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The maize actin-depolymerizing factor, ZmADF3, redistributes to the growing tip of elongating root hairs and can be induced to translocate into the nucleus with actin.

The maize actin depolymerizing factor, ZmADF3, binds G- and F-actin, and increases in vitro actin dynamics. Polyclonal antibodies have been raised against ZmADF3 and these detect a single band of approximately 17 kDa in all maize tissues examined, with the exception of pollen. In the development of root hairs, the distribution of ZmADF3 is related to actin reorganization. In the early stages of hair development, ZmADF3 is distributed throughout the cytoplasm. As the hair emerges and the microfilament bundles redirect to the outgrowth there is a simultaneous redistribution of ZmADF3 which now concentrates at the tip of the emerging hair and remains in this position as elongation proceeds. These observations show that ZmADF3 localizes to a region where actin is being remodelled during tip growth. After cytochalasin D treatment which disrupts actin filaments, short rods of ZmADF3 and actin appear in the nucleus suggesting that ZmADF3 may function by guiding actin to sites of actin polymerization.

Actin Depolymerizing Factors↗

Root establishment of perennial ryegrass (L. perenne) in diesel contaminated subsurface soil layers.

The efficiency of rhizosphere biodegradation of petroleum hydrocarbons heterogeneously distributed in soils is dependent on the ability of plant roots to prospect into contaminated zones. Rhizobox experiments were conducted to study the influence of diesel contaminated layers on the spatial distribution and the development of the roots of perennial ryegrass. Root distribution and root and shoot development were monitored over time. The final root and above ground biomass and the final TPH concentration were determined. The spatial distribution of the contaminant as well as the irrigation method used affected root distribution, plant development and TPH degradation and therefore ryegrass remediation potential. The results show that roots colonise fully uncontaminated soil and grow preferentially between zones of contamination. Conversely, when no immediate uncontaminated soil is available, roots grow through contaminated zones in order to prospect for uncontaminated soil.

Biodegradation, Environmental↗

Short root mutant of Lotus japonicus with a dramatically altered symbiotic phenotype.

Legume plants carefully control the extent of nodulation in response to rhizobial infection. To examine the mechanism underlying this process we conducted a detailed analysis of the Lotus japonicus hypernodulating mutants, har1-1, 2 and 3 that define a new locus, HYPERNODULATION ABERRANT ROOT FORMATION (Har1), involved in root and symbiotic development. Mutations in the Har1 locus alter root architecture by inhibiting root elongation, diminishing root diameter and stimulating lateral root initiation. At the cellular level these developmental alterations are associated with changes in the position and duration of root cell growth and result in a premature differentiation of har1-1 mutant root. No significant differences between har1-1 mutant and wild-type plants were detected with respect to root growth responses to 1-aminocyclopropane1-carboxylic acid, the immediate precursor of ethylene, and auxin; however, cytokinin in the presence of AVG (aminoetoxyvinylglycine) was found to stimulate root elongation of the har1-1 mutant but not the wild-type. After inoculation with Mesorhizobium loti, the har1 mutant lines display an unusual hypernodulation (HNR) response, characterized by unrestricted nodulation (hypernodulation), and a concomitant drastic inhibition of root and shoot growth. These observations implicate a role for the Har1 locus in both symbiotic and non-symbiotic development of L. japonicus, and suggest that regulatory processes controlling nodule organogenesis and nodule number are integrated in an overall mechanism governing root growth and development.

Mutation↗

Detection of hunter heterozygotes by enzymatic analysis of hair roots.

We have developed a procedure for testing iduronate sulfatase, the enzyme deficient in Hunter syndrome, in single hair roots. Beta-Hexosaminidase was used as the reference enzyme. The ratio of iduronate sulfatase to beta--hexosaminidase, expressed in arbitrary units of activity, is near zero for Hunter patients and greater than 0.6 in almost all roots of normal individuals. Hair roots of Hunter heterozygotes show a characteristic continuum of activity ratios, ranging from totally deficient up to and including the normal range. The results are consistent with the origin of hair roots from a small number of progenitor cells which obey the Lyon hypothesis. The proportion of roots with low activity can be used to discriminate between normal and heterozygous individuals.

Adolescent↗

The binding pattern of peanut lectin associated with sclerotome migration and the formation of the vertebral axis in the chick embryo.

Lectins have been used extensively to detect changes in carbohydrate moieties on the surfaces of embryonic cells during early development. Peanut agglutinin (PNA) in particular has been used to investigate changes related to cell differentiation. PNA has also been used to differentiate between the rostral and caudal sclerotome halves which have been shown to be functionally different, with neural crest cells and neurites traversing only the rostral half during their migration. In this study, we have sectioned and stained chick embryos between 3 and 8 days of age with PNA to examine the distribution of PNA binding sites associated with the vertebral column during this period and also to determine the fates of the rostral and caudal sclerotome halves. Ultrastructural localisation of PNA-gold conjugate showed that binding sites for this lectin were present intracellularly and extracellularly both on cell surfaces and in the matrix. At the light microscope level, a clear banding pattern emerged after staining with PNA which consisted of alternating light and dark staining along the entire length of the vertebral axis of the embryo. In the younger embryos, a simple banding pattern emerged where the rostral sclerotome half of each segment stained only lightly while the caudal half stained darkly. This banding pattern was present throughout the 6 day period of development and could be traced continuously but grew more complex as the sclerotome cells migrated to surround the notochord and neural tube and as the dorsal root ganglia developed. The rostral sclerotome half was found to contribute to the caudal part of one vertebral body and its neural arch, while the caudal sclerotome half was found to contribute to the intervertebral disc, the rostral half of the next caudal vertebra, and part of its neural arch.

Animals↗

Developmental regulation of the gibberellin biosynthetic gene GA1 in Arabidopsis thaliana.

The GA1 gene of Arabidopsis thaliana encodes ent-kaurene synthase A (KSA), which catalyzes the first committed step in the biosynthetic pathway of the plant hormone gibberellin (GA). Its location in the GA biosynthetic pathway has led to speculation that KSA regulation is one of the controlling steps. However, because KSA activity is so low that it is only measurable in Arabidopsis siliques, GA1 promoter-GUS reporter gene fusions and quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) were used to examine the expression pattern of GA1. The results from this study indicate that GA1 gene expression is highly regulated during growth and development, and it is restricted to specific cell types at the sites of expression. GA1 promoter activity is highest in rapidly growing tissues, e.g. shoot apicas, root tips, developing flowers and seeds. It is also active in the vascular tissue of some non-growing organs, such as expanded leaves, suggesting that these leaves may be a site of GA synthesis for transport to other organs. It was also found that the first one or two introns in the GA1 gene are required for proper expression. Because of the high degree of regulation, GA1 may act as a gatekeeper, controlling the flow of metabolites into the GA biosynthetic pathway, while the levels of specific bioactive GAs are controlled by other downstream steps.

Alkyl and Aryl Transferases↗

Native valve endocarditis due to Corynebacterium group JK.

We report a case of a 32-yr-old woman on chronic intermittent haemodialysis, who developed endocarditis due to a Corynebacterium group JK, involving both the native aortic and mitral valves. Despite a four-week treatment with vancomycin, an aortic root abscess developed. The diagnosis was confirmed on autopsy.

Adult↗

Root-surface caries formation: effect of in vitro APF treatment.

To determine the effect of one and four-minute acidulated phosphate fluoride, or APF (Oral-B Minute Gel, Oral-B Laboratories) treatments on artificial caries development in root surfaces, an in vitro study was conducted. The authors created a window of exposed root surface on extracted teeth by applying acid-resistant varnish to the remaining surface. APF treatment was administered and followed by a 24-hour deionized-distilled water rinse. A window of sound root surface that had been protected from the APF treatment was exposed and served as a control. Acidified gel was then applied to the exposed root surfaces to create the artificial caries. The authors found that APF treatment significantly enhanced resistance to demineralization of root surfaces from an in vitro acidogenic challenge. However, there was little difference in effectiveness between the one- and four-minute APF treatment times.

Acidulated Phosphate Fluoride↗

Inhibitory role of root hairs on transport within root culture bioreactors.

An experimental system was developed to produce root cultures of Hyoscyamus muticus with and without the profuse root hairs. Growth in the presence of 7.6 microM pyrene butyric acid (PBA) and 2.2 mM phosphate virtually eliminated root hairs, whereas growth rate, general morphology and nutrient yields remained unchanged in well-mixed flask culture. These root cultures were used to demonstrate decreased flow resistance in a tubular reactor as a result of root hair removal. To assess the impact on bioreactor performance, hairy and hairless root cultures were grown in a highly characterized 15-L bubble column bioreactor. In the absence of root hairs, the mixing was greatly enhanced; mixing times became shorter for the hairless culture at roughly 100 g (fresh weight)/L. By the end of the 3-week culture period, the mixing time of the hairy culture was 29 times longer than that of the hairless culture. The growth rate of the hairless culture in the bioreactor was as much as 2.4 times greater than growth of the hairy culture under the same conditions. The improved reactor performance was reflected in greater biomass accumulation and respiratory activity. These results show that the root hairs-which facilitate nutrient uptake in a static soil environment-are detrimental to growth in a liquid environment as an effect of their stagnating fluid flow and limiting oxygen availability.

Biological Transport↗

Does ethylene mediate cluster root formation under iron deficiency?

Casuarina glauca develops proteoid (cluster) roots in response to Fe deficiency. This study set out to investigate the possible involvement of ethylene in the initiation and/or the morphogenesis of cluster roots (CR). For this purpose, the effect of Ag+ added as silver thiosulfate, an inhibitor of ethylene action has been studied in plants growing hydroponically. No CR formation was observed in these growth conditions. Inhibition of ethylene biosynthesis by aminoethoxyvinylglycine, 1- aminoisobutyric acid, aminoxyacetic acid or cobalt chloride also eliminated the positive effect of Fe deficiency on CR formation in C. glauca. CR were not formed in Fe- deficient roots in the presence of ethylene inhibitors, suggesting a role for ethylene in the morphological responses to Fe deficiency. Interestingly, treatment of Casuarina plants with the ethylene precursor 1-aminocyclopropane-1-carboxylic acid stimulated significantly the formation of CR, even if plants are supplied with Fe. However, this stimulation did not reach the level of CR obtained in Fe-deficient plants. These results suggest that an ethylene-mediated signalling pathway is involved in CR formation process in C. glauca.

Chlorides↗

The Runx3 transcription factor regulates development and survival of TrkC dorsal root ganglia neurons.

The RUNX transcription factors are important regulators of linage-specific gene expression in major developmental pathways. Recently, we demonstrated that Runx3 is highly expressed in developing cranial and dorsal root ganglia (DRGs). Here we report that within the DRGs, Runx3 is specifically expressed in a subset of neurons, the tyrosine kinase receptor C (TrkC) proprioceptive neurons. We show that Runx3-deficient mice develop severe limb ataxia due to disruption of monosynaptic connectivity between intra spinal afferents and motoneurons. We demonstrate that the underlying cause of the defect is a loss of DRG proprioceptive neurons, reflected by a decreased number of TrkC-, parvalbumin- and beta-galactosidase-positive cells. Thus, Runx3 is a neurogenic TrkC neuron-specific transcription factor. In its absence, TrkC neurons in the DRG do not survive long enough to extend their axons toward target cells, resulting in lack of connectivity and ataxia. The data provide new genetic insights into the neurogenesis of DRGs and may help elucidate the molecular mechanisms underlying somatosensory-related ataxia in humans.

Afferent Pathways↗

Patterns of dental development in Homo, Australopithecus, Pan, and Gorilla.

Smith ([1986] Nature 323:327-330) distinguished patterns of development of teeth of juvenile fossil hominids as being "more like humans" or "more like apes" based on statistical similarity to group standards. Here, this central tendency discrimination (CTD) is tested for its ability to recognize ape and human patterns of dental development in 789 subadult hominoids. Tooth development of a modern human sample (665 black southern Africans) was scored entirely by an outside investigator; pongid and fossil hominid samples (59 Pan, 50 Gorilla, and 14 fossil hominids) were scored by the author. The claim of Lampl et al. ([1993] Am. J. Phys. Anthropol. 90:113-127) that Smith's 1986 method succeeds in only 8% of human cases was not sustained. Figures for overall success of classification (87% humans, 68% apes) mask important effects of teeth sampled and age class. For humans, the power of CTD varied between 53% and 92% depending on the number and kind of teeth available--nearly that of a coin toss when data described only two nearby teeth, but quite successful with more teeth or distant teeth. For apes, only age class affected accuracy: "Infant" apes (M1 development < or = root cleft complete, unemerged) were usually classed as humans, probably because the present developmental standard for great apes is in substantial error under 3 years of age. "Juvenile" apes (M1 > or = root 1/4), however, were correctly discriminated in 87% of cases. Overall, CTD can be considered reliable (accuracy of 92% for humans and 88% for apes) when data contrast development of distant dental fields and subjects are juveniles (not infants). Restricting analysis of fossils to specimens satisfying these criteria, patterns of dental development of gracile australopithecines and Homo habilis remain classified with African apes. Those of Homo erectus and Neanderthals are classified with humans, suggesting that patterns of growth evolved substantially in the Hominidae. To standardize future research, the computer program that operationalizes CTD is now available.

Aging↗

Response of bladder, urethral and intracavernous pressure to ventral lumbosacral root stimulation in Sprague-Dawley and Wistar rats.

Six Sprague-Dawley and six Wistar rats were used for electrostimulation of the L5 to S2 ventral roots. Landmarks for identification of the roots were developed; bladder, urethral and intracavernous pressures were recorded; and tail and leg movements were checked. Urethral sphincter contraction was elicited by stimulation of the L5-L6 ventral roots, while bladder contraction and penile erection were mediated by the L6-S1 ventral roots. The best sphincteric response and intracavernous pressure rise were obtained by stimulation of the L6 ventral root, and the highest bladder pressures by stimulation of the S1 ventral root. Stimulation of the S1-S2 ventral roots provoked ipsilateral tail movement; of L6, tail movement, hindleg muscle twitch, and slight toe spread; and of L5, hindleg stretch and plantar flexion. No significant differences were found between the two strains of rats, although a higher bladder pressure was recorded during stimulation of the L6 ventral root in Sprague-Dawley rats, which might be explained by a small caudal shift of the sacral parasympathetic nucleus in the Wistar strain.

Animals↗

Transduction of an Ethylene Signal Is Required for Cell Death and Lysis in the Root Cortex of Maize during Aerenchyma Formation Induced by Hypoxia.

Ethylene has been implicated in signaling cell death in the lysigenous formation of gas spaces (aerenchyma) in the cortex of adventitious roots of maize (Zea mays) subjected to hypoxia. Various antagonists that are known to modify particular steps in signal transduction in other plant systems were applied at low concentrations to normoxic and hypoxic roots of maize, and the effect on cell death (aerenchyma formation) and the increase in cellulase activity that precedes the appearance of cell degeneration were measured. Both cellulase activity and cell death were inhibited in hypoxic roots in the presence of antagonists of inositol phospholipids, Ca2+- calmodulin, and protein kinases. By contrast, there was a parallel promotion of cellulase activity and cell death in hypoxic and normoxic roots by contact with reagents that activate G-proteins, increase cytosolic Ca2+, or inhibit protein phosphatases. Most of these reagents had no effect on ethylene biosynthesis and did not arrest root extension. These results indicate that the transduction of an ethylene signal leading to an increase in intracellular Ca2+ is necessary for cell death and the resulting aerenchyma development in roots of maize subjected to hypoxia.

Journal Article↗

Chemical composition and ultrastructure of broad bean (Vicia faba L.) nodule endodermis in comparison to the root endodermis.

Ultrastructure and development of apoplastic barriers within indeterminate root nodules formed by Vicia faba L. were examined by light and electron microscopy. The nodule outer cortex is separated from the inner cortex by a heavily suberized nodule endodermis, which matures in submeristematic regions and possesses suberin lamellae. Unsuberized passage cells are present near vascular strands, which are surrounded by a vascular endodermis attached on the inner side of the nodule endodermal cell walls. The vascular endodermis appears immediately below the meristematic apex in developmental state I (Casparian bands), gradually develops suberin lamellae, and attains developmental state II at the base of the nodule. For chemical analysis apoplastic barrier tissues were dissected after enzymatic digestion of non-impregnated tissues. Root epidermal and endodermal cell walls as well as nodule outer cortex could be isolated as pure fractions; nodule endodermal cell walls could not be separated from vascular endodermal cell walls and enclosed xylem vessels. Gas chromatography-flame ionization detection and gas chromatography-mass spectrometry were applied for quantitative and qualitative analysis of suberin and lignin in isolated cell walls of these tissues. The suberin content of isolated endodermal cell walls of nodules was approximately twice that of the root endodermal cell walls. The suberin content of the nodule outer cortex and root epidermal cell walls was less than one-tenth of that of the nodule endodermal cell wall. Substantial amounts of lignin could only be found in the nodule endodermal cell wall fraction. Organic solvent extracts of the isolated tissues revealed long-chain aliphatic acids, steroids, and triterpenoid structures of the lupeol type. Surprisingly, extract from the outer cortex consisted of 89% triterpenoids whereas extracts from all other cell wall isolates contained not more than 16% total triterpenoids. The results of ultrastructural and chemical composition are in good correspondence and underline the important role of the examined tissues as apoplastic barriers.

Biopolymers↗

Developmental anatomy and branching of roots of four Zeylanidium species (podostemaceae), with implications for evolution of foliose roots.

Podostemaceae have markedly specialized and diverse roots that are adapted to extreme habitats, such as seasonally submerged or exposed rocks in waterfalls and rapids. This paper describes the developmental anatomy of roots of four species of Zeylanidium, with emphasis on the unusual association between root branching and root-borne adventitious shoots. In Z. subulatum and Z. lichenoides with subcylindrical or ribbon-like roots, the apical meristem distal (exterior) to a shoot that is initiated within the meristem area reduces and loses meristematic activity. This results in a splitting into two meristems that separate the parental root and lateral root (anisotomous dichotomy). In Z. olivaceum with lobed foliose roots, shoots are initiated in the innermost zone of the marginal meristem, and similar, but delayed, meristem reduction usually occurs, producing a parenchyma exterior to shoots located between root lobes. In some extreme cases, due to meristem recovery, root lobing does not occur, so the margin is entire. In Z. maheshwarii with foliose roots, shoots are initiated proximal to the marginal meristem and there is no shoot-root lobe association. Results suggest that during evolution from subcylindrical or ribbon-like roots to foliose roots, reduction of meristem exterior to a shoot was delayed and then arrested as a result of inward shifting of the sites of shoot initiation. The evolutionary reappearance of a protective tissue or root cap in Z. olivaceum and Z. maheshwarii in the Zeylanidium clade is implied, taking into account the reported molecular phylogeny and root-cap development in Hydrobryum.

Biological Evolution↗

25S rDNA-based molecular monitoring of glomalean fungi in sewage sludge-treated field plots.

Recycling of sewage wastes in agriculture is likely to affect the biological activity of soils through contamination of ecosystems by pathogens and metallic or organic micropollutants. The impact of sewage sludge spreading under field conditions on arbuscular mycorrhiza (AM) formation by a community of glomalean fungi was evaluated using a nested polymerase chain reaction (PCR) and discriminating primers based on 25S rDNA polymorphisms to detect different fungal species within root systems. Medicago truncatula was grown in soil of field plots amended or not with a composted sewage sludge, spiked or not with organic or metallic micropollutants. Overall AM development in roots decreased with sewage sludge application, and the relative abundance of five AM fungal morphotypes in root fragments was modified by the input of composted sludges. Sewage sludge spiked or not with organic pollutants had a generally positive effect on the relative diversity of AM fungal populations in planta, whereas after spreading of the sludge spiked with metallic pollutants, no variation was observed in the abundance of different species.

Agriculture↗