Effect of thallium fractions in the soil and pollution origins on Tl uptake by hyperaccumulator plants: a key factor for the assessment of phytoextraction

Phytoremediation is often discussed as a means of extracting trace metals in excess in the soil, but to increase its efficiency a better understanding of the factors controlling plant uptake is required. The main objective of this study was to examine the effect of origin (anthropogenic vs. geogenic) and mobility of thallium (Tl) in the rhizosphere on Tl uptake. Two Tl-hyperaccumulating Brassicaceae species, kale (Brassica oleracea acephala L. cv. Winterbor F1) and candytuft (Iberis intermedia Guers.), were grown in a rhizobox system to investigate the dynamics of Tl in the rhizosphere soil. Four different soils were used. Two soils contained high Tl amounts due to anthropogenic sources (emissions from a cement plant and mining activities). High Tl content in the two other soils was due to a high rock content (geogenic origin). On completion of growth in the rhizoboxes, the depletion of Tl in seven different chemical fractions, determined by sequential extraction, was compared to the plant uptake. Most of the Tl taken up was derived from the so-called "easily accessible" fractions in both soils with geogenic Tl as well as in the soils polluted by mining activities. Due to the small amounts of easily accessible Tl in the geogenic soils, Tl uptake by Brassicaceae was low. On the other hand, for the air emission-polluted soil, a high depletion of Tl from "less accessible" fractions was observed in addition to depletion of the easily accessible fractions. Hence, the latter soil demonstrated the highest potential for effective soil decontamination by phytoextraction within an appropriate time frame.     

Effects of arsenate and phosphate on their accumulation by an arsenic-hyperaccumulator Pteris vittata L.

Arsenate and phosphate interactions are important for better understanding their uptake and accumulation by plant due to their similarities in chemical behaviors. The present study examined the effects of arsenate and phosphate on plant biomass and uptake of arsenate and phosphate by Chinese brake (Pteris vittata L.), a newly-discovered arsenic hyperaccumulator. The plants were grown for 20 weeks in a soil, which received the combinations of 670, 2670, or 5340 mol kg^–1 arsenate and 800, 1600, or 3200 mol kg^–1 phosphate, respectively. Interactions between arsenate and phosphate influenced their availability in the soil, and thus plant growth and uptake of arsenate and phosphate. At low and medium arsenate levels (670 and 2670 mol kg^–1), phosphate had slight effects on arsenate uptake by and growth of Chinese brake. However, phosphate substantially increased plant biomass and arsenate accumulation by alleviating arsenate phytotoxicity at high arsenate levels (5340 mol kg^–1). Moderate doses of arsenate increased plant phosphate uptake, but decreased phosphate concentrations at high doses because of its phytotoxicity. Based on our results, the minimum P/As molar ratios should be at least 1.2 in soil solution or 1.0 in fern fronds for the growth of Chinese brake. Our findings suggest that phosphate application may be an important strategy for efficient use of Chinese brake to phytoremediate arsenic contaminated soils. Further study is needed on the mechanisms of interactive effects of arsenate and phosphate on Chinese brake in hydroponic systems.     

Ammonium and nitrate uptake rates and rhizosphere pH in non-mycorrhizal roots of Norway spruce [Picea abies (L.) Karst.]

Summary Relationships between root zone temperature, concentrations and uptake rates of NH ₄ ⁺ and NO ₃ ^– were studied in non-mycorrhizal roots of 4-year-old Norway spruce under controlled environmental conditions. Additionally, in a forest stand NH ₄ ⁺ and NO ₃ ^– uptake rates along the root axis and changes in the rhizosphere pH were measured. In the concentration (C_min) range of 100–150 M uptake rates of NH ₄ ⁺ were 3–4 times higher than those of NO ₃ ^– The preference for NH ₄ ⁺ uptake was also reflected in the minimum concentration (C_min) values. Supplying NH₄NO₃, the rate of NO ₃ ^– uptake was very low until the NH ₄ ⁺ concentrations had fallen below about 100 M. The shift from NH ₄ ⁺ to NO ₃ ^– uptake was correlated with a corresponding shift from net H⁺ production to net H⁺ consumption in the external solution. The uptake rates of NH ₄ ⁺ were correlated with equimolar net production of H⁺. With NO ₃ ^– nutrition net consumption of H⁺ was approximately twice as high as uptake rates of NO ₃ ^– In the forest stand the NO ₃ ^– concentration in the soil solution was more than 10 times higher than the NH ₄ ⁺ concentration (<100 M), and the rhizosphere pH of non-mycorrhizal roots considerably higher than the bulk soil pH. The rhizosphere pH increase was particularly evident in apical root zones where the rates of water and NO ₃ ^– uptake and nitrate reductase activity were also higher. The results are summarized in a model of water and nutrient transport to, and uptake by, non-mycorrhizal roots of Norway spruce in a forest stand. Model calculations indicate that delivery to the roots by mass flow may meet most of the plant demand of nitrogen and calcium, and that non-mycorrhizal root tips have the potential to take up most of the delivered nitrate and calcium.     

Growth response of Atriplex nummularia to inoculation with arbuscular mycorrhizal fungi at different salinity levels

Chenopods are generally regarded as non-host plants for mycorrhizal fungi and are believed not to benefit from colonization by mycorrhizal fungi. Perennial Atriplex nummularia Lindl., growing under field conditions, showed a relatively high level of colonization by mycorrhizal fungi (10–30% of root length colonized) in spring and summer. Accordingly, two glasshouse experiments were designed to assess the effects of inoculation with mycorrhizal fungi (with a single species or a mixture of different species) on growth, nutrient uptake, and rhizosphere bacterial community composition of A. nummularia at high and low salinity levels (2.2 and 12 dSm^–1). Only low and patchy colonization by mycorrhizal fungi (1–2 of root length colonized) was detected in inoculated plants under glasshouse conditions which was unaffected by salinity. Despite the low colonization, inoculation increased plant growth and affected nutrient uptake at both salinity levels. The effects were higher at an early stage of plant development (6weeks) than at a later stage (9–10 weeks). Salinity affected the bacterial community composition in the rhizosphere as examined by ribosomal intergenic spacer amplification (RISA) of 16S rDNA, digitization of the band patterns and multivariate analysis. The effects of inoculation with mycorrhizal fungi on growth of A. nummularia may be attributed to (i) direct effects of mycorrhizal fungi on plant nutrient uptake and/or (ii) indirect effects via mycorrhizal-induced changes in the bacterial community composition.     

Effects of a vesicular-arbuscular mycorrhizal fungus and other soil microorganisms on growth,mineral nutrient acquisition and root exudation of soil-grown maize plants

Maize (Zea mays L. cv. Alize) plants were grown in a calcareous soil in pots divided by 30-m nylon nets into three compartments, the central one for root growth and the outer ones for hyphal growth. Sterle soil was inoculated with either (1) rhizosphere microorganisms other than vesicular-arbuscular mycorrhizal (VAM) fungi, (2) rhizosphere microorganisms together with a VAM fungus [Glomus mosseae (Nicol. and Gerd.) Gerdemann and Trappel], or (3) with a gamma-irradiated inoculum as control. Plants were grown under controlled-climate conditions and harvested after 3 or 6 weeks. VAM plants had higher shootroot ratios than non-VAM plants. After 6 weeks, the concentrations of P, Zn and Cu in roots and shoots had significantly increased with VAM colonization, whereas Mn concentrations had significantly decreased. Root exudates were collected on agar sheets placed on the interface between root and hyphal compartments. Six-week-old VAM and non-VAM plants had similar root exudate compositions of 72–73% reducing sugars, 17–18% phenolics, 7% organic acids and 3% amino acids. In another experiment in which root exudates were collected on agar sheets with or without antibiotics, the amounts of amino acids and carbohydrates recovered were similar in VAM and non-VAM plants. However, threeto sixfold higher amounts of carbohydrates, amino acids and phenolics were recovered when antibiotics were added to the agar sheets. Thus, the high microbial activity in the rhizosphere and on the rhizoplane limits the exudates recovered from roots.     

Heavy metal tolerance and accumulation of Cd, Cr and Ni by Cannabis sativa L.

Experiments in semi-natural conditions were undertaken to assess hemp metal tolerance and its ability to accumulate cadmium, nickel and chromium. Cannabis sativa was grown in two soils, S1 and S2, containing 27, 74, 126 and 82, 115, 139 g g^–1 of Cd, Ni and Cr, respectively. After two months from germination and at ripeness, no significant alteration in plant growth or morphology was detected. On the contrary, a high hemp reactivity to heavy metal stress with an increase in phytochelatin and DNA content was observed during development, suggesting the Cannabis sativa ability to avoid cell damage by activating different molecular mechanisms. Metals were preferentially accumulated in the roots and only partially translocated to the above-ground tissues. The mean shoot Cd content was 14 and 66 g g^–1 for S1 and S2 soil, respectively. Although not negligible concentrations they were about 100 times lower than those calculated for the hyperaccumulator Thlaspi caerulescens. Similarly Ni uptake was limited if compared with that of the Ni-hyperaccumulator Alyssum murale. Chromium uptake was negligible. As expected on the base of the metal concentration detected in ripe plants, no statistically significant variation in soil metal content was detected after one crop of hemp. Nevertheless, a consistent amount (g) of Cd and Ni is expected to be extracted by 1 ha biomass of hemp (about 10 t) per year and along the time a slow restoration of deeper soil portions can be obtained by its wide root system (at least 0,5 m deep). In addition, the possibilities of growing hemp easily in different climates and using its biomass in non-food industries can make heavy metal contaminated soils productive. This means economical advantage along with a better quality of soil.     

Changes of Ni biogeochemistry in the rhizosphere of the hyperaccumulator Thlaspi goesingense

Processes in the rhizosphere of metal hyperaccumulator species are largely unknown. We investigated root-induced changes of Ni biogeochemistry in the rhizosphere of Thlaspi goesingense Hálácsy in a rhizobox experiment and in related soil chemical and Ni uptake studies. In the rhizobox, a root monolayer was separated from rhizosphere soil by a nylon membrane. Rhizosphere soil was then sliced into 0.5 mm layers and analyzed for changes in soluble (water-extractable, Ni _S) and labile (1 M NH ₄NO ₃-extractable, Ni _L) Ni pools. Ni _L in the rhizosphere was depleted due to excessive uptake in T. goesingense. Ni _S in the rhizosphere increased in contrast to expectations based on the experimental Ni desorption isotherm. Mathematical simulations following the Tinker–Nye–Barber approach overestimated the depletion of the Ni _L and predicted a decrease of Ni _S in the rhizosphere. In a hydroponic experiment, we demonstrated that T. goesingense takes up Ni ²⁺ but excludes metal–organic complexes. The model output was then improved in later versions considering this finding. A sensitivity analysis identified I _max and K _m, derived from the Michaelis–Menten uptake kinetics experiment to be the most sensitive of the model parameters. The model was also sensitive to the accuracy of the estimate of the initial Ni concentration (C _Si) in soil solution. The formation of Ni–DOM complexes in solution could not explain the poor fit as in contrast to previous field experiments, the correlation between soluble Ni and dissolved organic carbon (DOC) was weak. Ion competition of Ni with Ca and Mg could be ruled out as explanation of enhanced Ni solubility in the rhizosphere as the molar ratio of Ni/(Ca + Mg) in solution was not affected. However, a decreased Vanselov coefficient Kv near the root plane indicated (an apparent) lower selectivity of the exchange complex for Ni, possibly due to adsorption of oxalate exuded by T. goesingense roots or associated rhizosphere microbes. This conclusion is supported by field data, showing enhanced oxalate concentrations in the rhizosphere of T. goesingense on the same experimental soil. The implications for phytoextraction and bio-available contaminant stripping (BCS) as well as for future modeling and experimental work are discussed.     

Effects of Cd and Zn on the development of annual xylem rings of young Norway spruce (Picea abies) plants

Three-year-old spruce (Picea abies) saplings were planted and cultivated for 2 years in pots with 3 1 substrate, consisting of a homogenized mixture of sand, peat and forest soil with a high organic content (volume ratio 11.52). This substrate was amended with 10–180 mol Cd [kg soil dry weight (DW)]^–1, 50–7500 mol Zn (kg soil DW)^–1 (determined with 1 M ammonium acetate extracts) or combinations of both elements. Annual xylem growth rings in stems of plants treated with 50 mol Cd (kg soil DW)^–1 or 7500 mol Zn (kg soil DW)^–1 were significantly narrower than in control plants. Growth reductions were more pronounced in the second year of the experiment. The contents of Cd and Zn in stem wood and needles were positively correlated with the substrate concentrations. The Mg contents of the spruce needles were inversely correlated with soil concentrations of Cd and Zn. Root development was impeded at moderate concentrations of Cd (50 mol kg^–1) or Zn (1000 mol kg^–1) in the substrate. The adverse effects of potentially toxic trace elements, like Cd or Zn, on xylem growth of spruce plants are discussed with regard to possible growth reductions in forest trees under field conditions.     

Ethylene and carbon dioxide concentrations of soils as influenced by rhizosphere of crops under field and pot conditions

A method for collecting low volumes of soil gas from a small region, and a technique for determining small concentrations of ethylene using an enrichment process are described. Using these methods, it was found that ethylene and carbon dioxide (CO₂) concentrations of soils varied considerably depending on the presence or absence of a rhizosphere. Ethylene was much higher (31–375 nL L^–1; mean: 207) in non-cropped areas (i.e., soils without rhizosphere) than in the rhizosphere region (8–136 nL L^–1; mean: 38) of a field in which maize or soybean were grown. On the other hand, CO₂ concentrations were higher in rhizosphere than in non-rhizosphere soil, especially in pot experiments. The rate of ethylene decomposition was, however, much greater in rhizosphere soil (55 nL g^–1 day^–1) than in non-rhizosphere soil (34 nL g^–1 day^–1). Higher microbial activity was presumed to result in the decrease of ethylene concentration and the increase in CO₂ in rhizosphere regions. The implications of these results in relation to the influence of ethylene in rhizosphere on plant growth, and the role of soil microbes on decomposition of ethylene is discussed.     

Phytoavailability of Cd and Zn in soil estimated by stable isotope exchange and chemical extraction

The distribution of labile Cd and Zn in two contrasting soils was investigated using isotopic exchange techniques and chemical extraction procedures. A sewage sludge amended soil from Great Billings (Northampton, UK) and an unamended soil of the Countesswells Association obtained locally (Aberdeen, UK) were used. ¹¹⁴Cd and ⁶⁷Zn isotopes were added to a water suspension of each soil and the labile metal pool (E-value) determined from the isotope dilution. Samples were obtained at 13 time points from 1h to 50 days. For the sewage sludge amended soil, 29 g Cd g^–1 (86% of total) and 806 g Zn g^–1 (65% of total) were labile and for the Countesswells soil the value was 8.6 g Zn g^–1 (13% of total); limits of detection prevented a Cd E-value from being measured in this soil. The size of the labile metal pool was also measured by growing plants for 90 days and determining the isotopic content of the plant tissue (L-value). Thlaspi caerulescensJ. & C. Presl (alpine penny cress), a hyperaccumulator of Zn and Cd, Taraxacum officinale Weber (dandelion) and Hordeum vulgare L. (spring barley) were used. L-values were similar across species and lower than the E-values. On average the L-values were 23±0.8 g Cd g^–1 and 725±14 g Zn g^–1 for the Great Billings soil and 0.29±0.16 g Cd g^–1 and 7.3±0.3 g Zn g^–1 for the Countesswells soil. The extractable metal content of the soils was also quantified by extraction using 0.1 M NaNO₃, 0.01 M CaCl₂, 0.5 M NaOH, 0.43 M CH₃COOH and 0.05 M EDTA at pH 7.0. Between 1.3 and 68% of the total Cd and between 1 and 50% of the total Zn in the Great Billings soil was extracted by these chemicals. For the Countesswells soil, between 6 and 83% of the total Cd and between 0.1 and 7% of the total Zn was extracted. 0.05 M EDTA and 0.43 M CH₃COOH yielded the greatest concentrations for both soils but these were less than the isotopic estimates. On the whole, E-values were numerically closer to the L-values than the chemical extraction values. The use of isotopic exchange provides an alternative estimate of the labile metal pool within soils compared to existing chemical extraction procedures. No evidence was obtained that T. caerulescens is able to access metal within the soil not freely available to the other plants species. This has implications for long term remediation strategies using hyperaccumulating plant species, which are unlikely to have any impact on non-labile Cd and Zn in contaminated soil.     

Short-term studies of NO 3 − uptake in Pisum using 13NO 3 −

Influx, efflux and net uptake of NO ₃ ^– was studied in Pisum sativum L. cv. Marma in short-term experiments where ¹³NO ₃ ^– was used to trace influx. The influx rate in N-limited plants was similar both during net uptake at external concentrations of around 50 M, and at low external NO ₃ ^– concentrations (4–6 M) when net uptake was practically zero. Efflux could be inferred from discrepancies between influx and net uptake but was never very high in the N-limited plants during net uptake. Close to the threshold concentration for not NO ₃ ^– uptake, efflux was high and equalled influx. Thus, the threshold concentration can be regarded as a NO ₃ ^– compensation point. The inclusion of NH ₄ ⁺ in the outer medium decreased influx by about 40% but did not significantly affect efflux. The roles of NO ₃ ^– fluxes and nitrate-reductase activity in regulating/limiting NO ₃ ^– utilization are discussed.Abbreviations DW dry weight - FW fresh weight - R_N relative nitrogen addition rate     

Distribution of soil fractions and location of soil bacteria in a vertisol under cultivation and perennial grass

Effects of soil management on soil characteristics were investigated on the rhizosphere (RPP) and the nonrhizosphere (NRPP) soil of a re-grass vertisol underDigitaria decumbens and in the soil under continuous cultivation (CC). A low energy technique allowed to separate eight size and density fractions, including macro- and micro-aggregates while preserving soil bacteria. Organic C and N, microbial biomass C and the number of total bacteria (AODC) and ofAzospirillum brasilense and their distribution were determined in soil fractions isolated from the CC, NRPP and RPP soils. Soil macroaggregates (>2000 m) were similarly predominant in the NRPP and RPP soils when the dispersible clay size fraction (<2 m) respresented more than 25% of the CC soil mass. The main increase of C content in RPP originated from the macroaggregates (> 2000 m) and from the root fraction, not from the finer separates. The proportion of organic C as microbial biomass C revealed the low turnover of microbial C in the PP situations, especially in the clay size fraction of the NRPP soil. A common shift of AODC toward the finer separates from planted soils (CC and RPP) revealed the influence of living plants on the distribution of soil bacteria. The relative abundance ofA. brasilense showed the presence of the active roots ofDigitaria in the macroaggregates and their contact with the dispersible clay size fraction of the rhizosphere soil.     

Uptake of Metals by Plants Sharing a Rhizosphere with the Hyperaccumulator Thlaspi caerulescens

The hyperaccumulator Thlaspi caerulescens was grown with Hordeum vulgare and Lepidium heterophyllum in a split pot experiment to examine the effect of rhizosphere interactions on metal uptake. The objective was to assess the viability of such intercropping as either (1) a system of ‘phytoprotection’ for nonaccumulating plants or (2) a means of enhancing phytoextraction with large-biomass crops through increased metal mobilization within the shared rhizosphere. The plants were separated by (1) an impermeable barrier, (2) a permeable root barrier, or (3) no physical barrier to allow different degrees of root interaction. Studies of rhizosphere effects using split pot experiments are subject to considerable uncertainty by the need to relate test results to appropriate control plants. This was resolved by comparing plant metal concentrations to ‘equivalent’ control plants, with the same yield, based on the observed variation in metal concentration with yield under similar growing conditions.

Cadmium concentration in H. vulgare was increased by a factor of 2.4 when it was grown alongside T. caerulescens without a barrier. In contrast, the uptake of zinc by H. vulgare was significantly decreased, probably through metal depletion within the zone of the Zn-hyperaccumulator's rhizosphere. T. caerulescens also apparently increased the concentration of Cd in H. vulgare by a factor of 1.4 when the roots of the two plants were separated by a permeable barrier that allowed movement of soil solution but prevented physical mixing of roots. The concentrations of all the metals studied (Cd, Zn, Cu, Pb, Ni) were greater in T. caerulescens when the hyperaccumulator was grown alongside either L. heterophyllum or H. vulgare without a root barrier — probably through successful exploitation of a greater volume of soil. However, this effect was not seen in the presence of a partial barrier, except in the case of Cu when T. caerulescens was grown alongside H. vulgare.

These results suggest that T. caerulescens may alter conditions in shared rhizospheres and thereby affect the availability of selected metals to neighboring plants. Thus, it is possible that under-sowing some plants with small hyperaccumulators may potentially offer an alternative form of management for marginally contaminated soils. There was limited evidence of an intercropped hyperaccumulator mobilizing selected metals and restricting the availability of others. However, changes in uptake of selected metals by the larger plant may be quite small compared with the requirements of crop protection or the short-term requirements of many land remediation programs.     

Metal-humic complexes and plant micronutrient uptake: a study based on different plant species cultivated in diverse soil types

There are several studies in the literature dealing with the effect of metal-humic complexes on plant metal uptake, but none of them correlate the physicochemical properties of the complexes with agronomic results. Our study covers both aspects under various experimental conditions. A humic extract (SHE) obtained from a sapric peat was selected for preparing the metal–humic complexes used in plant experiments. Fe–, Zn– and Cu–humic complexes with a reaction stoichiometry of 2:0.25 (humic:metal, w/w) were chosen after studying their stability and solubility with respect to pH (6–9) and the humic:metal reaction stoichiometry. Wheat and alfalfa plants were greenhouse cultured in pots containing one of three model soils: an acid, sandy soil and two alkaline, calcareous soils. Treatments were: control (no additions), SHE (53 mg kg^–1 of SHE), and metal (Cu, Zn and Fe)–SHE complexes (2.5 and 5 mg kg^–1 of metal rate and a SHE concentration to make 53 mg kg ^–1). Cu- and Zn–humic complexes significantly (p0.05) increased the plant uptake and the DTPA-extractable soil fraction of complexed micronutrients in most plant–soil systems. However, these effects were associated with significant increases (p0.05) of shoot and root dry weight only in alfalfa plants. In wheat, significant increases of root and shoot dry matter were only observed in the Cu–humic treated plants growing in the acid soil, where Cu deficiency was more intense. The Fe–humic complex did not increase Fe plant assimilation in any plant–soil system, but SHE increased Fe-uptake and/or DTPA-extractable soil Fe in the wheat–calcareous soil systems. These results, taken together with those obtained from the study of the pH- and SHE:metal ratio-dependent SHE complex solubility and stability, highlight the importance of the humic:Fe complex stoichiometry on iron bioavailability as a result of its influence on complex solubility.     

Influence of clay mineral type and organic matter content on the uptake of239Pu and241Am by plants

Summary The uptake of²³⁹Pu and²⁴¹Am from different clay mineral-organic matter-sand mixtures simulating contrasting soil types was examined in growth chamber experiments. The mixtures represented various combinations of organic matter (0, 5 and 10%), kaolinite (11 type) and montomorillonite (21 type) clay minerals, each at the levels of 5, 10 and 25%, and purified quartz sand (as filler).Results indicated a marked reduction in uptake of both²³⁹Pu and²⁴¹Am with increase in organic matter as well as clay content of the mixtures. The Pu Concentration Ratios (CRs) ranged from (2.5–7.0)×10^–3 in the case of kaolinite-organic matter mixtures, and from (0.9–5.5)×10^–3 in the case of montmorillonite-organic matter mixtures. The corresponding values of Am Concentration Ratios (CRs) obtained were (1.9–725.4)×10^–3 in the case of kaolinite-organic matter mixtures, and between (0.7–3.5)×10^–3 for the montmorillonite-organic matter mixtures.Reduction in the uptake of²⁴¹Am with increasing clay content was more pronounced in the montmorillonite clay-organic matter mixtures as compared to that in the case of kaolinite-organic matter mixtures. While similar qualitative reduction in²³⁹Pu CRs with increasing clay content was observed, the reduction was less marked than in the case of²⁴¹Am. The values for Am CRs were higher than the corresponding Pu CRs in kaolinite based mixtures whereas in the case of montmorillonite-organic matter mixtures Pu CRs exceeded the Am CRs.Increasing organic matter content and its interaction with both kaolinite and montmorillonite clay minerals were found to be equally effective in reducing the uptake of²³⁹Pu as well as²⁴¹Am by plants.     

Changes in phosphorus concentrations and pH in the rhizosphere of some agroforestry and crop species

The aim of this work was to assess whether agroforestry species have the ability to acquire P from pools unavailable to maize. Tithonia diversifolia(Hemsley) A. Gray, Tephrosia vogelii Hook f., Zea mays and Lupinus albusL. were grown in rhizopots and pH change and depletion of inorganic and organic P pools measured in the rhizosphere. Plants were harvested at the same growth stage, after 56 days for maize and white lupin and 70 days for tithonia and tephrosia, and the rhizosphere sampled. The rhizosphere was acidified by tithonia (pH change –0.3 units to pH 4.8) and lupins (–0.2 units to 4.9), alkalinised by tephrosia (+0.4 units to pH 5.4), and remained unchanged with maize growth. Concurrent with acidification in the rhizosphere of tithonia there was a decline in resin-P (0.8 g P g^–1). However, there was also a decline in NaOH extractable inorganic P (NaOH-P_i) (5.6 g P g^–1 at the root surface) and organic P pools (NaOH-P_o) (15.4 g P g^–1 at 1.5 mm from the root), which would not be expected without specific P acquisition mechanisms. Alkalinisation of tephrosia rhizosphere was accompanied by changes in all measured pools, although the large depletion of organic P (21.6 g P g^–1 at 5 mm from the root) suggests that mineralisation, as well as desorption of organic P, was stimulated. The size of changes of both pH and P pools varied with distance away from the rhizoplane. Decline of more recalcitrant P pools with the growth of the agroforestry species contrasted with the effect of maize growth, which was negligible on resin-P and NaOH-P_i, but led to an accumulation of P as NaOH-P_o (14.2 g P g^–1 at 5 mm from the root). Overall the depletion of recalcitrant P pools, particularly P_o, suggests that the growth of tithonia and tephrosia enhance desorption and dissolution of P, while also enhancing organic P mineralisation. Both species appear to have potential for agroforestry technologies designed to enhance the availability of P to crops, at least in the short term.     

Nitrogen fixation and metabolism by groundwater-dependent perennial plants in a hyperarid desert

The Central Asian Taklamakan desert is characterized by a hyperarid climate with less than 50 mm annual precipitation but a permanent shallow groundwater table. The perched groundwater (2–16 m) could present a reliable and constant source of nitrogen throughout the growing season and help overcome temporal nitrogen limitations that are common in arid environments. We investigated the importance of groundwater and nitrogen fixation in the nitrogen metabolism of desert plants by assessing the possible forms and availability of soil N and atmospheric N and the seasonal variation in concentration as well as isotopic composition of plant N. Water availability was experimentally modified in the desert foreland through simulated flooding to estimate the contribution of surface water and temporally increased soil moisture for nutrient uptake and plant–water relations. The natural vegetation of the Taklamakan desert is dominated by plants with high foliar nitrogen concentrations (2–3% DM) and leaf nitrate reductase activity (NRA) (0.2–1 mol NO₂^– g^–1 FW h^–1). There is little evidence that nitrogen is a limiting resource as all perennial plants exhibited fast rates of growth. The extremely dry soil conditions preclude all but minor contributions of soil N to total plant N so that groundwater is suggested as the dominant source of N with concentrations of 100 M NO₃^–. Flood irrigation had little beneficial effect on nitrogen metabolism and growth, further confirming the dependence on groundwater. Nitrogen fixation was determined by the ¹⁵N natural abundance method and was a significant component of the N-requirement of the legume Alhagi, the average contribution of biologically fixed nitrogen in Alhagi was 54.8%. But nitrogen fixing plants had little ecological advantage owing to the more or less constant supply of N available from groundwater. From our data we conclude that the perennial species investigated have adapted to the environmental conditions through development of root systems that access groundwater to satisfy demands for both water and nutrients. This is an ecologically favourable strategy since only groundwater is a predictable and stable resource.     

Changes of tomato rhizosphere microflora following application of the herbicide diphenamid to soil infested with Fusarium oxyspomm f.sp. lycopersici

Significant variations were detected in species composition between untreated rhizosphere and nonrhizosphere soils of tomato plants. Application of different concentrations of active ingredient of the herbicide diphenamid (5–250 ppm) to these soils caused significant alterations in species assemblages as compared with untreated soils. Also variations in species composition were denoted between treated rhizosphere and non-rhizosphere soils.Diphenamid concentrations of 10–100 ppm significantly affected microbial counts in soil and rhizosphere of tomato plants. Counts have been stimulated at diphenamid concentrations ranging from 10–50 ppm for fungi and 10–100 ppm for bacteria. At concentrations higher than the upper limits of these ranges, R/S values were not significantly affected.The results also indicated that Fusarium oxyspomm f.sp. lycopersici populations were unaffected by diphenamid at the recommended field rate (10 ppm). Above this concentration and within the conditions of the experiment, the pathogen maintained its population at detectable inocula. Population counts of Aspergillus candidus, a species reported to be able to degrade diphenamid, were high in both treated rhizosphere and non-rhizosphere soils.     

A technique for studying rhizosphere processes in tree crops: soil phosphorus depletion around camellia (Camellia japonica L.) roots

Rhizosphere studies on tree crops have been hampered by the lack of a satisfactory method of sampling soils at various distances in the rhizosphere. A modified root study container (RSC) technique developed for annual crops, grasses and legumes was used to study the mechanisms by which camellia plants (Camellia japonica L.) utilise soil P in the glasshouse and field. Plants belonging to the Camellia family (e.g. tea) have the ability to utilise P from relatively unavailable native P sources and for this reason camellia plants were selected for this study.In the glasshouse trial, the RSCs were filled with a Recent soil, treated with P fertilisers; North Carolina phosphate rock (NCPR), diammonium phosphate (DAP), mono calcium phosphate (MCP) and single superphosphate (SSP) at 200 g P g^-1 soil. A planar mat of roots was physically separated by a 24 m polyester mesh and the soil on the other side of this mesh was cut into thin slices parallel to the rhizoplane and analysed for pH, and different forms of P (organic, P_o and inorganic, P_i) to understand P depletion at different distances from camellia roots. In the field trial this technique was modified and used to study the rhizosphere processes in mature camellia trees fertilised with only SSP and NCPR.In both field and glasshouse trials, all P fertilisers increased all the bulk soil P fractions except NaOH-P_o over unfertilised soil with the greatest increases being in the H₂SO₄-P_i fraction in the NCPR treatment and NaOH-P_i in the SSP treatment. Resin-P, NaOH-P_i and H₂SO₄-P_i were significantly lower in the rhizosphere soil compared to the bulk soil whereas NaOH-P_o was higher in the rhizosphere soil than in the bulk soil. Plant and microbial P uptake were thought to be the major causes for the low resin-P rather than P fixation by Fe and Al because the NaOH-P_i fraction which is a measure of Fe-P and Al-P, also decreased in the rhizosphere soil. The rhizo-deposition of NaOH-P_o suggests that labile inorganic P was immobilized by rhizosphere microbes which were believed to have multiplied as a result of carbon exudates from the roots. A marked reduction in pH (about 0.2–0.4 in the glasshouse and 0.2 in the field trial) was observed near the rhizoplane compared to that in the bulk soil in all treatments. The pH near the rhizoplane as well as in the bulk soil was highest for NCPR treated soil. The increase in pH in the NCPR treatment over the control was consistent with the number of protons consumed during the dissolution of NCPR. In both trials, the dissolution of NCPR in the rhizosphere was higher than in the bulk soil due to lower pH and plant uptake of solution P in the rhizosphere. The RSC technique proved to be a viable aid to study the rhizosphere processes in tree crops.     

Anion-dependent transport of thallous ions through human erythrocyte membrane

Summary Unidirectional fluxes of ²⁰⁴Tl⁺ through the human red blood cell membrane were measured. The inward rate coefficient measured in a K⁺-free saline was 15.6±0.6 hr^–1. The influx of Tl⁺ could be partially inhibited with 0.1mm ouabain (by 28%), 0.1mm DIDS (by 50%) or 1mm furosemide (by 51%). The inhibitory effects of ouabain and DIDS or furosemide were additive. Half-maximal responses were seen at 0.72 m and 0.22mm concentrations of DIDS and furosemide, respectively. A similar action of these blockers on Tl⁺ influx was observed in the erythrocytes incubated in MgCl₂-sucrose media. The outward rate coefficient of ²⁰⁴Tl was also inhibited by DIDS and furosemide (by 65 and 52%, respectively). Rate coefficients of ²⁰⁴Tl influx and efflux decreased significantly in the red cells exposed to Cl^–-free media (NaNO₃ or Mg(NO₃)₂-sucrose). Under these conditions addition of DIDS and furosemide led to only a small inhibition of Tl⁺ fluxes. There was a linear increase in Tl⁺ influx with rising of external Cl^– concentration within 80–155mm or HCO ₃ ^– concentration from 20 to 40mm when the sum of anions was kept constant (155mm) with NO ₃ ^– . The HCO ₃ ^– -stimulated Tl⁺ influx was completely blocked by 0.05mm DIDS but only 67% by 1mm furosemide. The present study provides direct evidence for the occurrence of Cl^– (HCO ₃ ^– )-dependent, DIDS-sensitive movement of Tl⁺ across the human erythrocyte membrane in both directions. Under physiological conditions, about half of net Tl⁺ fluxes occurs due to an anion exchange mechanism. Our data fail to detect a contribution of the Na-K-Cl cotransport system to Tl⁺ transport in human erythrocytes.