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Putative inhibitory extracellular matrix molecules at the dorsal root entry zone of the spinal cord during development and after root and sciatic nerve lesions.

The dorsal root entry zone (DREZ) of the spinal cord is the interface between the central and peripheral nervous systems and is the pathway through which sensory afferents enter the central nervous system during development. However, in the rat, the DREZ becomes a boundary to regenerating sensory axons after Postnatal Days 2-3. The cellular and molecular mechanisms that cause regenerative failure at the DREZ after the critical period for regeneration are unknown. Recent studies demonstrate that two extracellular matrix molecules, Cytotactin/tenascin (CT) and chondroitin 6-sulfate-containing proteoglycans (C-6S-PG) are present in normal boundary regions of the brain and spinal cord during development. In the present study we sought to visualize the expression of these two putative inhibitory molecules in the DREZ of normally developing and adult animals, and also in animals after injury. CT and C-6S-PG spread laterally from the midline to the DREZ by Postnatal Day 3, correlating exactly with the end of the critical period. The staining intensity for these two molecules increases further in the DREZ after root lesions, but not sciatic lesions, at ages when axons cannot regenerate into the spinal cord. Following root lesion CT and C-6S-PG were mostly present in association with reactive glia at the DREZ and in white matter, rather than with reactive glia in grey matter of the dorsal horn, suggesting that astroglia are heterogeneous in their response to root lesion. The coexpression of CT and C-6S-PG may create a molecular barrier which might channel or deflect axons at the DREZ during CNS development and inhibit their growth during regeneration.

Aging↗

Transgenic proteoid roots of white lupin: a vehicle for characterizing and silencing root genes involved in adaptation to P stress.

White lupin (Lupinus albus L.) has become an illuminating model for the study of plant adaptation to phosphorus (P) deficiency. It adapts to -P stress with a highly coordinated modification of root development and biochemistry resulting in short, densely clustered secondary roots called proteoid (or cluster) roots. In order to characterize genes involved in proteoid root formation and function in a homologous system, we have developed an Agrobacterium rhizogenes-based transformation system for white lupin roots that allows rapid analysis of reporter genes as well as RNA interference (RNA(i))-based gene silencing. We used this system to characterize a lupin multidrug and toxin efflux (Lupinus albus MULTIDRUG AND TOXIN EFFLUX, LaMATE) gene previously shown to have enhanced expression under -P stress. Here, we show that LaMATE had high expression in proteoid roots not only under -P, but also under -Fe, -N, -Mn and +Al stress. A portion containing the putative LaMATE promoter was fused to GUS and enhanced green fluorescence protein (EGFP) reporter genes, and a translational LaMATE::EGFP fusion was constructed under control of the LaMATE promoter. The LaMATE promoter directed P-dependent GUS and EGFP expression to proteoid roots. Confocal microscopy in white lupin and Arabidopsis point to the plasma membrane as the likely location of the LaMATE protein. LaMATE displayed homology to FRD3 in Arabidopsis, but did not complement an Arabidopsis ferric reductase defective 3 (FRD3) mutant. RNA(i)-based gene silencing was shown to effectively reduce LaMATE expression in transformed white lupin roots. LaMATE RNAi-silenced plants displayed an about 20% reduction in dry weight.

Adaptation, Physiological↗

Distinct cell morphotypes of Aureobasidium melanogenum ZN exhibit differential functional profiles in promoting maize growth.

Black yeast-like fungi of the genus Aureobasidium exhibit morphological plasticity, but whether distinct cellular states within the same genetic background are associated with different plant growth-promoting functions remains unclear. Here, yeast-like cells (YL), swollen cells (SC), and chlamydospores (CH) of Aureobasidium melanogenum ZN were characterized. YL was associated mainly with siderophore production and laccase activity, SC with extracellular polysaccharide accumulation, and CH with phosphate mobilization and higher ammonia and IAA production. Whole-genome and comparative genomic analyses revealed a shared repertoire related to nutrient acquisition, auxin-associated metabolism, extracellular oxidation, and carbohydrate remodeling, with expansions in nutrient- and cell-surface-related gene families. Transcriptomic and metabolomic analyses showed distinct deployment of these capacities, with CH exhibiting broad reprogramming of tryptophan-associated, nitrogen, phosphate, central-carbon, and amino-acid metabolism. In maize, CH at the optimal inoculation concentration of 105 CFU·mL-1 produced the strongest growth promotion, increasing plant height, dry biomass, root length, root surface area, and root volume by 58.6%, 365.1%, 191.0%, 194.3%, and 222.4%, respectively. Consistent with this pronounced growth phenotype, maize root transcriptomics showed coordinated CH-induced responses involving root development, nutrient transport, redox regulation, and root-interface remodeling. Root-zone tracking showed greater short-term stability and persistence of CH. These findings identify cellular state as an important functional dimension of Aureobasidium-plant interactions and provide a basis for developing fungal inoculants with defined beneficial cellular states.

Zea mays↗

Prevalence of delayed emergence of permanent teeth as a result of local factors.

Delayed emergence of permanent teeth as a result of local etiologic factors was found in 45 of 1,032(4.3%) persons ages 8 through 18 in a dentally indigent population. In 36 persons (3.4%) delayed emergence resulted from causes other than those associated with premature primary molar extraction. This statistic probably approximates the potential for this problem in a treated population group. Teeth most commonly involved were mandibular second premolars, maxillary canines, and maxillary central incisors. The most common causes, respectively, were space loss, palatal position, and mesiodens. Supernumerary, malformed, and congenitally missing teeth were more frequent in persons with delayed emergence than in the rest of the population sampled. The difference was statistically significant. No correlation was found between delayed emergence and sex of the patient. In this study, teeth delayed in emergence behaved as other teeth in that they exhibited normal root development and did not contribute to resorption of adjacent roots. Except for maxillary canines, enlarged follicular spaces did not develop after eruption

Adolescent↗

Smear layer production by 3 rotary reamers with different cutting blade designs in straight root canals: a scanning electron microscopic study.

OBJECTIVES: The design of the cutting blade of rotary instruments may affect the outcome of root canal instrumentation in terms of cleanliness. The aim of this scanning electron microscopic study was to compare the quality and amount of smear layer generated in the apical third of straight root canals by 2 rotary nickel-titanium reamers and 1 rotary steel reamer with different cutting blade designs. STUDY DESIGN: Seventy intact, single-rooted human mandibular premolars with straight, fully developed roots were selected for this study. Before instrumentation, the cervical portion of all teeth was removed by using a microtome (Isomet), leaving 13-mm-long roots. Automated preparation was performed with ProFile (n = 20) and Hero 642 (n = 20) reamers by using the crown-down technique and with a stainless steel engine reamer (Mani; n = 20) by using a reaming motion. All root canals were instrumented to No. 40. A control group (pulp extirpation with barbed broaches; n = 10) was also included. Irrigation with 3 mL of a 1% sodium hypochlorite (NaOCl) solution was performed after each instrumentation. After the instrumentation, each root was split longitudinally, and a scanning electron microscope was used to examine the selected areas of the canal walls at the apical third from 2 different perspectives. A 4-category scoring system for smear layer was used, and the resulting scores were statistically analyzed. RESULTS: The least smear layer remained in the Hero 642 group at the selected apical third of straight root canals (P < .05). However, all instruments left a smear layer. The surface texture of the smear layer, in addition to the depth and the frequency of packed materials into the dentinal tubules, varied with instrument type. CONCLUSION: These data revealed that the design of the cutting blade of rotary instruments can affect root canal cleanliness in straight root canals. This information may be useful in the selection of nickel-titanium rotary reamers.

Dental Alloys↗

A prospective study of factors affecting survival of replanted permanent incisors in children.

OBJECTIVE: To examine the factors associated with the onset of resorption in replanted avulsed permanent incisor teeth and to demonstrate their patterns of survival free of resorption. SETTING: Department of Paediatric Dentistry, Belfast. DESIGN: Prospective recording of the timing of the injury and of replantation, the storage media, the degree of root development and contamination, and the completion of root treatment. Clinical and radiographic reviews at 3-month intervals. Root resorption classified as replacement and inflammatory. Logistic regression and survival curves with freedom from resorption as outcomes. RESULTS: In 50 avulsed incisors in children aged 6-16 years the best predictor of overall resorption was total time of dryness, and that for replacement resorption was total extra-oral time. The timing of detection of resorption varied from 102 days to 997 days. CONCLUSION: Both total extra-oral time and time stored dry are important factors for the onset of resorption in replanted avulsed teeth in children.

Adolescent↗

Developmental expression and biochemical analysis of the Arabidopsis atao1 gene encoding an H2O2-generating diamine oxidase.

A copper amine oxidase encoding gene, atao1, has been isolated and characterized from Arabidopsis thaliana. Sequence analysis reveals that atao1 encodes a 668 amino acid polypeptide (ATAO1) with 48% identity to copper amine oxidases from pea and lentil. The promoter region of atao1 was transcriptionally fused with the reporter genes encoding beta-glucuronidase and modified green fluorescent protein. Analysis of transgenic Arabidopsis together with in situ hybridization of wild-type plants reveals temporally and spatially discrete patterns of gene expression in lateral root cap cells, vascular tissue of roots, developing leaves, the hypocotyl, and in the style/stigmatal tissue. Enzyme activity assays show that ATAO1 preferentially oxidizes the aliphatic diamine putrescine with production of the corresponding aldehyde, ammonia and hydrogen peroxide, a recognized plant signal molecule and substrate for peroxidases. Histochemical analysis reveals that atao1 expression in developing tracheary elements precedes and overlaps with lignification and therefore is a good marker for vascular development. In both vascular tissue and the root cap, atao1 expression occurs in cells destined to undergo programmed cell death.

Amine Oxidase (Copper-Containing)↗

Genetic ablation of root cap cells in Arabidopsis.

The root cap is increasingly appreciated as a complex and dynamic plant organ. Root caps sense and transmit environmental signals, synthesize and secrete small molecules and macromolecules, and in some species shed metabolically active cells. However, it is not known whether root caps are essential for normal shoot and root development. We report the identification of a root cap-specific promoter and describe its use to genetically ablate root caps by directing root cap-specific expression of a diphtheria toxin A-chain gene. Transgenic toxin-expressing plants are viable and have normal aerial parts but agravitropic roots, implying loss of root cap function. Several cell layers are missing from the transgenic root caps, and the remaining cells are abnormal. Although the radial organization of the roots is normal in toxin-expressing plants, the root tips have fewer cytoplasmically dense cells than do wild-type root tips, suggesting that root meristematic activity is lower in transgenic than in wild-type plants. The roots of transgenic plants have more lateral roots and these are, in turn, more highly branched than those of wild-type plants. Thus, root cap ablation alters root architecture both by inhibiting root meristematic activity and by stimulating lateral root initiation. These observations imply that the root caps contain essential components of the signaling system that determines root architecture.

Arabidopsis↗

The xylem of rice (Oryza sativa) is colonized by Azorhizobium caulinodans.

Following inoculation with Azorhizobium caulinodans ORS571 (pXLGD4), lateral root development of rice and colonization of lateral root cracks by bacteria were shown to be stimulated by the flavonoid naringenin. Rice seedlings growing aseptically in the presence of naringenin were inoculated with ORS571 (pXLGD4), carrying the lacZ reporter gene. By microscopic analysis of sections of inoculated rice roots, it has been demonstrated that the xylem of rice roots can be colonized by Azorhizobium caulinodans. We discuss whether this colonization of the xylem of rice roots by azorhizobia could provide a suitable niche for endophytic nitrogen fixation.

Azorhizobium caulinodans↗

Efficient virus-induced gene silencing in roots using a modified tobacco rattle virus vector.

Due to their capability of eliciting a form of posttranscriptional gene silencing (termed virus-induced gene silencing or VIGS), plant viruses are increasingly used as reverse-genetics tools for functional characterization of plant genes. RNA viruses have been shown to trigger silencing in a variety of host plants, including members of Solanacae and Arabidopsis (Arabidopsis thaliana). Several factors affect the silencing response, including host range and viral tropism within the plant. The work presented here demonstrates that a modified tobacco rattle virus (TRV) vector retaining the helper protein 2b, required for transmission by a specific vector nematode, not only invades and replicates extensively in whole plants, including meristems, but also triggers a pervasive systemic VIGS response in the roots of Nicotiana benthamiana, Arabidopsis, and tomato (Lycopersicon esculentum). This sustained VIGS response was exemplified by the silencing of genes involved in root development (IRT1, TTG1 [transparent testa glabra], RHL1 [root hairless1], and beta-tubulin), lateral root-meristem function (RML1 [root meristemless1]), and nematode resistance (Mi). Roots of silenced plants exhibit reduced levels of target mRNA and phenocopy previously described mutant alleles. The TRV-2b vector displays increased infectivity and meristem invasion, both key requirements for efficient VIGS-based functional characterization of genes in root tissues. Our data suggest that the TRV helper protein 2b may have an essential role in the host regulatory mechanisms that control TRV invasion.

Animals↗

Higher activity of an aldehyde oxidase in the auxin-overproducing superroot1 mutant of Arabidopsis thaliana.

Aldehyde oxidase (AO; EC 1.2.3.1) activity was measured in seedlings of wild type or an auxin-overproducing mutant, superroot1 (sur1), of Arabidopsis thaliana. Activity staining for AO after native polyacrylamide gel electrophoresis separation of seedling extracts revealed that there were three major bands with AO activity (AO1-3) in wild-type and mutant seedlings. One of them (AO1) had a higher substrate preference for indole-3-aldehyde. This AO activity was significantly higher in sur1 mutant seedlings than in the wild type. The difference in activity was most apparent 7 d after germination, the same time required for the appearance of the remarkable sur1 phenotype, which includes epinastic cotyledons, elongated hypocotyls, and enhanced root development. Higher activity was observed in the root and hypocotyl region of the mutant seedlings. We also assayed the indole-3-acetaldehyde oxidase activity in extracts by high-performance liquid chromatography detection of indole-3-acetic acid (IAA). The activity was about 5 times higher in the extract of the sur1 seedlings, indicating that AO1 also has a substrate preference for abscisic aldehyde. Treatment of the wild-type seedlings with picloram or IAA caused no significant increase in AO1 activity. This result suggested that the higher activity of AO1 in sur1 mutant seedlings was not induced by IAA accumulation and, thus, strongly supports the possible role of AO1 in IAA biosynthesis in Arabidopsis seedlings.

Aldehyde Oxidase↗

Seasonal adaptations of the tuberous roots of Ranunculus asiaticus to desiccation and resurrection by changes in cell structure and protein content.

The annual developmental cycle of tuberous roots of Ranunculus asiaticus was studied with respect to structure and content of their cells, to understand how these roots are adapted to desiccation, high temperature and rehydration. Light microscopy, histochemical analysis, and protein analyses by SDS-PAGE were employed at eight stages of annual root development. During growth and maturation of the roots, cortical cells increased in size and their cell walls accumulated pectin materials in a distinct layer to the inside of the primary walls, with pits between adjoining cells. The number of starch granules and protein bodies also increased within the cells. Several discrete proteins accumulated. Following quiescence and rehydration of the roots there was a loss of starch and proteins from the cells, and cell walls decreased in thickness. The resurrection geophyte R. asiaticus possesses desiccation-tolerant annual roots. They store carbon and nitrogen reserves within their cells, and pectin within the walls to support growth of the plant following summer quiescence and rehydration.

Adaptation, Physiological↗

Heavy metals in white lupin: uptake, root-to-shoot transfer and redistribution within the plant.

The translocation of manganese (Mn), nickel (Ni), cobalt (Co), zinc (Zn) and cadmium (Cd) in white lupin (Lupinus albus cv. Amiga) was compared considering root-to-shoot transport, and redistribution in the root system and in the shoot, as well as the content at different stages of cluster roots and in other roots. To investigate the redistribution of these heavy metals, lupin plants were labelled via the root for 24 h with radionuclides and subsequently grown hydroponically for several weeks. 54Mn, 63Ni and 65Zn were transported via the xylem to the shoot. 63Ni and 65Zn were redistributed afterwards via the phloem from older to younger leaves, while 54Mn remained in the oldest leaves. A strong retention in the root was observed for 57Co and 109Cd. Cluster roots contained higher concentrations of all heavy metals than noncluster roots. Concentrations were generally higher at the beginning of cluster root development (juvenile and immature stages). Mature cluster roots also contained high levels of 54Mn and 57Co, but only reduced concentrations of 63Ni, 65Zn and 109Cd.

Autoradiography↗

Idiopathic, late, slow, and irregular dental development: report of case.

Although reports of generalized idiopathic diminished root formation have been identified by a few authors, only one such case has been associated with the condition of Stevens-Johnson syndrome. The present clinical report provides additional evidence to implicate the condition of Stevens-Johnson syndrome in a case of generalized abnormal root development.

Child, Preschool↗

Effects of aluminum on growth and nutrient status of Douglas-fir seedlings grown in culture solution.

Effects of Al on growth, nutrient uptake and proton efflux were studied in Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedlings grown for about nine months in culture solutions with a pH between 3.4 and 3.6 and with both calcium and magnesium (Ca + Mg) at a concentration of 0.1, 0.5 or 2.5 mM. In the absence of Al, plant dry matter production and root development increased with increasing concentrations of (Ca + Mg) in the culture solution. At the low and intermediate (Ca + Mg) concentrations, optimal root and shoot development were observed at an Al concentration of 4 mg l(-1). At the highest (Ca + Mg) concentration, Al up to 4 mg l(-1) did not affect growth, but at higher concentrations, it significantly reduced both shoot and root growth. As the concentration of (Ca + Mg) in the nutrient solution increased, the concentrations of Ca and Mg increased in shoots and roots also. The concentrations of Ca and Mg in the roots were unaffected by the presence of Al, whereas in the shoots they were either unaffected, or increased, by Al. Concentrations of Al in, or on, roots, or in shoots, did not change in response to changing concentrations of Ca + Mg in the nutrient medium. In general, concentrations of P and K in shoots and roots were higher in seedlings grown in nutrient solutions containing Al. Stimulation of growth by moderate concentrations of Al, which was observed at suboptimal (Ca + Mg) concentrations, was associated with a low NH(4) preference and a low root proton efflux. The Al-induced increase in internal K concentration and reduction in NH(4) preference may be explained by a lower efflux of K and NO(3), respectively.

Journal Article↗

An auxin transport independent pathway is involved in phosphate stress-induced root architectural alterations in Arabidopsis. Identification of BIG as a mediator of auxin in pericycle cell activation.

Arabidopsis (Arabidopsis thaliana) plants display a number of root developmental responses to low phosphate availability, including primary root growth inhibition, greater formation of lateral roots, and increased root hair elongation. To gain insight into the regulatory mechanisms by which phosphorus (P) availability alters postembryonic root development, we performed a mutant screen to identify genetic determinants involved in the response to P deprivation. Three low phosphate-resistant root lines (lpr1-1 to lpr1-3) were isolated because of their reduced lateral root formation in low P conditions. Genetic and molecular analyses revealed that all lpr1 mutants were allelic to BIG, which is required for normal auxin transport in Arabidopsis. Detailed characterization of lateral root primordia (LRP) development in wild-type and lpr1 mutants revealed that BIG is required for pericycle cell activation to form LRP in both high (1 mm) and low (1 microm) P conditions, but not for the low P-induced alterations in primary root growth, lateral root emergence, and root hair elongation. Exogenously supplied auxin restored normal lateral root formation in lpr1 mutants in the two P treatments. Treatment of wild-type Arabidopsis seedlings with brefeldin A, a fungal metabolite that blocks auxin transport, phenocopies the root developmental alterations observed in lpr1 mutants in both high and low P conditions, suggesting that BIG participates in vesicular targeting of auxin transporters. Taken together, our results show that auxin transport and BIG function have fundamental roles in pericycle cell activation to form LRP and promote root hair elongation. The mechanism that activates root system architectural alterations in response to P deprivation, however, seems to be independent of auxin transport and BIG.

Arabidopsis↗

The transcription factor NO TRANSMITTING TRACT/WIP2 modulates cytokinin homeostasis in Arabidopsis.

The transcription factor WIP2/NO TRANSMITTING TRACT (WIP2/NTT) belongs to the WIP zinc finger family. Loss of WIP/NTT function in Arabidopsis thaliana causes alterations in specific tissues in the gynoecium. It also impairs root development, but only when combined with the loss of WIP4 and WIP5 function, due to redundancy. Certain mutant loss-of-function phenotypes can be recovered by cytokinin application, NTT interacts with cytokinin signaling components, and the phenotypes displayed by plants with increased WIP2/NTT expression also suggest a possible interaction with this pathway. Therefore, the objective of this study was to investigate the relationship between WIP2/NTT and the cytokinin pathway. To overcome the issue of genetic redundancy, we used a commonly used inducible system. We found that WIP2/NTT induction alters cytokinin levels and signaling in a tissue-specific manner, as shown by cytokinin content measurements and TCSn::GFP reporter analysis. Transcriptome analyses revealed candidate target genes related to the cytokinin pathway. Yeast one-hybrid and transactivation assays demonstrated direct NTT binding to regulatory regions of the cytokinin genes ISOPENTENYL TRANSFERASE 5 (IPT5), ARABIDOPSIS HISTIDINE PHOSPHOTRANSFER PROTEIN 6 (AHP6), and CYTOKININ OXIDASE/DEHYDROGENASE 7 (CKX7) involved in cytokinin biosynthesis, signaling, and degradation, respectively. Moreover, immunolocalization assays revealed that cytokinin distribution was altered in loss of function mutants and after NTT induction. The results of this work indicate that WIP2/NTT modulates cytokinin homeostasis.

Cytokinins↗

Cell ablation studies in plant development.

As an alternative and complementary approach to genetic targeted ablation, laser ablation has been applied to a variety of plant systems to investigate the basics of development in plants. The recent relevant literature is reviewed and provides concrete examples of how root development in Arabidopsis and cell polarization in Fucus have been studied using laser microsurgery. Guidelines are proposed for researchers who would like to apply laser ablation techniques to a given developmental problem.

Arabidopsis↗