activity of bacterial populations in the rhizosphere was examined by using culture-independent methods. Nucleic acids were extracted from rhizosphere soil samples associated with the bases of roots or root tips of plants harvested at different stages of development. PCR and reverse transcriptase (RT) PCR were used to amplify 16S ribosomal DNA (rDNA) and 16S rRNA, respectively, and the products were subjected to denaturing gradient gel electrophoresis (DGGE). Prominent DGGE bands were excised and sequenced to gain insight into the identities of predominantly present (PCR) and predominantly active (RT-PCR) bacterial populations. The majority of DGGE band sequences were related to bacterial genera previously associated with the rhizosphere, such as Pseudomonas, Comamonas, Variovorax, and Acetobacter, or typical of root-free soil environments, such as Bacillus and Arthrobacter. The PCR-DGGE patterns observed for bulk soil were somewhat more complex than those obtained from rhizosphere samples, and the latter contained a subset of the bands present in bulk soil. DGGE analysis of RT-PCR products detected a subset of bands visible in the rDNA-based analysis, indicating that some dominantly detected bacterial populations did not have high levels of metabolic activity. The sequences detected by the RT-PCR approach were, however, derived from a wide taxonomic range, suggesting that activity in the rhizosphere was not determined at broad taxonomic levels but rather was a strain- or species-specific phenomenon. Comparative analysis of DGGE profiles grouped all DNA-derived root tip samples together in a cluster, and within this cluster the root tip samples from young plants formed a separate subcluster. Comparison of rRNA-derived bacterial profiles showed no grouping of root tip samples versus root base samples. Rather, all profiles derived from 2-week-old plant rhizosphere soils grouped together regardless of location along the root.

Bacteria↗

Developmental changes induced by gamma irradiation in Ipomoea batatas L. Lam (sweet potato).

Storage roots of Ipomoea batatas L. Lam were exposed to gamma irradiation at 90, 180, and 240 Gy. The two highest doses caused a delay in the initiation of rooting and especially of shooting; they also caused formation of rosette-like, stemless shoots without apical meristems. The storage roots irradiated with 90 Gy did form stems, but 50% of them had no apical meristem. The irradiated storage roots produced absorbent roots which developed shoots after 70 days of growth. Nonirradiated storage roots did not differentiate shoots from the absorbent roots. When grown in vitro, phytomers, stem segments with leaves and an axillary meristem, separated from the shoots of irradiated storage roots and exhibited growth aberrations, very intense rooting, and a delay in shooting. Phytomers from nonirradiated normal plants were irradiated with 10, 20, 30, 40, and 90 Gy and grown in a hormone-free medium. The 40- and 90-Gy doses delayed shooting as well as rooting. Only phytomers exposed to 40 and 90 Gy differentiated shoots from the absorbent roots. A stimulation of growth revealed in the accumulation of dry mass was found in the shoots of phytomers irradiated with 10 to 30 Gy. The long after-effects of irradiation as well as possible causes of growth stimulation are discussed.

Dose-Response Relationship, Radiation↗

GFRalpha-3, a protein related to GFRalpha-1, is expressed in developing peripheral neurons and ensheathing cells.

We report here the identification of a gene, termed GFRalpha-3 (glial cell line-derived neurotrophic factor family receptor alpha-3), related to GFRalpha-1 and GFRalpha-2 (also known as GDNFR-alpha and GDNFR-beta), and describe distribution of GDNFalpha-3 in the nervous system and other parts of the mouse body during development and in the adult. GFRalpha-3 in situ hybridization signals were found mainly in the peripheral nervous system, with prominent signals in developing dorsal root and trigeminal ganglia. Sympathetic ganglia were also positive. Developing nerves manifested strong GFRalpha-3 mRNA signals, presumably generated by the Schwann cells. Olfactory ensheathing cells were also positive. Other non-neuronal cells appearing positive during development included chromaffin cells in the adrenal gland and small clusters of cells in the intestinal epithelium. In the central nervous system no robust signals could be detected at any stage investigated with the present probes. Compared with the previously described GFRalpha-1 and GFRalpha-2 mRNAs, which are widely distributed in the central nervous system and peripheral organs, the expression of GFRalpha-3 mRNA is much more restricted. The prominent expression in Schwann cells during development suggests a key role for GFRalpha-3 in the development of the peripheral nervous system. As Schwann cells are known to lack expression of the transducing RET receptor, we propose that a possible function of GFRalpha-3 during development could be to bind Schwann cell-derived GDNF-like ligands, thus presenting such molecules to growing axons.

Amino Acid Sequence↗

Regulated heterogeneity in 12-kDa P-protein phosphorylation and composition of ribosomes in maize (Zea mays L.).

Maize (Zea mays L.) possesses four distinct approximately 12-kDa P-proteins (P1, P2a, P2b, P3) that form the tip of a lateral stalk on the 60 S ribosomal subunit. RNA blot analyses suggested that the expression of these proteins was developmentally regulated. Western blot analysis of ribosomal proteins isolated from various organs, kernel tissues during seed development, and root tips deprived of oxygen (anoxia) revealed significant heterogeneity in the levels of these proteins. P1 and P3 were detected in ribosomes of all samples at similar levels relative to ribosomal protein S6, whereas P2a and P2b levels showed considerable developmental regulation. Both forms of P2 were present in ribosomes of some organs, whereas only one form was detected in other organs. Considerable tissue-specific variation was observed in levels of monomeric and multimeric forms of P2a. P2b was not detected in root tips, accumulated late in seed embryo and endosperm development, and was detected in soluble ribosomes but not in membrane-associated ribosomes that copurified with zein protein bodies of the kernel endosperm. The phosphorylation of the 12-kDa P-proteins was also developmentally and environmentally regulated. The potential role of P2 heterogeneity in P-protein composition in the regulation of translation is discussed.

Phosphorylation↗

The COW1 locus of arabidopsis acts after RHD2, and in parallel with RHD3 and TIP1, to determine the shape, rate of elongation, and number of root hairs produced from each site of hair formation.

Two recessive mutant alleles at CAN OF WORMS1 (COW1), a new locus involved in root hair morphogenesis, have been identified in Arabidopsis thaliana L. Heynh. Root hairs on Cow1- mutants are short and wide and occasionally formed as pairs at a single site of hair formation. The COW1 locus maps to chromosome 4. Root hairs on Cow1- plants form in the usual positions, suggesting that the phenotype is not the result of abnormal positional signals. Root hairs on Cow1- roots begin hair formation normally, forming a small bulge, or root hair initiation site, of normal size and shape and in the usual position on the hair-forming cell. However, when Cow1- root hairs start to elongate by tip growth, abnormalities in the shape and elongation rate of the hairs become apparent. Genetic evidence from double-mutant analysis of cow1-1 and other loci involved in root hair development supports our conclusion that COW1 is required during root hair elongation.

Alleles↗

Canopy and environmental control of root dynamics in a long-term study of Concord grape.

Below-ground carbon allocation represents a substantial fraction of net photosynthesis in plants, yet timing of below-ground allocation and endogenous and exogenous factors controlling it are poorly understood. Minirhizotron techniques were used to examine root populations of Vitis labruscana Bailey cv. Concord under two levels of dormant-season canopy removal and irrigation. Root production, pigmentation, death and disappearance to a depth of 110 cm were determined over two wet and two dry years (1997-2000). There was continual root production and senescence, with peak root production rates occurring by midseason. Later in the season, when reproductive demands for carbon were highest and physical conditions limiting, few roots were produced, especially in dry years in nonirrigated vines. Root production under minimal canopy pruning was generally greater and occurred several weeks earlier than root production under heavy pruning, corresponding to earlier canopy development. Initial root production occurred in shallow soils, likely due to temperatures at shallow depths being warmer early in the season. Our study showed intricate relationships between internal carbon demands and environmental conditions regulating root allocation.

Agriculture↗

[Phosphoric stress and antioxidative power of wheat roots].

The change of antioxydative power of metabolites in the wheat root system (cv Polesskaya 90 and cv Polesskaya 92) under effect of stress doses of phosphoric nutrition (4-fold dose and phosphoric deficit in nutrient mixture of water culture) proves that phosphoric stress causes peroxidation of membrane lipids. These changes are a nonspecific reaction of membranes in root epithelial cells. Stress and responsive changes in the membrane system are recorded by an antioxydative power decreasing root metabolites. This is the first stage of plant biochemical adaptation (7 to 14 days of growth). The second stage (21 to 28 days of growth) is characterised by the increasing of antioxydative power. However, nonspecific character of the primary reaction of membrane system in root cells at high doses and phosphoric deficite in nutrient mixture changes into clear specific reaction at physiological adaptation level. This is revealed in the change of growth function and kinetics of phosphoric accumulation. Phosphoric deficiency at critical phosphoric concentration in plant roots (0.2% P2O5 of dry mass) results in the increase of growth processes. This is revealed in accumulation of root biomass. At high phosphoric concentrations (1.04% P2O3) the development of root system is inactivated and phosphoric absorption is decreased. The similar specificity in the changes of growth and absorption function of root system is considered to be adapting. The changes of antioxydative power and hence the destroying apparatus in epithelial cells of root system advance the damage at physiological and morphological level and can define wheat adaptation ability to extreme conditions of phosphoric nutrition.

Adaptation, Biological↗

Altered growth of transgenic tobacco lacking leaf cytosolic pyruvate kinase.

Previously, we reported that transformation of tobacco (Nicotiana tabacum L.) with a vector containing a potato cytosolic pyruvate kinase (PKc) cDNA generated two plant lines specifically lacking leaf PKc (PKc-) as a result of co-suppression. PKc deficiency in these primary transformants did not appear to alter plant development, although root growth was not examined. Here we report a striking reduction in root growth of homozygous progeny of both PKc- lines throughout development under moderate (600 microE m-2 s-1) or low (100 microE m-2 s-1) light intensities. When both PKc- lines were cultivated under low light, shoot and flower development were also delayed and leaf indentations were apparent. Leaf PK activity in the transformants was significantly decreased at all time points examined, whereas root activities were unaffected. Polypeptides corresponding to PKc were undetectable on immunoblots of PKc- leaf extracts, except in 6-week-old low-light-grown PKc- plants, in which leaf PKc expression appeared to be greatly reduced. The metabolic implications of the kinetic characteristics of partially purified PKc from wild-type tobacco leaves are discussed. Overall, the results suggest that leaf PKc deficiency leads to a perturbation in source-sink relationships.

Cytosol↗

Comparative aspects of the inner root sheath in adult and developing hairs of mammals in relation to the evolution of hairs.

The inner root sheath (IRS) allows the exit of hairs through the epidermal surface. The fine structure of monotreme and marsupial IRS and trichohyalin is not known. Using electron microscopy and immunocytochemistry, the localization of trichohyalin and transglutaminase have been studied in monotreme and marsupial hairs, and compared with trichohyalin localization in placental hairs. Trichohyalin in all mammalian species studied here is recognized by a polyclonal antibody against sheep trichohyalin. This generalized immunoreactivity suggests that common epitopes are present in trichohyalin across mammals. In differentiating IRS cells, trichohyalin granules of variable dimensions are composed of an immunolabelled amorphous matrix associated with a network of 10-12-nm-thick keratin filaments. Transglutaminase labelling is present among keratin bundles and trichohyalin granules, and in condensed nuclei of terminally differentiating cells of the inner root sheath. The IRS in monotreme hairs is multistratified but lacks a distinguishable Henle layer. Cornification of IRS determines the sculpturing of the fibre cuticle and later shedding from the follicle for the exit of the hair fibre on the epidermal surface. It is hypothesized that the stratification of IRS in Henle, Huxley and IRS cuticle layers is derived from a simpler organization, like that present in the IRS of monotremes. The IRS is regarded as a localized shedding/sloughing layer needed for the exit of hairs without injury to the epidermis. The formation of the IRS during the evolution of mammalian epidermis allowed the physiological exit of hairs produced inside the skin. The peculiar morphogenesis of hairs in possible primitive skins, such as those of the monotremes (mammals with some reptilian characteristics) or the tails of some rodents (a scaled skin), may elucidate the evolution of hairs. In monotreme and rodent tail skin, the dermal papilla remains localized on the proximal side of the hair peg and forms a hair placode with bilateral symmetry. The papilla is progressively surrounded by the down-growing hair peg until a dermal papilla with radial symmetry is formed. It is speculated that the progressive reduction of the extended dermal papilla of reptilian scales into small and deep papillae of therapsid reptiles produced hairs in mammals.

Animals↗

Development of an in situ root caries model. A. In vitro investigations.

OBJECTIVES: The paper describes preliminary in vitro investigations, the objectives of which were to examine the influence of certain experimental parameters on artificial carious lesion formation in root hard tissues, and their remineralisation. These experiments formed part of a wider study that aimed to develop an in situ model of root caries, based on the existing coronal caries model used in Liverpool. The present studies examined the effects (a) of the anatomical origin of the dentine, the presence or absence of cementum, the exposure time and the type of demineralising system, on lesion development, and (b) of baseline lesion size on the extent and location of mineral re-precipitation. METHODS: Mineral content parameters in plano-parallel sections taken from dentine lesions were determined by computer-controlled transverse microradiography. RESULTS: The importance of the anatomical origin of the dentine on lesion formation was investigated by comparing in vitro lesion formation in premolar and molar dentine, and in dentine from apical, middle and coronal thirds of the root: no difference was observed between these sites. Lesions formed more rapidly in acid buffer solutions than in acid gel systems, and were more reliably produced when cementum was removed. The effect of baseline lesion size on subsequent in vitro remineralisation demonstrated that a small baseline mineral content was associated with a larger percentage mineral gain. The location of mineral deposition throughout the lesion was also influenced by baseline mineral content parameters. CONCLUSIONS: The results form a basis for the further development of an in situ dentinal caries model, providing data to suggest that manipulation of parameters involved in the preparation of artificial carious lesions has a significant effect on the behaviour of the lesion, particularly the phenomenon of remineralisation. Further work is needed to investigate the behaviour of the model in situ.

Bicuspid↗

[Different methods of forming root systems in plants using maize and flax seedlings as examples].

Comparison of the appearance and development of lateral roots in the flax and maize seedlings has shown the way of root branching in the flax, as distinct from that in most plants. Some primordia in the flax main root did not develop immediately into lateral roots, but remained quiescent, which determines different reactions of the maize and flax root systems to experimental influences. Decapitation of the main root in the maize did not leads to a significant increase in the number of lateral roots, while in the flax, their number noticeably increased due to the development of previously quiescent primordia into lateral roots. The treatment with synthetic auxin did not induce the formation of additional primordia and lateral roots in the maize roots. In the flax, the number of primordia increased significantly and that of lateral roots increased to a somewhat lesser extent. Apparently, the development of a primordium into a lateral root proceeds in two stages and they have different regulation.

Flax↗

Automatic pH-cycling caries model applied on root hard tissue.

The development of a caries lesion is a dynamic process that includes frequently alternating periods of de- and remineralization, a process for which no in vitro models currently exists. The aim of this study was to develop a model which could repeatedly generate identical pH versus time curves over extended periods of time. To test the model demineralization studies on cementum/dentin were performed. Cementum/dentin blocks were repeatedly subjected to de- and remineralizing solutions delivered by a computerized pump system connected to a pH-meter and a recorder. pH versus time curves similar to those produced in plaque after carbohydrate intake were generated 18 times during 15 h followed by a remineralizing period of 9 h every day for 16 days. The intra-day and inter-day variation of the pH-cycling model were found to be low. The mineral change of the blocks was followed by 125I absorptiometry and the transmission through the blocks during the experimental period increased in average by 1.4 +/- (SE) 0.18%. Microradiographic analysis of the transversal sections of the blocks confirmed the development of lesions resembling root caries.

Absorptiometry, Photon↗

[Changes with time in incomplete root and periodontal tissues of young dog teeth with torque loading applied].

We investigated, using labeling techniques (tetracycline, calcein) and routine histopathological methods (H. E. and Azan stainings), the histological changes with passage of time in incomplete root and the periodontium of young dog teeth with torque loading applied. The maxillary left first, second and third incisor teeth were used as anchorage teeth, and the maxillary right second incisor was loaded with either 3 degrees or 9 degrees torque. The orthodontic forces of these are 20-30 g and 80-90 g, respectively. Some temporal disturbances (such as mild, moderate, or severe bendings of labial and palatal apexes) were observed in certain regions of the apex of the incomplete root. However, with subsequent development of the root, the disturbances were repaired and pulp of the teeth was normally formed. Furthermore, although periodontium was also temporarily disturbed, it developed normally with the passage of time.

Alveolar Bone Loss↗