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PubMed2006

Root meristems in Medicago truncatula tissue culture arise from vascular-derived procambial-like cells in a process regulated by ethylene.

Leaf explants of Medicago truncatula were used to investigate the origins of auxin-induced root formation. On the application of auxin there is some callus formation (not the massive amount that occurs in response to auxin plus cytokinin) and roots appear shortly after the first visible callus. Histological examination reveals morphologically distinctive sheets of callus cells that emanate from the veins of the leaf explants and, within this cell type, root primordia are produced as well as some vascular tissue cells. What is suggested is that the vein-derived cells (VDCs) are procambial-like and function as pluripotent stem cells with a propensity to form root meristems or vascular tissues in response to added auxin. The development of root primordia from these pluripotent cells was clearly up-regulated by the use of the sickle (skl) mutant, which is a mutant impaired in ethylene signal transduction while the wild type and the sunn mutant, defective in auxin polar transport, produced similar numbers of roots. The skl mutant in generating many more roots concomitantly formed fewer vascular tissues. The root meristems differentiate similarly to normal roots producing a central cylinder of vascular tissue, which connects with the leaf explant veins. The VDCs appear to be derived from the cells of or near the phloem. The leaf observations suggest that a pool of stem cells exist in vascular tissue that, in combination with auxin and perhaps other factors, drive a diversity of plant development outcomes that is species specific. The way auxin interacts with other hormones is a key factor in determining the stem cell fate. The histological data in this study also assist in the interpretation of the molecular analysis of auxin-induced root formation in cultured leaves of M. truncatula.

Ethylenes↗
PubMed2004

Ground plan of the polychaete brain--I. Patterns of nerve development during regeneration in Dorvillea bermudensis (Dorvilleidae).

The nervous systems of adult specimens and regenerating fragments of Dorvillea bermudensis ("Polychaeta," Dorvilleidae) were stained with antisera directed against acetylated alpha-tubulin and analyzed by indirect fluorescence and confocal laser scanning microscopy. The anlage of each circumesophageal connective is initially doubled in regenerates and starts with the outgrowth of two nerves from each side of the ventral cord of the amputee. The inner nerves on each side, which become the ventral roots, fuse medially to form the ventral cerebral commissure, whereas the outer nerves become the dorsal roots. As development proceeds, both roots split again, to form the four cerebral commissures. In later stages, the two circumesophageal connectives of each side merge, the point of fusion progressing from the ventral cord toward the brain. In D. bermudensis, this process stops halfway along the connective, thus producing the typical polychaete pattern according to Orrhage ([ 1995] Acta Zool 79:215-234): Each single circumesophageal connective divides near the brain into dorsal and ventral roots, which themselves split into two branches to form the four cerebral commissures. From the results presented here, we conclude that each circumesophageal connective is basically a paired structure but is partially fused in species possessing dorsal roots and completely fused in species lacking them. This may also be true for Clitellata, in which dorsal roots have hitherto not been found. At the posterior end, the outgrowing fibers form five connectives, of which the two outermost of each side fuse in an anteroposterior direction, forming the main connectives. Outgrowing fibers of the stomatogastric nerve stumps soon form the stomatogastric ring, which subsequently is linked with the new brain via stomatogastric connectives.

Acetylation↗
PubMed1984

Prenatal development of the rat dorsal root ganglia. A scanning electron-microscopic study.

This study describes three-dimensional aspects of the development and pseudo-unipolarization of neuroblasts and the maturation of satellite cells in prenatal rat dorsal root ganglia, using scanning electron microscopy, after removal of extracellular connective tissue components by trypsin digestion and HC1 hydrolysis. At 14 days of gestation, the vast majority of neurons are spindle-shaped or bipolar and only 3% are unipolar, while at 16 and 18 days this percentage has increased to 30% and 91%, respectively. The initial portions of the central and peripheral neuronal processes gradually approach each other and form a common initial portion. Finally, the cytoplasm of this common initial portion becomes thinner and elongates to form the stem process of the mature cell. Satellite cells are present from the beginning of the period studied, but intricate networks of branching satellite cell processes only develop after about day 17.

Animals↗
PubMed1998

The tropic response of plant roots to oxygen: oxytropism in Pisum sativum L.

Plant roots are known to orient growth through the soil by gravitropism, hydrotropism, and thigmotropism. Recent observations of plant roots that developed in a microgravity environment in space suggested that plant roots may also orient their growth toward oxygen (oxytropism). Using garden pea (Pisum sativum L. cv. Weibul's Apollo) and an agravitropic mutant (cv. Ageotropum), root oxytropism was studied in the controlled environment of a microrhizotron. A series of channels in the microrhizotron allowed establishment of an oxygen gradient of 0.8 mmol mol-1 mm-1. Curvature of seedling roots was determined prior to freezing the roots for subsequent spectrophotometric determinations of alcohol dehydrogenase activity. Oxytropic curvature was observed all along the gradient in both cultivars of pea. The normal gravitropic cultivar showed a maximal curvature of 45 degrees after 48 h, while the agravitropic mutant curved to 90 degrees. In each cultivar, the amount of curvature declined as the oxygen concentration decreased, and was linearly related to the root elongation rate. Since oxytropic curvature occurred in roots exposed to oxygen concentrations that were not low enough to induce the hypoxically responsive protein alcohol dehydrogenase, we suspect that the oxygen sensor associated with oxytropism does not control the induction of hypoxic metabolism. Our results indicate that oxygen can play a critical role in determining root orientation as well as impacting root metabolic status. Oxytropism allows roots to avoid oxygen-deprived soil strata and may also be the basis of an auto-avoidance mechanism, decreasing the competition between roots for water and nutrients as well as oxygen.

Alcohol Dehydrogenase↗
PubMed2003

Metabolic profiling of root exudates of Arabidopsis thaliana.

In addition to accumulating biologically active chemicals, plant roots continuously produce and secrete compounds into their immediate rhizosphere. However, the mechanisms that drive and regulate root secretion of secondary metabolites are not fully understood. To enlighten two neglected areas of root biology, root secretion and secondary metabolism, an in vitro system implementing root-specific elicitation over a 48-day time course was developed. After roots of Arabidopsis thaliana had been elicited with salicylic acid, jasmonic acid, chitosan, and two fungal cell wall elicitors, the secondary metabolites subsequently secreted were profiled. High-performance liquid chromatography was used to metabolically profile compounds in the root exudates, and 289 possible secondary metabolites were quantified. The chemical structures of 10 compounds were further characterized by (1)H and (13)C NMR: butanoic acid, trans-cinnamic acid, o-coumaric acid, p-coumaric acid, ferulic acid, p-hydroxybenzamide, methyl p-hydroxybenzoate, 3-indolepropanoic acid, syringic acid, and vanillic acid. Several of these compounds exhibited a wide range of antimicrobial activity against both soil-borne bacteria and fungi at the concentration detected in the root exudates.

Anti-Bacterial Agents↗
PubMed2005

Osmotic regulation of root system architecture.

Although root system architecture is known to be highly plastic and strongly affected by environmental conditions, we have little understanding of the underlying mechanisms controlling root system development. Here we demonstrate that the formation of a lateral root from a lateral root primordium is repressed as water availability is reduced. This osmotic-responsive regulatory mechanism requires abscisic acid (ABA) and a newly identified gene, LRD2. Mutant analysis also revealed interactions of ABA and LRD2 with auxin signaling. Surprisingly, further examination revealed that both ABA and LRD2 control root system architecture even in the absence of osmotic stress. This suggests that the same molecules that mediate responses to environmental cues can also be regulators of intrinsic developmental programs in the root system.

Abscisic Acid↗
PubMed1988

Structural and microanalytical changes in dentition after radiotherapy applied in cases of tumour in the oral cavity region.

Morphological observations by means of the scanning electron and optical microscopes as well as microanalytical qualitative and quantitative tests concerning content and distribution of calcium, phosphorus and magnesium in the teeth being in irradiation field were made in patients treated for larynx cancer by methods of radiotherapy. The teeth enamel was in some places thinner and in some others it was lacking. The characteristic defects took place in the form of notches, cracks and of superficial destruction. In X-ray microanalyser the enamel showed some features of demineralization. In dentine, features of demineralization and on the other hand features of remineralization were seen. The root dentin, in which destructive processes but also reparation processes developed by root cement could be seen, was rather characteristic. Clear structural changes and disturbances of mineralization in hard tooth tissues being under therapeutic X-ray irradiation were observed.

Adult↗
PubMed2001

Growth of AM fungi on in vitro root organ culture of Sorghum vulgare and Saccharum officinarum.

Spores of Gl mosseae and Gig gigantea germinated on minimal medium produced extraradical mycelium. Gl. mosseae infected roots of S. officinarum in in vitro condition were inoculated in M medium with in vitro cultured roots of Sorghum vulgare (test roots). From the infected root of S. officinarum, the mycelium developed and it infected the test roots. The roots developed new mycelia and further the mycelia produced a few hyaline spores. In MS medium combined with soil extract, root exudate, thiamine HCl and inositol combination, spore germination and germ tube growth were higher when compared with other media.

Culture Techniques↗
PubMedDate not supplied

[Growth-regulating activity of N-benzyl- and O-benzyl-containing compounds belonging to a new group of synthetic analogues of natural auxins].

We studied the effect of benzylamine, benzyl alcohol, and their derivatives (constituting a new group of synthetic analogues of natural auxins) on rooting of leaf and stem cuttings, rhizogenesis and growth of barley plantlets and tomato seedlings, and tomato plant productivity. These compounds promoted rooting of leaf and stem bean cuttings, increased rhizogenic activity, and stimulated the development of root systems in barley and tomato seeds. The activity of the compounds studied was similar to that of standard substances (3-indoleacetic acid potassium salt and 2-naphthylacetic acid). The benzyl group attached to the oxygen or nitrogen atom was shown to be the smallest molecular structure which provided auxin activity of the compounds. Derivatives of benzyl alcohol containing the quaternary ammonium fragment possessed auxin and anti-gibberellin (retardant) properties. They were selected by chemical synthesis of low-molecular-weight bioregulators with desired properties (a combination of chemical fragments with complementary physiological activity in the molecule). Auxin and anti-gibberellin (retardant) activities produced a synergistic effect. Germination of seeds treated with these compounds was accompanied by a more significant increase in the weight and length of roots (compared to standard auxins). The rate of seedling establishment reached 100%. The development of fruits and accumulation of reserve nutrient substances were synchronized and accelerated after spraying vegetating plants with solutions of studied compounds. The synergistic effect underlay a significant increase in the amount and quality of the crop (e.g., tomatoes).

Benzene Derivatives↗
PubMed1992

Molecular cloning of a cDNA encoding aspartate aminotransferase-P2 from lupin root nodules.

Two isoenzymic forms of aspartate aminotransferase are present in the plant fraction of developing lupin root nodules. One of these forms, aspartate aminotransferase-P2 (AAT-P2), increases dramatically with the onset of biological nitrogen fixation and is associated with the assimilation of ammonia by the plant in the Rhizobium-legume symbiosis. A day 18 lupin nodule cDNA library in the lambda ZapII vector was immunoscreened with a monoclonal antibody specific for AAT-P2 and yielded two near-full-length 1700 bp clones. These clones were sequenced. Amino acid sequences from three peptides derived from immunopurified AAT-P2 were aligned, and showed 100% homology with the amino acid sequence deduced from the cDNA clones. The DNA sequence showed 50% homology with AAT sequences from a range of animal sources. Conversion of the clones to the phagemid form allowed their expression in Escherichia coli where both exhibited enzyme activity that could be immunoprecipitated with AAT-P2-specific monoclonal antibodies. Western blot analysis revealed protein moieties with molecular masses of 39, 43, 45 and 55 kDa. The 5' end of the clones coded for a hydrophobic leader sequence of about 50 amino acids indicative of a targeting sequence and consistent with the plastid localisation of nodule AAT-P2.

Amino Acid Sequence↗
PubMed2004

Citrate-permeable channels in the plasma membrane of cluster roots from white lupin.

White lupin (Lupinus albus) is well adapted to phosphorus deficiency by developing cluster roots that release large amounts of citrate into the rhizosphere to mobilize the sparingly soluble phosphorus. To determine the mechanism underlying citrate release from cluster roots, we isolated protoplasts from different types of roots of white lupin plants grown in phosphorus-replete (+P) and phosphorus-deficient (-P) conditions and used the patch-clamp technique to measure the whole-cell currents flowing across plasma membrane of these protoplasts. Two main types of anion conductance were observed in protoplasts prepared from cluster root tissue: (1) an inwardly rectifying anion conductance (IRAC) activated by membrane hyperpolarization, and (2) an outwardly rectifying anion conductance (ORAC) that became more activated with membrane depolarization. Although ORAC was an outward rectifier, it did allow substantial inward current (anion efflux) to occur. Both conductances showed citrate permeability, with IRAC being more selective for citrate3- than Cl- (PCit/PCl = 26.3), while ORAC was selective for Cl- over citrate (PCl/PCit = 3.7). Both IRAC and ORAC were sensitive to the anion channel blocker anthracene-9-carboxylic acid. These currents were also detected in protoplasts derived from noncluster roots of -P plants, as well as from normal (noncluster) roots of plants grown with 25 microm phosphorus (+P). No differences were observed in the magnitude or frequency of IRAC and ORAC currents between the cluster roots and noncluster roots of -P plants. However, the IRAC current from +P plants occurred less frequently than in the -P plants. IRAC was unaffected by external phosphate, but ORAC had reduced inward current (anion efflux) when phosphate was present in the external medium. Our data suggest that IRAC is the main pathway for citrate efflux from white lupin roots, but ORAC may also contribute to citrate efflux.

Calcium↗
PubMed1995

Role of the Differentiation of Root Epidermal Cells in Nod Factor (from Rhizobium meliloti)-Induced Root-Hair Depolarization of Medicago sativa.

The stage of differentiation of epidermal cells and the development of root hairs was found to be important for the induction of depolarization in root hairs of Medicago sativa by Nod factor [NodRm-IV(S)] isolated from the bacterium Rhizobium meliloti. The electrical membrane response was concentration dependent, having its major effect (amplitude of the depolarization and number of root hairs that responded) at 10-8 and 10-7 M Nod factor. This response was correlated with a morphological effect of Nod factor in the root-hair-deformation bioassay at similar concentrations. The effect of Nod factor on depolarization and root-hair deformation showed specificity with respect to the structure, since unsulfated Nod molecules were inactive, as was the synthetic N,N',N",N"'- tetraacetylchitotetraose. The Nod factor that is O-acetylated at the nonreducing sugar was as efficient in root-hair deformation and membrane depolarization as the sulfated Nod factor.

Journal Article↗
PubMed2003

Prolactin and its receptor are expressed in murine hair follicle epithelium, show hair cycle-dependent expression, and induce catagen.

Here, we provide the first study of prolactin (PRL) and prolactin receptor (PRLR) expression during the nonseasonal murine hair cycle, which is, in contrast to sheep, comparable with the human scalp and report that both PRL and PRLR are stringently restricted to the hair follicle epithelium and are strongly hair cycle-dependent. In addition we show that PRL exerts functional effects on anagen hair follicles in murine skin organ culture by down-regulation of proliferation in follicular keratinocytes. In telogen follicles, PRL-like immunoreactivity was detected in outer root sheath (ORS) keratinocytes. During early anagen (III to IV), the developing inner root sheath (IRS) and the surrounding ORS were positive for PRL. In later anagen stages, PRL could be detected in the proximal IRS and the inner layer of the ORS. The regressing (catagen) follicle showed a strong expression of PRL in the proximal ORS. In early anagen, PRLR immunoreactivity occurred in the distal part of the ORS around the developing IRS, and subsequently to a restricted area of the more distal ORS during later anagen stages and during early catagen. The dermal papilla (DP) stayed negative for both PRL and PRLR throughout the cycle. Telogen follicles showed only a very weak PRLR staining of ORS keratinocytes. The long-form PRLR transcript was shown by real-time polymerase chain reaction to be transiently down-regulated during early anagen, whereas PRL transcripts were up-regulated during mid anagen. Addition of PRL (400 ng/ml) to anagen hair follicles in murine skin organ culture for 72 hours induced premature catagen development in vitro along with a decline in the number of proliferating hair bulb keratinocytes. These data support the intriguing concept that PRL is generated locally in the hair follicle epithelium and acts directly in an autocrine or paracrine manner to modulate the hair cycle.

Animals↗
PubMed2005

Expression patterns of erythropoietin and its receptor in the developing spinal cord and dorsal root ganglia.

Recombinant human erythropoietin (EPO) is neuroprotective in animal models of adult spinal cord injury, and reduces apoptosis in adult dorsal root ganglia after spinal nerve crush. The present work demonstrates that spinal cord and dorsal root ganglia share dynamic expression patterns of EPO and its receptor (EPOR) during development. C57Bl mice from embryonic days (E) 8 (E8) to E19 were studied. In spinal cord and dorsal root ganglia, EPOR expression in all precursor cells preceded the expression of EPO in subsets of neurons. On E11, EPO-immunoreactive spinal motoneurons and ganglionic sensory neurons resided adjacent to EPOR-expressing radial glial cells and satellite cells, respectively. From E12 onwards, EPOR-immunoreactivity decreased in radial glial cells and, transiently, in satellite cells. Simultaneously, large-scale apoptosis of motoneurons and sensory neurons started, and subsets of neurons were labelled by antibodies against EPOR. Viable neurons expressed EPO and EPOR. Up to E12.5, apoptotic cells were EPOR-immunopositive, but variably EPO-immunonegative or EPO-immunopositive. Thereafter, EPO-immunonegative and EPOR-immunopositive apoptotic cells predominated. Our findings suggest that EPO-mediated neuron-glial and, later, neuron-neuronal interactions promote the differentiation and/or the survival of subsets of neurons and glial cells in central as well as in peripheral parts of the embryonic nervous system. Correspondingly, expression of phospho-Akt-1/protein-kinase B extensively overlapped expression sites of EPO and EPOR, but was absent from apoptotic cells. Identified other sites of EPO and/or EPOR expression include radial glial cells that transform to astrocytes, cells of the floor plate and notochord as well as neural crest-derived boundary cap cells at motor exit points and cells of the primary sympathetic chain.

Animals↗
PubMed2000

[Influence of root module design on growth and development of plants].

Wheat crop was grown in various root modules (RM) in which nutritional solution was injected through porous membrane (2-6 microns). Seeds were laid on the surface of membrane or in pearlite covering the membrane. Root modules 1 and 3 contained ceramic porous tubes 10 mm and 22 mm in diameter, respectively. RM 2 was outfitted with a porous titanium plate, RM 4--with a porous ceramic tube (Ø 10 mm) buried in pearlite, and RM 5--with a porous titanium plate under the 2.5 cm thick layer of pearlite. The area of membrane surface per one plant was equal to 3 cm2 in RM 1 and about 17 cm2 RM 2 and 3. Pearlite volume per a plant made up about 40 cm2 in RM 4.5. The object of study was Triticum aestivum L., sp. Super Dwarf. The plants grew for 49 days under the white light of luminescent lamps at water potential (WP) = -0.4, -3.0 and -5.0 kPa. WP of pearlite equal, dry mass and anticipated productivity of the crop were much higher as compared to RM without pearlite. Significant reduction of these parameters was reflective of WP drop no matter the RM type. However, it was more expressed at -5 kPa. In RM filled with pearlite the mass and productivity of crop reduced at -5.0 kPa. Design of RM essentially altered the volumetric spread of roots. Thus, root math was formed immediately on flat porous plates, commonly below the bare tubes and both on top and on the bottom of RM with pearlite. These data can help development of RM for space greenhouses.

Botany↗
PubMed2001

Molecular cloning, characterization and expression analysis of a catalase cDNA from hot pepper (Capsicum annuum L.).

We report the cloning of a catalase cDNA from hot pepper (Capsicum annuum L.) and its expression patterns. CaCat1 is consisted of 1837 bp containing one open reading frame (ORF) of 1479 and 45 bp/313 bp of 5'/3'-untranslated region. Deduced amino acid sequence of CaCat1 showed the 95% and 78% identity with that of Nicotiana tabacum Cat1 and Nicotiana plumbaginifolia Cat2, respectively. Northern hybridization shows that CaCat1 transcripts are more abundant in stems than in leaves and roots, and in early stages than in mature stage of fruit development. In roots its transcripts are induced in response to aluminum and NaCl. In addition, its transcription levels under light (12 h)/dark (12 h) cycle and constant light conditions exhibit circadian rhythm, reaching a maximum at late in the dark period or early in the light period. The morning specific circadian regulation of CaCat1 was also observed in NpCat1, but not in NtCat1, suggesting difference in evolutionary rates between coding region and regulatory region of the catalase genes.

Journal Article↗
PubMed1996

Requirements for in vitro rooting of Quercus robur and Q. rubra shoots derived from mature trees.

Stabilized shoot cultures initiated from crown material of six adult Quercus robur L. trees and from basal epicormic shoots of a Quercus rubra L. tree showed good in vitro rooting capacity. An initial five-day dark period generally improved the rooting response but was detrimental to plantlet quality. There were clonal differences in rooting capacity. The concentration and exposure time of the indolebutyric acid (IBA) treatment were critical for root induction. In both species, best rooting efficiency was achieved by culture in medium containing 25 mg l(-1) IBA for 24 h and subsequent transfer to an auxin-free medium containing 1% activated charcoal. For all clones tested, the charcoal benefited both shoot quality and root system development, the latter being enhanced by the formation of many lateral roots. Total root system area and length, measured with a digital image analyzer, were significantly greater in medium containing charcoal than in medium lacking charcoal. Because darkening the basal part of the shoots with aluminum foil during the rooting phase only caused a small increase in rooting, we conclude that the large effect of charcoal on rooting was the result of adsorption of inhibitory compounds from the medium or the explant or both, rather than of basal darkening. Other factors affecting the rooting response of Q. robur were: (a) the position on the tree of the material from which cultures were initiated (the topophysical effect); and (b) shoot quality. Recycling the same horizontally placed explant on multiplication medium allowed three successive crops of shoots to be obtained, and rootability was typically maintained from crop to crop.

Journal Article↗
PubMed1999

Effect of salt stress on nodulation and nitrogen fixation in legumes.

It is now well established that almost all phases of root nodule development in legumes are adversely affected by saline conditions in the rooting medium. There is also a general agreement that the rhizobia are more tolerant to salt stress than the host plant, but they show considerable strain variability in growth and survival under saline conditions. Inhibitory effect of salinity on nodulation has been attributed to decrease in rhizobial colonisation and shrinkage and lack of root hair formation. Salt stress also induces premature senescence of already formed nodules. Both N2-fixation activity and nodule respiration are inhibited sharply on exposure of plants to saline conditions. The decrease in N2-fixation has been ascribed to direct effect on nitrogenase activity or an indirect effect through decrease in leghemoglobin content, respiratory rate, malate concentrations in nodules and photosynthate availability. Salinity increases oxygen diffusion resistance in the nodules and alters their ultrastructure. Decrease in N2-fixation in nodules under salinity is also accompanied by parallel decrease in the activity of H2O2-scavenging enzymes like catalase, ascorbate peroxidase and the level of antioxidants like ascorbic acid. Nodules appear to undergo osmoregulation under saline conditions by accumulating physiologically compatible solutes like proline, sugars (pinnitol) and lactic acid. The intensity of the adverse effects of salinity on nodule functioning depends on plant species, rhizobial strain, duration of exposure to saline conditions, nature, concentration and mode of salt application.

Fabaceae↗