Evaluation of a novel tool for sampling root exudates from soil-grown plants compared to conventional techniques

The release of low molecular weight (LMW) organic compounds (e.g. organic acids, amino acids, sugars, etc.) by living plant roots significantly contributes to the development of chemical, physical as well as microbial rhizosphere gradients. Suitable and accurate sampling procedures are crucial for enhancing our understanding of the dynamics of related rhizosphere processes. Here we compare common sampling techniques with a novel tool for root exudate collection that allows non-destructive and repetitive sampling from soil-grown roots. Root exudates from Zea mays L. were collected using the following techniques: (i) hydroponic growth and sampling, (ii) soil growth and hydroponic sampling and (iii) rhizoboxes fitted with a novel in situ root exudate collecting tool. Furthermore, rhizosphere soil solution for the analysis of exudates and microbial metabolites was sampled using micro-suction cups (iv). The effect of different sampling solutions (deionised water and 0.5 mM CaCl₂) on organic acid and amino acid exudation patterns was also investigated. The novel exudate collecting tool was successfully tested for root exudate sampling. Results showed that particularly amino acid exudation rates were significantly affected by growth conditions and sampling procedures, while organic acid exudation patterns varied less across the different sampling setups. Despite qualitative and quantitative differences, exudation rates were in the same order of magnitude across the different sampling procedures. Soil solution concentrations obtained from micro-suction-cup sampling at defined distance to the root surface showed no distinct gradient, highlighting the importance of soil microorganisms in regulating the soil solution concentration of LMW C compounds either via microbial degradation or the release of microbial metabolites. The exudate collector offers new opportunities to assess root exudation rates and composition from soil-grown plants and thus enhances our knowledge of fundamental rhizosphere processes.     

Characterization of Root Exudates at Different Growth Stages of Ten Rice (Oryza sativa L.) Cultivars

Abstract: Plant root exudates play important roles in the rhizosphere. We tested three media (nutrient solution, deionized water and CaSO₄ solution) for three periods of time (2, 4 and 6 h) for collecting root exudates of soil‐grown rice plants. Nutrient culture solution created complications in the analyses of exudates for total organic C (TOC) by the wet digestion method and of organic acids by HPLC due to the interference by its components. Deionized water excluded such interference in analytical analyses but affected the turgor of root cells; roots of four widely different rice cultivars excreted 20 to 60 % more TOC in deionized water than in 0.01 M CaSO₄. Furthermore, the proportion of carbohydrates in TOC was also enhanced. Calcium sulfate solution maintained the osmotic environment for root cells and did not interfere in analytical procedures. Collection for 2 h avoided under‐estimation of TOC and its components exuded by rice roots, which occurred during prolonged exposure. By placing plants in 0.01 M CaSO₄ for 2 h, root exudates of soil‐grown traditional, tall rice cultivars (Dular, B40 and Intan), high‐yielding dwarf cultivars (IR72, IR52, IR64 and PSBRc 20), new plant type cultivars (IR65598 and IR65600) and a hybrid (Magat) were collected at seedling, panicle initiation, flowering and maturity and characterized for TOC and organic acids. The exudation rates were, in general, lowest at seedling stage, increased until flowering but decreased at maturity. Among organic acids, malic acid showed the highest concentration followed by tartaric, succinic, citric and lactic acids. With advancing plant growth, exudation of organic acids substituted exudation of sugars. Root and shoot biomass were positively correlated with carbon exudation suggesting that it is driven by plant biomass. As root exudates provide substrates for methanogenesis in rice fields, large variations in root exudation by cultivars and at different growth stages could greatly influence CH₄ emissions. Therefore, the use of high‐yielding cultivars with lowest root excretions, for example IR65598 and IR65600, would mediate low exudate‐induced CH₄ production. The screening of exciting rice cultivars and breeding of new cultivars with low exudation rates could offer an important option for mitigation of CH₄ emission from rice agriculture to the atmosphere.     

锌营养状况对小麦根细胞膜透性的影响

小麦缺锌不仅导致根系K~ 和NO_3~-泌出量增加,而且低分子量有机化合物如氨基酸、糖类化合物和酚类化合物的泌出量也明显提高。重新供锌(ZnSO_4)12h后,根系K~ 、NO_3~-、氨基酸和碳水化合物的泌出量迅速减少,随着时间的延长,泌出量接近对照水平。结果说明锌对根细胞膜结构的稳定性及膜功能的完整性是必不可少的。     

Low-level herbivory by root-knot nematodes (Meloidogyne incognita) modifies root hair morphology and rhizodeposition in host plants (Hordeum vulgare)

Low amounts of root infestation by plant parasitic nematodes are suggested to increase nutrient supply and in turn enhance microbial activity and net mineralization rate in the rhizosphere. These effects are generally related to “leakage” of plant-derived metabolites from damaged roots. Besides leakage, the present study examines other nematode–host interactions such as alterations in root exudation and morphology, which were almost not considered yet. This includes undamaged root parts in order to assess systemic plant response. The root-knot nematode Meloidogyne incognita (Kofoid and White 1919; Chitwood 1949) and barley (Hordeum vulgare L. cv. Europa) was used as model system. Host plants were grown in mini-rhizotrons inoculated with 0, 2,000, 4,000 or 8,000 M. incognita for 4 weeks. Root morphology, rhizodeposition (sugars, carboxylates, amino acids), and rhizosphere microbial communities (PLFAs) were assessed. In treatments with 4,000 nematodes, shoot biomass, total N and P content increased by the end of the experiment. Generally, an enhanced release of plant metabolites (sugars, carboxylates, amino acids) from the apical root zone occurred 1 week after inoculation with 4,000 and 8,000 M. incognita, indicating root leakage. Low levels of root herbivory stimulated root hair elongation in both infected and uninfected roots. These systemic changes in root morphology likely contributed to the increased sugar exudation in uninfected roots in all nematode treatments at 3 weeks after inoculation. Root-knots formed a separate microhabitat within the root-system. They were characterised by decreased rhizodeposition and increased fungal to bacterial ratio in the adhering rhizosphere soil. The present study provides the first evidence that, apart from leakage, nematode root herbivory at background levels induces local and systemic effects on root morphology and exudation, which in turn may affect plant performance.     

Root Exudation of Phytochemicals in Arabidopsis Follows Specific Patterns That Are Developmentally Programmed and Correlate with Soil Microbial Functions

Plant roots constantly secrete compounds into the soil to interact with neighboring organisms presumably to gain certain functional advantages at different stages of development. Accordingly, it has been hypothesized that the phytochemical composition present in the root exudates changes over the course of the lifespan of a plant. Here, root exudates of in vitro grown Arabidopsis plants were collected at different developmental stages and analyzed using GC-MS. Principle component analysis revealed that the composition of root exudates varied at each developmental stage. Cumulative secretion levels of sugars and sugar alcohols were higher in early time points and decreased through development. In contrast, the cumulative secretion levels of amino acids and phenolics increased over time. The expression in roots of genes involved in biosynthesis and transportation of compounds represented in the root exudates were consistent with patterns of root exudation. Correlation analyses were performed of the in vitro root exudation patterns with the functional capacity of the rhizosphere microbiome to metabolize these compounds at different developmental stages of Arabidopsis grown in natural soils. Pyrosequencing of rhizosphere mRNA revealed strong correlations (p<0.05) between microbial functional genes involved in the metabolism of carbohydrates, amino acids and secondary metabolites with the corresponding compounds released by the roots at particular stages of plant development. In summary, our results suggest that the root exudation process of phytochemicals follows a developmental pattern that is genetically programmed.     

Exometabolites of bread wheat and tomato affecting the plant-microbe interactions in the rhizosphere

The composition of organic acids, carbohydrates, and amino acids released by the roots of wheat (Triticum aestivum L., cv. Obelisk) and tomato (Lycopersicon esculentum L., cv. Carmello) was examined. Tomato roots released threefold more organic acids, sevenfold more tyrosine, 3.8-fold more threonine, and 2.8-fold more arginine that wheat roots; in contrast, wheat roots released 4.1-fold more tryptophan than tomato roots. Differences in the ratio between organic acids and sugars and also in the intensity of L-tryptophan release could affect the interaction between tested plants and microorganisms in their rhizosphere.     

Effect of Pseudomonas fluorescens on the root exudates of two tomato mutants differently sensitive to Fe chlorosis

Plants with different Fe-mobilization properties are known to differ in the amount and kind of Fe-reducing and Fe-chelating compounds exuded by their roots. Although rhizosphere bacteria are known to affect the exudation of organic compounds by the plant roots, their effect on the root exudates of plants differing in Fe-mobilization properties is not known. We studied the effect of Pseudomonas fluorescens, on the exudation of sugars and organic and amino acids by roots of an iron chlorosis-resistant (T3238FER) and a chlorosis-susceptible (T3238fer) tomato mutant. Under sterile conditions two tomato mutants grew equally well and did not differ in the total amount of sugars and organic acid exuded by their roots. More amino acids, however, were exuded by the roots of T3238FER than T323fer. Mutants differed in the amount of oxalic acid and the amino acids Ala, Asp, Gaba, Gln, Gly, His, Hyl, Ile, Leu, Lys, Phe, Pro, and Val exuded by their roots into sterile rooting media. Addition of P. fluorescens to the rooting medium did not affect the growth of T3238FER but stimulated the root growth of chlorosis-susceptible T3238fer, reduced the amounts of glucose, arabinose and fructose but increased the amount of sucrose, reduced the amounts of fumaric, malic and oxalic acid but increased the amounts of citric and succinic acid in the rooting media of both mutants. P. fluorescens resulted in the following changes in the amino acids in the rooting media: reduced the amounts of Gly, Leu, and Lys in T3238FER, and of Asp, Gln, Hyp, and Ile in T3238fer, and increased the amounts of Cys, Glu, His, Hyp, Ile, Phe and Tyr in T3238FER and of Ala, Glu, His, Phe, and Ser in T323fer—in cases more than 40-fold. These differential effects of P. fluorescens in altering the pattern of organic and amino acids compounds with some Fe-chelating properties detected in the rooting medium of these two mutants may indicate that the differences in Fe-chlorosis susceptibility of these tomato mutants may be the result of, or modified by, the interactions between plant roots and rhizosphere microorganisms. We postulate that the Fe-chlorosis susceptibility in plants may be the product of the interactions between soil microorganisms and plant roots, and may not be solely related to the plant per se.     

Anaerobic stimulation of root exudates and disease of peas

Summary The relationships between root exudation, root disease and anaerobic root stresses were investigated. Sand culture and mist chamber studies demonstrated that low O₂ and high CO₂ reduced plant growth and increased the exudation of ethanol, amino acids, and sugars by pea roots. The relative loss of ethanol by roots was much greater in treatments with atmospheres of N₂ containing 30% CO₂ than in treatments of air containing 30% CO₂ or N₂. Ethanol was not detected in the nutrient solution of aerated plant roots. Atmospheres of N₂ plus 30% CO₂ caused 500% greater mycelial growth ofFusarium solani f. sp.pisi and 400% more disease of inoculated pea roots. Relative losses of four amino acids and four sugars were much greater in atmospheres of N₂ plus 30% CO₂ than in N₂ or air.     

Environmental Control of Root Exudation of Low-Molecular Weight Organic Acids in Tropical Rainforests

The availability of phosphorus (P) can limit net primary production (NPP) in tropical rainforests growing on highly weathered soils. Although it is well known that plant roots release organic acids to acquire P from P-deficient soils, the importance of organic acid exudation in P-limited tropical rainforests has rarely been verified. Study sites were located in two tropical montane rainforests (a P-deficient older soil and a P-rich younger soil) and a tropical lowland rainforest on Mt. Kinabalu, Borneo to analyze environmental control of organic acid exudation with respect to soil P availability, tree genus, and NPP. We quantified root exudation of oxalic, citric, and malic acids using in situ methods in which live fine roots were placed in syringes containing nutrient solution. Exudation rates of organic acids were greatest in the P-deficient soil in the tropical montane rainforest. The carbon (C) fluxes of organic acid exudation in the P-deficient soil (0.7?mol?C?m^?2?month^?1) represented 16.6% of the aboveground NPP, which was greater than those in the P-rich soil (3.1%) and in the lowland rainforest (4.7%), which exhibited higher NPP. The exudation rates of organic acids increased with increasing root surface area and tip number. A shift in vegetation composition toward dominance by tree species exhibiting a larger root surface area might contribute to the higher organic acid exudation observed in P-deficient soil. Our results quantitatively showed that tree roots can release greater quantities of organic acids in response to P deficiency in tropical rainforests.     

Root excretion of carboxylic acids and protons in phosphorus-deficient plants

Phosphorus deficiency-induced metabolic changes related to exudation of carboxylic acids and protons were compared in roots of wheat (Triticum aestivum L. cv Haro), tomato (Lycopersicon esculentum L., cv. Moneymaker), chickpea (Cicer arietinum) and white lupin (Lupinus albus L. cv. Amiga), grown in a hydroponic culture system. P deficiency strongly increased the net release of protons from roots of tomato, chickpea and white lupin, but only small effects were observed in wheat. Release of protons coincided with increased exudation of carboxylic acids in roots of chickpea and white lupin, but not in those of tomato and wheat. P deficiency-induced exudation of carboxylic acids in chickpea and white lupin was associated with a larger increase of carboxylic acid concentrations in the roots and lower accumulation of carboxylates in the shoot tissue compared to that in wheat and tomato. - Citric acid was one of the major organic acids accumulated in the roots of all investigated species in response to P deficiency, and this was associated with increased activity and enzyme protein levels of PEP carboxylase, which is required for biosynthesis of citrate. Accumulation of citric acid was most pronounced in the roots of P-deficient white lupin, chickpea and tomato. Increased PEP carboxylase activity in the roots of these plants coincided with decreased activity of aconitase, which is involved in the breakdown of citric acid in the TCA cycle. In the roots of P-deficient wheat plants, however, the activities of both PEP carboxylase and aconitase were enhanced, which was associated with little accumulation of citric acid. The results suggest that P deficiency-induced exudation of carboxylic acids depends on the ability to accumulate carboxylic acids in the root tissue, which in turn is determined by biosynthesis, degradation and partitioning of carboxylic acids or related precursors between roots and shoot. In some plant species such as white lupin, there are indications for a specific transport mechanism (anion channel), involved in root exudation of extraordinary high amounts of citric acid. This revised version was published online in June 2006 with corrections to the Cover Date.     

Aluminum resistance in two cultivars of Zea mays L.: Root exudation of organic acids and influence of phosphorus nutrition

Root exudation of organic acids as Al-chelating compounds and P nutrition have been suggested to play a major role in Al-resistance in higher plants. Effects of Al exposure on maize plant growth, and organic acid root content and root exudation under various levels of P nutrition were examined. Sikuani, a Colombian maize cultivar tolerant to acid soils with high Al saturation, and Corso, a Swiss cultivar, were grown in sterile hydroponic conditions for 21 days. Al-caused inhibition of root growth was lower in Sikuani than in Corso. Al effect on plant growth was decreased with increasing P content in roots. Al content in roots increased with increasing P content and was higher in Sikuani than in Corso. When exposed to Al, the contents in root apices as well as the root exudation of citric and malic acids in Corso and citric, malic and succinic acids in Sikuani increased, and were higher in Sikuani than in Corso. Increased PEP carboxylase (PEPC) activity in root apices after Al exposure partially explained the variations of organic acid content in the roots. These Al-induced changes in PEPC activity, organic acid content and exudation were reduced in plants supplied with higher P concentrations during the 21 days prior to treatment. Increased secretion of organic acids after exposure to Al appeared to be specific to Al and was not totally explained by increased root content in organic acids.     

Organic acids, sugars, and L-tryptophane in exudates of vegetables growing on stonewool and their effects on activities of rhizosphere bacteria

The influence of stonewool substrate on the exudation of the major soluble carbon nutrients and of the auxin precursor tryptophane for Pseudomonas biocontrol agents was studied. To this end, the composition of the organic acids and sugars, as well that of tryptophane, of axenically collected exudates of seed, seedlings, and roots of tomato, cucumber, and sweet pepper was determined. The major results were as follows. i) The total amount of organic acid is much higher than that of total sugar. ii) Exudation of both organic acids and sugars increases during plant growth. iii) Citric, succinic, and malic acids represent the major organic acids, whereas fructose and glucose are the major sugars. iv) Compared with glass beads as a neutral substrate, stonewool substantially stimulates exudation of organic acids and sugars. v) It appeared that enhanced root-tip-colonizing bacteria isolated previously from the rhizosphere of tomato and cucumber grow much better in minimal medium with citrate as the sole carbon source than other, randomly selected rhizobacteria do. This indicates that the procedure which selects for excellent root-tip colonizers enriches for strains which utilize the major exudate carbon source citrate. vi) The content of L-tryptophane, the direct precursor of auxin, is approximately 60-fold higher in seedling exudates of tomato and sweet pepper than in cucumber seedling exudates, indicating a higher possibility of plant growth stimulation after inoculation with auxin-producing rhizobacteria for tomato and sweet pepper crops than for cucumber. However, the biocontrol strain Pseudomonas fluorescens WCS365, which is able to convert tryptophane into auxin, did not stimulate growth of these three crops. In contrast, this strain did stimulate growth of roots of radish, a plant which exudes nine times more tryptophane than tomato does.     

Quantitative and qualitative effects of phosphorus on extracts and exudates of sudangrass roots in relation to vesicular-arbuscular mycorrhiza formation

A comparison was made of water-soluble root exudates and extracts of Sorghum vulgare Pers. grown under two levels of P nutrition. An increase in P nutrition significantly decreased the concentration of carbohydrates, carboxylic acids, and amino acids in exudates, and decreased the concentration of carboxylic acids in extracts. Higher P did not affect the relative proportions of specific carboxylic acids and had little effect on proportions of specific amino acids in both extracts and exudates. Phosphorus amendment resulted in an increase in the relative proportion of arabinose and a decrease in the proportion of fructose in exudates, but did not have a large effect on the proportion of individual sugars in extracts. The proportions of specific carbohydrates, carboxylic acids, and amino acids varied between exudates and extracts. Therefore, the quantity and composition of root extracts may not be a reliable predictor of the availability of substrate for symbiotic vesicular-arbuscular mycorrhizal fungi. Comparisons of the rate of leakage of compounds from roots with the growth rate of vesicular-arbuscular mycorrhizal fungi suggest that the fungus must either be capable of using a variety of organic substrates for growth, or be capable of inducing a much higher rate of movement of specific organic compounds across root cell membranes than occurs through passive exudation as measured in this study.     

Prolonged root hypoxia induces ammonium accumulation and decreases the nutritional quality of tomato fruits

Here we examined the effects of root hypoxia (1-2% oxygen) on the physiology of the plant and on the biochemical composition of fruits in tomato (Solanum lycopersicum cv. Micro-Tom) plants submitted to gradual root hypoxia at first flower anthesis. Root hypoxia enhanced nitrate absorption with a concomitant release of nitrite and ammonium into the medium, a reduction of leaf photosynthetic activity and chlorophyll content, and an acceleration of fruit maturation, but did not affect final fruit size. Quantitative metabolic profiling of mature pericarp extracts by (1)H NMR showed that levels of major metabolites including sugars, organic acids and amino acids were not modified. However, ammonium concentration increased dramatically in fruit flesh, and ascorbate and lycopene concentrations decreased. Our data indicate that the unfavorable effects of root hypoxia on fruit quality cannot be explained by two of the well-known effects of root hypoxia on the plant, namely a decrease in photosynthesis or an excess in ethylene production, but may instead result from disturbances in the supply of either growth regulators or ammonium, by the roots.     

Exudation of amino acids by intact and damaged roots of wheat and peas

Summary Wheat and pea seedlings were grown aseptically in solution-culture and the total free amino nitrogen released by the roots was determined by a quantitative ninhydrin test. Amino nitrogen from wheat plants after 14 days growth was not detected by the test, indicating the release of less than 3 µg of amino nitrogen from a culture of 15 plants. Pea plants of the same age released from 2 to 7 µg per plant. Paper chromatograms of highly concentrated undisturbed solution-cultures revealed up to 13 amino compounds from wheat and 11 from pea. The pattern of amino acids in exudates was similar to that in crushed roots, except for an unidentified amino compound which was detected only in exuded material. The total amino nitrogen and relative proportions of several amino acids in the root exudates of sand-grown peas was influenced by several ratios of oxygen and carbon dioxide supplied to the root zone. Roots, experimentally damaged by swirling and rinsing in sand, released in 1 hour amino nitrogen of from 73 to 120 per cent of that released by normal exudation over a 2-week period. Our findings suggest that experimental and environmental root damage may be responsible for a large proportion of organic materials released by growing plant roots.Trade names are used in this publication to provide specific information. Their use does not constitute a guarantee of the products named and does not signify that they are approved by the U.S. Department of Agriculture to the exclusion of others of suitable composition.     

Membrane-mediated decrease in root exudation responsible for phorphorus inhibition of vesicular-arbuscular mycorrhiza formation

The mechanism responsible for phosphorus inhibition of vesicular-arbuscular mycorrhiza formation in sudangrass (Sorghum vulgare Pers.) was investigated in a phosphorus-deficient sandy soil (0.5 micrograms phosphorus per gram soil) amended with increasing levels of phosphorus as superphosphate (0, 28, 56, 228 micrograms per gram soil). The root phosphorus content of 4-week-old plants was correlated with the amount of phosphorus added to the soil. Root exudation of amino acids and reducing sugars was greater for plants grown in phosphorus-deficient soil than for those grown in the phosphorus-treated soils. The increase in exudation corresponded with changes in membrane permeability of phosphorus-deficient roots, as measured by K⁺ (⁸⁶Rb) efflux, rather than with changes in root content of reducing sugars and amino acids. The roots of phosphorus-deficient plants inoculated at 4 weeks with Glomus fasciculatus were 88% infected after 9 weeks as compared to less than 25% infection in phosphorus-sufficient roots; these differences were correlated with root exudation at the time of inoculation. For plants grown in phosphorus-deficient soil, infection by vesicular-arbuscular mycorrhizae increased root phosphorus which resulted in a decrease in root membrane permeability and exudation compared to nonmycorrhizal plants. It is proposed that, under low phosphorus nutrition, increased root membrane permeability leads to net loss of metabolites at sufficient levels to sustain the germination and growth of the mycorrhizal fungus during pre- and postinfection. Subsequently, mycorrhizal infection leads to improvement of root phosphorus nutrition and a reduction in membrane-mediated loss of root metabolites.     

Microbial products trigger amino acid exudation from plant roots

Plants naturally cycle amino acids across root cell plasma membranes, and any net efflux is termed exudation. The dominant ecological view is that microorganisms and roots passively compete for amino acids in the soil solution, yet the innate capacity of roots to recover amino acids present in ecologically relevant concentrations is unknown. We find that, in the absence of culturable microorganisms, the influx rates of 16 amino acids (each supplied at 2.5 microm) exceed efflux rates by 5% to 545% in roots of alfalfa (Medicago sativa), Medicago truncatula, maize (Zea mays), and wheat (Triticum aestivum). Several microbial products, which are produced by common soil microorganisms such as Pseudomonas bacteria and Fusarium fungi, significantly enhanced the net efflux (i.e. exudation) of amino acids from roots of these four plant species. In alfalfa, treating roots with 200 microm phenazine, 2,4-diacetylphloroglucinol, or zearalenone increased total net efflux of 16 amino acids 200% to 2,600% in 3 h. Data from (15)N tests suggest that 2,4-diacetylphloroglucinol blocks amino acid uptake, whereas zearalenone enhances efflux. Thus, amino acid exudation under normal conditions is a phenomenon that probably reflects both active manipulation and passive uptake by microorganisms, as well as diffusion and adsorption to soil, all of which help overcome the innate capacity of plant roots to reabsorb amino acids. The importance of identifying potential enhancers of root exudation lies in understanding that such compounds may represent regulatory linkages between the larger soil food web and the internal carbon metabolism of the plant.     

Biochemical Changes in Root Exudate and Xylem Sap of Tomato Plants Infected with Meloidogyne incognita

Under two monoxenic culture techniques of growing plants (filter paper and silica sand cultures), sugar in root exudate from Meloidogyne incognita-infected tomato increased 133 to 836% over controls. In contrast, amino acids were moderately reduced 52 to 56%. Chromatographic analysis showed that galled root exudate contained three sugars, twelve amino acids, and three organic acids, whereas healthy root exudate contained four sugars, fifteen amino acids, and four organic acids. Polysaccharide was responsible for the large increase of sugars in galled root exudates. The concn and the absolute amount of total sugars in the infected plant xylem sap were greater than in healthy plant xylem sap up to 6 wk after inoculation, whereas amino acids were moderately lower than in controls throughout the test period. Chromatographic analysis showed that xylem sap from both healthy and infected plants at 4 wk after inoculation contained four sugars and five organic acids. We identified 18 and 17 amino acids in the healthy and infected plant xylem sap, respectively. The concn of sugar increased as the nematode inoculum increased at 2, 4 and 6 wk after inoculation. The amino acids in all samples from the infected plant moderately decreased with an increase of nematode inoculum. We suggest that changes in total sugars and amino acids, of infected plant xylem sap and root exudate are a probable mechanism by which tomato plants are predisposed to Fusarium wilt.     

Distribution of phenolic compounds within seed and seedlings of two Vicia faba cvs differing in their seed tannin content,and study of their seed and root phenolic exudations

The present study analysed the physiology of phenolic exudation by the seed and the root of two Vicia faba cultivars, one rich in condensed tannin (cv. Alfred) and one free-tannin cultivar (cv. Blandine). We first analysed the phenolic exudation from the intact seeds and from the different seed tissues (coat and cotyledon separately). Results indicated that the whole seed exudation in water after 24 h was reduced when the seed was incubated at 4°C in comparison with 30°C. The characterisation and separation of phenolic compounds was achieved by HPLC procedures. The two cultivars of Vicia faba, cv. Blandine and cv. Alfred, were containing in the seed two different chemical patterns. The phenolic patterns of the cv. Alfred seed coat was dominated by catechin derivatives, condensed tannins and flavones. The phenolic pattern of cotyledon and whole seed are very closed and namely made of phenolic acids, and catechin, and flavones as minor compounds. For cv. Blandine, the seed coats contain phenolic acids, flavones, flavonols and dihydroflavonols; the phenolic profile of whole seeds and cotyledons were mainly based on phenolic acids. The kinetic of root phenolic exudation was analysed on the first 21 days of the root growth; the data indicated that phenolic compounds were rapidly released from the emerging root as the amounts of phenolic compounds in exudates were maximum the first day after the seed germination.The HPLC analysis of the phenolic compounds exudated by the root, depicted catechin, phenol acids derivatives and various flavones and flavanones.     

Root exudates of plants

Summary The effect of temperature on the exudation of sugars and amino acids from germinating seeds of maize and cucumber and the effect of the so called cold shock on root exudation of plant seedling was studied. After the first forty-eight hours the exudation of these compounds from germinating seeds generally rises in proportion to the temperature. However, certain relative changes were observed in the composition of the exudates. Exudation of maltose and fructose with arabinose, for example is lower at 28°C than at 19°C.When plants previously growing under favourable temperature conditions were exposed for three days to a lower temperature, there was a several times higher exudation from the roots of maize and a marked rise in exudation from roots of cucumber in comparison with control plants. In the exudates of maize affected by cold shock 3 new oligosaccharides and fructose with saccharose in addition to previously detected substances were found.