Effect of associative bacteria on element composition of barley seedlings grown in solution culture at toxic cadmium concentrations

The response of barley seedlings to inoculation with associative rhizobacteria Azospirillum lipoferum 137, Arthrobacter mysorens 7, Agrobacterium radiobacter 10 and Flavobacterium sp. L30 was studied in hydroponic and quartz sand cultures in the presence of 50 microM CdCl2. Cadmium caused severe inhibition in the growth and uptake of nutrient elements by the plants. Inoculation with the bacteria slightly stimulated root length and biomass of hydroponically grown Cd-treated seedlings. The bacteria increased the content of nutrients such as P, Mg, Ca, Fe, Mn and Na in roots and or shoots of the plants grown in the absence of Cd. Positive changes in the element composition caused by the bacteria were less pronounced in Cd-treated plants, whereas the total amount of nutrients taken by the inoculated plants was generally increased significantly. The content of Cd in the inoculated plants was unchanged, except increased in roots upon addition of A. lipoferum 137. Inoculation did not affect the activity of peroxidase, alpha-mannosidase, phosphodiesterae, alpha-galactosidase, and concentration of sulfhydryl compounds used as biochemical markers of stress in plant roots. The results showed that associative bacteria were capable of decreasing partially the toxicity of Cd for the barley plants through the improvement in uptake of nutrient elements.     

The isochorismate pathway is negatively regulated by salicylic acid signaling in O<Subscript>3</Subscript>-exposed <Emphasis Type="Italic">Arabidopsis</Emphasis>

Phytochelatins (PCs) are heavy metal binding peptides that play an important role in sequestration and detoxification of heavy metals in plants. In this study, our goal was to develop transgenic plants with increased tolerance for and accumulation of heavy metals from soil by expressing an Arabidopsis thaliana AtPCS1 gene, encoding phytochelatin synthase (PCS), in Indian mustard (Brassica juncea L.). A 35S promoter fused to a FLAG–tagged AtPCS1 cDNA was expressed in Indian mustard, and transgenic lines, designated pc lines, were evaluated for tolerance to and accumulation of Cd and Zn. Transgenic plants with moderate AtPCS1 expression levels showed significantly higher tolerance to Cd and Zn stress, but accumulated significantly less Cd and Zn than wild type plants in both shoot and root tissues. However, transgenic plants with highest expression of the transgene did not exhibit enhanced Cd and Zn tolerance. Shoots of Cd-treated pc plants had significantly higher levels of phytochelatins and thiols than wild-type plants. Significantly lower concentrations of gluthatione in Cd-treated shoot and root tissues of transgenic plants were observed. Moderate expression levels of phytochelatin synthase improved the ability of Indian mustard to tolerate certain levels of heavy metals, but at the same time did not increase the accumulation potential for Cd and Zn.     

Iron plaque decreases cadmium accumulation in Oryza sativa L. and serves as a source of iron

• Cadmium (Cd) contamination occurs in paddy soils; hence it is necessary to reduce Cd content of rice. Application and mode of action of ferrous sulphate in minimizing Cd in rice was monitored in the present study.
• Pot culture with Indian rice variety Swarna (MTU 7029) was maintained in Cd‐spiked soil containing ferrous sulphates, which is expected to reduce Cd accumulation in rice. Responses in rhizosphere pH, root surface, metal accumulation in plant and molecular physiological processes were monitored.
• Iron plaque was induced on root surfaces after FeSO4 application and the amount of Fe in plaque reduced with increases in Cd in the soil. Rhizosphere pH decreased during plaque formation and became more acidic due to secretion of organic acids from the roots under Cd treatment. Moreover, iron chelate reductase activity increased with Cd treatment, but in the absence of Cd, activity of this enzyme increased in plaque‐induced plants. Cd treatment caused expression of OsYSL18, whereas OsYSL15 was expressed only in roots without iron plaque. Fe content of plants increased during plaque formation, which protected plants from Cd‐induced Fe deficiency and metal toxicity. This was corroborated with increased biomass, chlorophyll content and quantum efficiency of photo‐synthesis among plaque‐induced plants.
• We conclude that ferrous sulphate‐induced iron plaque prevents Cd accumulation and Fe deficiency in rice. Iron released from plaque via organic acid mediated dissolution during Cd stress.

     

矿区分离丛枝菌根真菌对万寿菊吸Cd潜力影响

盆栽试验研究土壤不同施Cd水平(0、5、20、50μg/g)下,接种矿区污染土壤中丛枝菌根真菌对万寿菊根系侵染率、植株生物量及Cd吸收与分配的影响。结果表明:接种丛枝菌根真菌显著提高Cd胁迫下万寿菊的根系侵染率和植株生物量;随着施Cd水平提高,各处理植株Cd浓度显著增加。各施Cd水平下万寿菊地上部Cd吸收量远远高于根系Cd吸收量,在土壤施Cd量达到50μg/g时,接种处理地上部Cd吸收量是根系的3.48倍,对照处理地上部Cd吸收量是根系的1.67倍;同一施Cd水平下接种处理植株Cd吸收量要显著高于对照。总体上,试验条件下污染土壤中分离的丛枝菌根真菌促进了万寿菊对土壤中Cd的吸收,并在一定程度上增加Cd向地上部分的运转,表现出植物提取的应用潜力。     

Mechanisms of Cadmium Mobility and Accumulation in Indian Mustard

Indian mustard (Brassica juncea L.), a high biomass crop plant, accumulated substantial amounts of cadmium, with bioaccumulation coefficients (concentration of Cd in dry plant tissue/concentration in solution) of up to 1100 in shoots and 6700 in roots at nonphytotoxic concentrations of Cd (0.1 [mu]g/mL) in solution. This was associated with a rapid accumulation of phytochelatins in the root, where the majority of the Cd was coordinated with sulfur ligands, probably as a Cd-S4 complex, as demonstrated by x-ray absorption spectroscopy. In contrast, Cd moving in the xylem sap was coordinated predominantly with oxygen or nitrogen ligands. Cd concentrations in the xylem sap and the rate of Cd accumulation in the leaves displayed similar saturation kinetics, suggesting that the process of Cd transport from solution through the root and into the xylem is mediated by a saturable transport system(s). However, Cd translocation to the shoot appeared to be driven by transpiration, since ABA dramatically reduced Cd accumulation in leaves. Within leaves, Cd was preferentially accumulated in trichomes on the leaf surface, and this may be a possible detoxification mechanism.     

Rhizosphere Bacteria Enhance Selenium Accumulation and Volatilization by Indian Mustard

Indian mustard (Brassica juncea L.) accumulates high tissue Se concentrations and volatilizes Se in relatively nontoxic forms, such as dimethylselenide. This study showed that the presence of bacteria in the rhizosphere of Indian mustard was necessary to achieve the best rates of plant Se accumulation and volatilization of selenate. Experiments with the antibiotic ampicillin showed that bacteria facilitated 35% of plant Se volatilization and 70% of plant tissue accumulation. These results were confirmed by inoculating axenic plants with rhizosphere bacteria. Compared with axenic controls, plants inoculated with rhizosphere bacteria had 5-fold higher Se concentrations in roots (the site of volatilization) and 4-fold higher rates of Se volatilization. Plants with bacteria contained a heat-labile compound in their root exudate; when this compound was added to the rhizosphere of axenic plants, Se accumulation in plant tissues increased. Plants with bacteria had an increased root surface area compared with axenic plants; the increased area was unlikely to have caused their increased tissue Se accumulation because they did not accumulate more Se when supplied with selenite or selenomethionine. Rhizosphere bacteria also possibly increased plant Se volatilization because they enabled plants to overcome a rate-limiting step in the Se volatilization pathway, i.e. Se accumulation in plant tissues.     

Rhizoplane colonization of pea seedlings by Rhizobium leguminosarum and a deleterious root colonizing Pseudomonas sp. and effects on plant growth

Some pseudomonads produce a toxin that specifically inhibits winter wheat (Triticum aestivum L.) root growth and the growth of several microorganisms. The toxin does not inhibit pea (Pisum sativum) root growth, but the organisms are aggressive root colonizers and their effect on Rhizobium leguminosarum growth, colonization, and nodulation of peas was not known. Peas were grown in Leonard jars in the greenhouse. Pea roots were inoculated with R. leguminosarum, a toxin-producing Pseudomonas sp., both, or neither (control). The Pseudomonas sp. colonized pea roots more rapidly and in greater number than R. leguminosarum after ten days. In the presence of the Pseudomonas sp., the R. leguminosarum population on the rhizoplane was less at ten days. When the roots were inoculated with both R. leguminosarum and Pseudomonas sp., the number of nodules were greater than when R. leguminosarum was inoculated alone, but nodule dry weight and pea shoot biomass were similar to plants inoculated with only R. leguminosarum. Although these results need confirmation with non-sterile soil and field studies, these preliminary results indicate that peas will not be affected by wheat root-inhibitory rhizobacteria.     

Regulation of plasma membrane aquaporins by inoculation with a Bacillus megaterium strain in maize (Zea mays L.) plants under unstressed and salt-stressed conditions

It is documented that some plant-growth-promoting rhizobacteria (PGPR) enhance plant salt tolerance. However, as to how PGPR may influence two crucial components of plant salt tolerance such as, root hydraulic characteristics and aquaporin regulation has been almost unexplored. Here, maize (Zea mays L.) plants were inoculated with a Bacillus megaterium strain previously isolated from a degraded soil and characterized as PGPR. Inoculated plants were found to exhibit higher root hydraulic conductance (L) values under both unstressed and salt-stressed conditions. These higher L values in inoculated plants correlated with higher plasma membrane type two (PIP2) aquaporin amount in their roots under salt-stressed conditions. Also, ZmPIP1;1 protein amount under salt-stressed conditions was higher in inoculated leaves than in non-inoculated ones. Hence, the different regulation of PIP aquaporin expression and abundance by the inoculation with the B. megaterium strain could be one of the causes of the different salt response in terms of root growth, necrotic leaf area, leaf relative water content and L by the inoculation treatment.     

Rhizobacteria improve sugarcane growth and photosynthesis under well‐watered conditions

Morpho‐physiological changes caused by particular plant growth‐promoting rhizobacteria were evaluated in sugarcane plants under varying water availability. Under well‐watered conditions, we have found one rhizobacteria isolate (IAC‐RBcr5) able to enhance root dry matter and photosynthesis of sugarcane plants. The IAC‐RBcr5 genome was sequenced and high similarity was found with Pseudomonas putida GB‐1. Based on increased root system size of inoculated plants, we hypothesised that sugarcane plants inoculated with IAC‐RBcr5 would have improved performance under water deficit. Although IAC‐RBcr5 had improved plant leaf CO₂ assimilation under water shortage, inoculation caused reduction of biomass accumulation in sugarcane. The negative influence of water deficit on shoot growth rate and root traits such as volume, area, diameter, length and specific root area was higher in plants treated with IAC‐RBcr5 as compared to non‐inoculated ones. However, rhizobacteria‐induced improvements in leaf and root proline contents would represent a strategy for storing carbon and nitrogen during low water availability and helping both organisms to resume their metabolism after rehydration. In conclusion, we found and identified a rhizobacterium able to improve growth and photosynthesis of sugarcane plants. Such benefit for plant growth was lost under low water availability as a likely consequence of increased carbon‐energy demand by rhizobacteria and their sensitivity to drought.     

Bacteria associated with yellow lupine grown on a metal‐contaminated soil: in vitro screening and in vivo evaluation for their potential to enhance Cd phytoextraction

In order to stimulate selection for plant‐associated bacteria with the potential to improve Cd phytoextraction, yellow lupine plants were grown on a metal‐contaminated field soil. It was hypothesised that growing these plants on this contaminated soil, which is a source of bacteria possessing different traits to cope with Cd, could enhance colonisation of lupine with potential plant‐associated bacteria that could then be inoculated in Cd‐exposed plants to reduce Cd phytotoxicity and enhance Cd uptake. All cultivable bacteria from rhizosphere, root and stem were isolated and genotypically and phenotypically characterised. Many of the rhizobacteria and root endophytes produce siderophores, organic acids, indole‐3‐acetic acid (IAA) and aminocyclopropane‐1‐carboxylate (ACC) deaminase, as well as being resistant to Cd and Zn. Most of the stem endophytes could produce organic acids (73.8%) and IAA (74.3%), however, only a minor fraction (up to 0.7%) were Cd or Zn resistant or could produce siderophores or ACC deaminase. A siderophore‐ and ACC deaminase‐producing, highly Cd‐resistant Rhizobium sp. from the rhizosphere, a siderophore‐, organic acid‐, IAA‐ and ACC deaminase‐producing highly Cd‐resistant Pseudomonas sp. colonising the roots, a highly Cd‐ and Zn‐resistant organic acid and IAA‐producing Clavibacter sp. present in the stem, and a consortium composed of these three strains were inoculated into non‐exposed and Cd‐exposed yellow lupine plants. Although all selected strains possessed promising in vitro characteristics to improve Cd phytoextraction, inoculation of none of the strains (i) reduced Cd phytotoxicity nor (ii) strongly affected plant Cd uptake. This work highlights that in vitro characterisation of bacteria is not sufficient to predict the in vivo behaviour of bacteria in interaction with their host plants.     

Comparison of tolerance of <Emphasis Type="Italic">Brassica juncea</Emphasis> and <Emphasis Type="Italic">Vigna radiata</Emphasis> to cadmium

The effect of different cadmium concentrations (6–120 μM) on Hill reaction activity (HRA) of isolated chloroplasts, contents of chlorophylls (Chls) and carotenoids (Cars), and Cd uptake and accumulation in plant organs of Indian mustard (Brassica juncea L. cv. Vitasso) and mung bean [Vigna radiata (L.) Wilczek] were determined. The Cd stress inhibited photochemical activity of isolated chloroplasts of both species and in both tested developmental stages. On the basis of EC₅₀ values, the mung bean showed a higher sensitivity to Cd treatment than Indian mustard. The higher sensitivity of both species was determined in the earlier than in the older developmental stage. The leaves of Cd-treated plants possessed lower contents of Chls and Cars in both species and the negative effect increased with Cd concentration. A difference between species was also found in Cd uptake and accumulation. In both species, Cd was accumulated more in roots than in shoots, with higher accumulation in Indian mustard than in mung bean.     

Anatomy and morphology of photinia (Photinia × fraseri Dress) in vitro plants inoculated with rhizobacteria

Photinia × fraseri Dress (photinia) is a woody plant with high ornamental value. The anatomy and morphology of micropropagated photinia inoculated with the plant growth-promoting rhizobacteria Azospirillum brasilense and Azotobacter chroococcum, in combination with pulses of 49.2 μM indole-3-butyric acid during rhizogenesis, were characterized using light and electron microscopy. Leaves of inoculated in vitro plants showed better development than those subjected to auxin control only. All inoculated treatments, independent of the bacterial strain used, had leaves with two layers of palisade parenchyma, a thick cuticle and linear unicellular trichomes. There was no proliferation of undifferentiated tissue in any treatment and the plants showed shoot–root vascular connections. Ex vitro leaves and in vitro plants inoculated with Azospirillum brasilense Cd and Azotobacter chroococcum 42 had large stomata with elliptic aperture radially surrounded by small stomata on the abaxial foliar surface. In addition, plants of these treatments had a large root hair zone over the root surface. Bacteria were only observed on surfaces of root hairs. The results suggest that the structural changes induced by bacterial inoculation of photinia in vitro plants could lead to better adaptation to ex vitro conditions after transplanting.     

Transgenic Indian mustard (<Emphasis Type="Italic">Brassica juncea</Emphasis>) plants expressing an <Emphasis Type="Italic">Arabidopsis</Emphasis> phytochelatin synthase (<Emphasis Type="Italic">AtPCS1</Emphasis>) exhibit enhanced As and Cd tolerance

Phytochelatins (PCs) are post-translationally synthesized thiol reactive peptides that play important roles in detoxification of heavy metal and metalloids in plants and other living organisms. The overall goal of this study is to develop transgenic plants with increased tolerance for and accumulation of heavy metals and metalloids from soil by expressing an Arabidopsis thaliana AtPCS1 gene, encoding phytochelatin synthase (PCS), in Indian mustard (Brassica juncea L.). A FLAG-tagged AtPCS1 gDNA, under its native promoter, is expressed in Indian mustard, and transgenic pcs lines have been compared with wild-type plants for tolerance to and accumulation of cadmium (Cd) and arsenic (As). Compared to wild type plants, transgenic plants exhibit significantly higher tolerance to Cd and As. Shoots of Cd-treated pcs plants have significantly higher concentrations of PCs and thiols than those of wild-type plants. Shoots of wild-type plants accumulated significantly more Cd than those of transgenic plants, while accumulation of As in transgenic plants was similar to that in wild type plants. Although phytochelatin synthase improves the ability of Indian mustard to tolerate higher levels of the heavy metal Cd and the metalloid As, it does not increase the accumulation potential of these metals in the above ground tissues of Indian mustard plants.     

Early changes in root characteristics of maize (Zea mays) following seed inoculation with the PGPR Azospirillum lipoferum CRT1

The effect of direct inoculation of seeds with the plant growth promoting rhizobacteria (PGPR) Azospirillum lipoferum CRT1 was assessed on maize (Zea mays) grown for 35 days after sowing (d.a.s.) in controlled conditions (greenhouse) in a luvisol soil from south-eastern France. WhinRhizo^® software was used to describe the following changes in the root system morphology for each plant: distribution and average root diameter, root surface and the number of tips. The stress at breakage and stiffness of the roots in tension were also determined. Evaluation of biochemical components of roots was achieved by direct Attenuated Total Reflectance (or reflection) (ATR)-Fourier transform infrared (FTIR) on root section. Inoculated roots exhibited significantly larger numbers of tips and extending surface to rhizosphere when compared to controls. Measured mechanical parameters of inoculated roots showed a slight increase in rupture stress up to the largest diameter (1.2 mm) when compared to controls. Stiffness (Young’s modulus) values were nearly constant for inoculated plants with higher values than for non-inoculated plants at day 26 and day 35. Using Principal Components Analysis of ATR-FTIR profiles, the polysaccharide enrichment of inoculated roots compared to controls was found at day 35. Noticeable absorbance at wavenumber specific to aromatic ether (lignin) was observed in control plants. All these data had a pattern of immature root properties, when maize was inoculated with Azospirillum lipoferum CRT1. Observed modifications of root development are possibly conducive to unseen beneficial effects, like water retention, resistance to mechanical stress, or root litter quality. Studies on more mature plants are required to assess if the differences between inoculated and control plants would persist or become accentuated with time until harvest.     

Alleviation of Cadmium Toxicity in Brassica juncea L. (Czern. & Coss.) by Calcium Application Involves Various Physiological and Biochemical Strategies

Calcium (Ca) plays important role in plant development and response to various environmental stresses. However, its involvement in mitigation of heavy metal stress in plants remains elusive. In this study, we examined the effect of Ca (50 mM) in controlling cadmium (Cd) uptake in mustard (Brassica juncea L.) plants exposed to toxic levels of Cd (200 mg L⁻¹ and 300 mg L⁻¹). The Cd treatment showed substantial decrease in plant height, root length, dry weight, pigments and protein content. Application of Ca improved the growth and biomass yield of the Cd-stressed mustard seedlings. More importantly, the oil content of mustard seeds of Cd-stressed plants was also enhanced with Ca treatment. Proline was significantly increased in mustard plants under Cd stress, and exogenously sprayed Ca was found to have a positive impact on proline content in Cd-stressed plants. Different concentrations of Cd increased lipid peroxidation but the application of Ca minimized it to appreciable level in Cd-treated plants. Excessive Cd treatment enhanced the activities of antioxidant enzymes superoxide dismutase, ascorbate peroxidase and glutathione reductase, which were further enhanced by the addition of Ca. Additionally, Cd stress caused reduced uptake of essential elements and increased Cd accumulation in roots and shoots. However, application of Ca enhanced the concentration of essential elements and decreased Cd accumulation in Cd-stressed plants. Our results indicated that application of Ca enables mustard plant to withstand the deleterious effect of Cd, resulting in improved growth and seed quality of mustard plants.     

A Study on Cadmium Phytoremediation Potential of Indian Mustard,Brassica juncea

The objective of this study was to investigate Cd phytoremediation ability of Indian mustard, Brassica juncea. The study was conducted with 25, 50, 100, 200 and 400 mg Kg^?1 CdCl₂ in laboratory for 21 days and Cd concentrations in the root, shoot and leaf tissues were estimated by atomic absorption spectroscopy. The plant showed high Cd tolerance of up to 400 mg Kg^?1 but there was a general trend of decline in the root and shoot length, tissue biomass, leaf chlorophyll and carotenoid contents. The tolerance index (TI) of plants were calculated taking both root and shoot lengths as variables. The maximum tolerance (TI _shoot = 87.4 % and TI _root = 89.6 %) to Cd toxicity was observed at 25 mg Kg^?1, which progressively decreased with increase in dose. The highest shoot (10791 μg g^?1 dry wt) and root (9602 μg g^?1 dry wt) Cd accumulation was achieved at 200 mg kg^?1 Cd treatment and the maximum leaf Cd accumulation was 10071.6 μg g^?1 dry wt achieved at 100 mg Kg^?1 Cd, after 21 days of treatment. The enrichment coefficient and root to shoot translocation factor were calculated, which, pointed towards the suitability of Indian mustard for removing Cd from soil.     

Ultrastructural localization and identification ofAzospirillum brasilense Cd on and within wheat root by immuno-gold labeling

Azospirillum brasilense Cd localization in wheat roots was studied by light microscopy, by scanning, and by transmission electron microscopy.A. brasilense Cd cells were specifically identified immunocytochemically around and within root tissues.A. brasilense Cd cells found both outside and inside inoculated roots were intensively labeled with colloidal gold. In non-axenic cultures other bacterial strains or plant tissue were not labeled, thereby providing a non-interfering background. The roots of axenic grown wheat plants were colonized both externally and internally byA. brasilense Cd after inoculation, whereas non-axenic cultures were colonized by other bacterial strains as well.A. brasilense Cd cells were located on the root surface along the following zones: the root tip, the elongation, and the root-hair zone. However, bacteria were located within the cortex only in the latter two zones. In a number of observations, an electron dense material mediated the binding of bacterial cells to outer surfaces of epidermal cells, or between adjacent bacterial cells.A. brasilense Cd were found in root cortical intercellular spaces, but were not detected in either the endodermal layer or in the vascular system. This study proposes that in addition to root surface colonization,A. brasilense Cd forms intercellular associations within wheat roots.     

Biofumigation: Isothiocyanates released frombrassica roots inhibit growth of the take-all fungus

The presence of root tissue of the brassicas canola and Indian mustard inhibited growth of pure cultures of the fungal pathogen which causes take-all of wheat [Gaeumannomyces graminis (Sacc.) Arx and Oliver var.tritici, abbreviated as Ggt]. Ggt growth was generally inhibited more in the presence of Indian mustard roots than canola roots. Dried irradiated roots were consistently effective in reducing Ggt growth, but growth inhibition by young live roots and macerated roots was not consistent. The inhibitory compound(s) were shown to be volatile because the symmetry of Ggt growth was not affected by the proximity of theBrassica tissue. Volatile breakdown products from maceratedBrassica roots were identified using a gas chromatograph-mass spectrometer. The major compounds found were isothiocyanates (ITCs). Canola roots released mostly methyl ITC and Indian mustard roots released mostly phenylethyl ITC. Low concentrations of these and related compounds inhibited growth of Ggt in pure culture when supplied as the vapour of pure chemicals in concentrations within the range expected during breakdown ofBrassica roots in soil.     

Role of the modification in root exudation induced by arbuscular mycorrhizal colonization on the intraradical growth of Phytophthora nicotianae in tomato

We studied the role of modification in root exudation induced by colonization with Glomus intraradices and Glomus mosseae in the growth of Phytophthora nicotianae in tomato roots. Plants were grown in a compartmentalized plant growth system and were either inoculated with the AM fungi or received exudates from mycorrhizal plants, with the corresponding controls. Three weeks after planting, the plants were inoculated or not with P. nicotianae growing from an adjacent compartment. At harvest, P. nicotianae biomass was significantly reduced in roots colonized with G. intraradices or G. mosseae in comparison to non-colonized roots. Conversely, pathogen biomass was similar in non-colonized roots supplied with exudates collected from mycorrhizal or non-mycorrhizal roots, or with water. We cannot rule out that a mycorrhiza-mediated modification in root exudation may take place, but our results did not support that a change in pathogen chemotactic responses to host root exudates may be involved in the inhibition of P. nicotianae.     

Alterations in membrane potential and in proton efflux in plant roots induced byAzospirillum brasilense

Inoculation of soybean seedlings withAzospirillum brasilense Cd significantly reduced the membrane potential in every root part and was being maximal in the root elongation zone. Monitoring the proton efflux pattern of inoculated wheat roots by severalA. brasilense strains and byPseudomonas sp. for prolonged periods (up to 200h) revealed a change from the bimodal pattern of proton efflux of non inoculated roots. This change was not related to root colonization ability but to bacterial capacity to induce changes in root surface area. Continuous perfusion of the plant nutrient solution with a fresh solution (from inoculation time), eliminated the enhancing effect of inoculation on proton efflux. We propose thatA. brasilense inoculation influences membrane activity and subsequently proton efflux in roots, probably through the release of an as yet unidentified bacterial signal.