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PubMed2003

An improved reversed-phase liquid chromatographic method for the analysis of low-molecular mass organic acids in plant root exudates.

Reversed-phase column liquid chromatography (RPLC) was used for the separation and quantification of 10 low-molecular-mass organic acids (malic, malonic, lactic, acetic, maleic, citric, cis-aconitic, succinic, fumaric, and trans-aconitic) in plant root exudates. A mobile phase of 93% 25 mM KH2PO4 at pH 2.5 and 7% methanol at a flow-rate of 1 ml min(-1) resolved all 10 acids in 15 min on a C18 column. Experiments demonstrated a significant (P < 0.05) effect of sample pH on detector response, with peak heights being significantly lower at pH 6.0 compared with pH 2.5, but peak area showed no significant difference. At pH 8.0 and above, both peak height and area differed significantly from injections made at pH 2.5. Limits of detection (LOD) for the 10 acids ranged from 0.05 to 24 microM. Finally, the improved method was applied for the analysis of root exudates from soil cultured field pea, Banksia attenuata, white lupin, and chickpea.

Acids↗
PubMed2001

Ability of Agrogyron elongatum to accumulate the single metal of cadmium, copper, nickel and lead and root exudation of organic acids.

Agrogyron elongatum were grown in nutrient solution containing moderate to high amounts of separate heavy metal of Cd, Cu, Ni and Pb in a greenhouse for a 9-day. Cd, Cu, Ni and Pb generally led to decrease in the elongation of roots although the length of seedlings exposed to Cd and Pb at 0.05 and 0.5 mg/L showed to be slightly greater than that of controls. Of the four metals in the experiment, Pb was absorbed and accumulated to the highest level, with the concentrations of 92,754 mg/kg dry weight (DW) in roots and 11,683 mg/kg DW in shoots. Cd was moderately accumulated in Agrogyron elongatum, but the maximum bioaccumulation coefficients (BCs) for roots and shoots were observed. The patterns for Cu and Ni uptake and distribution in plants differed from those of Pb and Cd, as it was showed that the shoot accumulation of Cu and Ni was significantly higher than in roots. A. elongatum had the highest Ni concentration in shoots (30,261 mg/kg DW) at the external concentration of 250 mg/L. Cu ranked second, with a shoot concentration of 12,230 mg/kg DW when 50 mg/L Cu in solution was applied. For the four trace elements tested, the highest concentrations in shoots decreased by the order of Ni > Cu > Pb > Cd (mg/kg DW), and those in roots were Pb > Cd > Ni > Cu (mg/kg DW). Malic, oxalic and citric acids exuded by roots exposed to 1 and 50 mg/L of the metals were detected. Release of organic acids from plants significantly differed among the metal treatments. Cu was most effectively in inducing root exudation of the three types of organic acids. Cd, and Ni were also the inducers of secretion of malic and oxalic acids. With reference of Pb, a small amounts of malic and oxalic acids were detected in the root exudates, but few quantities of citric acid were found. However, no correlation between alternations in root exudation of organic acids and metal accumulation could be established.

Cadmium↗
PubMed2005

Differential gene expression by Metarhizium anisopliae growing in root exudate and host (Manduca sexta) cuticle or hemolymph reveals mechanisms of physiological adaptation.

Like many other fungal pathogens Metarhizium anisopliae is a facultative saprophyte with both soil-dwelling and insect pathogenic life-stages. In addition, as M. anisopliae traverses the cuticle and enters the hemolymph it must adapt to several different host environments. In this study, we used expressed sequence tags and cDNA microarray analyses to demonstrate that physiological adaptation by M. anisopliae to insect cuticle, insect hemolymph, bean root exudate (a model for life in the soil), and nutrient rich Sabouraud dextrose broth (SDB) involves different subsets of genes. Overall, expression patterns in cuticle and hemolymph clustered separately from expression patterns in root exudates and SDB, indicative of critical differences in transcriptional control during pathogenic and saprophytic growth. However, there were differences in gene expression between hemolymph and cuticle and these mostly involved perception mechanisms, carbon metabolism, proteolysis, cell surface properties, and synthesis of toxic metabolites. These differences suggest previously unsuspected stratagems of fungal pathogenicity that can be tested experimentally. Examples include the switch-off of cuticle-degrading proteases and a dramatic cell wall reorganization during growth in hemolymph.

Adaptation, Physiological↗
PubMed2005

Effect of corn root exudates on the degradation of atrazine and its chlorinated metabolites in soils.

DIMBOA (3,4-dihydro-2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3-one), a major benzoxazinone of Poaceae plants, was isolated and purified from corn seedlings. The effect of isolated and purified DIMBOA on the degradation of atrazine [2-chloro-4-(ethylamino)-6-(isopropylamino)-s-triazine], and its toxic breakdown products, desethylatrazine [2-chloro-4-amino-6-(isopropylamino)-s-triazine; DEA] and desisopropylatrazine [2-chloro-4-(ethylamino)-6-amino-s-triazine; DIA], was studied in the absence of plants using batch experiments, while the effect of corn root exudates on these compounds was determined in hydroponic experiments. Degradation experiments were performed in the presence and absence of 50 microM, 1 mM, or 5 mM DIMBOA resulting in ratios of DIMBOA to pesticide of 1:1, 20:1, and 100:1. We observed a 100% degradation of atrazine to hydroxyatrazine within 48 h at a ratio of DIMBOA to atrazine of 100:1. DIMBOA had the largest effect on atrazine, while it was about three times less effective on DEA and DIA. Corn (Zea mays L. cv. LG 2185) was exposed to 10 mg L(-1) of either atrazine, DEA, or DIA for 11 d in a growth chamber experiment. Up to 4.3 micromol L(-1) d(-1) of hydroxyatrazine were formed in the nutrient solutions by plants exposed to atrazine, while the formation of hydroxylated metabolites from plants exposed to DEA and DIA was smaller and also delayed. The formation of hydroxylated metabolites increased in the solution with plant age in all atrazine, DEA, and DIA treatments. HMBOA (3,4-dihydro-2-hydroxy-7-methoxy-2H-1,4-benzoxazin-3-one), the lactam precursor of DIMBOA, and a tentatively identified derivative of MBOA (2,3-dihydro-6-methoxy-benzoxazol-2-one) were detected in the corn root exudates. Mass balance calculations revealed that up to 30% of the disappearance of atrazine and DEA, and up to 10% of DIA removal from the solution medium in our study could be explained by the formation of hydroxylated metabolites in the solution itself. Our results show that higher plants such as corn have the potential to promote the hydrolysis of triazine residues in soils by exudation of benzoxazinones.

Atrazine↗
PubMed1974

Assimilation, Distribution, and Root Exudation of C by Ponderosa Pine Seedlings under Induced Water Stress.

The effect of specific levels of induced water stress on the root exudation of (14)C from 9-month-old and 12-month-old ponderosa pine (Pinus ponderosa Laws.) seedlings was examined. Polyethylene glycol (PEG-4000) was used to decrease root solution water potentials by 0, -1.9, -2.6, -5.5, -9.6 and -11.9 bars in either aerated 0.25X Hoagland's nutrient solution or aerated distilled water.Assimilation of (14)CO(2) by plants under stress and subsequent translocation of (14)C label to the roots were both inhibited by a decrease in substrate water potential. Six days after (14)CO(2) introduction essentially no (14)C was detected in the roots of plants maintained at solution potentials of -5.5 bars or below. In subsequent studies (14)CO(2) was introduced 4 days prior to induction of stress. This allowed sufficient time for distribution of (14)C label throughout the root system.Root exudation of (14)C-labeled sugars, amino acids, and organic acids from plants in nutrient solution showed an increase from 0 to -1.9 bars, a decline from -1.9 to about -5.5 bars, and then an increase again from -5.5 to -11.9 bars. As substrate potential decreased, sugars as a percentage of total exudate increased, organic acids decreased and amino acids showed a slight decrease. Marked changes in percentages occurred between 0 and -2.6 bars. The exudation of sugars, amino acids, and organic acids from plants in distilled water showed similar trends in response to water stress as those in nutrient solution, but the quantity of total (14)C exuded was greater.

Journal Article↗
PubMed1989

Fungal Growth Stimulation by CO(2) and Root Exudates in Vesicular-Arbuscular Mycorrhizal Symbiosis.

Transformed roots of carrot were used to determine the effects of root metabolites on hyphal development from spores of the vesicular-arbuscular mycorrhizal fungus Gigaspora margarita. Hyphal growth of this obligately biotrophic symbiont was greatly stimulated by a synergistic interaction between volatile and exudated factors produced by roots. Root volatiles alone provided little stimulation, and root exudates alone had no effect. For the first time, carbon dioxide was demonstrated to be a critical root volatile involved in the enhancement of hyphal growth. C-labeled root volatiles were fixed by the fungus and thus strongly suggested that CO(2) served as an essential carbon source.

Journal Article↗
PubMed2003

Correlation between phytotoxicity on annual ryegirass (Lolium rigidum) and production dynamics of allelochemicals within root exudates of an allelopathic wheat.

An improved allelopathic correlation between phytotoxicity measured in root growth bioassay upon annual ryegrass (Lolium rigidum Gaud.) and the concentrations of a selection of dynamically produced allelochemicals quantified in the root exudates of cv. Khapli wheat (Triticum turgidum ssp. durum (Desf.) Husn.) monitored during the first 15 days of wheat seedling growth in a sterile, agar-water medium, has been established. Changes over the 15-day growth period in the quantities of five exuded benzoxazinones and seven phenolic acids were measured simultaneously using GC/MS/MS. Substantiating pure compound dose-response measurements were conducted over a range of concentrations for the putative allelochemicals within the wheat exudates. One synergism-based proposal using the monitored compounds to explain the observed low-exudate-concentration phytotoxicity was explored, but was found to be experimentally inadequate.

Gas Chromatography-Mass Spectrometry↗
PubMed2005

Investigation of selenium-containing root exudates of Brassica juncea using HPLC-ICP-MS and ESI-qTOF-MS.

Selenium-containing root exudates were investigated in a known selenium accumulator model plant. Indian mustard (Brassica juncea) plants were grown hydroponically and supplemented with selenite (SeO(3)(2-)) in a 25% Hoagland's nutrient solution. Additive concentrations were 0, 1, 5 and 20 microg mL(-1) Se with five replicate plants per treatment level. Plants were exposed to the respective Se solutions for two weeks, then placed in deionized water for two more weeks. The hydroponic solutions were collected for analysis after the first two weeks of selenium supplementation (day 14) and twice during the deionized water period (days 21 and 28). Separation by ion-pairing high performance liquid chromatography was followed by inductively coupled plasma-mass spectrometry (ICP-MS) for selenium specific detection. Chromatographic peaks unable to be identified by retention-time matching were collected for analysis by electrospray ionization mass spectrometry (ESI-MS). Additional chemical experiments were performed for structural elucidation. Several selenium-containing compounds were identified in the exudate-containing solution and two were identified as selenocystine and the selenosulfate (SSeO(3)(2-)) ion. The presence of dimethylselenide (CH(3)SeCH(3)) is also observed but cannot be attributed exclusively to plant exudation because plants were not grown in sterile conditions. Further, the incorporation of fortified selenoamino acids into peptide structures was found to occur under neutral pH conditions, suggesting that exuded enzymes might facilitate such a reaction. Finally, physiological differences resulting from selenium supplementations were noted and discussed.

Chromatography, High Pressure Liquid↗
PubMedDate not supplied

Low molecular weight organic acids and fatty acids in root exudates of two Lupinus cultivars at flowering and fruiting stages.

Low molecular weight organic acids (LOAs) and fatty acids in root exudates of two lupin cultivars, Lupinus albus cv. Multolupa and L. luteus cv. Tremosilla, were determined at flowering and fruiting stages. LOAs were analysed by capillary electrophoresis. Acetic and citric acids were the most abundant, especially the latter in L. luteus at the flowering stage (5922.79 micrograms/g dry root). The significant decrease in acid content of both cultivars from flowering to fruiting stages was also striking. The highest levels of acetic acid were detected in L. luteus at fruiting stage (1542.03 micrograms/g dry root). The significant citrate production in L. luteus could be related to the low phosphorus concentration in the studied soils but not to proteoid roots, which were detected only in L. albus. The source of the LOAs detected in these exudates is also discussed, since they may be produced either by the plant or by the associated rhizobacteria. The profile of phospholipid fatty acids was determined by high-resolution GC. A high level of 18:2 omega 6 (a fatty-acid specific to fungi) was found in exudates of L. luteus (a mycorrhizal plant) in contrast to L. albus (a non-mycorrhizal plant).

Acids↗
PubMed2005

[Terpenoids in root exudates of different allelopathic rice varieties].

In this paper, a strong allelopathic rice variety PI312777 and a weak allelopathic rice variety Lemont were used as test materials, and the root exudates of two rice varieties grown in the media of sand and soil were respectively collected by using circulation system approach. The terpenoids in ether-extracts were detected by GC-MS, and characterized by spectrum database. The results showed that some terpenoids such as limonene oxide, limonene dioxide, carvone oxide, carveol, and cedrol were detected in the ether-extracts. Different rice varieties had the similar kinds of terpenoids, but their existing modes and quantities were different. The differences between different various cultural methods and between rice varieties were further discussed.

Oryza↗
PubMedDate not supplied

Uptake and exudation of phenolic compounds by wheat and antimicrobial components of the root exudate.

In the course of our study, it was found that phenyl propenoic acid derivatives were readily taken up by wheat. Leaf leachate components were chosen for the feeding experiments and p-coumaric acid, ferulic acid and caffeic acid were found to be quickly taken up into the plants via the roots. The analytical study revealed that the exudate contained potent antimicrobial compounds together with amino acids and sugars. Besides the primary metabolites, 4-hydroxystyrene, 3-methoxy-4-hydroxystyrene and 3-methoxy-4-acetoxystyrene were identified as exudate components from wheat roots in sterile hydroponic culture. This indicates that these antimicrobial components may play a significant role in the defense system as allelochemicals for the rhizosphere.

Anti-Bacterial Agents↗
PubMed2001

Modification of the protein expression pattern induced in the nitrogen-fixing actinomycete Frankia sp. strain ACN14a-tsr by root exudates of its symbiotic host Alnus glutinosa and cloning of the sodF gene.

Two-dimensional (2-D) polyacrylamide gel electrophoresis was used to detect proteins induced in Frankia sp. strain ACN14a-tsr by root exudates of its symbiotic host, Alnus glutinosa. The 5 most prominent proteins were purified from 2-D gels and characterized by N-terminal sequencing. All of these proteins had a high percentage of similarity with known stress proteins. One protein match was the Fe superoxide dismutase (Fe-SOD), another was a tellurite resistance protein (Ter), the third was a bacterioferritin comigratory protein (Bcp); and two matches, differing only by their isoelectric point, were the same small heat shock protein (Hsp), a major immune reactive protein found in mycobacteria. This suggests that the symbiotic microorganism Frankia, first responds with a normal stress response to toxic root products of its symbiotic host plant. To confirm its identity, the gene corresponding to the Fe-SOD protein, sodF was isolated from a genomic library by a PCR-approach and sequenced. It is the first stress response gene characterized in Frankia.

Actinomycetales↗
PubMed2004

Antifungal compounds from the root and root exudate of Zea mays.

A maize plant (Zea mays) planted in a test tube was found to inhibit the growth of the soil-borne plant pathogen, Fusarium oxysporum f. sp. melongenae. The antifungal compounds, 6-methoxybenzoxazolinone and 6,7-dimethoxybenzoxazolinone, were isolated from an ethanol extract of Zea mays roots, and (6R)-7,8-dihydro-3-oxo-alpha-ionone and (6R,9R)-7,8-dihydro-3-oxo-alpha-ionol were isolated from the root exudate.

Antifungal Agents↗
PubMed2004

Use of sucrose-agar globule with root exudates for mass production of vesicular arbuscular mycorrhizal fungi.

A sucrose-agar globule (SAG) was newly introduced to increase production of the vesicular arbuscular mycorrhizal (VAM) fungal spores, Gigaspora gigantea and Glomus fasciculatum. An SAG inoculum and a sucrose-agar globule with root exudates (SAGE) inoculum were prepared, and their spore productions were compared with a soil inoculum. When the SAGE was used as the inoculum on sucrose-agar medium plates the number of spores was increased (35% more than the soil inoculum). After the soil inoculum and SAGE were inoculated on an experimental plant, Zingiber officinale, the percentage root colonization, number of VAM spores, and dry matter content were analyzed. It was observed that the SAGE showed a higher percentage of root colonization (about 10% more), and increases in the number of spores (about 26%) and dry matter (more than 13%) for the two VAM fungal spores than the soil inoculum. The results of this study suggested that the SAGE inoculum may be useful for the mass production of VAM fungi and also for the large scale production of VAM fungal fertilizer.

Colony Count, Microbial↗
PubMed2004

Anti-fungal sesquiterpenoid from the root exudate of Solanum abutiloides.

The Solanum abutiloides plant is highly resistant to soil-borne pathogens such as Fusarium oxysporum f. sp. melongenae, Verticillium dahliae, and Ralstonia solanacearum. This species is utilized as a mating source of resistant cultivars and is also used as a rootstock. The root exudate of Solanum abutiloides was extracted from a soil system composed of charcoal and vermiculite. Anti-fungal activity was found in the extract, and an active ingredient was isolated. The chemical structure of the active compound was determined to be 3-beta-acetoxysolavetivone, a new sesquiterpenoid. The anti-fungal activity of 3-beta-acetoxysolavetivone examined by the inhibition of spore germination of Fusarium oxysporum was close to that of lubimin, and higher than that of solavetivone.

Antifungal Agents↗
PubMed2002

[Impact of root exudates from transgenic plants on soil micro-ecosystems].

With the commercialization of transgenic plants, ecological risk assessment of transgenic plants has been scheduled. Many problems such as gene transfer from transgenic plants to related wild species, production of super weeds and super virus, tolerance to insect-resistant transgenic plants, and distruption of biodiversity have been taken place in some transgenic plants. The influences of root exudates from transgenic plants on soil micro-ecosystems were reviewed.

Aspergillus niger↗
PubMed2004

Engineering root exudation of Lotus toward the production of two novel carbon compounds leads to the selection of distinct microbial populations in the rhizosphere.

The culture of opine-producing transgenic Lotus plants induces the increase in the rhizosphere of bacterial communities that are able to utilize these molecules as sole carbon source. We used transgenic Lotus plants producing two opines, namely mannopine and nopaline, to characterize the microbial communities directly influenced by the modification of root exudation. We showed that opine-utilizers represent a large community in the rhizosphere of opine-producing transgenic Lotus. This community is composed of at least 12 different bacterial species, one third of which are able to utilize the opine mannopine and two thirds the opine nopaline. Opine utilizers are diverse, belonging to the Gram-positive and -negative bacteria. We described two novel mannopine-utilizing species, Rhizobium and Duganella spp., and five novel nopaline-utilizing species, Duganella, Afipia, Phyllobacterium, Arthrobacter, and Bosea spp. Although opine utilizers mostly belong to the alpha-Proteobacteria, Rhizobiaceae family, there is little overlap between the populations able to utilize each of the two opines produced by the plants. Noticeably, in the rhizosphere of transgenic Lotus, only the opine mannopine favors the growth of Agrobacterium tumefaciens, the bacterium from which opines have been characterized. The diversity of opine utilizers from the rhizosphere of Lotus plants is greater than that observed from any other environment. Therefore, transgenic plants with engineered exudation constitute an excellent tool to isolate and characterize specific microbial populations.

Arginine↗