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Strigol: biogenesis and physiological activity.

The role played by molecules of the strigolactone family in stimulating the germination of seeds of parasitic weeds of the genera Striga, Orobanche and Alectra has never been clearly elucidated. The biogenesis of these unusual terpenoid lactones, originally identified in minute quantities in the root exudates of a small number of host plants and two or three "false hosts", also remains obscure. These lactones, as the chemical signals which initiate the life cycle of Striga, are consequently at the forefront of the Striga research effort. This paper reviews recent key discoveries relating to the biosynthesis and mode of action of strigolactones, and summarises the evidence suggesting that these molecules may be far more widely distributed and have a greater physiological significance than has hitherto been appreciated.

Germination↗

Retrotransfer of DNA in the rhizosphere.

Retrotransfer of DNA refers to the phenomenon by which a plasmid travels from a host strain to a recipient one and returns to the original host, bringing with it DNA from the recipient. The resultant host strain with DNA from the recipient is called a retrotransconjugant. The retrotransfer phenomenon mediated by the TOL plasmid pWW0 and other plasmids has been documented on plates under optimal laboratory culture conditions, but never under natural conditions. In this work, we show that retrotransfer mediated by the IncP9 TOL pWW0 plasmid occurs in the rhizosphere, a niche in which the continuous supply of nutrients via root exudates allows cells to reach a high density. This suggests that this unusual sexual fertilization may be of great importance in lateral gene transfer. We also show that retrotransfer of DNA seems to require co-integration of the plasmid and the host chromosome and subsequent resolution, because a TOL plasmid with a mutation in the tnpR gene, encoding the resolvase of the Tn4653 of the TOL plasmid, was self-transferred between Pseudomonas strains, but unable to mobilize chromosome.

Blotting, Southern↗

Cytokinins in the Phloem Sap of White Lupin (Lupinus albus L.).

Cytokinin-like activity in samples of xylem and phloem sap collected from field-grown plants of white lupin (Lupinus albus L.) over a period of 9 to 24 weeks after sowing was measured using the soybean hypocotyl callus bioassay following paper chromatographic separation. The phloem sap was collected from shallow incisions made at the base of the stem, the base of the inflorescence (e.g. stem top), the petioles, and the base and tip of the fruit. Xylem sap was collected as root exudate from the stump of plants severed a few centimeters above ground level. Concentration of cytokinin-like substances was highest in phloem sap collected from the base of the inflorescence and showed an increase over the entire sampling period (from week 10 [61 nanogram zeatin equivalents] to week 24 [407 nanogram zeatin equivalents]). Concentrations in the xylem sap and in the other phloem saps were generally lower. Relatively high concentrations of cytokinin-like substances in petiole phloem sap (70 to 130 nanogram zeatin equivalents per milliliter) coincided in time with high concentrations in sap from the base of the inflorescence (see above). Concentrations in sap (phloem or xylem) from the base of the stem were very much lower. This finding is consistent with movement of cytokinins from leaves into the developing inflorescence and fruit, rather than direct input to the fruit from xylem sap. However, an earlier movement of cytokinins from roots into leaves via the xylem cannot be ruled out. Sap collected at an 18-week harvest was additionally separated by sequential C(18) reversed-phase high performance liquid chromatography --> NH(2) normal phase high performance liquid chromatography, bioassayed, and then analyzed by electron impact gas chromatography-mass spectrometry. Identification of zeatin riboside and dihydrozeatin as two of the major cytokinins in combined sap samples was accomplished by gas chromatography-mass spectrometry-selected ion monitoring.

Journal Article↗

Perception and modification of plant flavonoid signals by rhizosphere microorganisms.

Flavonoids are a diverse class of polyphenolic compounds that are produced as a result of plant secondary metabolism. They are known to play a multifunctional role in rhizospheric plant-microbe and plant-plant communication. Most familiar is their function as a signal in initiation of the legume-rhizobia symbiosis, but, flavonoids may also be signals in the establishment of arbuscular mycorrhizal symbiosis and are known agents in plant defence and in allelopathic interactions. Flavonoid perception by, and impact on, their microbial targets (e.g. rhizobia, plant pathogens) is relatively well characterized. However, potential impacts on 'non-target' rhizosphere inhabitants ('non-target' is used to distinguish those microorganisms not conventionally known as targets) have not been thoroughly investigated. Thus, this review first summarizes the conventional roles of flavonoids as nod gene inducers, phytoalexins and allelochemicals before exploring questions concerning 'non-target' impacts. We hypothesize that flavonoids act to shape rhizosphere microbial community structure because they represent a potential source of carbon and toxicity and that they impact on rhizosphere function, for example, by accelerating the biodegradation of xenobiotics. We also examine the reverse question, 'how do rhizosphere microbial communities impact on flavonoid signals?' The presence of microorganisms undoubtedly influences the quality and quantity of flavonoids present in the rhizosphere, both through modification of root exudation patterns and microbial catabolism of exudates. Microbial alteration and attenuation of flavonoid signals may have ecological consequences for below-ground plant-microbe and plant-plant interaction. We have a lack of knowledge concerning the composition, concentration and bioavailability of flavonoids actually experienced by microbes in an intact rhizosphere, but this may be addressed through advances in microspectroscopic and biosensor techniques. Through the use of plant mutants defective in flavonoid biosynthesis, we may also start to address the question of the significance of flavonoids in shaping rhizosphere community structure and function.

Biosensing Techniques↗

Regulation of pssA and pssB gene expression in Rhizobium leguminosarum bv. trifolii in response to environmental factors.

Exopolysaccharide (EPS) produced by Rhizobium leguminosarum plays an important role in indeterminate-type nodulation. The pssA gene, encoding the first glycosyl transferase and pssB, encoding inositol monophosphatase, are involved in EPS synthesis and regulation in R. leguminosarum bv. trifolii. Using pssA-lacZ and pssB-lacZ plasmid-borne transcriptional fusions, a stimulating effect of phosphate on the transcription of both genes was observed. The increase in the transcription of pssA and pssB genes was correlated with a significant enhancement in EPS production in rhizobia grown in the high phosphate concentrations. In the presence of ammonium, transcription of the pss-lacZ fusions was repressed, especially the pssB-lacZ fusion, but EPS production still remained enhanced. Root exudate positively influenced the level of pss-lacZ transcription. These results indicate a complex regulation of pss gene transcription by several environmental factors. In addition, the expression of pssA and pssB genes in the symbiotic state was examined using pss-gusA plasmid-borne transcriptional fusions. It was found that the pssA-gusA fusion was not expressed in the nodules. The pssB-gusA fusion was expressed mainly in the infection zone and interzone of the nodule and the pattern of expression was dependent on the plant phosphate availability.

Bacterial Proteins↗

Short-term effects of rhizosphere microorganisms on fe uptake from microbial siderophores by maize and oat.

Effects of rhizosphere microorganisms on Fe uptake by oat (Avena sativa) and maize (Zea mays) were studied in short-term (10 h) nutrient solution experiments. Fe was supplied either as microbial siderophores (pseudobactin [PSB] or ferrioxamine B [FOB]) or as phytosiderophores obtained as root exudates from barley (epi-3-hydroxy-mugineic acid [HMA]) under varied population densities of rhizosphere microorganisms (axenic, uninoculated, or inoculated with different microorganism cultures). When maize was grown under axenic conditions and supplied with FeHMA, Fe uptake rates were 100 to 300 times higher compared to those in plants supplied with Fe siderophores. Fe from both sources was taken up without the involvement of an extracellular reduction process. The supply of FeHMA enhanced both uptake rate and translocation rate to the shoot (more than 60% of the total uptake). However, increased density of microorganisms resulted in a decrease in Fe uptake rate (up to 65%), presumably due to microbial degradation of the FeHMA. In contrast, when FeFOB or FePSB was used as the Fe source, increased population density of microorganisms enhanced Fe uptake. The enhancement of Fe uptake resulted from the uptake of FeFOB and FePSB by microorganisms adhering to the rhizoplane or living in the free space of cortical cells. The microbial apoplastic Fe pool was not available for root to shoot transport or, thus, for utilization by the plants. These results, in addition to the low uptake rate under axenic conditions, are in contrast to earlier hypotheses suggesting the existence of a specific uptake system for Fe siderophores in higher plants. The bacterial siderophores PSB and FOB were inefficient as Fe sources for plants even when supplied by stem injection. It was concluded that microorganisms are involved in degradation processes of microbial siderophores, as well as in competition for Fe with higher plants.

Journal Article↗

Soil plant microbe interactions in phytoremediation.

Use of vegetation in remediation of soil and groundwater contaminated with organic materials is a promising, cost-effective alternative to the more established treatment methods used at hazardous waste sites. Plants can transpire groundwater and lower the concentrations of organic contaminants in soils and groundwater. The evapotranspirational activity of vegetation acts as a natural pump-and-treatment system. Plants have shown the capacity to absorb, uptake, and convert organic contaminants to less toxic metabolites in laboratory and field studies. Vegetation also plays a significant role in bioremediation. This is because plants stimulate the degradation of organic compounds in the rhizosphere by the release of root exudates and enzymes. Success of any plant-based remediation system depends on the interaction of plants with the surrounding soil medium and the contaminant. Knowing the fate of an organic contaminant in the soil can help determine the persistence of the contaminant in the terrestrial environment and ultimately the success of any remediation method. Also, an understanding is needed of soil-plant-microbe interactions that determine the fate of organic contaminants in the soil-plant ecosystem. This paper presents an overview of the subsurface environment and fate and transport processes of organic contaminants as affected by soil-plant-microbe interactions.

Biodegradation, Environmental↗

Chromatographic separation and in vitro activity of sorgoleone congeners from the roots of sorghum bicolor.

Sorgoleone, 2-hydroxy-5-methoxy-3-[(8'Z,11'Z)-8',11',14'-pentadecatriene]-p-benzoquinone (1), and its corresponding hydroquinone are the major components of the root exudate of Sorghum bicolor. The name sorgoleone includes minor analogues differing in the length or degree of unsaturation of the 3-alkyl side chain. These compounds are known to be phytotoxic, probably through inhibition of photosystem II (PSII) driven oxygen evolution, as previously demonstrated for 1. Isolation of these sorgoleone congeners was achieved by C(8) column chromatography and argentation thin-layer chromatography, and the purified compounds were structurally characterized. The abilities of the minor sorgoleones to inhibit PSII were similar to that of the major compound, suggesting that all of these sorgoleone congeners contribute to the overall allelopathy of sorghum.

Benzoquinones↗

Utilising the synergy between plants and rhizosphere microorganisms to enhance breakdown of organic pollutants in the environment.

BACKGROUND: Phytoremediation is a promising technology for the cleanup of polluted environments. The technology has so far been used mainly to remove toxic heavy metals from contaminated soil, but there is a growing interest in broadening its applications to remove/degrade organic pollutants in the environment. Both plants and soil microorganisms have certain limitations with respect to their individual abilities to remove/breakdown organic compounds. A synergistic action by both rhizosphere microorganisms that leads to increased availability of hydrophobic compounds, and plants that leads to their removal and/or degradation, may overcome many of the limitations, and thus provide a useful basis for enhancing remediation of contaminated environments. MAIN FEATURES: The review of literature presented in this article provides an insight to the nature of plant-microbial interactions in the rhizosphere, with a focus on those processes that are relevant to the breakdown and/or removal of organic pollutants. Due consideration has been given to identify opportunities for utilising the plant-microbial synergy in the rhizosphere to enhance remediation of contaminated environments, RESULTS AND DISCUSSION: The literature review has highlighted the existence of a synergistic interaction between plants and microbial communities in the rhizosphere. This interaction benefits both microorganisms through provision of nutrients by root exudates, and plants through enhanced nutrient uptake and reduced toxicity of soil contaminants. The ability of the plant-microbial interaction to tackle some of the most recalcitrant organic chemicals is of particular interest with regard to enhancing and extending the scope of remediation technologies. CONCLUSIONS: Plant-microbial interactions in the rhizosphere offer very useful means for remediating environments contaminated with recalcitrant organic compounds. OUTLOOK: A better knowledge of plant-microbial interactions will provide a basis for improving the efficacy of biological remediations. Further research is, however, needed to investigate different feedback mechanisms that select and regulate microbial activity in the rhizosphere.

Biodegradation, Environmental↗

Expression of a Pseudomonas putida aminotransferase involved in lysine catabolism is induced in the rhizosphere.

Using a transposon carrying a promoterless lux operon to generate transcriptional fusions by insertional mutagenesis, we have identified a Pseudomonas putida gene with increased expression in the presence of corn root exudates. Expression of the transcriptional fusion, induced by the amino acid lysine, was detected in P. putida in the rhizosphere of plants as well as in response to seed exudates. The mutant was unable to grow on lysine or delta-aminovalerate as carbon sources, which indicates that the affected function is involved in the pathway for lysine catabolism. However, the mutant strain grew with glutaric acid, the product of delta-aminovalerate metabolism via glutaric acid semialdehyde, as a C source. The translated sequence of the interrupted gene showed high levels of similarity with aminotransferases. These sets of data suggest that the product of this gene has delta-aminovalerate aminotransferase activity. This is the first direct genetic evidence correlating a DNA sequence with such activity in Pseudomonadaceae.

DNA Transposable Elements↗

Ginsenosides stimulate the growth of soilborne pathogens of American ginseng.

Ginseng saponins (ginsenosides) were isolated from soil associated with the roots of commercially grown American ginseng (Panax quinquefolius L.), identified via LC-MS and quantified via analytical HPLC. The ginsenosides, including F(11), Rb(1), Rb(2), Rc, Rd, Re and Rg(1), represented between 0.02 and 0.098% (average 0.06%) of the mass of the soil collected from roots annually between 1999 and 2002. The same ginsenosides were also isolated from run-off of undisturbed plants grown in pots in a greenhouse using a root exudate trapping system. To investigate (1) whether these saponins could influence the growth of pythiaceous fungi pathogenic to ginseng, and (2) whether soil levels of ginsenosides were sufficient to account for any effects, bioassays were completed using a crude saponin extract and an ecologically relevant concentration of purified ginsenosides. Thus, when cultured on media containing crude saponins, the colony weight of both Phytophthora cactorum and Pythium irregulare was significantly greater than that of control, indicating a strong growth stimulation by ginsenosides. The growth of Pythium irregulare was also significantly stimulated after addition of an ecologically relevant, low concentration (i.e. 0.06%) of purified ginsenosides to culture medium. By contrast, growth of the saprotrophic fungus Trichoderma hamatum was slightly (but not significantly) inhibited under the same conditions. These results imply that ginsenosides can act as allelopathic stimulators of the growth of pythiaceous fungi in the rhizosphere, and this may contribute to the disease(s) of this crop.

Dose-Response Relationship, Drug↗

[QTLs mapping of leaf traits and root vitality in a recombinant inbred line population of rice].

By employing a recombinant inbred line (RIL) population of 247 lines derived from an inidca-indica cross Zhenshan 97B x Milyang 46, a linkage map consisting of 158 DNA markers was constructed and used for the determination of QTLs conditioning five leaf traits and root exudates. The leaf traits analyzed were leaf area, leaf length, leaf width, leaf perimeter and leaf length/width ratio measured on top first leaf, top second and top third leaves. The RIL population showed transgressive segregation on each trait, and highly significant or significant positive correlations were observed between all traits except between leaf length and width, and between leaf perimeter and length/width ratio. A total of 24 QTLs located in 9 intervals were detected to have significant additive effects for leaf traits analyzed, with LOD scores ranging 2.9-11.8 and 4.0%-32.5% phenotypic variation explained for a single QTL. Clustering of QTLs for leaf traits was evident. In interval RM197-RZ516 on chromosome 6, 2 QTLs for leaf length, 2 QTLs for leaf width and 3 QTLs for leaf length/width ratio of different leaves were detected, among which the alleles for increasing trait values were from Zhenshan 97B for leaf length and leaf length/width ratio, and from Milyang 46 for leaf width. In interval RM1-RG532 on chromosome 1,2 QTLs for leaf length and 2 QTLs for leaf perimeter were detected on top first leaf and top second leaf, respectively, and all the alleles for increasing trait values were from Milyang 46. In this interval, larger additive effects were observed for QTLs detected on top first leaf than at top second leaf. In interval RZ667-B10B on chromosome 6, a QTL for leaf perimeter of top second leaf was detected, accounting for 8.0% phenotypic variation. In intervals RZ66-RM264 and RG81-RM313 on chromosome 8 and 12, each QTL was detected for leaf length of top third leaf and explained for 9.0% and 15.3% phenotypic variation, respectively. Epistasis analysis detected 56 and 4 significant additive-by-additive interactions for leaf traits and root vitality, explaining 2.7% to 13.7% and 6.8% to 14.9% of the total phenotypic variation, respectively. On comparison with QTLs for yield traits detected in the same population previously, it was found that the majority of QTLs for leaf traits and root vitality and those for yield traits were located in similar intervals. Fine mapping of QTLs for both leaf and yield traits in these chromosomal regions would facilitate investigations of the source-sink relationship in rice.

Chromosome Mapping↗

Impact of genetically modified crops and their management on soil microbially mediated plant nutrient transformations.

One of the potential environmental effects of the recent rapid increase in the global agricultural area cultivated with transgenic crops is a change in soil microbially mediated processes and functions. Among the many essential functions of soil biota are soil organic matter decomposition, nutrient mineralization and immobilization, oxidation-reduction reactions, biological N fixation, and solubilization. However, relatively little research has examined the direct and indirect effects of transgenic crops and their management on microbially mediated nutrient transformations in soils. The objectives of this paper are to review the available literature related to the environmental effects of transgenic crops and their management on soil microbially mediated nutrient transformations, and to consider soil properties and climatic factors that may affect the impact of transgenic crops on these processes. Targeted genetic traits for improved plant nutrition include greater plant tolerance to low Fe availability in alkaline soils, enhanced acquisition of soil inorganic and organic P, and increased assimilation of soil N. Among the potential direct effects of transgenic crops and their management are changes in soil microbial activity due to differences in the amount and composition of root exudates, changes in microbial functions resulting from gene transfer from the transgenic crop, and alteration in microbial populations because of the effects of management practices for transgenic crops, such as pesticide applications, tillage, and application of inorganic and organic fertilizer sources. Possible indirect effects of transgenic crops, including changes in the fate of transgenic crop residues and alterations in land use and rates of soil erosion, deserve further study. Despite widespread public concern, no conclusive evidence has yet been presented that currently released transgenic crops, including both herbicide and pest resistant crops, are causing significant direct effects on stimulating or suppressing soil nutrient transformations in field environments. Further consideration of the effects of a wide range of soil properties, including the amount of clay and its mineralogy, pH, soil structure, and soil organic matter, and variations in climatic conditions, under which transgenic crops may be grown, is needed in evaluating the impact of transgenic crops on soil nutrient transformations. Future environmental evaluation of the impact of the diverse transgenic crops under development could lead to an improved understanding of soil biological functions and processes.

Agriculture↗

An improved axenic system for studying pre-infection development of the parasitic plant Orobanche ramosa.

BACKGROUND AND AIMS: Broomrapes (Orobanche spp.) are holoparasitic weeds that cause devastating losses in many economically important crops. The molecular mechanisms that control early stages of host infection in Orobanche are poorly understood, partly due to the lack of experimentally tractable in vitro systems that allow the efficient application of molecular tools. Here an improved axenic system for the analysis of pre-infection stages in O. ramosa in the absence of the host plant is described. METHODS: An optimized protocol for seed disinfection, based on formaldehyde, was developed. Orobanche ramosa seeds were conditioned in Petri dishes with filter paper, stimulated by addition of the synthetic strigol analogue GR24, and the percentage of germination as well as attachment-organ formation was determined. KEY RESULTS: Treatment of O. ramosa seeds with tobacco-root exudate or with GR24 resulted in highly reproducible germination rates around 70 %. A conditioning period of 8 d was both necessary and sufficient to allow optimal germination in response to GR24. Conditioned seeds that were dehydrated for several months remained fully responsive to GR24 without the need of a new conditioning period. Treatments as short as 5 min with GR24 were sufficient to fully and irreversibly induce the seed germination response. Approximately half of the germinated seeds initiated attachment-organ development. Similar rates of attachment organ induction were also detected in the rare cases of seeds that had germinated spontaneously on water. CONCLUSIONS: The results suggest that the conditioning period produces persistent changes in the seeds required for responsiveness to external stimulants. The rapid action of GR24 suggests that it may act via a receptor-mediated signalling mechanism. While germination in O. ramosa is induced by exogenous stimuli, attachment organ differentiation appears to be triggered by unknown endogenous signals. The new in vitro culture system will have useful applications for the molecular analysis of early stages of parasitic development in Orobanche.

Disinfectants↗

Characterization of a sugar-binding protein from Azospirillum brasilense mediating chemotaxis to and uptake of sugars.

Our approach to the isolation of plant-inducible bacterial genes of Azospirillum brasilense, based on the analysis of protein patterns of bacteria grown in the presence and in the absence of plant root exudates, led to the identification of an acidic 40 kDa protein. Cloning and sequencing analysis of the corresponding coding DNA region revealed the presence of two open reading frames transcribed in the same orientation. The deduced ORF1 protein, which corresponds to the 40 kDa protein, is very similar to the periplasmic ChvE protein, identified in Agrobacterium tumefaciens and involved in enhanced virulence. The deduced ORF2 protein shows homology to members of the LysR family of transcriptional regulators. The function of the ChvE-like protein in A. brasilense was investigated further. The protein, designated as SbpA (sugar binding protein A), is involved in the uptake of D-galactose and functions in the chemotaxis of A. brasilense towards several sugars, including D-galactose, L-arabinose and D-fucose. Expression of the sbpA gene requires the presence of the same sugars in the growth medium and is enhanced further in combination with carbon starvation of A. brasilense cells.

Agrobacterium tumefaciens↗

Simultaneous growth on citrate reduces the effects of iron limitation during toluene degradation in Pseudomonas.

Rhizoremediation has been suggested as an attractive bioremediation strategy for the effective breakdown of pollutants in soil. The presence of plant root exudates such as organic acids, sugars, and amino acids that may serve as carbon sources or biosynthetic building blocks and the limited bioavailability of iron may influence the degradation of pollutants in the rhizosphere. To test the effect of such compounds on hydrocarbon degradation, trace concentrations of yeast extract or mixtures of organic acids and amino acids were added to continuous cultures of Pseudomonas putida mt2 and P. putida WCS358 (TOL) growing on toluene. By addition of these compounds increased growth yields and higher specific growth rates on toluene were obtained. The effects of iron limitation on the substrate utilization pattern of both strains were tested by growing the strains on a mixture of toluene and the readily degradable carbon source citrate while the iron concentration was varied. Simultaneous use of both substrates under carbon-limited as well as iron-limited conditions was observed. Growth yields were less reduced and iron requirement was lower during iron-limited growth in the toluene + citrate grown cultures compared to cultures in which toluene was used as the sole carbon source. The kinetic properties of the cells for toluene degradation were less hampered by the lack of iron when citrate was used as an additional carbon source. The results indicate that the availability of low concentrations of natural organic compounds, such as produced in the rhizosphere, may positively influence the degradative performance of hydrocarbon-degrading bacteria.

Amino Acids↗

Carbon assimilation and turnover in grassland vegetation using an in situ (13)CO(2) pulse labelling system.

A mobile laboratory was developed to administer a controlled flow of (13)C labelled CO(2) at ambient concentrations ( approximately 350 ppm) in the field. The stable isotope delivery (SID) system consists of an isotope-mixing unit with flow control to a series of 12 independent labelling chambers. In-line CPU controlled infrared gas analysers allow automated measurement of chamber CO(2) concentrations and gas flow management. A preliminary experiment was established on an upland pasture located at the NERC Soil Biodiversity experimental site, Sourhope, UK, in August 1999. The objective of this investigation was to determine the magnitude of pulse-derived C incorporation into a typical upland plant community. To achieve this, the SID system was deployed to pulse-label vegetation with CO(2) enriched with (13)C (50 atom %) at ambient concentrations ( approximately 350 ppm) on two consecutive days in August 1999. Samples of headspace CO(2), shoot and root were taken on four occasions over a period of 28 days after (13)C labelling. These materials were then prepared for (13)C/(12)C ratio determination by continuous-flow/combustion/isotope ratio mass spectrometry (CF-C-IRMS). Results showed that pulse derived CO(2)-C was assimilated at a rate of 128 +/- 32 microg g shoot-C hour(-1). Dynamic samplings showed that pulse-derived (13)C concentrations in the labelled plant tissues declined by 77.4 +/- 6% after 48 hours. The rapid decline in (13)C concentrations in plant matter was the result of C loss from the plant in the form of respired CO(2) and root exudates, and dilution by subsequent unlabelled C assimilates. This novel system offers considerable potential for in situ tracer investigations.

Carbon↗

Clovers secrete specific phenolic compounds which either stimulate or repress nod gene expression in Rhizobium trifolii.

Rhizobium trifolii mutants containing Escherichia coli lac gene fusions to specific nodulation (nod) genes were used to characterise phenolic compounds secreted from the roots of white clover (Trifolium repens) plants. These compounds either had stimulatory or inhibitory effects upon the induction of the nod genes. The stimulatory compounds were hydroxylated flavones and the most active compound was 7,4'-dihydroxyflavone. The inhibitory compounds present in white clover root exudates were umbelliferone (a coumarin) and formononetin (an isoflavone). Transcriptional activation of nod gene promoters in response to short exposures (3 h) of 7,4'-dihydroxyflavone was growth phase dependent; cells in early log phase were highly responsive to flavone additions in vitro and nod gene induction could be detected within 20 min of exposure at 5 x 10 M. Cells in other growth phases were generally unresponsive. A 10-fold molar excess of umbelliferone to 7,4'-dihydroxyflavone resulted in complete inhibition of nod gene induction. Some commercially-obtained flavones were found to have weak stimulatory activity but could also inhibit nod gene induction by more effective stimulatory compounds. Strong stimulatory and inhibitory compounds all possessed a 7-hydroxy moiety and showed other structural similarities. This suggested that there was one binding site for these compounds. Because the response to these compounds was rapid, we propose that these phenolics act at the bacterial membrane or that an active uptake system is involved.

Journal Article↗