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Accumulation and transport of abscisic Acid and its metabolites in ricinus and xanthium.

When intact plants of Xanthium strumarium L. were water stressed, the youngest leaves accumulated the highest levels of abscisic acid (ABA). On the other hand, when leaves of different ages were detached and then stressed, the capacity to produce ABA was highest in the mature leaves. Radioactive ABA was transported from mature leaves to the shoot tips and young leaves, as well as to the roots, as evidenced by the presence of radioactive ABA and phaseic acid in the xylem exudate coming from the roots. Thus, ABA was recirculated in the plant, moving down the stem in the phloem and back up in the transpiration stream to the mature leaves. Phloem exudate collected by the use of the EDTA technique had a high concentration of ABA and phaseic acid which increased several-fold after water stress. The high ABA levels in immature leaves and apical buds are, therefore, mainly due to import from older leaves, rather than to in situ synthesis.In Ricinus communis L. cv gibsonii labeled ABA was rapidly exported from a mature leaf as measured by the appearance of radioactive ABA in the phloem collected from incisions in the stem below the treated leaf. After 2 h, small amounts of radioactive phaseic acid and dihydrophaseic acid were also present in the phloem exudate. The glucosyl ester of abscisic acid, beta-d-glucopyranosyl abscisate, was hydrolyzed before entering the phloem and moved as free ABA. It is concluded that the glucosyl ester of ABA itself cannot cross cell membranes and is, therefore, not transported.

Journal Article↗

Aluminum resistance of cowpea as affected by phosphorus-deficiency stress.

Plants growing in acid soils suffer both phosphorus (P) deficiency and aluminum (Al) toxicity stresses. Selection of genotypes for adaptation to either P deficiency or Al toxicity has sometimes been unsuccessful because these two soil factors often interact. Two experiments were conducted to evaluate eight cowpea genotypes for Al resistance and to study the combined effect of P deficiency and Al toxicity stress on growth, P uptake, and organic acid anion exudation of two genotypes of contrasting Al resistance selected from the first experiment. Relative root inhibition by 30 microM Al ranged from 14% to 60% and differed significantly among the genotypes. Al significantly induced callose formation, particularly in Al-sensitive genotypes. P accumulation was significantly reduced (28% and 95%) by Al application for both the Al-resistant and the Al-sensitive genotypes. Al supply significantly enhanced malate release of root apices of both genotypes. However, the exudation rate was significantly higher in the Al-resistant genotype. P deprivation induced an enhanced malate exudation in the presence of Al only in the Al-resistant genotype IT89KD-391. Citrate exudation rate of the root apices was lower than malate exudation by a factor of about 10, and was primarily enhanced by P deficiency in both genotypes. Al treatment further enhanced citrate exudation in P-sufficient, but not in P-deficient plants. The level of citrate exudation was consistently higher in the Al-resistant genotype IT89KD-391 particularly in presence of Al. It is concluded that the Al-resistant genotype is better adapted to acid Al-toxic and P-deficient soils than the Al-sensitive genotype since both malate and citrate exudation were more enhanced by combined Al and P-deficiency stresses.

Aluminum↗

Bacterial genetic loci implicated in the Pseudomonas putida GR12-2R3--canola mutualism: identification of an exudate-inducible sugar transporter.

Pseudomonas putida GR12-2R3 promotes the emergence and growth of diverse plant species. Analyses of TnphoA insertion mutations are revealing bacterial characteristics pertinent to the plant-microbe interaction. Pseudomonas putida PG269 is a TnphoA insertion derivative of GR12-2R3 that expresses canola seed exudate-inducible alkaline phosphatase (PhoA) activity. It promoted the growth of canola roots, as well as strain GR12-2R3, and outgrew its parent when they were cocultured in the presence of canola roots or in liquid seed exudate medium. (In contrast, mutant PG126 failed to promote canola root growth and was outgrown by its parent strain.) The PhoA activity of strain PG269 was induced by glucosamine and other sugars; glucosamine inhibited the growth of strain GR12-2R3 and stimulated the growth of strain PG269. Strain PG269 contained two TnphoA insertions: seiA1::TnphoA and seiB1::TnphoA. Strain PG312, which contained only insertion seiA1::TnphoA, shared all aspects of the PG269 phenotype, except the ability to outcompete strain GR12-2R3 during coculture. Insertion seiA1::TnphoA interrupted an open reading frame related in sequence to members of the MalF family of sugar transporter subunits. The PhoA-inducing fraction of canola seed exudate was hydrophilic, low in molecular weight, and heat stable. It cochromatographed with basic amino acids and amino sugars, and was inactivated by strains GR12-2R3 and PG269. Gene seiA may encode a subunit of an ABC transporter with broad specificity for glucose and related sugars whose expression can be induced by exudate sugars.

Amino Acid Sequence↗

Moving Waves of Bacterial Populations and Total Organic Carbon along Roots of Wheat.

> Abstract To determine if spatial variation in soluble carbon sources along the root coincides with different trophic groups of bacteria, copiotrophic and oligotrophic bacteria were enumerated from bulk soil and rhizosphere samples at 2 cm intervals along wheat roots 2, 3, and 4 weeks after planting. There was a moderate rhizosphere effect in one experiment with soil rich in fresh plant debris, and a very pronounced rhizosphere effect in the second experiment with soil low in organic matter. We obtained wavelike patterns of both trophic groups of bacteria as well as water-soluble total organic carbon (TOC) along the whole root length (60 or 90 cm). TOC concentrations were maximal at the root tip and base and minimal in the middle part of the roots. Oscillations in populations of copiotrophic and oligotrophic bacteria had two maxima close to the root tip and at the root base, or three maxima close to the tip, in the middle section, and at the root base. The location and pattern of the waves in bacterial populations changed progressively from week to week and was not consistently correlated with TOC concentrations or the location of lateral root formation. Thus, the traditional view that patterns in bacterial numbers along the root directly reflect patterns in exudation and rhizodeposition from several fixed sources along the root may not be true. We attributed the observed wavelike patterns in bacterial populations to bacterial growth and death cycles (due to autolysis or grazing by predators). Considering the root tip as a moving nutrient source, temporal oscillations in bacterial populations at any location where the root tip passed would result in moving waves along the root. This change in concept about bacterial populations in the rhizosphere could have significant implications for plant growth promotion and bioremediation.

Journal Article↗

Organic acids, sugars, and L-tryptophane in exudates of vegetables growing on stonewool and their effects on activities of rhizosphere bacteria.

The influence of stonewool substrate on the exudation of the major soluble carbon nutrients and of the auxin precursor tryptophane for Pseudomonas biocontrol agents was studied. To this end, the composition of the organic acids and sugars, as well that of tryptophane, of axenically collected exudates of seed, seedlings, and roots of tomato, cucumber, and sweet pepper was determined. The major results were as follows. i) The total amount of organic acid is much higher than that of total sugar. ii) Exudation of both organic acids and sugars increases during plant growth. iii) Citric, succinic, and malic acids represent the major organic acids, whereas fructose and glucose are the major sugars. iv) Compared with glass beads as a neutral substrate, stonewool substantially stimulates exudation of organic acids and sugars. v) It appeared that enhanced root-tip-colonizing bacteria isolated previously from the rhizosphere of tomato and cucumber grow much better in minimal medium with citrate as the sole carbon source than other, randomly selected rhizobacteria do. This indicates that the procedure which selects for excellent root-tip colonizers enriches for strains which utilize the major exudate carbon source citrate. vi) The content of L-tryptophane, the direct precursor of auxin, is approximately 60-fold higher in seedling exudates of tomato and sweet pepper than in cucumber seedling exudates, indicating a higher possibility of plant growth stimulation after inoculation with auxin-producing rhizobacteria for tomato and sweet pepper crops than for cucumber. However, the biocontrol strain Pseudomonas fluorescens WCS365, which is able to convert tryptophane into auxin, did not stimulate growth of these three crops. In contrast, this strain did stimulate growth of roots of radish, a plant which exudes nine times more tryptophane than tomato does.

Acids↗

Carbon Dioxide Fixation in Soybean Roots and Nodules: I. CHARACTERIZATION AND COMPARISON WITH N(2) FIXATION AND COMPOSITION OF XYLEM EXUDATE DURING EARLY NODULE DEVELOPMENT.

These studies demonstrate that soybean (Merr) roots and nodules possess an active system for fixing CO(2). The maximum rates of CO(2) fixation observed for roots and nodules of intact plants were 120 and 110 nanomoles CO(2) fixed per milligram dry weight per hour, respectively. Results of labeling studies suggest a primary role for phosphoenolpyruvate carboxylase in CO(2) assimilation in these tissues. After pulse-labeling with (14)CO(2) for 2 minutes, 70% of the total radioactivity was lost within 18 minutes via respiration and/or translocation out of nodules. During the vegetative stages of growth of soybeans grown symbiotically, CO(2) fixation in nodules increased at the onset of N(2) fixation but declined to a lower level prior to the decrease in N(2) fixation. This decrease coincided with a decrease in the transport of amino acids, especially asparagine, and an increase in the export of ureides. These findings are consistent with a dual role for CO(2) fixation, providing substrates for energy-yielding metabolism and supplying carbon skeletons for NH(4) (+) assimilation and amino acid biosynthesis.

Journal Article↗

Factors which affect the amount of inorganic phosphate, phosphorylcholine, and phosphorylethanolamine in xylem exudate of tomato plants.

Phosphate in the xylem exudate of tomato (Lycopersicon esculentum) plants was 70 to 98% inorganic phosphate (Pi), 2 to 30% P-choline, and less than 1% P-ethanolamine. Upon adding (32)Pi to the nutrient, Pi in xylem exudate had the same specific activity within 4 hours. P-choline and P-ethanolamine reached the same specific activity only after 96 hours. The amount of Pi in xylem exudate was dependent on Pi concentration in the nutrient and decreased from 1700 to 170 micromolar when Pi in the nutrient decreased from 50 to 2 micromolar. The flux of 0.4 nmoles organic phosphate per minute per gram fresh weight root into the xylem exudate was not affected by the Pi concentration in the nutrient solution unless it was below 1 micromolar. During 7 days of Pi starvation, Pi in the xylem exudate decreased from 1400 to 130 micromolar while concentrations of the two phosphate esters remained unchanged.The concentration of phosphate esters in the xylem exudate was increased by addition of choline or ethanolamine to the nutrient solution, but Pi remained unchanged. Upon adding [(14)C]choline to the nutrient, 10 times more [(14)C]P-choline than [(14)C]choline was in the xylem exudate and 85 to 90% of the ester phosphate was P-choline. When [(14)C]ethanolamine was added, [(14)C]P-ethanolamine and [(14)C]ethanolamine in the xylem sap were equal in amount. P-choline and P-ethanolamine accumulated in leaves of whole plants at the same time and the same proportion as observed for their flux into the xylem exudate. No relationship between the transport of P-choline and Pi in the xylem was established. Rather, the amount of choline in xylem exudate and its incorporation into phosphatidylcholine in the leaf suggest that the root is a site of synthesis of P-choline and P-ethanolamine for phospholipid synthesis in tomato leaves.

Journal Article↗

[The effect of tryptophan of plant root metabolites on the phyto stimulating activity of rhizobacteria ].

Aseptic tomato and radish roots were found to exude 2.8-5.3 and 290-390 ng tryptophan per seedling per day. The inoculation of radish plants with rhizosphere pseudomonads increased the root biomass by 1.4 times. The inoculation of tomato plants with the same pseudomonads was ineffective. The beneficial effect of bacterial inoculation on the radish plants can be explained by the fact that the introduced rhizobacteria produce the plant growth-stimulating hormone indole-3-acetic acid. In pot experiments, the addition of this phytohormone to the soil increased the mass of radish roots by 36%. The phytohormonal action of the rhizosphere microflora was found to be efficient provided that the concentration of tryptophan in the rhizosphere is sufficiently high.

Indoleacetic Acids↗

Expression of a ferredoxin-dependent glutamate synthase gene in mesophyll and vascular cells and functions of the enzyme in ammonium assimilation in Nicotiana tabacum (L.).

GLU1 encodes the major ferredoxin-dependent glutamate synthase (Fd-GOGAT, EC 1.4.7.1) in Arabidopsis thaliana (ecotype Columbia). With the aim of providing clues on the role of Fd-GOGAT, we analyzed the expression of Fd-GOGAT in tobacco (Nicotiana tabacum L. cv. Xanthi). The 5' flanking element of GLU1 directed the expression of the uidA reporter gene in the palisade and spongy parenchyma of mesophyll, in the phloem cells of vascular tissue and in the roots of tobacco. White light, red light or sucrose induced GUS expression in the dark-grown seedlings in a pattern similar to the GLU1 mRNA accumulation in Arabidopsis. The levels of GLU2 mRNA encoding the second Fd-GOGAT and NADH-glutamate synthase (NADH-GOGAT, EC 1.4.1.14) were not affected by light. Both in the light and in darkness, (15)NH4(+) was incorporated into [5-(15)N]glutamine and [2-(15)N]glutamate by glutamine synthetase (GS, EC 6.3.1.2) and Fd-GOGAT in leaf disks of transgenic tobacco expressing antisense Fd-GOGAT mRNA and in wild-type tobacco. In the light, low level of Fd-glutamate synthase limited the [2-(15)N]glutamate synthesis in transgenic leaf disks. The efficient dark labeling of [2-(15)N]glutamate in the antisense transgenic tobacco leaves indicates that the remaining Fd-GOGAT (15-20% of the wild-type activity) was not the main limiting factor in the dark ammonium assimilation. The antisense tobacco under high CO2 contained glutamine, glutamate, asparagine and aspartate as the bulk of the nitrogen carriers in leaves (62.5%), roots (69.9%) and phloem exudates (53.2%). The levels of glutamate, asparagine and aspartate in the transgenic phloem exudates were similar to the wild-type levels while the glutamine level increased. The proportion of these amino acids remained unchanged in the roots of the transgenic plants. Expression of GLU1 in mesophyll cells implies that Fd-GOGAT assimilates photorespiratory and primary ammonium. GLU1 expression in vascular cells indicates that Fd-GOGAT provides amino acids for nitrogen translocation.

Amino Acid Oxidoreductases↗

Root xylem embolisms and refilling. Relation To water potentials of soil, roots, and leaves, and osmotic potentials of root xylem Sap

Embolism and refilling of vessels was monitored directly by cryomicroscopy of field-grown corn (Zea mays L.) roots. To test the reliability of an earlier study showing embolism refilling in roots at negative leaf water potentials, embolisms were counted, and root water potentials (Psiroot) and osmotic potentials of exuded xylem sap from the same roots were measured by isopiestic psychrometry. All vessels were full at dawn (Psiroot -0.1 MPa). Embolisms were first seen in late metaxylem vessels at 8 AM. Embolized late metaxylem vessels peaked at 50% at 10 AM (Psiroot -0.1 MPa), fell to 44% by 12 PM (Psiroot -0.23 MPa), then dropped steadily to zero by early evening (Psiroot -0.28 MPa). Transpiration was highest (8.5 μg cm-2 s-1) between 12 and 2 PM when the percentage of vessels embolized was falling. Embolized vessels were refilled by liquid moving through their lateral walls. Xylem sap was very low in solutes. The mechanism of vessel refilling, when Psiroot is negative, requires further investigation. Daily embolism and refilling in roots of well-watered plants is a normal occurrence and may be a component of an important hydraulic signaling mechanism between roots and shoots.

Journal Article↗

A novel function of abscisic acid in the regulation of rice (Oryza sativa L.) root growth and development.

Plant roots retain developmental plasticity and respond to environmental stresses or exogenous plant growth regulators by undergoing profound morphological and physiological alteration. In this study, we investigated the effects of exogenous ABA on root growth and development in Taichung native 1 (TN1) rice. Exogenous application of 10 microM ABA leads to swelling, root hair formation and initiation of lateral root primodia in the tips of young, seminal rice roots. Cortex cells increased in size and were irregularly shaped. ABA treatment significantly increased 2, 3, 5-triphenyl tetrazolium chloride (TTC) reductase ability in the root tips and the exudation rate of xylem sap. In addition, the K(+) ion content in xylem sap increased nearly 2-fold, but not that of Ca(2+) or Mg(2+). Analysis of proteins expressed in the root tips identified several ABA-induced or -repressed proteins, including actin depolymerization factor (ADF), late embryo abundant protein (LEA), putative steroid membrane-binding protein, ferredoxin thionine reductase and calcium-binding protein. The effects of ABA on root morphogenesis change were Ca(2+) dependent and required the participation of calmodulin and de novo protein synthesis. A model is presented that illustrates how ABA acts through a potential cellular and signal transduction mechanism to induce morphological and physiological changes in rice roots.

Abscisic Acid↗

Calcium hydroxide barrier over the apical root-end of a type III dens invaginatus after endodontic and surgical treatment.

AIM: To report the simultaneous endodontic and surgical treatment of a tooth associated with Oehlers type III dens invaginatus and a persistent periapical lesion, which comprised root-end resection, root-end filling and application of a calcium hydroxide barrier placed on the resected dentine surface. SUMMARY: Three root canals were identified in a tooth with a type III dens invaginatus, which presented with a necrotic pulp, wide foraminal opening and extensive periapical lesion, and with a previous history of acute abscess, intracanal exudate and fistula. After root canal preparation followed by intracanal application of calcium hydroxide pastes, the clinical-pathological status persisted. After periapical curettage and root-end resection, the root canals were filled, followed by root-end filling with Sealer 26 mixed with zinc oxide powder to a clay-like consistency. Calcium hydroxide paste was then applied over the exposed dentinal surface forming a covering over the root apex. At the 20-month follow-up examination the patient had no symptoms and no fistula; advanced periapical bone repair was obvious on the radiograph. KEY LEARNING POINTS: Because of the variable morphology and extent of invagination, type III dens invaginatus represents a challenge for conventional treatment, often leading to the need for a surgical approach. Sealer 26 thickened with zinc oxide powder provided satisfactory clinical properties for use as a root-end filling material. Application of a calcium hydroxide barrier over the resected root-end is a potential treatment option to encourage tissue repair.

Adolescent↗

Oxalate exudation by taro in response to Al

Roots of taro (Colocasia esculenta [L.] Schott cvs Bun-long and Lehua maoli) exuded increasing concentrations of oxalate with increasing Al stress. This exudation was a specific response to excess Al and not to P deficiency. Addition of oxalate to Al-containing solutions ameliorated the toxic effect of Al.

Journal Article↗

Interleukin-1beta and tumour necrosis factor-alpha levels in periapical exudates.

AIM: The aim of this study was to determine IL-1beta and TNF-alpha levels in periapical exudates and to evaluate their relationship with clinical and radiological findings. METHODOLOGY: Periapical exudates were collected from root canals of 35 single-rooted teeth using absorbent paper points. IL-1beta and TNF-alpha levels were determined by enzyme-linked immunosorbent assays. The samples were divided into two groups according to the presence or absence of clinical signs including swelling and/or fistula, pain on palpation and/or percussion, and pus discharge from canals. Periapical exudate samples were divided into two groups according to size of periapical radiolucent area. RESULTS: The mean concentration of IL-1beta (72.79 ng mL-1) in periapical exudates was approximately 12-fold higher than TNF-alpha(6.17 ng mL-1). There was no significant correlation between these cytokines (P > 0.05). IL-1beta levels in canals with larger radiolucent areas (long axis > or =1 cm) were significantly higher than those with small areas (P < 0.05). There was a tendency toward higher levels of IL-1beta in groups with clinical signs, but the differences were not significant (P > 0.05). CONCLUSIONS: Periapical exudate levels of both cytokines failed to reflect periapical disease state.

Alveolar Bone Loss↗

Phytoextraction of weathered p,p'-DDE by zucchini (Cucurbita pepo) and cucumber (Cucumis sativus) under different cultivation conditions.

Previous studies have shown that zucchini (Cucurbita pepo) and cucumber (Cucumis sativus) under field conditions are good and poor accumulators, respectively, of persistent organic pollutants from soil. Here, each species was grown under three cultivation regimes: dense (five plants in 5 kg soil): nondense (one plant in 80 kg soil): and field conditions (two to three plants in approximately 789 kg soil). p,p'-DDE and inorganic element content in roots, stems, leaves, and fruit were determined. In addition. rhizosphere, near-root, and unvegetated soil fractions were analyzed for concentrations of 11 low-molecular-weight organic acids (LMWOA) and 14 water-extractable inorganic elements. Under field conditions, zucchini phytoextracted 1.3% of the weathered p,p'-DDE with 98% of the contaminant in the aerial tissues. Conversely, cucumber removed 0.09% of the p,p'-DDE under field conditions with 83% in the aerial tissues. Under dense cultivation, cucumber produced a fine and fibrous root system not observed in our previous experiments and phytoextracted 0.78% of the contaminant, whereas zucchini removed only 0.59% under similar conditions. However. cucumber roots translocated only 5.7% of the pollutant to the shoot system, while in zucchini 48% of the p,p'-DDE in the plant was present in the aerial tissue. For each species, the concentrations of LMWOA in soil increased with increasing impact by the root system both within a given cultivation regime (i.e., rhizosphere > near-root > unvegetated) and across cultivation regimes (i.e., dense > nondense > field conditions). Under dense cultivation, the rhizosphere concentrations of LMWOAs were significantly greater for cucumber than for zucchini; no species differences were evident in the other two cultivation regimes. To enable direct comparison across cultivation regimes, total in planta p,p'-DDE and inorganic elements were mass normalized or multiplied by the ratio of plant mass to soil mass. For cucumber, differences in total p,p'-DDE and inorganic element content among the cultivation regimes largely disappear upon mass normalization, indicating that greater uptake of both types of constituents in the dense condition is due to greater plant biomass per unit soil. Conversely, for zucchini the mass normalized content of p,p'-DDE and inorganic elements is up to two orders of magnitude greater under field conditions than under dense cultivation, indicating a unique physiological response of C. pepo in the field. The role of cultivation conditions and nutrient availability in controlling root morphology, organic acid exudation, and contaminant uptake is discussed.

Biodegradation, Environmental↗

Accumulation of lipochitin oligosaccharides and NodD-activating compounds in an efficient plant--Rhizobium nodulation assay.

During legume plant--Rhizobium spp. interactions, leading to the formation of nitrogen-fixing root nodules, the two major determinants of host plant-specificity are plant-produced nod gene inducers (NodD protein activating compounds) and bacterial lipochitin oligosaccharides (LCOs or Nod factors). In a time course, we describe the accumulation of LCOs in an efficient nodulation assay with Vicia sativa subsp. nigra and Rhizobium leguminosarum, in connection with the presence of NodD-activating compounds in the exudate of V. sativa roots. Relatively small amounts of both LCOs and NodD-activating compounds were found to be required for initiation of nodulation during the first days after inoculation. A strong increase in the amount of NodRlv-V[18:4,Ac] LCOs preceded root infection and nodule primordium formation. In contrast to the situation with non-nodulating rhizobia and nonmitogenic LCOs, the amount of NodD-activating compounds in the culture medium remained small after addition of nodulating rhizobia or mitogenic LCOs. Furthermore, addition of nodulating rhizobia or mitogenic LCOs resulted in nearly complete inhibition of root hair formation and elongation, whereas nonmitogenic LCOs stimulated root hair growth. Retention of NodD-activating compounds in the root may inhibit root hair growth.

Bacterial Proteins↗

Glucose-induced activation of rubidium transport and water flux in sunflower root systems.

Excised 20-d-old sunflower roots (Helianthus annuus L. cv. Sun-Gro 393) were used to study the effect of different sugars on rubidium and water fluxes. The roots sensed and absorbed glucose from the external medium inducing the activation of rubidium accumulated in the root (Rb(+) root), the flux of exuded rubidium (J(Rb)) and, to a lesser degree, the exudation rate (J(v)). These effects were also triggered by fructose, but not by 6-deoxyglucose (6-dG), a glucose analogue which is not a substrate for hexokinase (HXK). The effect of 2-deoxyglucose (2-dG), an analogue that is phosphorylated but not further metabolized, was complex, suggesting an inhibitory effect on solute transport to the xylem. The amounts of glucose required to activate rubidium and water fluxes were similar to those previously reported to regulate different processes in other plants (0.5--10 mM). When sorbitol was used instead of glucose, neither rubidium uptake (Rb(+) root plus J(Rb)) nor J(v) was activated. It is proposed that glucose present in the root plays an important signalling role in the regulation of Rb(+) (K(+)) and water transport in plant roots.

Carbon Radioisotopes↗