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PubMed1998

Altered Zn compartmentation in the root symplasm and stimulated Zn absorption into the leaf as mechanisms involved in Zn hyperaccumulation in thlaspi caerulescens

We investigated Zn compartmentation in the root, Zn transport into the xylem, and Zn absorption into leaf cells in Thlaspi caerulescens, a Zn-hyperaccumulator species, and compared them with those of a related nonaccumulator species, Thlaspi arvense. 65Zn-compartmental analysis conducted with roots of the two species indicated that a significant fraction of symplasmic Zn was stored in the root vacuole of T. arvense, and presumably became unavailable for loading into the xylem and subsequent translocation to the shoot. In T. caerulescens, however, a smaller fraction of the absorbed Zn was stored in the root vacuole and was readily transported back into the cytoplasm. We conclude that in T. caerulescens, Zn absorbed by roots is readily available for loading into the xylem. This is supported by analysis of xylem exudate collected from detopped Thlaspi species seedlings. When seedlings of the two species were grown on either low (1 &mgr;M) or high (50 &mgr;M) Zn, xylem sap of T. caerulescens contained approximately 5-fold more Zn than that of T. arvense. This increase was not correlated with a stimulated production of any particular organic or amino acid. The capacity of Thlaspi species cells to absorb 65Zn was studied in leaf sections and leaf protoplasts. At low external Zn levels (10 and 100 &mgr;M), there was no difference in leaf Zn uptake between the two Thlaspi species. However, at 1 mM Zn2+, 2.2-fold more Zn accumulated in leaf sections of T. caerulescens. These findings indicate that altered tonoplast Zn transport in root cells and stimulated Zn uptake in leaf cells play a role in the dramatic Zn hyperaccumulation expressed in T. caerulescens.

Journal Article↗
PubMed2003

Use of a promoter trap to identify Bacillus cereus genes regulated by tomato seed exudate and a rhizosphere resident, Pseudomonas aureofaciens.

The goal of this study was to identify genes in Bacillus cereus, a bacterium commonly associated with plant seeds and roots, that are affected by compounds originating from a host plant, tomato, or another rhizosphere resident, Pseudomonas aureofaciens. We constructed a B. cereus chromosomal DNA library in a promoter-trap plasmid, pAD123, which contains a promoterless version of the green fluorescent protein (GFP) gene, gfpmut3a. The library was screened by using fluorescence-activated cell sorting for clones showing a change in GFP expression in response to either tomato seed exudate or culture supernatant of P. aureofaciens strain 30-84. We identified two clones carrying genes that were induced by the presence of tomato seed exudate and nine clones carrying genes that were repressed by P. aureofaciens culture supernatant. A clone chosen for further study contained an open reading frame, designated lipA, that encodes a deduced protein with a lipoprotein signal peptide sequence similar to lipoproteins in B. subtilis. Expression of gusA under control of the lipA promoter increased twofold when cells were exposed to tomato seed exudate and in a concentration-dependent manner when exposed to a mixture of amino acids. When the wild type and a 10-fold excess of a lipA mutant were applied together to tomato seeds, 2 days after planting, the wild type displayed medium-dependent culturability, whereas the lipA mutant was unaffected. This study demonstrates the power of a promoter trap to identify genes in a gram-positive bacterium that are regulated by the biotic environment and resulted in the discovery of lipA, a plant-regulated gene in B. cereus.

Bacillus cereus↗
PubMed2001

Identification and effects of interaction phytotoxic compounds from exudate of Cistus ladanifer leaves.

Eleven allelochemicals (ferulic acid, cinnamic acid, 4-hydroxybenzoic acid, hydroxycinnamic acid, methyl propionate, oxalic acid, methylmalonic acid, p-anisic acid, butyric acid, 3-hydroxybutyric acid, and azulene) were identified in the exudate of Cistus ladanifer L. We studied the effect of each on germination, cotyledon emergence, root length, and cotyledon length of Rumex crispus. Three groups were distinguished with respect to phytotoxic activity: compounds with low activity (ferulic acid, 4-hydroxybenzoic acid, oxalic acid, methylmalonic acid, p-anisic acid, hydroxybutyric acid, and azulene), with intermediate activity (cinnamic acid and hydroxycinnamic acid), and with high activity (methyl propionate and butyric acid). The effect of the interaction of the compounds was studied. When acting conjointly, all combinations tested produced a more negative effect on both germination and seedling growth than when acting alone. The interaction affected cotyledon emergence and root length more negatively than germination and cotyledon length. When hydroxycinnamic acid and cinnamic acid were added to these mixtures there was an enhancement in the phytotoxic activity, accentuating the effect of the other allelochemicals.

3-Hydroxybutyric Acid↗
PubMed1986

Enzymes of ammonia assimilation and ureide biosynthesis in soybean nodules: effect of nitrate.

The effect of nitrate on N(2) fixation and the assimilation of fixed N(2) in legume nodules was investigated by supplying nitrate to well established soybean (Glycine max L. Merr. cv Bragg)-Rhizobium japonicum (strain 3I1b110) symbioses. Three different techniques, acetylene reduction, (15)N(2) fixation and relative abundance of ureides ([ureides/(ureides + nitrate + alpha-amino nitrogen)] x 100) in xylem exudate, gave similar results for the effect of nitrate on N(2) fixation by nodulated roots. After 2 days of treatment with 10 millimolar nitrate, acetylene reduction by nodulated roots was inhibited by 48% but there was no effect on either acetylene reduction by isolated bacteroids or in vitro activity of nodule cytoplasmic glutamine synthetase, glutamine oxoglutarate aminotransferase, xanthine dehydrogenase, uricase, or allantoinase. After 7 days, acetylene reduction by isolated bacteroids was almost completely inhibited but, except for glutamine oxoglutarate aminotransferase, there was still no effect on the nodule cytoplasmic enzymes. It was concluded that, when nitrate is supplied to an established symbiosis, inhibition of nodulated root N(2) fixation precedes the loss of the potential of bacteroids to fix N(2). This in turn precedes the loss of the potential of nodules to assimilate fixed N(2).

Journal Article↗
PubMed1985

Quantifying Apoplastic Flux through Red Pine Root Systems Using Trisodium, 3-hydroxy-5,8,10-pyrenetrisulfonate.

The fluorescent compound trisodium, 3-hydroxy-5,8,10-pyrenetrisulfonate (PTS) was used to quantify the apoplastic flux through red pine (Pinus resinosa Ait.) root systems-that portion of the total water flux reaching the xylem without ever crossing a semipermeable membrane. Flow was induced by pressure through detopped root systems, and by transpiration through intact seedlings. Apoplastic flux was determined by multiplying total flux by the ratio of PTS concentration in the xylem exudate to PTS concentration in the bathing medium.Under aeration, apoplastic flux was less than 1% of total flux. Under anaerobic conditions, up to 50% of total flux was apoplastic suggesting that anaerobic conditions change the pathway of water flow into root xylem. The change under anaerobic conditions was reversible. Detopped root systems under pressure and intact seedlings under transpiration gave similar results. In detopped root systems, the magnitude of the pressure gradient may alter the apoplastic contribution to total flux.

Journal Article↗
PubMed2003

Detection of Filifactor alocis in endodontic infections associated with different forms of periradicular diseases.

Recent molecular studies have expanded the list of suspected endodontic pathogens. The aim of this study was to evaluate the occurrence of Filifactor alocis in primary endodontic infections associated with different forms of periradicular diseases. Identification by nested polymerase chain reaction was performed in root canal samples from teeth associated with either asymptomatic periradicular lesions or acute apical periodontitis. Samples were also taken by aspiration of purulent exudate associated with acute apical abscesses. DNA extracted from the samples was initially amplified using universal 16S rDNA primers followed by a second round of amplification using the first PCR products to detect a specific fragment of F. alocis 16S rDNA. F. alocis was detected in 12/21 (57.1%) root canal samples from teeth showing asymptomatic periradicular lesions and in 3/10 (30%) samples taken from root canals associated with acute apical periodontitis. This species occurred in 8/19 (42.1%) pus aspirates obtained from abscessed teeth. In general, F. alocis was detected in 23/50 (46%) samples taken from endodontic infections. These findings suggest that F. alocis is involved in the etiology of different forms of periradicular diseases and has the potential to be an endodontic pathogen.

Adolescent↗
PubMed1973

The velocities of ion transport into and through the xylem of roots: findings with a two-point application pulse-chase technique.

Excised corn roots, Zea mays, had radioactively labeled solution applied at two points along their length, for 1 minute, and were then kept in dilute, unlabeled nutrient solution. During this "chase" period, exudate was collected at 1-minute intervals, and its content of radioions was determined. Two pulses of label appeared in succession, originating at the points of application near the cut end of the root and farther from the cut end, respectively. Calculation yields the velocity at which the ions moved radially across the root into the xylem (v(r)) and the velocity at which they moved longitudinally within the xylem (v(l)). For Rb(+) labeled with (86)Rb, v(r) was 1.8 and v(l), 35 cm/hr. For Br(-) labeled with (82)Br, v(r) was 1.4 and v(l), 103 cm/hr.

Journal Article↗
PubMed2006

Incidence and antimicrobial susceptibility of Porphyromonas gingivalis isolated from mixed endodontic infections.

AIM: To investigate the prevalence of Porphyromonas gingivalis in root canals of infected teeth with periapical abscesses and to investigate the antimicrobial susceptibility of this species to some frequently prescribed antibiotics. METHODOLOGY: Samples were obtained from 70 root canals of abscessed teeth. Microbial sampling, isolation and bacterial identification were accomplished using appropriate culture methods for anaerobic species. The antimicrobial susceptibility of the 20 strains of P. gingivalis isolated was determined by using the E-test. The antimicrobial agents tested were amoxicillin, amoxicillin + clavulanate, azythromycin, benzylpenicillin, cephaclor, clindamycin, erythromycin, metronidazole and tetracycline. RESULTS: A total of 352 individual strains, belonging to 69 different species, were isolated. Eighty three percent of the strains were strict anaerobes and 47.5% of the isolated bacteria were Gram-negative. Porphyromonas gingivalis was found in 20 root canals and was most frequently found in symptomatic cases. Statistically, the presence of P. gingivalis was related to purulent exudates and pain on palpation (both P < 0.05). All P. gingivalis strains were sensitive to amoxicillin, amoxicillin + clavulanate, cephaclor, clindamycin, benzylpenicyllin, metronidazole and tetracycline. The lowest range of minimum inhibitory concentration (MIC) (0.026-0.125 microg mL(-1)) was observed against amoxicillin + clavulanate and clindamycin. The lowest MIC 90 was observed against clindamycin (0.064 microg mL(-1)). One strain was resistant to erythromycin and eight strains were resistant to azythromycin. CONCLUSION: Porphyromonas gingivalis pathogen is isolated with frequency from root canals of infected teeth with periapical abscesses. Amoxicillin, as well as amoxicillin-clavulanic acid and benzylpenicillin were effective against P. gingivalis.

Adolescent↗
PubMed2004

Apex locators: update and prospects for the future.

The accuracy of working length determination in root canal treatment plays a crucial role in the treatment and prevention of periapical disease. Since the introduction of the first apex locator by Sunada in 1961, new and more accurate devices have been developed. Electronic apex locators reduce the number of radiographs required and may detect root canal perforations and fractures. Third and fourth generation apex locators can operate in the presence of intact or necrotic tissue, blood, inflammatory exudates, and various irrigants. The Raypex 4 and the Apex NRG novel apex locators are discussed in detail.

Dental Instruments↗
PubMed2005

The early stages of three Palaearctic species of saproxylic hoverflies (Syrphidae, Diptera).

The third stage larva and puparium of the cerioidine hoverflies, Ceriana vespiformis (Latreille, 1804) and Sphiximorpha subsessilis (Illiger in Rossi, 1807) and the milesine, Spilomyia digitata (Rondani, 1865) (Diptera, Syrphidae) are described from specimens collected in the field. Ceriana vespiformis and S. digitata larvae were obtained from wet, decaying roots and tree holes of Fraxinus angustifolius L., respectively, in Spain and S. subsessilis from an exudation of sap on an Abies alba Miller tree in France.

Abies↗
PubMed1995

Studies on antiinflammatory effect of Japanese Oriental medicines (kampo medicines) used to treat inflammatory diseases.

This study was carried out to examine the antiinflammatory effect of aqueous extracts of Kakkon-to (K), Kakkon-oren-ogon-to (KO), Kikyo-to (KK), Haino-to (H), Haino-san (HS), Mao-to (M) and Senkinkeimei-san (SK), which have been used for the treatment of stomatitis, tonsillitis, cold and chronic inflammatory diseases, and to elucidate the mode of their effects. Oral administration of K, KO, KK, H, HS, M and SK inhibited dose-dependently the increase of dye leakage induced by acetic acid in mice. Further antiinflammatory study was carried out on KK, H and HS which showed potent inhibition. All three extracts significantly inhibited the carrageenin-induced edema and the cotton pellet-induced granuloma formation. From these results, it is suggested that KK, H and HS may inhibit both the early exudative stage and the late proliferative stage in inflammatory processes. These extracts are comprised of Platycodon root as do other crude drugs, and the root may be partly responsible for the antiinflammatory effects induced.

Acetates↗
PubMedDate not supplied

Paranasal sinusitis complicated by inspissated exudate in the ventral conchal sinus.

Primary paranasal sinusitis complicated by inspissated exudate within the ventral conchal sinus was diagnosed in five horses. Clinical signs included a unilateral, foul-smelling, mucopurulent nasal exudate of 2 to 7 months' duration. Two of the horses had partial nasal obstruction from distortion of the ventral concha. Radiographs of the skull showed a mass of soft tissue density dorsal to the roots of the superior third and fourth or fourth and fifth cheek teeth. Treatment included bone flap maxillary sinusotomy with exposure of the ventral nasal concha and removal of inspissated exudate. Resolution of the sinusitis occurred in all five horses.

Animals↗
PubMed2006

Comparative studies on the effect of a protein-synthesis inhibitor on aluminium-induced secretion of organic acids from Fagopyrum esculentum Moench and Cassia tora L. roots.

Aluminium (Al)-induced secretion of organic acids from plant roots is considered a mechanism of Al resistance, but the processes leading to the secretion of organic acids are still unknown. In the present study, a protein-synthesis inhibitor, cycloheximide (CHM), was used to investigate its effect on Al-induced organic acid secretion in a pattern I (rapid exudation of organic acids under Al stress) plant buckwheat (Fagopyrum esculentum Moench) and a pattern II (exudation of organic acids was delayed by several hours under Al stress) plant Cassia tora L. A dose-response experiment showed that the secretion of oxalate by buckwheat roots was not affected by CHM when added in the range from 0 to 50 microM, with or without exposure to 100 microm Al, but the secretion of citrate was completely inhibited by 30 microM CHM in C. tora. A time-course experiment showed that even prolonged exposure to 20 microM CHM did not affect oxalate secretion in buckwheat, but significantly inhibited citrate secretion in C. tora. However, citrate synthase (CS) activity in C. tora was not affected during 12 h exposure to 100 microM Al when compared with that in control roots, although CHM can inhibit CS activity effectively. These results indicated that CS activity was not related to Al-regulated citrate efflux in C. tora. The total protein was decreased by 14.0% and 32.3% in C. tora and buckwheat root tip, respectively, after 3-h treatment with 20 microM CHM. A 3-h pulse with 20 microM CHM completely inhibited citrate efflux in C. tora during the next 6-h exposure to Al, although a small amount of citrate was exuded after 9-h exposure. However, oxalate efflux in buckwheat was not influenced by a similar treatment. In buckwheat, a 3-h pulse with 100 microM Al maintained oxalate secretion at a high level during the next 9 h, with or without CHM treatment. Conversely, in C. tora a 6-h pulse with 100 microM Al induced significant secretion of citrate which was inhibited by the CHM. Taken together, these findings suggest that both de novo synthesis and activation of an anion channel are needed for Al-induced secretion of citrate in C. tora, but in buckwheat the plasma membrane protein responsible for oxalate secretion pre-exists.

Aluminum↗
PubMed1966

Iron Translocation II. Citrate/Iron Ratios in Plant Stem Exudates.

Stem exudates of sunflower, soybean, cucumber, and tomato were analyzed for citrate and iron content. Generally, the lowest iron treatments given at decapitation were associated with lowest citrate levels in the exudate and intermediate treatments with the highest citrate levels. Citrate reached 6.2 x 10(-4)m in sunflower exudate. 7.3 x 10(-4)m in soybean, 9.4 x 10(-4)m in cucumber, and 1.8 x 10(-4)m in tomato. Some of these values represent a doubling or tripling of citrate when compared to the lowest ones obtained. High Fe depressed the citrate content of exudate in some cases.As iron was raised in the nutrient, the increases of iron in the exudate were proportionately greater than those of citrate. The highest Fe treatments led to exaggerated uptake of iron by sunflower. In some cases iron was translocated in molar excess of citrate. A time-response experiment with tomato showed a rapid flooding of iron into the roots but a relatively slow release of iron into xylem exudate.Similar electrophoretic patterns of iron were obtained despite changes in citrate and iron with time. Exudates from sunflower, cucumber, and tomato gave iron concentrations in the position of iron-citrate.

Journal Article↗
PubMed2001

Cluster roots: model experimental tools for key biological problems.

The cluster root is made up of a number of determinate rootlets tightly grouped along the parent root. Each rootlet grows for a limited time, and then the meristem stops dividing and differentiates. Following cessation of growth, an exudative burst occurs, wherein, over 2-3 d, large amounts of organic acids, as well as phosphatases and phenolics, are exuded from the rootlets. There is a concomitant acidification of the rhizosphere. It is suggested that the temporal and spatial predictability of developmental and functional events in these structures makes them valuable as experimental tools with which to investigate key issues in plant developmental biology, physiology, ecophysiology, evolutionary biology, and biotechnology.

Biological Evolution↗
PubMed2006

Contrasting soil ciliate species richness and abundance between two tropical plant species: a test of the plant effect.

We still have a rudimentary understanding about the mechanism by which plant roots may stimulate soil microbial interactions. A biochemical model involving plant-derived biochemical fractions, such as exudates, has been used to explain this "rhizosphere effect" on bacteria. However, the variable response of other soil microbial groups, such as protozoa, to the rhizosphere suggests that other factors could be involved in shaping their communities. Thus, two experiments were designed to: (1) determine whether stimulatory and/or inhibiting factors associated with particular plant species regulate ciliate diversity and abundance and (2) obtain a better understanding about the mechanism by which these plant factors operate in the rhizosphere. Bacterial and chemical slurries were reciprocally exchanged between two plant species known to differ in terms of ciliate species richness and abundance (i.e., Canella winterana and plantation Tectona grandis). Analysis of variance showed that the bacteria plus nutrients and the nutrients only treatment had no significant effect on overall ciliate species richness and abundance when compared to the control treatment. However, the use of only colpodean species increased the taxonomic resolution of treatment effects revealing that bacterial slurries had a significant effect on colpodean ciliate species richness. Thus, for particular rhizosphere ciliates, biological properties, such as bacterial diversity or abundance, may have a strong influence on their diversity and possibly abundance. These results are consistent with a model of soil bacteria-mediated mutualisms between plants and protozoa.

Plants↗
PubMed1987

Origin of growth-induced water potential : solute concentration is low in apoplast of enlarging tissues.

We developed a new method to measure the solute concentration in the apoplast of stem tissue involving pressurizing the roots of intact seedlings (Glycine max [L.] Merr. or Pisum sativum L.), collecting a small amount of exudate from the surface of the stem under saturating humidities, and determining the osmotic potential of the solution with a micro-osmometer capable of measuring small volumes (0.5 microliter). In the elongating region, the apoplast concentrations were very low (equivalent to osmotic potentials of -0.03 to -0.04 megapascal) and negligible compared to the water potential of the apoplast (-0.15 to -0.30 megapascal) measured directly by isopiestic psychrometry in intact plants. Most of the apoplast water potential consisted of a negative pressure that could be measured with a pressure chamber (-0.15 to -0.28 megapascal). Tests showed that earlier methods involving infiltration of intercellular spaces or pressurizing cut segments caused solute to be released to the apoplast and resulted in spuriously high concentrations. These results indicate that, although a small amount of solute is present in the apoplast, the major component is a tension that is part of a growth-induced gradient in water potential in the enlarging tissue. The gradient originates from the extension of the cell walls, which prevents turgor from reaching its maximum and creates a growth-induced water potential that causes water to move from the xylem at a rate that satisfies the rate of enlargement. The magnitude of the gradient implies that growing tissue contains a large resistance to water movement.

Journal Article↗