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Chemotaxis of Bradyrhizobium japonicum to soybean exudates.

The chemotactic response of Bradyrhizobium japonicum toward soybean seed and root exudates was examined. Assays using various isoflavones and fractionated exudate indicated that isoflavones are not the principal attractants in exudates. Likewise, induction of nod genes with isoflavones or seed exudate before assay did not enhance chemotaxis. Screening of numerous compounds revealed that only dicarboxylic acids and the amino acids glutamate and aspartate were strong attractants. The presence of glutamate, aspartate, and dicarboxylic acids in appreciable concentrations in soybean seed and root exudates indicates that these compounds likely represent natural chemoattractants for B. japonicum.

Amino Acids↗

[Capacity evaluation of different media for isolating bacteria populations from the rhizosphere of tomato plants with DGGE].

Bacteria from the rhizosphere of tomato plants were cultured on nutrient broth, YG, soil extract and root exudate media. The capabilities of these 4 media for recovering the bacterium communities were evaluated with denaturing gradient gel electrophoresis (DGGE) technique. Results showed that there were certain differences in the obtained bacterial communities when different media and culture tempitures were applied. Numbers of CFU on solid media indicated that the YG agar medium incubated at 20 degrees C could recover more populations than the high-nutrient-concentration Nutrient agar medium; while the root exudate based medium recovered the highest numbers of bacteria from the rhizosphere of tomato plants. 16S rDNA fragments amplified by PCR from different media and rhizosphere soil bacterium DNA were analyzed with DGGE. The DGGE fingerprints showed that there was a discrepancy between bacterial populations observed by cultivation techniques and molecular retrieval. The rhizosphere pattern consisted of more strong bands than the media. The DGGE patterns of the 4 different media incubated at 28 degrees C showed a relatively high level of similarity. In contrast, those of the 4 media incubated at 20 degrees C were significantly different from each other. Shannon-Wiener index (H), species richness (S), Cs values and Cluster dendrogram analysis of different samples revealed that the YG agar medium and the root exudate based medium recovered the highest proportion of predominant populations from the rhizosphere. In the study a new strategy has been proposed for evaluating the capacity of different media in terms of recovering the predominant populations.

Bacteria↗

Root Carbon Dioxide Fixation by Phosphorus-Deficient Lupinus albus (Contribution to Organic Acid Exudation by Proteoid Roots).

When white lupin (Lupinus albus L.) is subjected to P deficiency lateral root development is altered and densely clustered, tertiary lateral roots (proteoid roots) are initiated. These proteoid roots exude large amounts of citrate, which increases P solubilization. In the current study plants were grown with either 1 mM P (+P-treated) or without P (-P-treated). Shoots or roots of intact plants from both P treatments were labeled independently with 14CO2 to compare the relative contribution of C fixed in each with the C exuded from roots as citrate and other organic acids. About 25-fold more acid-stable 14C, primarily in citrate and malate, was recovered in exudates from the roots of -P-treated plants compared with +P-treated plants. The rate of in vivo C fixation in roots was about 4-fold higher in -P-treated plants than in +P-treated plants. Evidence from labeling intact shoots or roots indicates that synthesis of citrate exuded by -P-treated roots is directly related to nonphotosynthetic C fixation in roots. C fixed in roots of -P-treated plants contributed about 25 and 34% of the C exuded as citrate and malate, respectively. Nonphotosynthetic C fixation in white lupin roots is an integral component in the exudation of large amounts of citrate and malate, thus increasing the P available to the plant.

Journal Article↗

[Root excretion and plant resistance to metal toxicity].

Reducing the availability and toxicity of metal pollutants before they enter plant body is one of the main mechanisms of plant resistance to metals. Root system is the first entrance of metal pollutants from soil into plant. It can excrete organic acids, amino acids, sugars, growth substances etc. into rhizosphere. Root excretion affects the metal absorption by plant through changing the physical and chemical characters(such as pH, Eh) of rhizosphere. The metal pollutants are detained outside the roots by chelation, complexation and disposition with root exudates. The quantity and activity of metals in rhizosphere are changed by root exudates through changing the composition, activity and excretion of microbes there. After reviewing the research advances on root excretion and plant resistance to metals, the authors pointed out the existing problems and the major topics and significance of further research.

Environmental Pollutants↗

Rhizobium nod Gene Inducers Exuded Naturally from Roots of Common Bean (Phaseolus vulgaris L.).

Four compounds exuded from young roots of a black-seeded bean (Phaseolus vulgaris L., cv PI165426CS) induce transcription of nod genes in Rhizobium leguminosarum biovar phaseoli. The three most active nod gene inducers were identified by spectroscopic methods (ultraviolet/visible absorbance, proton nuclear magnetic resonance, and mass spectrometry) as being eriodictyol (5,7,3',4' -tetrahydroxyflavanone), naringenin (5,7,4' -trihydroxyflavanone), and a 7-O-glycoside of genistein (5,7,4' -trihydroxyisoflavone). Comparisons with authentic standards verified the chemical structures of the aglycones and their capacity to induce beta-galactosidase activity in R. leguminosarum strains containing nodA-lacZ or nodC-lacZ fusions controlled by R. leguminosarum biovar phaseoli nodD genes. Roots of 9-day-old seedlings released 42, 281, and 337 nanomoles per plant per day of genistein, eriodictyol, and naringenin, respectively. Genistein and naringenin induced higher maximum beta-galactosidase activities and required lower concentrations for half-maximum induction than eriodictyol. Comparing the nod gene-inducing activity of seed rinses with root exudate from PI165426CS bean showed that root flavonoids were released at about 6% the rate of those from seeds on a molar basis, but on average the individual compounds from roots were approximately three times more active than nod gene inducers from seeds.

Journal Article↗

Production of Tsr factor by Rhizobium meliloti.

The root exudates of alfalfa (Medicago sativa) and mungbean (Vigna radiata) induced the Tsr (thick and short roots) factor production in Rhizobium meliloti. The factor caused a 30-40% reduction of root length in alfalfa seedlings. Pea root exudate had no Tsr induction activity. The flavonoid naringenin could replace the roots in inducing Tsr production. Naringenin-induced Tsr factor caused 70% shortening of main roots. The Tsr inducing property of naringenin was specific since quercetin and syringaldehyde had no such effect.

Bacterial Proteins↗

Hairy roots of Helianthus annuus: a model system to study phytoremediation of tetracycline and oxytetracycline.

The release of antibiotics to the environment has to be controlled because of serious threats to human health. Hairy root cultures of Helianthus annuus (sunflower), along with their inherent rhizospheric activity, provide a fast growing, microbe-free environment for understanding plant-pollutant interactions. The root system catalyzes rapid disappearance of tetracycline (TC) and oxytetracycline (OTC) from aqueous media, which suggests roots have potential for phytoremediation of the two antibiotics in vivo. In addition, in vitro modifications of the two antibiotics by filtered, cell- and microbe-free root exudates suggest involvement of root-secreted compounds. The modification is confirmed from changes observed in UV spectra of exudate-treated OTC. Modification appears to be more dominant at the BCD chromophore of the antibiotic molecule. Kinetic analyses dismiss direct enzyme catalysis; the modification rates decrease with increasing OTC concentrations. The rates increase with increasing age of cultures from which root exudates are prepared. The decrease in modification rates upon addition of the antioxidant ascorbic acid (AA) suggests involvement of reactive oxygen species (ROS) in the antibiotic modification process.

Biodegradation, Environmental↗

Diurnal variations in hydraulic conductivity and root pressure can be correlated with the expression of putative aquaporins in the roots of lotus japonicus

The hydraulic conductivity of excised roots (Lp(r)) of the legume Lotus japonicus (Regel) K. Larsen grown in mist (aeroponic) and sand cultures, was found to vary over a 5-fold range during a day/night cycle. This behaviour was seen when Lp(r) was measured in roots exuding, either under root pressure (osmotic driving force), or under an applied hydrostatic pressure of 0.4 MPa which produced a rate of water flow similar to that in a transpiring plant. A similar daily pattern of variation was seen in plants grown in natural daylight or in controlled-environment rooms, in plants transpiring at ambient rates or at greatly reduced rates, and in plants grown in either aeroponic or sand culture. When detached root systems were connected to a root pressure probe, a marked diurnal variation was seen in the root pressure generated. After excision, this circadian rhythm continued for some days. The hydraulic conductivity of the plasma membrane of individual root cells was measured during the diurnal cycle using a cell pressure probe. Measurements were made on the first four cell layers of the cortex, but no evidence of any diurnal fluctuation could be found. It was concluded that the conductance of membranes of endodermal and stelar cells may be responsible for the observed diurnal rhythm in root Lp(r). When mRNAs from roots were probed with cDNA from the Arabidopsis aquaporin AthPIP1a gene, an abundant transcript was found to vary in abundance diurnally under high-stringency conditions. The pattern of fluctuations resembled closely the diurnal pattern of variation in root Lp(r). The plasma membranes of root cells were found to contain an abundant hydrophobic protein with a molecular weight of about 31 kDa which cross-reacted strongly to an antibody raised against the evolutionarily conserved N-terminal amino acid sequence of AthPIP1a.

Journal Article↗

At the maize/Agrobacterium interface: natural factors limiting host transformation.

BACKGROUND: Agrobacterium tumefaciens has been successfully harnessed as the only natural vector for the incorporation of foreign genes into higher plants, but its use in the grain crops is often limited. Low transformation efficiency has been partly attributed to a failure in the initial events in the transformation process, specifically in the capacity of the VirA/VirG two-component system to induce expression of the virulence genes. RESULTS: Here we show that the root exudate of Zea mays seedlings specifically inhibits virulence gene expression, determine that 2-hydroxy-4,7-dimethoxybenzoxazin-3-one (MDIBOA), which constitutes > 98% of the organic exudate of the roots of these seedlings, is the most potent and specific inhibitor of signal perception in A. tumefaciens-mediated gene transfer yet discovered, and develop a model that is able to predict the MDIBOA concentration at any distance from the root surface. Finally, variants of A. tumefaciens resistant to MDIBOA-mediated inhibition of vir gene expression have been selected and partially characterized. CONCLUSIONS: These results suggest a strategy in which a plant may resist pathogen invasion by specifically blocking virulence gene activation and yet ensure that the 'resistance factor' does not accumulate to levels sufficient to impose toxicity and selection pressure on the pathogen. The data further establish that naturally occurring inhibitors directed against signal perception by the VirA/VirG two-component regulatory system can play an important role in host defense. Finally, selected variants resistant to specific MDIBOA inhibition may now be used to extend the transformation efficiency of maize and possibly other cereals.

Agrobacterium tumefaciens↗

The Rhizobium meliloti putA gene: its role in the establishment of the symbiotic interaction with alfalfa.

Little is known about the energy sources used by rhizobia during colonization, invasion and root nodule formation on leguminous plants. We have recently reported that an impaired proline metabolism in rhizobium meliloti leads to a reduced nodulation efficiency and competitiveness on alfalfa roots. In the present study we have characterized the R. meliloti proline dehydrogenase gene (putA) and addressed the question of its role in symbiosis. This rhizobial gene encodes a 1224-amino-acid-long polypeptide which is homologous to enteric bacteria, Rhodobacter capsulatus and Bradyrhizobium japonicum PutA proteins. Like the situation in these bacteria, sequence analysis identified the proline dehydrogenase (PDH) and pyrroline-5-carboxylate dehydrogenase (P5CDH) domains in the R. meliloti putA-encoded protein. Beta-galactosidase assays performed with free-living cells carrying a putA-lacZ transcriptional fusion revealed that R. meliloti putA gene expression is induced by proline, autoregulated by its encoded product, and independent of the general nitrogen regulatory system (Ntr). In addition, analysis of putA expression during the different steps of the symbiotic interaction with alfalfa showed that expression of this gene is turned on by the root exudates (RE), during root invasion and nodule formation, but not in differentiated nitrogen-fixing bacteroids. Furthermore, we show that the PutA- phenotype leads to a significant reduction of alfalfa root colonization by R. meliloti.

Bacterial Proteins↗

Root mucilage from pea and its utilization by rhizosphere bacteria as a sole carbon source.

Plant roots secrete a complex polysaccharide mucilage that may provide a significant source of carbon for microbes that colonize the rhizosphere. High molecular weight mucilage was separated by high-pressure liquid chromatography gel filtration from low molecular weight components of pea root exudate. Purified pea root mucilage generally was similar in sugar and glycosidic linkage composition to mucilage from cowpea, wheat, rice, and maize, but appeared to contain an unusually high amount of material that was similar to arabinogalactan protein. Purified pea mucilage was used as the sole carbon source for growth of several pea rhizosphere bacteria, including Rhizobium leguminosarum 8401 and 4292, Burkholderia cepacia AMMD, and Pseudomonas fluorescens PRA25. These species grew on mucilage to cell densities of three- to 25-fold higher than controls with no added carbon source, with cell densities of 1 to 15% of those obtained on an equal weight of glucose. Micromolar concentrations of nod gene-inducing flavonoids specifically stimulated mucilage-dependent growth of R. leguminosarum 8401 to levels almost equaling the glucose controls. R. leguminosarum 8401 was able to hydrolyze p-nitrophenyl glycosides of various sugars and partially utilize a number of purified plant polysaccharides as sole carbon sources, indicating that R. leguminosarum 8401 can make an unexpected variety of carbohydrases, in accordance with its ability to extensively utilize pea root mucilage.

Amino Acids↗

[Oil-oxidizing potential of associative rhizobacteria of the genus Azospirillum].

The oil-oxidizing potential of associative rhizobacteria of the genus Azospirillum was studied under laboratory conditions. After screening, A. brasilense strain SR80 was chosen for further investigation. The strain was capable of degrading 56.5% of crude oil (added in a concentration of 1%) over 14 days in a medium containing malate as an additional source of carbon and energy. Studies of associative properties showed that the strain had positive chemotaxis to wheat root exudates, colonized wheat roots, and produced indolyl-3-acetic acid. The synthesis of indolyl-3-acetic acid was not inhibited by oil. Under hydroponic conditions, crude oil stimulated growth of A. brasilense SR80, which promoted development of the wheat root system in the presence of oil and enhanced the level of oil degradation by the plant-microbial association.

Azospirillum↗

The Rhizosphere Microbiome: A Key Mediator of Crop Responses to Fertilization Strategies.

The rhizosphere microbiome, the plant's "second genome" is pivotal for crop nutrient acquisition, health, and stress responses. While fertilization ensures high agricultural yields, a key challenge is reshaping this microbiome to boost crop performance. This review synthesizes how mineral, organic, and bio-organic/microbial inoculant fertilizers affect rhizosphere microbial structure, diversity, and function. Long-term excessive mineral fertilizers (especially nitrogen) reduce microbial diversity, diminish beneficial groups (e.g., diazotrophs, PGPR), and disrupt microbial networks via soil acidification and altered root exudates, causing continuous cropping obstacles. In contrast, organic fertilizers improve soil microenvironments, maintaining high microbial diversity, enriching beneficial taxa (e.g., Proteobacteria, Actinobacteria), and enhancing community complexity. Bio-organic fertilizers/microbial inoculants "engineer" the microbiome by introducing exogenous beneficial microbes (e.g., Bacillus, Pseudomonas, AMF), directly promoting growth, suppressing diseases, and "reconditioning" indigenous beneficial communities. We also clarify how fertilization regulates plant-microbe dialog via root exudates and rhizosphere chemistry (e.g., pH, ion balance), discuss current challenges (causality, lab-to-field translation, genotype-microbiome-fertilization interactions), and outline future directions. Integrating rhizosphere microbiome management into fertilization is crucial for reducing chemical fertilizer reliance and advancing agricultural green transformation.

fertilization strategies microbial community assem↗

Plant growth and cation composition of two cultivars of spring wheat (Triticum aestivum L.) differing in P uptake efficiency.

Phosphorus (P)-zinc (Zn) interactions were investigated in two wheat cultivars (Brookton versus Krichauff) differing in P uptake efficiency. The experiment was done in a growth chamber. Rock phosphate (RP) or CaHPO4 (CaP) were used as P sources, and ammonium nitrate (AN) or nitrate only (NO) were used as nitrogen sources. Two Zn levels were used, 0.22 mg x kg(-1) (LZ) and 2.2 mg ZnSO4.7H2O x kg(-1) (HZ), respectively. P availability significantly affected plant biomass production, but Zn supply had little effect. Plants fed ammonium nitrate had significantly lower concentrations of cations than those fed nitrate only. Cultivar Brookton (with higher P uptake efficiency) consistently had lower concentrations of cations than cv. Krichauff (with low P uptake efficiency) under limited P supply. The differences in concentrations of cations increased with the decrease in P availability, but were not affected by Zn supply. The ratio of potassium in roots to shoots of cultivar Brookton was always higher than in cultivar Krichauff. Based on these findings, it is postulated that the lower concentrations of cations in cultivar Brookton are related to root exudation of organic anions, and a conceptual model is established to describe the regulation of root exudation of organic anions and concentrations of cations.

Anions↗

Change in acceptability of barley plants to aphids after exposure to allelochemicals from couch-grass (Elytrigia repens).

The response of the bird cherry-oat aphid, Rhopalosiphum padi, to barley plants was investigated following exposure of the plants to root allelochemicals from the aggressive weed couch-grass, Elytrigia (Agropyron) repens. Plants were treated either with root exudates from living couch-grass plants or with previously identified couch-grass root compounds 15-hydroxyindole-3-acetic acid, DL-5-hydroxytryptophan, L-5-hydroxytryptophan hydrate, and 6-hydroxy-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (carboline)] either separately or in mixtures. In choice and no-choice settling tests, aphid acceptance of barley plants was significantly reduced following treatment with root exudates, and the carboline when tested alone or in combination with the other compounds. In contrast, the other compounds without the carboline were less active in reducing aphid acceptance. In a probing bioassay, individual substances were either neutral or stimulatory to aphids, indicating that the reduced settling was probably not due to direct effects on aphids, but rather due to effects on the plant. This was confirmed in olfactometer assays, in which aphids were repelled by odors from barley plants following treatment with a mixture containing all four chemicals.

Animals↗

Aluminum resistance in maize cannot be solely explained by root organic acid exudation. A comparative physiological study.

Root apical aluminum (Al) exclusion via Al-activated root citrate exudation is widely accepted as the main Al-resistance mechanism operating in maize (Zea mays) roots. Nonetheless, the correlation between Al resistance and this Al-exclusion mechanism has not been tested beyond a very small number of Al-resistant and Al-sensitive maize lines. In this study, we conducted a comparative study of the physiology of Al resistance using six different maize genotypes that capture the range of maize Al resistance and differ significantly in their genetic background (three Brazilian and three North American genotypes). In these maize lines, we were able to establish a clear correlation between root tip Al exclusion (based on root Al content) and Al resistance. Both Al-resistant genotypes and three of the four Al-sensitive lines exhibited a significant Al-activated citrate exudation, with no evidence for Al activation of root malate or phosphate release. There was a lack of correlation between differential Al resistance and root citrate exudation for the six maize genotypes; in fact, one of the Al-sensitive lines, Mo17, had the largest Al-activated citrate exudation of all of the maize lines. Our results indicate that although root organic acid release may play a role in maize Al resistance, it is clearly not the only or the main resistance mechanism operating in these maize roots. A number of other potential Al-resistance mechanisms were investigated, including release of other Al-chelating ligands, Al-induced alkalinization of rhizosphere pH, changes in internal levels of Al-chelating compounds in the root, and Al translocation to the shoot. However, we were unsuccessful in identifying additional Al-resistance mechanisms in maize. It is likely that a purely physiological approach may not be sufficient to identify these novel Al-resistance mechanisms in maize and this will require an interdisciplinary approach integrating genetic, molecular, and physiological investigations.

Aluminum↗

Effect of volatile substances released from Origanum majorana and Ocimum basilicum on the rhizosphere and phyllosphere fungi of Phaseolus vulgaris.

Differences were found in the counts and occurrence of fungi in the phyllosphere and thizosphere of two representatives of the Lamiacea family, Origanum majorana and Ocimum basilicum, and in the phyllosphere and rhizosphere of Phaseolus vulgaris growing separately or in coenosis with O. majorana or O. basilicum. Both the volatile substances released from ground leaves of the two latter plant species and the root exudates affected considerably spore germination of isolated phylospheric and rhizospheric fungi. The results indicated a possible role of root exudates and volatile substances released from leaves in colonization of rhizosphere and/or phyllosphere by fungi, especially in associations of various plants.

Air Microbiology↗

Phytoremediation potential of Myriophyllum aquaticum and Pistia stratiotes to modify antibiotic growth promoters, tetracycline, and oxytetracycline, in aqueous wastewater systems.

Antibiotics are frequently used in the United States as feed efficiency promoters and medicines for livestock that is destined for human consumption. These antibiotics are released into the environment through the runoff and wastewater streams from animal feedlots and land applications of manure. The exposure of microorganisms to these antibiotics has reportedly resulted in the development of resistant species of microorganisms, which in turn can lead to human health hazards. Phytoremediation of these antibiotics can be a useful tool for countering this problem. Aquatic plants, Myriophyllum aquaticum (parrot feather) and Pistia stratiotes (water lettuce), were used for studying phytoremediation of tetracycline (TC) and oxytetracycline (OTC) from aqueous media. TC and OTC are two of the most commonly used tetracyclines in veterinary medicine. M. aquaticum and P. stratiotes gave high antibiotic modification rates of both antibiotics. Kinetic analyses dismiss direct enzyme catalysis; the modification rates decreased with increasing OTC concentrations. Sterile, cell-free root exudates (filtered through 0.2 microm membranes) from both species also exhibited comparable antibiotic modification rates. The involvement of root-secreted metabolites in antibiotic modification is suggested. The changes in the UV absorbance spectra of OTC during treatment with the root exudates confirmed the modification.

Anti-Bacterial Agents↗