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Increased ecological amplitude through heterosis following wide outcrossing in Banksia ilicifolia R.Br. (Proteaceae).

To assess whether wide outcrossing (over 30 km) in the naturally fragmented Banksia ilicifolia R.Br. increases the ecological amplitude of offspring, we performed a comparative greenhouse growth study involving seedlings of three hand-pollinated progeny classes (self, local outcross, wide outcross) and a range of substrates and stress conditions. Outcrossed seedlings outperformed selfed seedlings, with the magnitude of inbreeding depression as high as 62% for seed germination and 37% for leaf area. Wide outcrossed seedlings outperformed local outcrossed seedlings, especially in non-native soils, facilitated in part by an improved capacity to overcome soil constraints through greater root carboxylate exudation. Soil type significantly affected seedling growth, and waterlogging and water deficit decreased growth, production of cluster roots, root exudation and total plant P uptake. Our results suggest that the interaction of narrow ecological amplitude and the genetic consequences of small fragmented populations may in part explain the narrow range of local endemics, but that wide outcrossing may provide opportunities for increased genetic variation, increased ecological amplitude and range expansion.

Ecology↗

Effects of external cations and respiratory inhibitors on electrical potential of the xylem exudate of excised corn roots.

Glass capillary microelectrodes were used to study the electrical potential difference (PD) between the xylem exudate of excised corn roots, Zea mays L. Golden Bantam hybrid, and the external solution. A survey of the effects of various ions on the PD was made. With 1 mm single salt solutions, the PD was between 25 and 50 mv, exudate negative. The PD responded to concentration differences in single salt solutions of K(+), Na(+), and Ca(2+) in a manner suggestive of cation selectivity and cation diffusion potentials. With Ca(2+) present, the PD was insensitive to concentration changes of other cations. Substitution of NO(3) (-) for Cl(-) in K(+) solutions increased the PD by 2 to 5 mv, although in general the PD showed little response to anion concentration changes. The PD was partially abolished by cyanide. The remaining fraction of the PD was sensitive to concentration changes in external K(+), and we postulate that the PD is the result of both a diffusion potential and an electrogenic pump.

Journal Article↗

Restricted nitrate influx and reduction in corn seedlings exposed to ammonium.

The effect of ambient ammonium (0.5 millimolar [(14)NH(4)](2)SO(4)) added to a nutrient solution containing 1.0 millimolar K(15)NO(3), 99 atom per cent (15)N, upon [(15)N]nitrate assimilation and utilization of previously accumulated [(14)N]nitrate was investigated. Corn seedlings, 5-day-old dark-grown decapitated (experiment I) and 10-day-old light-grown intact (experiment II), which had previously been grown on K(14)NO(3) nutrient solution, were used. In both experiments, the presence of ambient ammonium decreased [(15)N]nitrate influx (20% after 6 hours) without significantly affecting the efflux of previously accumulated [(14)N]nitrate. In experiment I, relative reduction of [(15)N]nitrate (reduction as a percentage of influx) was inhibited more than was [(15)N]nitrate influx. Nevertheless, in experiment I, where all reduction could be assigned to the root system, the absolute inhibition of reduction during the 12 hours (13 micromoles/root) was less than the absolute inhibition in influx (24 micromoles/root). The data suggest that the influence of ammonium on [(15)N]nitrate influx could not be totally accounted for by the decrease in the potential driving force which resulted from restricted reduction; an additional impact on the influx process is indicated. Reduction of [(15)N]nitrate in experiment II after 6 hours accounted for 30 and 18% of the tissue excess (15)N in the control and ammonium treatments, respectively. Relative distribution of (15)N between roots and exudate (experiment I), or between roots and shoots (experiment II) was not affected by ammonium. On the other hand, the accumulation of [(15)N]nitrate in roots, shoots, and xylem exudate was enhanced by ammonium treatment compared to the control, whereas the accumulation of reduced (15)N was inhibited.

Journal Article↗

Mediation of pathogen resistance by exudation of antimicrobials from roots.

Most plant species are resistant to most potential pathogens. It is not known why most plant-microbe interactions do not lead to disease, although recent work indicates that this basic disease resistance is multi-factorial. Here we show that the exudation of root-derived antimicrobial metabolites by Arabidopsis thaliana confers tissue-specific resistance to a wide range of bacterial pathogens. However, a Pseudomonas syringae strain that is both at least partly resistant to these compounds and capable of blocking their synthesis/exudation is able to infect the roots and cause disease. We also show that the ability of this P. syringae strain to block antimicrobial exudation is dependent on the type III secretory system.

Anti-Infective Agents↗

Developmental physiology of cluster-root carboxylate synthesis and exudation in harsh hakea. Expression of phosphoenolpyruvate carboxylase and the alternative oxidase.

Harsh hakea (Hakea prostrata R.Br.) is a member of the Proteaceae family, which is highly represented on the extremely nutrient-impoverished soils in southwest Australia. When phosphorus is limiting, harsh hakea develops proteoid or cluster roots that release carboxylates that mobilize sparingly soluble phosphate in the rhizosphere. To investigate the physiology underlying the synthesis and exudation of carboxylates from cluster roots in Proteaceae, we measured O2 consumption, CO2 release, internal carboxylate concentrations and carboxylate exudation, and the abundance of the enzymes phosphoenolpyruvate carboxylase and alternative oxidase (AOX) over a 3-week time course of cluster-root development. Peak rates of citrate and malate exudation were observed from 12- to 13-d-old cluster roots, preceded by a reduction in cluster-root total protein levels and a reduced rate of O2 consumption. In harsh hakea, phosphoenolpyruvate carboxylase expression was relatively constant in cluster roots, regardless of developmental stage. During cluster-root maturation, however, the expression of AOX protein increased prior to the time when citrate and malate exudation peaked. This increase in AOX protein levels is presumably needed to allow a greater flow of electrons through the mitochondrial electron transport chain in the absence of rapid ATP turnover. Citrate and isocitrate synthesis and accumulation contributed in a major way to the subsequent burst of citrate and malate exudation. Phosphorus accumulated by harsh hakea cluster roots was remobilized during senescence as part of their efficient P cycling strategy for growth on nutrient impoverished soils.

Carbon Dioxide↗

[Effects of 6-BA on cluster root formation and organic acid exudation in white lupin grown under phosphorus deficiency].

Phosphorus deficiency results in cluster root formation and increased organic acid exudation. The regulatory mechanisms for these processes, however, are not yet clear. In the present study, influences of 6-BA (6-benzyl aminopurine) on cluster root formation, exudation of citrate and malate and their concentrations in the root clusters of P-deficient white lupin plants were studied by using non-destructive localized collection method and high-performance liquid chromatography (HPLC) technique. The results showed that application of exogenous 6-BA to P-deficient plants did not influence plant growth and P distribution within the plant (Tables 2,3), whereas the cluster root formation (Fig. 1, Table 1) and organic acid exudation (Table 4) were inhibited. The inhibitory effects could be reversed by omitting 6-BA from the growth medium, and even some stimulatory effects was observed, when lower concentration of 6-BA (10(-8) mol/L) was applied (Fig. 1, Tables 1 and 4). The inhibitory effects of higher concentration of 6-BA (10(-7) mol/L) were not reversible. (Table 4). Treatment with 6-BA also had some influence on organic acid concentration in the tissue of cluster roots (Table 5). The possible reasons for the effects on cluster root formation and organic acid exudation by 6-BA are discussed.

Acids↗

Flagella-driven chemotaxis towards exudate components is an important trait for tomato root colonization by Pseudomonas fluorescens.

Motility is a major trait for competitive tomato root-tip colonization by Pseudomonas fluorescens. To test the hypothesis that this role of motility is based on chemotaxis toward exudate components, cheA mutants that were defective in flagella-driven chemotaxis but retained motility were constructed in four P. fluorescens strains. After inoculation of seedlings with a 1:1 mixture of wild-type and nonmotile mutants all mutants had a strongly reduced competitive root colonizing ability after 7 days of plant growth, both in a gnotobiotic sand system as well as in nonsterile potting soil. The differences were significant on all root parts and increased from root base to root tip. Significant differences at the root tip could already be detected after 2 to 3 days. These experiments show that chemotaxis is an important competitive colonization trait. The best competitive root-tip colonizer, strain WCS365, was tested for chemotaxis toward tomato root exudate and its major identified components. A chemotactic response was detected toward root exudate, some organic acids, and some amino acids from this exudate but not toward its sugars. Comparison of the minimal concentrations required for a chemotactic response with concentrations estimated for exudates suggested that malic acid and citric acid are among major chemo-attractants for P. fluorescens WCS365 cells in the tomato rhizosphere.

Amino Acids↗

The change of periapical exudate prostaglandin E2 levels during root canal treatment.

We have previously demonstrated that detectable levels of prostaglandin E2 exist in periapical exudates (PE-PGE2) from root canals of periapical periodontitis. Our data from a cross-sectional study also suggested the association of PE-PGE2 concentrations with the active phase of periapical lesions. The objective of this study was to investigate the longitudinal changes of PE-PGE2 levels during root canal treatment. Periapical exudate specimens were quantitatively sampled from root canals of 20 nonvital teeth at consecutive treatment visits to measure PGE2 concentrations. The mean PE-PGE2 levels significantly decreased following the endodontic therapy (158.0 +/- 54.1 pg/microliter in the first samples versus 48.4 +/- 37.9 pg/microliter in the second samples). A significant correlation was also found between the changes in PE-PGE2 levels and the clinical disease expression of periapical periodontitis (p < 0.01). Decreased PGE2 production is considered to reflect the remission of disease activity. These results suggest the central roles of PGE2 in regulatory mechanisms for the expression of periapical disease.

Dental Pulp Cavity↗

Multiple Aluminum-Resistance Mechanisms in Wheat (Roles of Root Apical Phosphate and Malate Exudation).

Although it is well known that aluminum (Al) resistance in wheat (Triticum aestivum) is multigenic, physiological evidence for multiple mechanisms of Al resistance has not yet been documented. The role of root apical phosphate and malate exudation in Al resistance was investigated in two wheat cultivars (Al-resistant Atlas and Al-sensitive Scout) and two near-isogenic lines (Al-resistant ET3 and Al-sensitive ES3). In Atlas Al resistance is multigenic, whereas in ET3 resistance is conditioned by the single Alt1 locus. Based on root- growth experiments, Atlas was found to be 3-fold more resistant in 20 [mu]M Al than ET3. Root-exudation experiments were conducted under sterile conditions; a large malate efflux localized to the root apex was observed only in Atlas and in ET3 and only in the presence of Al (5 and 20 [mu]M). Furthermore, the more Al-resistant Atlas exhibited a constitutive phosphate release localized to the root apex. As predicted from the formation constants for the Al-malate and Al-phosphate complexes, the addition of either ligand to the root bathing solution alleviated Al inhibition of root growth in Al-sensitive Scout. These results provide physiological evidence that Al resistance in Atlas is conditioned by at least two genes. In addition to the alt locus that controls Al-induced malate release from the root apex, other genetic loci appear to control constitutive phosphate release from the apex. We suggest that both exudation processes act in concert to enhance Al exclusion and Al resistance in Atlas.

Journal Article↗

Radial salt transport in corn roots.

Primary roots of solution-grown, 5-day-old or 6-day-old seedlings of corn (Zea mays L.) 10 to 14 cm in length were used to study radial salt transport. Measurements were made of the volume of root pressure exudation, salt concentration of the exudate, and rate of salt movement into the xylem exudate. The (32)P uptake, O(2) consumption, and dehydrogenase activity of the root cortex and stele also were studied.These roots produced copious root pressure exudate containing 4 to 10 times the concentration of (32)P in the external solution. Freshly separated stele from 5-day-old roots accumulated (32)P as rapidly as the cortex from which it was separated and the stele of intact roots also accumulated (32)P. Separated stele has a higher oxygen uptake than cortex. It also shows strong dehydrogenase activity with the tetrazolium test. The high oxygen consumption, (32)P uptake and strong dehydrogenase activity indicate that the cells of the stele probably play a direct role in salt transport.These data raise doubts concerning theories of radial salt transport into the xylem based on the assumption that the stele is unable to accumulate salt vigorously.

Journal Article↗

Root water flow and leaf stomatal conductance in aspen (Populus tremuloides) seedlings treated with abscisic acid.

Exogenous abscisic acid (ABA) applied to the roots and excised shoots of aspen (Populus tremuloides Michx.) inhibited stomatal conductance. However, the effect of ABA on stomatal conductance was more pronounced in the excised shoots compared with the intact seedlings. Approximately 10% of the ABA concentration applied to the roots was found in the xylem exudates of root systems exposed to a hydrostatic pressure of 0.3 MPa. A similar concentration of ABA applied to the excised shoots produced a faster and greater reduction of stomatal conductance. ABA applied to the roots had no effect on root steady-state flow rate over the 5-h experimental period. Moreover, pre-incubating root systems of intact seedlings for 12 h with 5 x 10(-5) M ABA did not significantly reduce volume flow density. Similarly, ABA had no effect on root hydraulic conductivity and the activation energy of root water flow rates.

Abscisic Acid↗

Electronic ohmmeter. An electronic device for the determination of the root canal length.

The electronic ohmmeter was devised for the accurate determination of root canal length. Fifty-one maxillary anterior teeth of 43 patients and 25 posterior root canals from 10 patients were selected for the investigation. The presence of a lesion in periapical tissue, wide root apex, and exudates in the root canal contributed to inaccuracy in the determination of the root canal length. Correct length can be found if precautions are taken. The inaccurate results were corrected by repeating the procedures and taking precautions during the estimation of root canal length. Measurement in the dry canal gave correct estimation of the root canal length. When distilled water or normal saline was used as conducting medium in root canals, accuracy was not as good as in dry canals. The electronic ohmmeter is a useful and economical device and an endodontic aid for determination of the root canal length.

Adolescent↗

Radial movement of oxygen in plant roots.

The radial movement of oxygen in excised corn and jack bean roots was measured with a platinum wire electrode embedded in the root tissue. Measurements were made with the roots exposed to air and with the roots immersed in nutrient solution in the presence and absence of millimolar sodium azide. Effective rates of oxygen diffusion in the root tissue were also measured from 5 to 30 C and compared to the respiration rates of similar root segments over the same temperature range. Under conditions which allow the roots to exude freely, the interior of the root operates under an oxygen deficit. Inhibition of respiratory oxygen uptake by low temperature or azide treatment increased the flux of oxygen to the root interior.

Journal Article↗

Plant growth promotion abilities and microscale bacterial dynamics in the rhizosphere of Lupin analysed by phytate utilization ability.

In the rhizosphere, phosphorus (P) levels are low because of P uptake into the roots. Rhizobacteria live on carbon (C) exuded from roots, and may contribute to plant nutrition by liberating P from organic compounds such as phytates. We isolated over 300 phytate (Na-inositol hexa-phosphate; Na-IHP)-utilizing bacterial strains from the rhizosheath and the rhizoplane of Lupinus albus (L.). Almost all of the isolates were classified as Burkholderia based on 16S rDNA sequence analysis. Rhizosheath isolates cultured with Na-IHP as the only source of C and P showed lower P uptake at the same extracellular phytase activity than rhizoplane strains, suggesting that bacteria from the rhizosheath utilized phytate as a C source. Many isolates also utilized insoluble phytate (Al-IHP and/or Fe-IHP). In co-culture with Lotus japonicus seedlings, some isolates promoted plant growth significantly.

Burkholderia↗

Rhizodeposition and the enhanced mineralization of 2,4-dichlorophenoxyacetic acid in soil from the Trifolium pratense rhizosphere.

Enhanced biodegradation of organic xenobiotic compounds in the rhizosphere is frequently recorded although the specific mechanisms are poorly understood. We have shown that the mineralization of 2,4-dichlorophenoxyacetic acid (2,4-D) is enhanced in soil collected from the rhizosphere of Trifolium pratense[e.g. maximum mineralization rate=7.9 days-1 and time at maximum rate (t1)=16.7 days for 12-day-old T. pratense soil in comparison with 4.7 days-1 and 25.4 days, respectively, for non-planted controls). The purpose of this study was to gain a better understanding of the plant-microbe interactions involved in rhizosphere-enhanced biodegradation by narrowing down the identity of the T. pratense rhizodeposit responsible for stimulating the microbial mineralization of 2,4-D. Specifically, we investigated the distribution of the stimulatory component(s) among rhizodeposit fractions (exudates or root debris) and the influence of soil properties and plant species on its production. Production of the stimulatory rhizodeposit was dependent on soil pH (e.g. t1 for roots grown at pH 6.5 was significantly lower than for those grown at pH 4.4) but independent of soil inorganic N concentration. Most strikingly, the stimulatory rhizodeposit was only produced by T. pratense grown in non-sterile soil and was present in both exudates and root debris. Comparison of the effect of root debris from plant species (three each) from the classes monocotyledon, dicotyledon (non-legume) and dicotyledon (legume) revealed that legumes had by far the greatest positive impact on 2,4-D mineralization kinetics. We discuss the significance of these findings with respect to legume-rhizobia interactions in the rhizosphere.

2,4-Dichlorophenoxyacetic Acid↗

Reduction of Hydraulic Conductivity during Inhibition of Exudation from Excised Maize and Barley Roots.

The uncoupler, carbonyl cyanide m-chlorophenyl hydrazone (CCCP) is shown to reduce the hydraulic conductivity of barley, maize, mung bean, and onion roots. In barley and maize, the reduction in exudation from excised roots is partly due to the reduction in the permeability of the root to water (I(p)), but it can be inferred that the rate of salt release to the xylem, is also inhibited. The action of CCCP on L(p) is suggested to be mainly in blocking the symplasmic pathway at the plasmodesmata.

Journal Article↗

Spatial distribution of bacterial communities and phenanthrene degradation in the rhizosphere of Lolium perenne L.

Rhizodegradation of organic pollutants, such as polycyclic aromatic hydrocarbons, is based on the effect of root-produced compounds, known as exudates. These exudates constitute an important and constant carbon source that selects microbial populations in the plant rhizosphere, modifying global as well as specific microbial activities. We conducted an experiment in two-compartment devices to show the selection of bacterial communities by root exudates and phenanthrene as a function of distance to roots. Using direct DNA extraction, PCR amplification, and thermal gradient gel electrophoresis screening, bacterial population profiles were analyzed in parallel to bacterial counts and quantification of phenanthrene biodegradation in three layers (0 to 3, 3 to 6, and 6 to 9 mm from root mat) of unplanted-polluted (phenanthrene), planted-polluted, and planted-unpolluted treatments. Bacterial community differed as a function of the distance to roots, in both the presence and the absence of phenanthrene. In the planted and polluted treatment, biodegradation rates showed a strong gradient with higher values near the roots. In the nonplanted treatment, bacterial communities were comparable in the three layers and phenanthrene biodegradation was high. Surprisingly, no biodegradation was detected in the section of planted polluted treatment farthest from the roots, where the bacterial community structure was similar to those of the nonplanted treatment. We conclude that root exudates and phenanthrene induce modifications of bacterial communities in polluted environments and spatially modify the activity of degrading bacteria.

Bacteria↗