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PubMed2006

Root growth and exudate production define the frequency of horizontal plasmid transfer in the Rhizosphere.

To identify the main drivers of plasmid transfer in the rhizosphere, conjugal transfer was studied in the rhizospheres of pea and barley. The donor Pseudomonas putida KT2442, containing plasmid pKJK5::gfp, was coated onto the seeds, while the recipient P. putida LM24, having a chromosomal insertion of dsRed, was inoculated into the growth medium. Mean transconjugant-to-donor ratios in vermiculite were 4.0+/-0.8 x 10(-2) in the pea and 5.9+/-1.4 x 10(-3) in the barley rhizospheres. In soil, transfer ratios were about 10 times lower. As a result of a 2-times higher root exudation rate in pea, donor densities in pea (1 x 10(6)-2 x 10(9) CFU g(-1) root) were about 10 times higher than in barley. No difference in recipient densities was observed. In situ visualization of single cells on the rhizoplane and macroscopic visualization of the colonization pattern showed that donors and transconjugants were ubiquitously distributed in the pea rhizosphere, while they were only located on the upper parts of the barley roots. Because the barley root elongated about 10 times faster than the pea root, donors were probably outgrown by the elongating barley root. Thus by affecting the cell density and distribution, exudation and root growth appear to be key parameters controlling plasmid transfer in the rhizosphere.

Gene Transfer, Horizontal↗
PubMed2004

Phosphatase under-producer mutants have altered phosphorus relations.

Phosphorus (P) acquisition and partitioning are essential for plant homeostasis. P is available for plant uptake when in its inorganic form (H2PO4-, or Pi), but Pi is often limiting in soils. Plants secrete acid phosphatases (APases) into the apoplastic space, which may be important for obtaining Pi from organic P sources; however, the relative importance of these enzymes for plant P nutrition has yet to be determined. We demonstrate that the root-associated APase pool is increased in Arabidopsis when Pi is limiting and document five APase isoforms secreted from Arabidopsis roots. Previously, we presented the identification of the phosphatase under-producer (pup) mutants, which have decreased in vivo root APase staining when grown under low P conditions. Here, we present the characterization of one of these, pup3, and further studies with pup1. pup3 has 49%, 38%, and 37% less specific APase activity in exudates, roots, and shoots, respectively. Root-associated APase activity is decreased by 16% in pup1 and 25% in pup3, regardless of P treatment. Two APase activity isoforms are reduced in pup3 exudates, and root and shoot isoforms are also affected. One of the two exudate isoforms is recognized by a polyclonal antibody raised to an Arabidopsis purple APase recombinant protein (AtPAP12); however, AtPAP12 transcript levels are unaffected in the mutant. The pup3 mutation was mapped to 68.4 +/- 6.0 centimorgans on chromosome 5. Although P concentrations were not altered in pup1 and pup3 tissues when grown in nutrient solution in which Pi was the sole source of P, the mutants had 10% (pup1) and 17% (pup3) lower shoot P concentrations when grown in a peat-vermiculite mix in which the majority of the total P was present as organic P. Therefore, the pup defects, which include secreted APases, are functionally important for plant P nutrition.

Acid Phosphatase↗
PubMed2006

Effects of plants on the removal of hexavalent chromium in wetland sediments.

The effect of two wetland plants, Typha latifolia L. (cattail) and Phragmites australis (Cav.) Trin. ex Steud (common reed), on the fate of Cr(VI) in wetland sediments was investigated using greenhouse bench-scale microcosm experiments. The removal of Cr(VI) was monitored based on the vertical profiles of aqueous Cr(VI) in the sediments. The Cr(VI) removal rates were estimated taking into account plant transpiration, which was found to significantly concentrate dissolved species in the sediments. After correcting for evapotranspiration, the actual Cr(VI) removal rates were significantly higher than would be inferred from uncorrected profiles. On average, the Cr(VI) removal rates were 0.005 to 0.017 mg L(-1) d(-1), 0.0003 to 0.08 mg L(-1) d(-1), and 0.004 to 0.13 mg L(-1) d(-1) for the control, T. latifolia, and P. australis microcosms, respectively. The fate of the removed Cr(VI) was examined by determining the quantity and chemical speciation of the Cr in the sediment and plant materials. Chromium(III) was the dominant form of Cr in both the sediment and plants, and precipitation of Cr(III) in the sediment was the major pathway responsible for the disappearance of aqueous Cr(VI) from the pore water. Incubation results showed that abiotic reduction was the primary mechanism underlying Cr(VI) removal in the microcosm sediments. Organic compounds produced by plants, including root exudates and mineralization products of dead roots, are thought to be the factor that is either directly or indirectly responsible for the gap between Cr(VI) removal efficiencies in the sediments of the vegetated and unvegetated microcosms.

Chromium↗
PubMed2003

[Chemical behavior and ecological effects of pollutants acting on root-soil interface].

Root-soil interface is the main entrance of pollutants intruding into plant tissues and the special micro-ecosphere that can result in a series of ecological safety problems. In this review, the concept of root-soil interface is presented and it is suggested that root-soil interface behavior and effects of chemical pollutants should be grouped into two aspects as rhizosphere chemical behavior and ecological effects and root chemical behavior and ecological effects. These two aspects are theoretically discussed according to the latest research advances. Rhizosphere chemical behavior and ecological effects included rhizosphere pH environment and adsorptive behavior, rhizosphere oxidation-reduction behavior, rhizosphere chemical toxicology effects, rhizosphere microbiological effects, and rhizosphere bioenzymatic reaction etc., and root chemical behavior and ecological effects included root exudation, influence of enzymatic systems in roots, disturbance of normal physiological processes, change of cellular structure and function, and interference of structure and function of biological macromolecules. Particular attention should be paid to the significance as well as some problems of research on chemical behavior and ecological effects of pollutants acting root-soil interface in pollution ecology.

Adsorption↗
PubMed2005

Benzo[a]pyrene co-metabolism in the presence of plant root extracts and exudates: Implications for phytoremediation.

Benzo[a]pyrene, a high molecular weight (HMW) polycyclic aromatic hydrocarbon (PAH) was removed from solution by Sphingomonas yanoikuyae JAR02 while growing on root products as a primary carbon and energy source. Plant root extracts of osage orange (Maclura pomifera), hybrid willow (Salix albaxmatsudana), or kou (Cordia subcordata), or plant root exudates of white mulberry (Morus alba) supported 15-20% benzo[a]pyrene removal over 24 h that was similar to a succinate grown culture and an unfed acetonitrile control. No differences were observed between the different root products tested. Mineralization of (14)C-7-benzo[a]pyrene by S. yanoikuyae JAR02 yielded 0.2 to 0.3% (14)CO(2) when grown with plant root products. Collectively, these observations were consistent with field observations of enhanced phytoremediation of HMW PAH and corroborated the hypothesis that co-metabolism may be a plant/microbe interaction important to rhizoremediation. However, degradation and mineralization was much less for root product-exposed cultures than salicylate-induced cultures, and suggested the rhizosphere may not be an optimal environment for HMW PAH degradation by Sphingomonas yanoikuyae JAR02.

Benzo(a)pyrene↗
PubMed2006

Isoflavonoid exudation from white lupin roots is influenced by phosphate supply, root type and cluster-root stage.

The internal concentration of isoflavonoids in white lupin (Lupinus albus) cluster roots and the exudation of isoflavonoids by these roots were investigated with respect to the effects of phosphorus (P) supply, root type and cluster-root developmental stage. To identify and quantify the major isoflavonoids exuded by white lupin roots, we used high-pressure liquid chromatography (HPLC) coupled to electrospray ionization (ESI) in mass spectrometry (MS). The major exuded isoflavonoids were identified as genistein and hydroxygenistein and their corresponding mono- and diglucoside conjugates. Exudation of isoflavonoids during the incubation period used was higher in P-deficient than in P-sufficient plants and higher in cluster roots than in noncluster roots. The peak of exudation occurred in juvenile and immature cluster roots, while exudation decreased in mature cluster roots.Cluster-root exudation activity was characterized by a burst of isoflavonoids at the stage preceding the peak of organic acid exudation. The potential involvement of ATP-citrate lyase in controlling citrate and isoflavonoid exudation is discussed, as well as the possible impact of phenolics in repelling rhizosphere microbial citrate consumers.

Chromatography, Liquid↗
PubMed2006

Identification of Sinorhizobium meliloti early symbiotic genes by use of a positive functional screen.

The soil bacterium Sinorhizobium meliloti establishes nitrogen-fixing symbiosis with its leguminous host plant, alfalfa, following a series of continuous signal exchanges. The complexity of the changes of alfalfa root structures during symbiosis and the amount of S. meliloti genes with unknown functions raised the possibility that more S. meliloti genes may be required for early stages of the symbiosis. A positive functional screen of the entire S. meliloti genome for symbiotic genes was carried out using a modified in vivo expression technology. A group of genes and putative genes were found to be expressed in early stages of the symbiosis, and 23 of them were alfalfa root exudate inducible. These 23 genes were further separated into two groups based on their responses to apigenin, a known nodulation (nod) gene inducer. The group of six genes not inducible by apigenin included the lsrA gene, which is essential for the symbiosis, and the dgkA gene, which is involved in the synthesis of cyclic beta-1,2-glucan required for the S. meliloti-alfalfa symbiosis. In the group of 17 apigenin-inducible genes, most have not been previously characterized in S. meliloti, and none of them belongs to the nod gene family. The identification of this large group of alfalfa root exudate-inducible S. meliloti genes suggests that the interactions in the early stages of the S. meliloti and alfalfa symbiosis could be complex and that further characterization of these genes will lead to a better understanding of the symbiosis.

Bacterial Proteins↗
PubMed1987

Errors in the Measurement of Root Pressure and Exudation Volume Flow Rate Caused by Damage during the Transfer of Unsupported Roots between Solutions.

Plants of Zea mays were grown with their roots confined to growing tubes, consisting of cylindrical or spherical glass tubes fitted at the bottom with a stopcock. Nutrient solution was circulated past the roots, and when a plant was 21 to 25 days old, the stopcock was closed, the root excised from the plant and connected to an apparatus which measured root pressure and exudation volume flow rate. The stopcock was opened and solution was again circulated through the growing tube without dropping the level of the liquid bathing the root in the process. Measurements of pressure and flow rate were made continuously during a period in which the solution was replaced, first by draining and refilling the tube in situ, and second by replacing the growing tube with a beaker of solution. Both these manipulations caused at least temporary and frequently permanent drops in root pressure and flow rate. Plants were also grown in cylindrical tubes with a support medium of either glass beads or Raschig rings which filled the growing tubes. It is shown that the solution bathing these roots could be repeatedly replaced by draining and refilling with no visible effect on the measurements. It is recommended, therefore, that in future, support be provided for the roots of all experimental plants grown by solution culture.

Journal Article↗
PubMed2004

Isolated root caps, border cells, and mucilage from host roots stimulate hyphal branching of the arbuscular mycorrhizal fungus, Gigaspora gigantea.

Unlike previous reports that have shown that water soluble and volatile compounds from roots or root exudates play an important role in precolonization events during arbuscular mycorrhizal (AM) fungus-host root interactions (Bécard & Piché 1989, Giovannetti et al. 1993), the results shown here deal with particulate and viscous fractions isolated from host roots. Root caps and a slow sedimenting particulate fraction (SSPF) were rapidly isolated and separated from Ri T-DNA transformed carrot roots (D. carota) grown in liquid culture. In addition, border cells (BC) and mucilage were isolated from aseptically grown corn seedlings (Zea mays). Root caps, SSPF (composed mainly of small root cap fragments and some BCs), BCs, and mucilage all had an associated AM fungus hyphal branching stimulator. Root caps stored for 5 d at 4 degrees C appeared to either synthesize or slowly release the branching stimulator. Also, isolated root caps from roots grown in the absence of P contained more branch stimulating activity than those isolated from roots grown in the presence of P. Although the branching stimulation activity in particulate fractions was low compared to that of the exudate, the particulate fractions can stick to the root surface at considerable distances from the root tip. This may be significant during the infection and colonization of host roots at sites far removed from the primary location of exudation.

Daucus carota↗
PubMed2002

Cluster roots--an underground adaptation for survival in extreme environments.

Cluster roots are a characteristic of members of the Proteaceae and of several other plant species that are adapted to habitats of extremely low soil fertility, usually without formation of mycorrhizal associations. Functionally linked with intense mobilization of nutrients (P, Fe, Zn, Mn) by root-induced chemical changes (pH, root exudates, redox potential) in the rhizosphere, cluster-rooted plant species can serve as model plants to study rhizosphere processes and regulatory aspects of plant adaptations for chemical mobilization of nutrients in the rhizosphere.

Adaptation, Physiological↗
PubMed2005

Physiological and biochemical characterization of quinclorac resistance in a false cleavers (Galium spurium L.) biotype.

The physiological and biochemical basis for quinclorac resistance in a false cleavers (Galium spurium L.) biotype was investigated. There was no difference between herbicide resistant (R) and susceptible (S) false cleavers biotypes in response to 2,4-D, clopyralid, glyphosate, glufosinate-ammonium, or bentazon. On the basis of GR(50) (growth reduction of 50%) or LD(50) (lethal dose to 50% of tested plants) values, the R biotype was highly resistant to the acetolactate synthase (ALS) inhibitor, thifensulfuron-methyl (GR(50) resistance ratio R/S = 57), and quinolinecarboxylic acids (quinclorac R/S = 46), resistant to MCPA (R/S = 12), and moderately resistant to the auxinic herbicides picloram (R/S = 3), dicamba (R/S = 3), fluroxypyr (R/S = 3), and triclopyr (R/S = 2). The mechanism of quinclorac resistance was not due to differences in [(14)C]quinclorac absorption, translocation, root exudation, or metabolism. Seventy-two hours after root application of quinclorac, ethylene increased ca. 3-fold in S but not R plants when compared to controls, while ABA increased ca. 14-fold in S as opposed to ca. 3-fold in R plants suggesting an alteration in the auxin signal transduction pathway, or altered target site causes resistance in false cleavers. The R false cleavers biotype may be an excellent model system to further examine the auxin signal transduction pathway and the mechanism of quinclorac and auxinic herbicide action.

Abscisic Acid↗
PubMed1998

Characterization of LeMir, a root-knot nematode-induced gene in tomato with an encoded product secreted from the root.

A tomato gene that is induced early after infection of tomato (Lycopersicon esculentum Mill.) with root-knot nematodes (Meloidogyne javanica) encodes a protein with 54% amino acid identity to miraculin, a flavorless protein that causes sour substances to be perceived as sweet. This gene was therefore named LeMir (L. esculentum miraculin). Sequence similarity places the encoded protein in the soybean trypsin-inhibitor family (Kunitz). LeMir mRNA is found in root, hypocotyl, and flower tissues, with the highest expression in the root. Rapid induction of expression upon nematode infection is localized to root tips. In situ hybridization shows that LeMir is expressed constitutively in the root-cap and root-tip epidermis. The LeMir protein product (LeMir) was produced in the yeast Pichia pastoris for generation of antibodies. Western-blot analysis showed that LeMir expression is up-regulated by nematode infection and by wounding. LeMir is also expressed in tomato callus tissue. Immunoprint analysis revealed that LeMir is expressed throughout the seedling root, but that levels are highest at the root/shoot junction. Analysis of seedling root exudates revealed that LeMir is secreted from the root into the surrounding environment, suggesting that it may interact with soil-borne microorganisms.

Amino Acid Sequence↗
PubMed1996

Osmoadaptation by rhizosphere bacteria.

The osmolality of rhizosphere soil water is expected to be elevated in relation to bulk-soil water osmolality as a result of the exclusion of solutes by plant roots during water uptake, the release of plant root exudates, and the production of exopolymers by plant roots and rhizobacteria. In contrast, the osmolality of water within highly hydrated bulk soil is low (less than 50 Osm/kg); thus the ability to adapt to elevated osmolality is likely to be important for successful rhizosphere colonization by rhizobacteria. The present review focuses on the osmoadaptive responses of three gram-negative rhizobacterial genera: Rhizobium, Azospirillum, and Pseudomonas. Specifically, we examine the compatible solutes and osmoprotectants utilized by various species within these genera. The adaptation of rhizobacteria to hypoosmotic environments is also examined in the present review. In particular, we focus on the biosynthesis and accumulation of periplasmic glucans by rhizobacteria. Finally, the relationship between rhizobacterial osmoadaptation and selected plant-microbe interactions is considered.

Adaptation, Biological↗
PubMedDate not supplied

Investigation on temporal variation in methane emission from different rice cultivars under the influence of weeds.

A study of temporal variation in methane efflux from the rice-fields indicated that weeds could modulate the CH4 emission by transporting atmospheric O2 more efficiently than rice plants to the rhizosphere, which suppressed CH4 formation in the oxic condition, inhibiting methanogenic activity. A more oxic environment in the sediment was reflected by the higher redox potential in the weed growing plots. Besides, cultivar differences in methane efflux might be attributed to various plant activities, more importantly root exudation, development of aerenchyma and the biomass. Peak emission of CH4 at the flowering stage in all the rice cultivars was associated with maximum extension of root mat, releasing exudates, which serve as carbon source for the methanogenic bacteria for CH4 formation.

Agriculture↗
PubMed1997

Induced Reporter Gene Activity, Enhanced Stress Resistance, and Competitive Ability of a Genetically Modified Pseudomonas fluorescens Strain Released into a Field Plot Planted with Wheat.

The fates of Pseudomonas fluorescens R2fR and its mutant derivative RIWE8, which contains a lacZ reporter gene responsive to wheat root exudate, were compared in a field microplot. Inoculant survival, root colonization, translocation, resistance to stress factors, and reporter gene activity were assessed in bulk and wheat rhizosphere soils. Populations of both strains declined gradually in bulk and wheat rhizosphere soils and on the wheat rhizoplane as determined by specific CFU and immunofluorescence (IF). In samples from both bulk soil and wheat rhizosphere, IF cell counts were up to 3 orders of magnitude greater than the corresponding numbers of CFU after 120 days, indicating the presence of nonculturable inoculant cells. Estimates of RIWE8-specific target DNA molecule numbers in bulk soil samples 3 and 120 days after inoculation by most-probable-number PCR coincided with the corresponding CFU values. Transport of both strains to deeper soil layers was observed by 3 days after introduction into the microplot. Both strains colonized wheat roots similarly, and cells were seen scattered on the surface of 1-month-old wheat seedling roots by immunogold labelling-scanning electron microscopy. On average, reporter gene activity was significantly higher in wheat rhizosphere soil containing RIWE8 cells than in bulk soil or in soils containing R2fR cells. For both strains, resistance to the four stress factors ethanol, high temperature, high osmotic tension, and oxidative stress increased progressively with residence in soil. Cells from the rhizosphere of 11-day-old seedlings showed similar levels of resistance to osmotic and oxidative stresses and enhanced resistance to ethanol and heat as compared to cells from bulk soil. By 37 days, populations of R2fR and RIWE8 in the rhizosphere were significantly more sensitive to osmotic stress than were populations in bulk soil, whereas differences in response to the other stress factors were less evident. Hence, except for the induction of reporter gene expression in strain RIWE8 in the wheat rhizosphere, the data indicated that there were no great differences in the ecological properties in soil between the lacZ-modified and parental strains.

Journal Article↗
PubMed2001

Allelochemicals in wheat (Triticum aestivum L.): production and exudation of 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one.

An analytical technique employing gas chromatography and tandem mass spectrometry (GC/MS/MS) was employed to systematically screen fifty-eight wheat accessions for their differential production of 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) from three consecutive sources, i.e., the shoots, roots, and in the associated agar growth medium (collected as root exudates) of 17-day-old wheat seedlings. DIMBOA content differed significantly in the shoots, roots, or in the agar growth medium between accessions. DIMBOA accumulated differentially within the plant, with roots containing more DIMBOA than the shoots. Only 19% of accessions were able to exude DIMBOA from living roots into their growth medium, indicating the exudation of DIMBOA is accession-specific. DIMBOA level in root tissues is expected to be high when a high level of DIMBOA content is detected in the shoots. Wheat seedlings did not release detectable amounts of DIMBOA when the DIMBOA level was low in the root tissues. The valuable genetic material with high levels of DIMBOA in the shoots or roots identified in the present research could be used to breed for wheat cultivars with elevated allelopathic activity.

Agriculture↗
PubMed2004

[Difference in oxalate content between buckwheat and soybean leaves and its possible cause].

Buckwheat (Fagopyrum esculentum M.) and soybean (Glycine max L.) seedlings with expanded primary leaves were grown in 1/5 Hoagland culture solution for 10 d, then oxalate content was determined in leaves, roots and root exudates. The result showed that its content in buckwheat was much higher than that in soybean, respectively. This clearly indicated that the oxalate content difference in the leaves was due to the metabolic difference rather than to the different rate in transport to the roots and/or in exudation. Activity of oxalate-degrading enzyme oxalate oxidase could be detected in buckwheat, but not in soybean leaves, implicating that buckwheat leaves had some oxalate-degrading capability. This further demonstrated that the oxalate difference was caused by the change in the oxalate biosynthetic process. Buckwheat leaves contained more glyoxylate, and also higher glyoxylate-oxidizing activity of glycolate oxidase (GO) than soybean. GO from buckwheat had a lower Km for glyoxylate than that from soybean. In combination with the higher level of glyoxylate, buckwheat leaves might have higher rate of oxalate formation from glyoxylate. It is suggested that the rate of glyoxylate oxidation by GO may be one of the critical steps which control oxalate accumulation in plants.

Alcohol Oxidoreductases↗