Mode-synthesizing atomic force microscopy for volume characterization of mixed metal nanoparticles.

Atomic force microscopy (AFM) and other techniques derived from AFM have revolutionized the understanding of materials and biology at the nanoscale, but mostly provide surface properties. The observation of subsurface nanoscale features and properties remains a great challenge in nanometrology. The operating principle of the mode-synthesizing AFM (MSAFM) is based on the interaction of two ultrasonic waves, one launched by the AFM probe fp , a second launched by the sample fs , and their resulting nonlinear frequency mixing. Recent developments highlighted the need for quantitative correlation between the role of the frequency actuation of the probe fp and the sample fs . Here we present the great potential of MSAFM for advanced volume characterization of metallic nanoparticles presenting a multilayered structure composed of a nickel core surrounded by a gold envelope.

Modification of a commercial DNA extraction kit for safe and rapid recovery of DNA and RNA simultaneously from soil, without the use of harmful solvents.

An optimized method, based on the coupling of two commercial kits, is described for the extraction of soil nucleic acids, with simultaneous extraction and purification of DNA and RNA following a cascade scheme and avoiding the use of harmful solvents. The protocol canmonitor the variations in the recovery yield of DNA and RNA from soils of various types.The quantitative version of the protocol was obtained by testing the starting soil quantity, the grinding parameters and the final elution volumes, in order to avoid saturation of both kits. •A first soil-crushing step in liquid nitrogen could be added for the assessment of fungal parameters.•The protocol was efficienton different tropical soils, including Andosol, while their high contents of clays, including poorly crystalline clays, and Fe and Al oxides usually make the nucleic acid extraction more difficult.•The RNA recovery yield from the previous tropical soils appeared to correlate better to soil respiration than DNA, which is positively influenced by soil clay content.

Phosphorus nutrition of mycorrhizal trees.

Globally, phosphorus (P) limits productivity of trees in many forests and plantations especially in highly weathered, acidic or calcareous profiles. Most trees form mycorrhizal associations which are prevalent in the organic and mineral soil horizons. This review critically examines mechanisms that enhance the acquisition of P by tree roots. Mycorrhizal roots have a greater capacity to take up phosphate (Pi) from the soil solution than non-mycorrhizal root tips. Factors that contribute to this include the extent of extraradical hyphal penetration of soil and the physiology and biochemistry of the fungal/soil and fungal/plant interfaces. Ectomycorrhizal (ECM) trees are likely to benefit from association with basidiomycetes that possess several high-affinity Pi transporters that are expressed in extraradical hyphae and whose expression is enhanced by P deficiency. To understand fully the role of these putative transporters in the symbiosis, data regarding their localization, Pi transport capacities and regulation are required. Some ECM fungi are able to effect release of Pi from insoluble mineral P through excretion of low-molecular-weight organic anions such as oxalate, but the relative contribution of insoluble P dissolution in situ remains to be quantified. How the production of oxalate is regulated by nitrogen remains a key question to be answered. Lastly, phosphatase release from mycorrhizas is likely to play a significant role in the acquisition of Pi from labile organic forms of P (Po). As labile forms of Po can constitute the major fraction of the total P in some tropical and temperate soils, a greater understanding of the forms of Po available to the phosphatases is warranted.

Pinus pinaster seedlings and their fungal symbionts show high plasticity in phosphorus acquisition in acidic soils.

Young seedlings of maritime pine (Pinus pinaster Soland in Aït.) were grown in rhizoboxes using intact spodosol soil samples from the southwest of France, in Landes of Gascogne, presenting a large variation of phosphorus (P) availability. Soils were collected from a 93-year-old unfertilized stand and a 13-year-old P. pinaster stand with regular annual fertilization of either only P or P and nitrogen (N). After 6 months of culture in controlled conditions, different morphotypes of ectomycorrhiza (ECM) were used for the measurements of acid phosphatase activity and molecular identification of fungal species using amplification of the ITS region. Total biomass, N and P contents were measured in roots and shoots of plants. Bicarbonate- and NaOH-available inorganic P (Pi), organic P (Po) and ergosterol concentrations were measured in bulk and rhizosphere soil. The results showed that bulk soil from the 93-year-old forest stand presented the highest Po levels, but relatively higher bicarbonate-extractable Pi levels compared to 13-year-old unfertilized stand. Fertilizers significantly increased the concentrations of inorganic P fractions in bulk soil. Ergosterol contents in rhizosphere soil were increased by fertilizer application. The dominant fungal species was Rhizopogon luteolus forming 66.6% of analysed ECM tips. Acid phosphatase activity was highly variable and varied inversely with bicarbonate-extractable Pi levels in the rhizosphere soil. Total P or total N in plants was linearly correlated with total plant biomass, but the slope was steep only between total P and biomass in fertilized soil samples. In spite of high phosphatase activity in ECM tips, P availability remained a limiting nutrient in soil samples from unfertilized stands. Nevertheless young P. pinaster seedlings showed a high plasticity for biomass production at low P availability in soils.

Adaptation of fine roots to annual fertilization and irrigation in a 13-year-old Pinus pinaster stand.

Effects of fertilization and irrigation on fine roots and fungal hyphae were studied in 13-year-old maritime pine (Pinus pinaster Aït. in Soland), 7 years after the initiation of the treatments. The fertilization trials consisted of a phosphorus treatment, a complete fertilizer treatment (N, P, K, Ca and Mg), and an unfertilized treatment (control). Fertilizers were applied annually and were adjusted according to foliar target values. Two irrigation regimes (no irrigation and irrigation of a set amount each day) were applied from May to October. Root samples to depths of 120 cm were collected in summer of 2005, and the biomass of small roots (diameter 2-20 mm) and fine roots (diameter

A comparison of ammonium, nitrate and proton net fluxes along seedling roots of Douglas-fir and lodgepole pine grown and measured with different inorganic nitrogen sources.

Significant spatial variability in NH4+, NO3- and H+ net fluxes was measured in roots of young seedlings of Douglas-fir (Pseudotsuga menziesii) and lodgepole pine (Pinus contorta) with ion-selective microelectrodes. Seedlings were grown with NH4+, NO3-, NH4NO3 or no nitrogen (N), and were measured in solutions containing one or both N ions, or no N in a full factorial design. Net NO3- and NH4+ uptake and H+ efflux were greater in Douglas-fir than lodgepole pine and in roots not exposed to N in pretreatment. In general, the rates of net NH4+ uptake were the same in the presence or absence of NO3-, and vice versa. The highest NO3- influx occurred 0-30 mm from the root apex in Douglas-fir and 0-10 mm from the apex in lodgepole pine. Net NH4+ flux was zero or negative (efflux) at Douglas-fir root tips, and the highest NH4+ influx occurred 5-20 mm from the root tip. Lodgepole pine had some NH4+ influx at the root tips, and the maximum net uptake 5 mm from the root tip. Net H+ efflux was greatest in the first 10 mm of roots of both species. This study demonstrates that nutrient uptake by conifer roots can vary significantly across different regions of the root, and indicates that ion flux profiles along the roots may be influenced by rates of root growth and maturation.

An overview of BORIS: Bioavailability of Radionuclides in Soils.

The ability to predict the consequences of an accidental release of radionuclides relies mainly on the level of understanding of the mechanisms involved in radionuclide interactions with different components of agricultural and natural ecosystems and their formalisation into predictive models. Numerous studies and databases on contaminated agricultural and natural areas have been obtained, but their use to enhance our prediction ability has been largely limited by their unresolved variability. Such variability seems to stem from incomplete knowledge about radionuclide interactions with the soil matrix, soil moisture, and biological elements in the soil and additional pollutants, which may be found in such soils. In the 5th European Framework Programme entitled Bioavailability of Radionuclides in Soils (BORIS), we investigated the role of the abiotic (soil components and soil structure) and biological elements (organic compounds, plants, mycorrhiza, and microbes) in radionuclide sorption/desorption in soils and radionuclide uptake/release by plants. Because of the importance of their radioisotopes, the bioavailability of three elements, caesium, strontium, and technetium has been followed. The role of one additional non-radioactive pollutant (copper) has been scrutinised in some cases. Role of microorganisms (e.g., K(d) for caesium and strontium in organic soils is much greater in the presence of microorganisms than in their absence), plant physiology (e.g., changes in plant physiology affect radionuclide uptake by plants), and the presence of mycorrhizal fungi (e.g., interferes with the uptake of radionuclides by plants) have been demonstrated. Knowledge acquired from these experiments has been incorporated into two mechanistic models CHEMFAST and BIORUR, specifically modelling radionuclide sorption/desorption from soil matrices and radionuclide uptake by/release from plants. These mechanistic models have been incorporated into an assessment model to enhance its prediction ability by introducing the concept of bioavailability factor for radionuclides.

Root system architecture and receptivity to mycorrhizal infection in seedlings of Cedrus atlantica as affected by nitrogen source and concentration.

Effects of nitrogen (N) source and concentration on root system architecture and receptivity to mycorrhizal infection were studied in seedlings of Atlas cedar (Cedrus atlantica Manetti) grown in root observation boxes in a controlled-environment chamber. Nitrogen was supplied in a solution containing either NO3-; or NH4+ at a concentration of either 0.25 or 5.0 mM. Root extension was recorded twice weekly by tracing the roots growing in contact with the transparent face of the root observation box. Among treatments, lateral root production and branching density were greatest with 5.0 mM NO3-. Inoculation with mycelium of Tricholoma cedrorum Malencon was carried out 3 months after the start of the N treatments. The highest percentage of mycorrhizal roots, and the greatest amounts of living mycelium (estimated by the ergosterol assay) were observed in the NO3- treatments. Differences in root branching density among the N treatments were insufficient to explain the observed differences among treatments in the extent of mycorrhizal infection of seedlings.

Effects of experimental conditions on mycorrhizal relationships between Pinus sylvestris and Lactarius deliciosus and unprecedented fruit-body formation of the Saffron milk cap under controlled soilless conditions.

The mycorrhizal relationships between pines and two edible species of Lactarius sect. Dapetes were investigated by optimizing the experimental conditions of mycelial growth and of mycorrhizal colonization of pine seedlings. In vitro mycelial growth of Lactarius deliciosus and L. sanguifluus was improved on a buffered medium containing glucose, amino acids, and vitamins. Two methods of mycorrhization of pines with Lactarius deliciosus were tested. The mycorrhizal colonization was rapid and intense under non-aseptic conditions with a low nutrient supply and without exogenous glucose. A positive influence of mycorrhizal colonization on Pinus sylvestris growth was subsequently observed. Under axenic conditions and with a high nutrient supply, mycorrhization was stimulated at 10 g/L of exogenous glucose, irrespective of the phosphorus concentration. At high phosphorus level (1 mM) and 0.1, 1.0, or 10.0 g/L glucose, growth of Pinus sylvestris was reduced by inoculation. Stability and development of Pinus spp./Lactarius deliciosus symbioses were assayed in a climatic chamber using containers filled with a synthetic substrate. Over a 2-year culture period, the root systems of the pine seedlings were heavily colonized by Lactarius deliciosus. One year following inoculation, Lactarius deliciosus fruit-body primordia appeared associated with Pinus sylvestris seedlings. Six months later, two mature basidiomata were obtained. This is the first report of soilless fruit-body formation of this edible mushroom.

Stimulation of the ionic transport system in Brassica napus by a plant growth-promoting rhizobacterium (Achromobacter sp.).

A plant growth-promoting rhizobacterium belonging to the genus Achromobacter was isolated from the oil-seed-rape (Brassica napus) root. Growth promotion bioassays were performed with oilseed rape seedlings in a growth chamber in test tubes containing attapulgite and mineral nutrient solution, containing NO3- as N source. The presence of this Achromobacter strain increased shoot and root dry weight by 22-33% and 6-21%, respectively. Inoculation of young seedlings with the Achromobacter bacteria induced a 100% improvement in NO3- uptake by the whole root system. Observations on the seminal root of seedlings 20 h after inoculation showed that there was an enhancement of both the number and the length of root hairs, compared to non-inoculated seedlings. Electrophysiological measurements of NO3- net flux with ion-selective microelectrodes showed that inoculation resulted in a specific increase of net nitrate flux in a root zone morphologically similar in inoculated and non-inoculated plants. The root area increased due to root hair stimulation by the Achromobacter bacteria, which might have contributed to the improvement of NO3- uptake by the whole root system, together with the enhancement of specific NO3- uptake rate. Moreover, inoculated plants showed increased potassium net influx and proton net efflux. Overall, the data presented suggest that the inoculation of oilseed-rape with the bacteria Achromobacter affects the mineral uptake.

Regulation of nitrate reductase in the ectomycorrhizal basidiomycete, Hebeloma cylindrosporum Romagn., cultured on nitrate or ammonium.

The growth of Hebeloma cylindrosporum Romagn. on nitrate medium is associated with nitrate reductase activity equivalent to that measured in herbaceous angiosperms. Nitrate is not indispensable for the induction of NR, as thalli cultivated in ammonium have the same NR activity as thalli cultured on nitrate. However, the simultaneous presence in the culture medium of ammonium with an organic acid such as malate, citrate or succinate causes a diminution of NR activity. Glutamine or asparagine, by-products in the assimilation of ammonium and malate, could intervene in the regulation of this enzyme. The data suggest that the absence of nitrogen or the presence of a nitrogen source, such as ammonium, inappropriate for growth derepresses nitrate reductase in this fungus.

Nitrate nutrition of maritime pine (Pinus pinaster Soland in Ait.) ectomycorrhizal with Hebeloma cylindrosporum Romagn.

In maritime pine seedlings (Pinus pinaster Soland in Ait.), nitrate absorption and reduction capacities were estimated by the incorporation of 15 N-labelled nitrogen. This method allowed us to work on the whole plant Without perturbing plant behaviour. For this purpose, xylem and phloem transports were quantified. After 48 h i nitrate nutrition, mycorrhizal symbiosis with Hebeloma cylindrosporum Romagn. modified neither nitrate accumulation nor absorption and reduction capacities of this anion. Xylem transport of reduced nitrogen was similar in mycorrhizal and non-mycorrhizal seedlings. Only phloem transport was modified by mycorrhizal symbiosis. The intensity of the phloem transport in mycorrhizal pines was approximately twice that in non-mycorrhizal plants.