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Dual pathways for regulation of root branching by nitrate.

Root development is extremely sensitive to variations in nutrient supply, but the mechanisms are poorly understood. We have investigated the processes by which nitrate (NO3-), depending on its availability and distribution, can have both positive and negative effects on the development and growth of lateral roots. When Arabidopsis roots were exposed to a locally concentrated supply of NO3- there was no increase in lateral root numbers within the NO3--rich zone, but there was a localized 2-fold increase in the mean rate of lateral root elongation, which was attributable to a corresponding increase in the rate of cell production in the lateral root meristem. Localized applications of other N sources did not stimulate lateral root elongation, consistent with previous evidence that the NO3- ion is acting as a signal rather than a nutrient. The axr4 auxin-resistant mutant was insensitive to the stimulatory effect of NO3-, suggesting an overlap between the NO3- and auxin response pathways. High rates of NO3- supply to the roots had a systemic inhibitory effect on lateral root development that acted specifically at the stage when the laterals had just emerged from the primary root, apparently delaying final activation of the lateral root meristem. A nitrate reductase-deficient mutant showed increased sensitivity to this systemic inhibitory effect, suggesting that tissue NO3- levels may play a role in generating the inhibitory signal. We present a model in which root branching is modulated by opposing signals from the plant's internal N status and the external supply of NO3-.

Ammonium Chloride↗

Functional characterization of NtCDPK1 in tobacco.

We previously showed that NtCDPK1, a tobacco cal-cium-dependent protein kinase, interacts with and phosphorylates the Rpn3 regulatory subunit of the 26S proteasome, and that both NtCDPK1 and Rpn3 are mainly expressed in rapidly proliferating tissues, in-cluding shoot and root meristem. In this study, we ex-amined NtCDPK1 expression in roots using GUS ex-pression in transgenic Arabidopsis plants, and investi-gated its function in root development by generating transgenic tobacco plants carrying a sense NtCDPK1 transgene. GUS activity was first detected in roots two days after sowing. In later stages, strong GUS expres-sion was detected in the root meristem and elongation zone, as well as the initiation sites and branch points of lateral roots. Transgenic tobacco plants in which NtCDPK1 expression was suppressed were smaller, and their root development was abnormal, with reduced lateral root formation and less elongation. These re-sults suggest that NtCDPK1 plays a role in a signaling pathway regulating root development in tobacco.

Arabidopsis↗

The ectomycorrhizal symbiosis between Lactarius deliciosus and Pinus sylvestris in forest soil samples: symbiotic efficiency and development on roots of a rDNA internal transcribed spacer-selected isolate of L. deliciosus.

The effect on plant growth of pre-inoculation of Pinus sylvestris with the ectomycorrhizal (ECM) edible basidiomycete Lactarius deliciosus (isolate D45) under controlled conditions, and the development on roots of this basidiomycete, were investigated in gamma-irradiated and unsterilized containers containing different forest soil cores or a perlite-vermiculite mixture. Five months after planting, L. deliciosus mycorrhizal plants exhibited greater growth than the non-mycorrhizal ones in all soil types, i.e. up to a 325% increase in shoot height in the sterilized soils. The experiment demonstrated the dependency of P. sylvestris seedlings upon ECM symbiosis for their survival in gamma-irradiated, microbiologically disturbed soil samples. Furthermore, in two soils, the growth of L. deliciosus-inoculated seedlings was greater in the sterilized soil samples than in the non-sterilized ones, i.e. 46% and 132% increase in shoot height under sterilized soil conditions. In containers randomly sampled from each soil type, the degree of root colonization by the inoculated isolate, calculated as the number of mycorrhizal root tips divided by the total number of root tips x100, ranged from 80% to 35%. Within the short term, the inoculated isolate developed rapidly on roots, dominated, and hampered ectomycorrhiza formation by various unidentified (but not Lactarius) resident ECM fungi in unsterilized soil types. Results indicate that the ECM species L. deliciosus is worth investigating to ascertain if other isolates benefit pine growth like the isolate D45, and are therefore also attractive candidates for forestry applications in the Mediterranean area.

Basidiomycota↗

Early root cap development and graviresponse in white clover (Trifolium repens) grown in space and on a two-axis clinostat.

White clover (Trifolium repens) was germinated and grown in microgravity aboard the Space Shuttle (STS-60, 1994; STS-63, 1995), on Earth in stationary racks and in a slow-rotating two-axis clinostat. The objective of this study was to determine if normal root cap development and early plant gravity responses were dependent on gravitational cues. Seedlings were germinated in space and chemically fixed in orbit after 21, 40, and 72 h. Seedlings 96 h old were returned viable to earth. Germination and total seedling length were not dependent on gravity treatment. In space-flown seedlings, the number of cell stories in the root cap and the geometry of central columella cells did not differ from those of the Earth-grown seedlings. The root cap structure of clinorotated plants appeared similar to that of seedlings from microgravity, with the exception of three-day rotated plants, which displayed significant cellular damage in the columella region. Nuclear polarity did not depend on gravity; however, the positions of amyloplasts in the central columella cells were dependent on both the gravity treatment and the age of the seedlings. Seedlings from space, returned viable to earth, responded to horizontal stimulation as did 1 g controls, but seedlings rotated on the clinostat for the same duration had a reduced curvature response. This study demonstrates that initial root cap development is insensitive to either chronic clinorotation or microgravity. Soon after differentiation, however, clinorotation leads to loss of normal root cap structure and plant graviresponse while microgravity does not.

Fabaceae↗

Nitric oxide and cyclic GMP are messengers in the indole acetic acid-induced adventitious rooting process.

This report describes part of the signaling pathway and some of the molecules involved in the auxin-induced adventitious root formation in cucumber (Cucumis sativus). Previous results showed that nitric oxide (NO) mediates the auxin response during adventitious root formation (Pagnussat et al., 2002). To determine the order of action of indole acetic acid (IAA) and NO within the signal transduction pathway and to elucidate the target molecules that are downstream of NO action, cucumber hypocotyl cuttings were submitted to a pretreatment leading to endogenous auxin depletion. The auxin depletion treatment provoked a 3-fold reduction of the root number in comparison to the nondepleted explants. The NO-donor sodium nitroprusside was able to promote adventitious rooting in auxin-depleted explants, whereas the specific NO scavenger cPTIO prevented the effect of sodium nitroprusside. The endogenous NO level was monitored in both control and auxin-depleted explants using a NO-specific fluorescent probe. The NO level was 3.5-fold higher in control (nondepleted) explants than in auxin-depleted ones. The exogenous application of IAA restored the NO concentration to the level found in nondepleted explants. Because NO activates the enzyme guanylate cyclase (GC), we analyzed the involvement of the messenger cGMP in the adventitious root development mediated by IAA and NO. The GC inhibitor LY83583 reduced root development induced by IAA and NO, whereas the cell-permeable cGMP derivative 8-Br-cGMP reversed this effect. The endogenous level of cGMP is regulated by both the synthesis via GC and its degradation by the phosphodiesterase activity. When assayed, the phosphodiesterase inhibitor sildenafil citrate was able to induce adventitious rooting in both nondepleted and auxin-depleted explants. Results indicate that NO operates downstream of IAA promoting adventitious root development through the GC-catalyzed synthesis of cGMP.

Cucumis sativus↗

Phospholipid composition of a plasma membrane-enriched fraction from developing soybean roots.

Phospholipid polar head group and fatty acid composition were determined for plasma membrane enriched fractions from developing soybean root (Glycine max [L.] Merr. cult. Wells II). Plasma membrane vesicles were isolated from meristematic and mature sections of four-day-old dark grown soybean roots at pH 7.8 and in the presence of 5 millimolar ethylenediaminetetraacetate, 5 millimolar ethyleneglycol-bis (beta-aminoethyl ether)N,N tetraacetic acid, and 10 millimolar NaF. Lipid extracts analyzed for phospholipid composition revealed two major phospholipid components: phosphatidylcholine and phosphatidylethanolmine. Minor phospholipid components identified were phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, and diphosphatidylglycerol. Lipid degradation by endogenous phospholipase D during membrane isolation at pH 6.5 and in the absence of chelating agents and NaF resulted in the recovery of large amounts of phosphatidic acid. Phosphatidylcholine was the principal substrate for phospholipase D.Fatty acid composition was determined for plasma membrane phosphatidylcholine and phosphatidylethanolamine from meristematic and mature root tissue. The fatty acids identified were 16:0, 18:0, 18:1, 18:2, and 18:3. Fatty acid composition varied with both phospholipid class and the developmental stage of the root. Results suggest that differences in the composition of the major phospholipids of plasma membrane from meristemaic and mature root sections occur in the fatty acids and not in the polar head groups. These differences and those found in the composition of the polar head groups of the minor phospholipid components, e.g. phosphatidylglycerol, may be significant for structure-function relationships within the membrane.

Journal Article↗

Dental developmental status of southern Chinese girls in Hong Kong.

Dental developmental status of the permanent canines, premolars and second molars were studied in one hundred and two twelve-year-old Southern Chinese girls born and brought up in Hong Kong. Eruption of these teeth were defined as gingival emergence during the clinical examination while the root development was assessed from orthopantomographs. Root resorption pattern of the primary canines and molars were also studied with the aid of the orthopantomographs. Two-thirds of girls presented with permanent dentition stage. The root resorption patterns of the primary teeth were different from their order of exfoliation. The mandibular permanent teeth were in advanced in both eruption and root development than the corresponding maxillary teeth, except the first premolars.

Asian People↗

Regulation of symbiotic root nodule development.

Symbiosis between rhizobia and leguminous plants leads to the formation of N2-fixing root nodules. The interaction of rhizobia and plants shows a high degree of host specificity based on the exchange of chemical signals between the symbiotic partners. The plant signals, flavonoids exuded by the roots, activate the expression of nodulation genes, resulting in the production of the rhizobial lipochitooligosaccharide signals (Nod factors). Nod factors act as morphogens that, under conditions of nitrogen limitation, induce cells within the root cortex to divide and to develop into nodule primordia. This review focuses on how the production of Nod factors is regulated, how these signals are perceived and transduced by the plant root, and the physiological conditions and plant factors that control the early events leading to root nodule development.

Fabaceae↗

Characterization of cDNA for nodulin-75 of soybean: A gene product involved in early stages of root nodule development.

Establishment of a nitrogen-fixing root nodule is accompanied by a developmentally regulated expression of nodulin genes, only some of which, the so-called early nodulin genes, are expressed in stages preceding actual nitrogen fixation. We have isolated soybean cDNA clones representing early nodulin genes and have studied clone pENOD2 in detail. The cDNA insert of this clone hybridizes to nodule-specific RNA of 1200 nucleotides in length. The RNA that was hybrid-selected by the cloned ENOD2 DNA was in vitro translated to produce two nodulins with an apparent M(r) of 75,000, the N-75 nodulins. These two nodulins differ slightly in charge and one does not contain methionine. The amino acid sequence deduced from the DNA sequence shows that proline accounts for 45% of the 240 residues in these nodulins and the sequence contains at least 20 repeating heptapeptide units. The amino acid composition of none of the (hydroxy)proline-rich (glyco)proteins described in plants resembles the composition of the N-75 nodulins, especially with respect to the high glutamic acid and the low serine content. This suggests that the N-75 nodulins belong to a hitherto unidentified class of presumably structural proteins. The genes encoding the N-75 nodulins were found to be expressed in nodule-like structures devoid of intracellular bacteria and infection threads, indicating that these nodulins do not function in the infection process but more likely function in nodule morphogenesis.

Journal Article↗

Canine eruption into grafted alveolar clefts: a retrospective study.

BACKGROUND: The contemporary treatment of cleft lip and palate involves a sequence of surgical procedures and orthodontic management. Alveolar bone grafting (ABG) is usually undertaken after orthodontic expansion of the maxillary segments between the ages of eight and 12 years. Two of the important goals of alveolar bone grafting are the provision of bony support for the eruption of the canine and the closure of residual oro-nasal fistulae. The purpose of this study was to retrospectively evaluate the root development and eruption of the canine following ABG. METHODS: Group 1: radiographic and clinical records of a sample of 19 cleft patients who underwent alveolar bone grafting procedures, performed between 1996 and 1999 were reviewed. Group 2: a random sample of 15 cleft patients attending for routine dental review were clinically examined. The age of patient, degree of root development and eruption status of the canine, and presence of oronasal fistulae pre and post alveolar bone grafting were evaluated. RESULTS: Most cleft canines had continued root development and descended in the alveolus towards eruption following ABG. Four canine teeth (8 per cent) were impacted and required surgical exposure and orthodontic treatment following failure of eruption. Closure of anterior oro-nasal fistulae at the time of grafting was maintained post-operatively. CONCLUSIONS: This study demonstrated that canine root development and eruption continued satisfactorily through grafted alveolar clefts in most cases and closure of anterior oro-nasal fistulae was achieved in all cases.

Adolescent↗

Auxin and ethylene response interactions during Arabidopsis root hair development dissected by auxin influx modulators.

The plant hormones auxin and ethylene have been shown to play important roles during root hair development. However, cross talk between auxin and ethylene makes it difficult to understand the independent role of either hormone. To dissect their respective roles, we examined the effects of two compounds, chromosaponin I (CSI) and 1-naphthoxyacetic acid (1-NOA), on the root hair developmental process in wild-type Arabidopsis, ethylene-insensitive mutant ein2-1, and auxin influx mutants aux1-7, aux1-22, and double mutant aux1-7 ein2. Beta-glucuronidase (GUS) expression analysis in the BA-GUS transgenic line, consisting of auxin-responsive domains of PS-IAA4/5 promoter and GUS reporter, revealed that 1-NOA and CSI act as auxin uptake inhibitors in Arabidopsis roots. The frequency of root hairs in ein2-1 roots was greatly reduced in the presence of CSI or 1-NOA, suggesting that endogenous auxin plays a critical role for the root hair initiation in the absence of an ethylene response. All of these mutants showed a reduction in root hair length, however, the root hair length could be restored with a variable concentration of 1-naphthaleneacetic acid (NAA). NAA (10 nM) restored the root hair length of aux1 mutants to wild-type level, whereas 100 nM NAA was needed for ein2-1 and aux1-7 ein2 mutants. Our results suggest that insensitivity in ethylene response affects the auxin-driven root hair elongation. CSI exhibited a similar effect to 1-NOA, reducing root hair growth and the number of root hair-bearing cells in wild-type and ein2-1 roots, while stimulating these traits in aux1-7and aux1-7ein2 roots, confirming that CSI is a unique modulator of AUX1.

Arabidopsis↗

Analysis of forces developed during root canal preparation with the balanced force technique.

The aim of this study was to examine the forces and torque developed during root canal preparation with the balanced force technique using a recently described force-analyser device. A tooth was placed in a holder within the Endograph and forces and torques exerted were recorded. These parameters, which can be studied during preparation (on-line) or stored and examinated subsequently (off-line) generated endograms, which showed the forces generated with time. In addition, the endograms of preparations performed by students and endodontists, as well as deliberately induced failures in preparation technique (broken instruments), were compared. The values for the forces and torques depended on the size of the instruments and were related to the phase of the preparation. For the endodontists, the vertical and horizontal forces varied, respectively, from 0.08 +/- 0.01 kg for a size 15 to 0.65 +/- 0.10 kg for a size 45, and from 0.01 +/- 0.005 kg for a size 15 to 0.4 +/- 0.1 kg for a size 40. The torque varied from 0.08 +/- 1 kg mm-1 for a size 15 to 1.6 +/- 0.4 kg mm-1 for a size 45. With the endograms used as a reference, the relation between the developed vertical forces and the torque became more similar between the groups of endodontists and students. The Endograph provides a new approach to the analysis of preparation technique because it depicts the relationships between the different parameters of the preparation.

Amplifiers, Electronic↗

Analysis of 1202 orthopantograms to evaluate the potential of forensic age determination based on third molar developmental stages.

A total of 1202 orthopantograms of young dental patients were analyzed to assess the correlation of third molar root development with chronological age. The investigated patients were treated at a university dental clinic on an outpatient basis, included a variety of demographic characteristics (600 males and 602 females, 28% of other than central European background), and were 15-24 years old when the radiographs were obtained. Radiographs in which more than one third molar were either missing due to agenesia or extraction, or not evaluable because of deep destruction or marked tilting, were not included in the analysis. Our findings show that the growth patterns of third molars, based on seven defined stages of root development, did correlate with chronological age; age estimation when applied to a specific individual would involve a margin of error of +/- 2-4 years. There were no significant differences between the left and right jaw segments, but the stage of root development was generally more advanced in the upper than in the lower third molars. Root development was also more advanced among boys than among girls of the same age. There were no apparent differences in growth patterns based on national/ethnic background. While only 2.5% of 18-year-olds revealed fully developed third molars in all four quadrants, this percentage leaped to 38.4% among the 21-year-olds. To summarize, while the developmental stage of third molars is not per se a highly useful indicator of chronological age in juveniles and young adults, it is nevertheless a valuable supplementary parameter given the scarcity of other available age indicators.

Adolescent↗

Initial net CO2 uptake responses and root growth for a CAM community placed in a closed environment.

To help understand carbon balance between shoots and developing roots, 41 bare-root crassulacean acid metabolism (CAM) plants native to the Sonoran Desert were studied in a glass-panelled sealable room at day/night air temperatures of 25/15 degrees C. Net CO(2) uptake by the community of Agave schottii, Carnegia gigantea, Cylindropuntia versicolor, Ferocactus wislizenii and Opuntia engelmannii occurred 3 weeks after watering. At 4 weeks, the net CO(2) uptake rate measured for south-east-facing younger parts of the shoots averaged 1.94 micro mol m(-2) s(-1) at night, considerably higher than the community-level nocturnal net CO(2) uptake averaged over the total shoot surface, primarily reflecting the influences of surface orientation on radiation interception (predicted net CO(2) uptake is twice as high for south-east-facing surfaces compared with all compass directions). Estimated growth plus maintenance respiration of the roots averaged 0.10 micro mol m(-2) s(-1) over the 13-week period, when the community had a net carbon gain from the atmosphere of 4 mol C while the structural C incorporated into the roots was 23 mol. Thus, these five CAM species diverted all net C uptake over the 13-week period plus some existing shoot C to newly developing roots. Only after sufficient roots develop to support shoot water and nutrient requirements will the plant community have net above-ground biomass gains.

Agave↗

Environmentally induced plasticity of root hair development in Arabidopsis.

Postembryonic development of plants is dependent on both intrinsic genetic programs and environmental factors. The plasticity of root hair patterning in response to environmental signals was investigated in the Columbia-0 wild type and 19 Arabidopsis mutants carrying lesions in various parts of the root hair developmental pathway by withholding phosphate or iron (Fe) from the nutrient medium. In the aging primary root and in laterals of the wild type, the number of root hairs increased in response to phosphate and Fe deficiency in a manner typical of each growth type. Although an increase in root hair density in -phosphorus plants was mainly achieved by the formation of extra hairs over both tangential and radial wall of underlying cortical cells, roots of -Fe plants were characterized by a high percentage of extra hairs with two tips. Root hair patterning and hair length was differentially affected by the presence or absence of phosphate and Fe among the genotypes under investigation, pointing to separate cascades of gene activation under all three growth conditions. Divergence in root hair patterning was most pronounced among mutants with defects in genes that affect the first stages of differentiation, suggesting that nutritional signals are perceived at an early stage of epidermal cell development. During elongation of the root hairs, no differences in the requirement of gene products between the growth types were obvious. The role of genes involved in root hair development in the aging primary root of Arabidopsis under the various growth conditions is discussed.

Arabidopsis↗

Cooperative action of SLR1 and SLR2 is required for lateral root-specific cell elongation in maize.

Lateral roots play an important role in water and nutrient uptake largely by increasing the root surface area. In an effort to characterize lateral root development in maize (Zea mays), we have isolated from Mutator (Mu) transposon stocks and characterized two nonallelic monogenic recessive mutants: slr1 and slr2 (short lateral roots1 and 2), which display short lateral roots as a result of impaired root cell elongation. The defects in both mutants act specifically during early postembryonic root development, affecting only the lateral roots emerging from the embryonic primary and seminal roots but not from the postembryonic nodal roots. These mutations have no major influence on the aboveground performance of the affected plants. The double mutant slr1; slr2 displays a strikingly different phenotype than the single mutants. The defect in slr1; slr2 does not only influence lateral root specific cell elongation, but also leads to disarranged cellular patterns in the primary and seminal roots. However, the phase-specific nature of the single mutants is retained in the double mutant, indicating that the two loci cooperate in the wild type to maintain the lateral root specificity during a short time of early root development.

Genes, Plant↗

A protein phosphatase 2C gene, LjNPP2C1, from Lotus japonicus induced during root nodule development.

Symbiotic interactions between legumes and compatible strains of rhizobia result in root nodule formation. This new plant organ provides the unique physiological environment required for symbiotic nitrogen fixation by the bacterial endosymbiont and assimilation of this nitrogen by the plant partner. We have isolated two related genes (LjNPP2C1 and LjPP2C2) from the model legume Lotus japonicus that encode protein phosphatase type 2C (PP2C). Expression of the LjNPP2C1 gene was found to be enhanced specifically in L. japonicus nodules, whereas the LjPP2C2 gene was expressed at a similar level in nodules and roots. A glutathione S-transferase-LjNPP2C1 fusion protein was shown to have Mg2+- or Mn2+-dependent and okadaic acid-insensitive PP2C activity in vitro. A chimeric construct containing the full-length LjNPP2C1 cDNA, under the control of the Saccharomyces cerevisiae alcohol dehydrogenase promoter, was found to be able to complement a yeast PP2C-deficient mutant (pct1Delta). The transcript level of the LjNPP2C1 gene was found to increase significantly in mature nodules, and its highest expression level occurred after leghemoglobin (lb) gene induction, a molecular marker for late developmental events in nodule organogenesis. Expression of the LjNPP2C1 gene was found to be drastically altered in specific L. japonicus lines carrying monogenic-recessive mutations in symbiosis-related loci, suggesting that the product of the LjNPP2C1 gene may function at both early and late stages of nodule development.

Amino Acid Sequence↗

Three-year root caries increments: implications for clinical trials.

Seven hundred and ninety-six adult subjects (mean age, 39.9 years) received visual-tactile examinations for root caries over a three-year period. All subjects were employed or were the spouses of employees and resided in fluoride-deficient communities on Long Island, New York. During the three-year observation period, 81.4 percent of the subjects did not develop root caries. The 18.6 percent who developed root caries averaged 0.8 DFS/year. The subjects' ages and baseline root DFS status were associated with the development of a root DFS increment. The older the patient, especially aged 45 and older, the greater was the risk of developing root lesions or having root fillings placed. Subjects who had a root DFS score at baseline also were more likely to experience a root DFS increment. It is recommended that when designing clinical trials of agents purported to inhibit root caries, preselection criteria for the study population should consider the subjects' ages and past history of root lesions.

Adult↗