Soil microbial community as bioindicator of the recovery of soil functioning derived from metal phytoextraction with sorghum

A three-month microcosm study was carried out in order to evaluate: (i) the capacity of sorghum plants to phytoextract Cd (50 mg kg⁻¹) and Zn (1000 mg kg⁻¹) from artificially polluted soil and (ii) the possibility of biomonitoring the efficiency of phytoremediation using parameters related to the size, activity and functional diversity of the soil microbial community. Apart from plant and soil (total and bioavailable) metal concentrations, the following parameters were determined: soil physicochemical properties (pH, OM content, electrical conductivity, total N, and extractable P and K), dehydrogenase activity, basal- and substrate-induced respiration (with glucose and a model rhizodeposit solution, both adjusted to 800 mg C kg⁻¹ DW soil and 45.2 mg N kg⁻¹ DW soil), microbial respiration quotient, functional diversity through community level physiological profiles and, finally, seed germination toxicity tests with Lepidium sativum. Sorghum plants were highly tolerant to metal pollution and capable of reaching high biomass values in the presence of metals. In the first two harvests, values of shoot Cd concentrations were higher than 100 mg Cd kg⁻¹ DW, the threshold value for hyperaccumulators. Nonetheless, in the third harvest, the bioconcentration factor was 1.34 and 0.35 for Cd and Zn, respectively, well below the threshold value of 10 considered for a phytoextraction process to be feasible. In general, microbial parameters showed lower values in metal polluted than in control non-polluted soils, and higher values in planted than in control unplanted pots. As a result of the phytoextraction process, which includes both plant growth and metal phytoextraction, the functioning of the phytoremediated soil, as reflected by the values of the different microbial parameters here determined, was restored. Most importantly, although the phytoextracted soil recovered its function, it was still more phytotoxic than the control non-polluted soil.     

Greenhouse evaluation of EDTA effectiveness at enhancing Cd,Cr, and Ni uptake in <Emphasis Type="Italic">Helianthus annuus</Emphasis> and <Emphasis Type="Italic">Thlaspi caerulescens</Emphasis>

Background, Aims and Scope  Phytoremediation is a promising means for the treatment of heavy metal contamination. Although several species have been identified as hyperaccumulators, most studies have been conducted with only one metal. Experiments were conducted to investigate the ability of Helianthus annuus and Thlaspi caerulescens to simultaneously uptake Cd, Cr and Ni. Materials and Methods  The efficiency of plants grown in a sandy-loam soil was investigated. The ability of two EDTA concentrations (0.1 and 0.3 g kg⁻¹) for enhancing the phytoremediation of Cd, Cr and Ni at two different metal concentrations (24.75 mg kg⁻¹ and 90 mg kg⁻¹) was studied. Results   Thlaspi hyperaccumulated Ni with 0.1 g kg⁻¹ EDTA. When the EDTA dosage was increased to 0.3 g kg⁻¹, Thlaspi was able to hyperaccumulate both Ni and Cr. Since Thlaspi is a low-biomass plant, it was considered insufficient for full-scale applications. Helianthus annuus hyperacummulated Cr (with 0.1 g kg⁻¹ EDTA) and Cd (0.3 g kg⁻¹ EDTA). Discussion  When the contamination was 8.25 mg kg⁻¹ per metal, the total metal uptake was 10–25% (1.35 to 2.12 mg) higher and had the same uptake selectivity (Cr>>Cd>Ni) for both EDTA levels. It was hypothesized that complexation with EDTA interfered with Ni translocation. For these experiments, the optimal results were obtained with the H. annuus-0.1 g kg⁻¹ EDTA combination. Conclusions  Although the use of EDTA did increase the amount of metal that could be extracted, care should be taken during in-situ field applications. Chelators can also increase the amount of metals that are leached past the root zone. Metal leaching and subsequent migration could lead to ground water contamination as well as lead to new soil contamination. Recommendations and Perspectives  Additional research to identify the optimal EDTA dosage for field applications is warranted. This is necessary to ensure that the metals do not leach past the root zone. Identification of a plant that can hyperaccumulate multiple metals is critical for phytoremediation to be a viable remediation alternative. In addition to being able to hyperaccumulate multiple metals, the optimal plant must be fast growing with sufficient biomass to sequester the heavy metals.     

Contribution of arbuscular mycorrhizal fungi and soil amendments to remediation of a heavy metal-contaminated soil using sweet sorghum

Owing to their potential advantages such as waste reduction,recycling,and economic attributes,fast-growing bioenergy crops have the capacity to e?ectively phytoremediate heavy metal-contaminated soils.However,little is known about the role of microbial and chemical amendments in phytoremediation using bioenergy crops.Here,we studied the contributions of inoculation with the arbuscular mycorrhizal fungus (AMF) Acaulospora mellea ZZ and three soil amendments,i.e.,hydroxyapatite (HAP),manure,and bi...     

Phytoremediation of Mixed Soil Contaminants

Tests were conducted to study the influence of non-ionic surfactants Triton X-100 and Tween 80 on the removal of mixed contaminants from a sandy soil using phytoremediation. Cd(II) and Pb(II) were used to form the inorganic contaminant, while used engine oil was selected to form the organic contaminant. The Indian mustard (Brassica juncea) plant was the plant chosen for phytoremediation of the sandy soil that contained the mixed contaminant. Thirty days after the plants were grown in the greenhouse, surfactants were applied to test pots in which the soil had been spiked with 50 mg kg⁻¹ of CdCl₂, 500 mg kg⁻¹ of PbCl₂ and 500 mg kg⁻¹ of used engine oil. Two control tests were conducted in this study. Planted and unplanted control tests were conducted using soil without surfactants. Following these tests, the tests were completed using the plants and surfactants at different concentrations. Test results showed that Triton X-100 and Tween 80 at concentrations higher than their critical micellar concentration enhanced Cd(II) and Pb(II) accumulation in the plant roots. Further, test data showed that translocation of contaminants to plant shoots occurred for Cd(II) but not for Pb(II). At the same concentrations, Tween 80 was more effective than Triton X-100 in facilitating rhizodegradation of used engine oil. This study demonstrates that simultaneous phytoremediation of Pb(II), Cd(II) and oil can be enhanced by using non-ionic surfactant Tween 80. Leaching test results indicated that the enhanced phytoremediation could remove the mixed contaminants safely from the point of view of limiting groundwater contamination.     

IAA and 4‐Cl‐IAA Increases the Growth and Nitrogen Fixation in Mung Bean

A commonly occurring auxin indole‐3‐aceticacid (IAA) and a rarely occurring chlorosubstituted auxin (4‐Cl‐IAA) were compared for their impact on growth and nitrogen metabolism in mung bean for the first time. The plants were generated from healthy and Rhizobium coated seeds in earthen pots. The seedlings at 7 and/or 14 days were percolated with 0, 10^?10, 10^?8, or 10^?6 M of IAA or 4‐Cl‐IAA. The plants were sampled at 30 days after sowing (DAS) to assess the growth and various biochemical characteristics. The auxins significantly enhanced the growth (length and dry mass of shoot and root), nodule fresh mass, nitrogenase activity in fresh nodules, leaf carbonic anhydrase activity, chlorophyll content, and rate of photosynthesis. The effect of the auxins lasted up to the harvest where the seed yield, 100 seed mass, and number of pods per plant were significantly affected by the auxins. At a moderate concentration (10^?8 M), 4‐Cl‐IAA generated the best response. However, a comparable response was generated by the higher concentration (10^?6 M) of 4‐Cl‐IAA. The application of the hormone twice (at 7 and 14 DAS) was much more effective than single application (at 7 or 14 DAS). It was concluded that IAA and 4‐Cl‐IAA improved the growth and nitrogen fixation in mung bean. The 4‐Cl‐IAA proved more effective than IAA.     

Cadmium accumulation and distribution in sunflower plant

Cadmium (Cd) accumulation and distribution was studied in sunflower (Helianthus annuus L., public line HA‐89) plant. From an uncontaminated sandy loam brown forest soil with 162 μg kg^‐1 HNO₃/H₂O₂ extractable Cd the HA‐89 sunflower public line accumulated 114 ug kg^‐1 Cd in its kernels under open field conditions. This value is rather low as compared to data found by others. Sandy loam brown forest soil was treated with 0, 1 or 10 mg kg^‐1 of Cd to study the interaction of this heavy metal with young sunflower plants in a greenhouse pot experiment. The fresh weight and dry matter accumulation of sunflower plant organs (roots, shoots, leaves or heads) was unaffected by cadmium treatment of soil. The nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), copper (Cu), iron (Fe), manganese (Mn), or zinc (Zn) uptake of sunflower plant organs was not influenced by lower or higher Cd‐doses, except sunflower heads where 10 mg kg^‐1 of Cd treatment of soil significantly reduced the uptake of Ca, Fe, and Mn. Although Cd reduced the Zn uptake of roots, its rate was statistically not significant. Cadmium was accumulated prevalently in roots (1.21 mg kg^‐1,4.97 mg kg^‐1, or 13.69 mg kg^‐1 depending on Cd‐dose), and its concentration increased also in shoots or leaves. In spite of the short interaction time, elevated concentrations of cadmium (0.78 mg kg^‐1, 1.34 mg kg^‐1, or 3.02 mg kg^‐1 depending on Cd‐dose) were detected in just emerged generative organs (heads) of young sunflower plants.     

Chemical composition of cover crops and soil organic matter pools in no-tillage systems in the Cerrado

In no-tillage systems (NTS), cover crops are recommended to increase the productivity of agricultural systems. Furthermore, a greater diversity of cover crops in NTS favours an increase in soil carbon (C) stocks. However, there are scarce published data on the relationship between the chemical composition of cover crops and the accumulation of labile and stable fractions of SOM. We evaluated the relationship between the chemical composition of cover crops and SOM fractions, C stocks and maize yield. Hemicellulose, cellulose and lignin contents were determined for Urochloa ruziziensis, Canavalia brasiliensis, Cajanus cajan and Sorghum bicolor, cultivated in the off-season of maize. Canavalia brasiliensis had high N (20.96 g kg⁻¹) and hemicellulose (185.67 g kg⁻¹) contents, lower lignin content (39.50 g kg⁻¹) and high dry matter yield (3,251 kg ha⁻¹). All these characteristics resulted in a better SOM quality. Urochloa ruziziensis, with higher hemicellulose and lower lignin contents, and low lignin/N ratio, was associated with accumulation of TOC (19.95 and 18.33 g kg⁻¹ in 0- to 10-cm and 10- to 20-cm layers, respectively) and mineral-associated organic C (on average, 16.68 g kg⁻¹) in the soil. Cover plants with N:lignin ratio lower than 2.0 are fundamental for soil C sequestration. In conclusion, it is recommended the adoption of Urochloa ruziziensis and Canavalia brasiliensis as cover plants improve maize production, soil organic matter quality and C sequestration in the Cerrado region.     

Crop residues and fertilizer nitrogen influence residue decomposition and nitrous oxide emission from a Vertisol

Crop residues with high C/N ratio immobilize N released during decomposition in soil, thus reducing N losses through leaching, denitrification, and nitrous oxide (N₂O) emission. A laboratory incubation experiment was conducted for 84 days under controlled conditions (24°C and moisture content 55% of water-holding capacity) to study the influence of sugarcane, maize, sorghum, cotton and lucerne residues, and mineral N addition, on N mineralization–immobilization and N₂O emission. Residues were added at the rate of 3 t C ha⁻¹ to soil with, and without, 150 kg urea N ha⁻¹. The addition of sugarcane, maize, and sorghum residues without N fertilizer resulted in a significant immobilization of soil N. Amended soil had significantly (P < 0.05) lower NO₃⁻–N, which reached minimum values of 2.8 mg N kg⁻¹ for sugarcane (at day 28), 10.3 mg N kg⁻¹ for maize (day 7), and 5.9 mg N kg⁻¹ for sorghum (day 7), compared to 22.7 mg N kg⁻¹ for the unamended soil (day 7). During 84 days of incubation, the total mineral N in the residues + N treatments were decreased by 45 mg N kg⁻¹ in sugarcane, 34 mg kg⁻¹ in maize, 29 mg kg⁻¹ in sorghum, and 16 mg kg⁻¹ in cotton amended soil compared to soil + N fertilizer, although soil NO₃⁻–N increased by 7 mg kg⁻¹ in lucerne amended soil. The addition of residues also significantly increased amended soil microbial biomass C and N. Maximum emissions of N₂O from crop residue amended soils occurred in the first 4–5 days of incubation. Overall, after 84 days of incubation, the cumulative N₂O emission was 25% lower with cotton + N fertilizer, compared to soil + N fertilizer. The cumulative N₂O emission was significantly and positively correlated with NO₃⁻–N (r = 0.92, P < 0.01) and total mineral N (r = 0.93, P < 0.01) after 84 days of incubation, and had a weak but significant positive correlation with cumulative CO₂ in the first 3 and 5 days of incubation (r = 0.59, P < 0.05).     

Soil microbial communities response to herbicide 2,4-dichlorophenoxyacetic acid butyl ester

2,4-Dichlorophenoxyacetic acid butyl ester (2,4-D butyl ester) is extensively applied for weed control in cultivation fields in China, but its effect on soil microbial community remains obscure. This study investigated the microbial response to 2,4-D butyl ester application at different concentrations (CK, 10, 100 and 1000 μg g^?1) in the soils with two fertility levels, using soil dilution plate method and phospholipid fatty acid (PLFA) analysis. Culturable microorganisms were affected by the herbicide in both soils, particularly at the higher concentration. After treating soil with 100 μg g^?1 herbicide, culturable bacteria and actinomycetes were significantly higher, compared to other treatments. Treatment of soil with 1000 μg g^?1 2,4-D butyl ester caused a decline in culturable microbial counts, with the exception of fungal numbers, which increased over the incubation time. PLFA profiles showed that fatty acids for Gram-negative (GN) bacteria, Gram-positive (GP) bacteria, total bacteria and total fungi, as well as total PLFAs, varied with herbicide concentration for both soil samples. As herbicide concentration increased, the GN/GP ratio decreased dramatically in the two soils. The higher stress level was in the treatments with high concentrations of herbicide (1000 μg g^?1) for both soils. Principal component analysis of PLFAs showed that the addition of 2,4-D butyl ester significantly shifted the microbial community structure in the two soils. These results showed that the herbicide 2,4-D butyl ester might have substantial effects on microbial population and microbial community structure in agricultural soils. In particular, the effects of 2,4-D butyl ester were greater in soil with low organic matter and fertility level than in soil with high organic matter and fertility level.     

Mineral uptake and growth of sorghum colonized with VA mycorrhiza at varied soil phosphorus levels

Mineral nutrient uptake can be enhanced in plants inoculated with vesicular‐arbuscular mycorrhizal fungi (VAMF). The effects of the VAMF Glomus fasciculatum on uptake of P and other mineral nutrients in sorghum [Sorghum bicolor (L.) Moench] were determined in greenhouse experiments for plants grown on a low P (3.6 mg kg^‐1) soil (Typic Argiudolls) with P added at 0, 12.5, 25.0, and 37.5 mg kg^‐1 soil. Enhancements of growth and mineral nutrient uptake because of the VAMF association decreased as soil applications of P increased above 12.5 nig kg^‐1 soil. Root colonization with VAMF without added soil P resulted in increased dry matter yield equivalent to 12.5 mg P kg^‐1 soil (25 kg P ha^‐1). Total root length colonized with VAMF decreased as soil P level increased. Regardless of P added to the soil, mycorrhizal plants had higher leaf P concentrations and contents than did nonmycorrhizal plants. Enhanced contents, but not necessarily concentrations, of the other mineral nutrients were noted in shoots of mycorrhizal compared to nonmycorrhizal plants. Mycorrhizal plants had enhanced shoot contents of P, K, Zn, and Cu which could not be accounted for by increased growth. The VAMF associations with sorghum roots enhanced mineral nutrient uptake when P was sufficiently low in the soil.     

Effects of humic acids from vermicomposts on plant growth

The interactions between earthworms and microorganisms can produce significant quantities of plant growth hormones and humic acids which act as plant regulators. Experiments were designed to evaluate the effects of humic acids extracted from vermicompost and compare them with the action of commercial humic acid in combination with a commercial plant growth hormone, indole acetic acid (IAA) which is a commonly found in vermicomposts. In the first experiments, humic acids were extracted from cattle, food and paper waste vermicomposts. They were applied to a plant growth medium, Metro-Mix360 (MM360), at rates of 0, 250 or 500 mg humates kg⁻¹ dry wt. of MM360, to marigold, pepper, and strawberry plants in the greenhouse. Substitution of humates ranging from 250 to 1000 mg kg⁻¹ MM360 increased the growth of marigold and pepper roots, and increased the growth of roots and numbers of fruits of strawberries significantly. In other experiments, humic acids extracted from food waste vermicomposts were applied at a rate of 500 mg kg⁻¹ dry wt. of MM360, singly or in combination with IAA at a rate of 10⁻⁵ μM, to pepper seedlings. This experiment was designed to compare the differences in effects between the most effective dosage rate of humic acid from food waste, a phytohormone (IAA), and a commercial source of humic acid. The numbers of pepper flowers and fruits increased significantly in response to treatment with humic acid, IAA and a combination of humic acid and IAA. Peppers treated with humic acids extracted from food waste vermicomposts produced significantly more fruits and flowers than those treated with commercially-produced humic acids.     

Bioavailability,fractionation of pb and Zn in the rhizosphere of sunflower in chelators-amended contaminated soil

Assisted phytoremediation procedures have been widely employed as soil removal instrument of heavy metals from contaminated soils. Rhizosphere processes have a major impact on pb and Zn availability and its fractions in soils. The present study evaluates the effects of EDTA, citric acid (CA) and poultry manure extract (PME) on bioavailability and fractionation of pb, Zn in both the rhizosphere of sunflower (Helianthus annuus L.) and bulk soil. EDTA and CA were added to soils at the rates of 0, 0.5 and 1 mmol kg^?1 soil and PME at 0, 0.5 and 1 g kg^?1 soil as factorial in a completely randomized pattern with three replicates in greenhouse condition. Results showed that chelator application had a significant impact (p < 0.05) on pb, Zn extraction by different extractants and its fractions in soils. The order of concentrations of pb, Zn present in different fractions in soil treated by chelators was: oxides-bounded fraction > residual fraction > OM-bounded fraction > carbonate-bounded fraction > exchangeable fraction. Biochemical soil characteristics in the sunflower rhizosphere change resulting from its roots contributing to pb, Zn decline in mobile soil fractions, and change in soil pb, Zn fractions that are generally regarded as more stable.     

Growth and Nitrogen Fixation in Dhaincha/Sorghum and Dhaincha/Sunflower Intercropping Systems Using 15Nitrogen and 13Carbon Natural Abundance Techniques

A field experiment on dhaincha, sunflower, and sorghum plants grown in monocropping and intercropping systems was conducted to evaluate growth and nitrogen (N₂) fixation using ¹³carbon (C) and ¹⁵N natural abundance techniques. Intercropping of sesbania/sorghum showed a greater efficiency than monocropping in producing dry matter during the entire growth period, whereas the efficiency of producing dry matter in the sesbania/sunflower intercropping was similar to that in the monocropping system. Moreover, sorghum plants (C4) were more competitive than sesbania (C3) for soil N uptake, whereas sesbania seemed to be more competitive than its associated sunflower (C3). Nitrogen uptake in the mixed stand of sesbania/sorghum was improved as a result of the increase in soil N uptake by the component sorghum and the greater root nodule activity of component sesbania without affecting the amount of N₂ fixed. The Δ ¹³C in plant materials was affected by plant species and the cropping system.     

Intercropping of Sonchus asper and Vicia faba affects plant cadmium accumulation and root responses

The cadmium (Cd) pollution of farmland soil is serious in the world. The present study investigated the effects of intercropping Vicia faba and the hyperaccumulator Sonchus asper on the Cd accumulation and root responses (morphology and secreted organic acids) of plants grown on soil from a mining area in Yunnan Province, China, under different Cd stress levels (0, 50, 100, and 200 mg kg^-1). Intercropping increased the biomass of both S. asper and V. faba, as well as the Cd accumulation and Cd transfer coefficient from roots to shoots of S. asper, but decreased those of V. faba in the 200 mg kg^-1 Cd treatment. The Cd concentrations in roots, shoots, and grains from intercropped V. faba plants were positively correlated (P<0.05) with the N,N-diethyl phenylacetamide-extractable Cd content in soil. In the 50 mg kg^-1 Cd treatment, intercropping decreased citric and malic acids in root secretions of S. asper. Intercropped V. faba secreted more citric, oxalic, acetic, and malic acid compared with S. asper. Intercropping also increased root length and root surface area of both S. asper and V. faba. At 50 mg kg^-1 Cd, root length for S. asper was higher than that at 0 mg kg^-1 Cd, whereas V. faba roots had significantly decreased length and mean diameter. Intercropping of S. asper and V. faba is suggested as an in situ phytoremediation strategy of Cd-contaminated soils and may improve the food quality of V. faba.     

N2-fixation by ammonium-excreting Azospirillum brasilense in auxine-induced root tumours of wheat (Triticum aestivum L.)

Summary Wheat seedlings, treated with the auxine 2,4-dichlor-phenoxy acetic acid (2,4-D) during germination developed only a residual root system. Root elongation was extremely restricted and root tips were deformed to thick club-shaped tumours. When 2,4-D was added in a later stage of plant growth the plants developed additional nodule-like knots along primary roots. Root and shoot dry-matter production was slightly repressed in all 2,4-D treatments and N translocation from roots to shoots was repressed as well. When transferred to an auxine-free growth medium, the 2,4-D-affected roots were not capable of complete recovery. In plants inoculated gnotobiotically with Azospirillum brasilense, either with the wild type or with the NH ₄ ⁺ -excreting mutant strain C3, a 2,4-D addition increased rhizosphere acetylene-reduction activity at pO₂ 1.5 kPa. The O₂ sensitivity of root-associated nitrogenase activity tended to be reduced. The number of root-colonizing bacteria, at approximately 10⁸ colony-forming units (cfu) per g dry root, was similar in the 2,4-D treatments and untreated controls. Plant treatment with high concentrations of the chemical isomer 3,5-dichlor-phenoxy acetic acid (3,5-D) did not have comparable effects, either on plant development or on rhizosphere-associated nitrogenase activity. Root-tumour tissue inhabited by A. brasilense showed purple staining when subjected to a tetrazolium chloride solution, which may indicate intensive local nitrogenase activity in this tissue. Exposed to an ¹⁵N₂-enriched atmosphere, plants treated with 2,4-D and with A. brasilense incorporated significantly higher amounts of ¹⁵N than untreated controls. In all cases the highest values of ¹⁵N enrichment were found following inoculation with the NH ₄ ⁺ -excreting mutant strain C3. Present address: F. A. Janssens Laboratory of Genetics, Catholic University of Leuven, Willem de Croylan 42, B-3001 Heverlee, Belgium     

Enhancement of arbuscular mycorrhizal fungus (Glomus versiforme) on the growth and Cd uptake by Cd-hyperaccumulator Solanum nigrum

Arbuscular mycorrhizal fungus (AMF) can enhance plant growth and resistance to toxicity produced by heavy metals (HMs), affect the bioavailability of HMs in soil and the uptake of HMs by plants, and thus has been emerged as the most prominent symbiotic fungus for contribution to phytoremediation. A greenhouse pot experiment was conducted to assess the effect of Glomus versiforme BGC GD01C (Gv) on the growth and Cd accumulation of Cd-hyperaccumulator Solanum nigrum in different Cd-added soils (0, 25, 50, 100 mg Cd kg⁻¹ soil). Mycorrhizal colonization rates were generally high (from 71% to 82%) in Gv-inoculated treatments at all Cd levels. Gv colonization enhanced soil acid phosphatase activity, and hence elevated P acquisition and growth of S. nigrum at all Cd levels. Moreover, the presence of Gv significantly increased DTPA-extractable (phytoavailable) Cd concentrations in 25 and 50 mg Cd kg⁻¹ soils, but did not affect phytoavailable Cd in 100 mg Cd kg⁻¹ soil. Similarly, inoculation with Gv significantly increased Cd concentrations of S. nigrum in 25 and 50 mg Cd kg⁻¹ soils, but decreased Cd concentrations of the plants in 100 mg Cd kg⁻¹ soil. Overall, inoculation with Gv greatly improved the total Cd uptakes in all plant tissues at all Cd levels. The present results indicated that S. nigrum associated with Gv effectively improved the Cd uptake by plant and would be a new strategy in microbe-assisted phytoremediation for Cd-contaminated soils.     

Internal Boron Requirement of Young Sunflower Plants: Proposed Diagnostic Criteria

Abstract

In a greenhouse study, a significant increase in sunflower (Helianthus annuus L., cv. Hysun 33) dry matter yield was observed with boron (B) application to a B-deficient (hot water-extractable, 0.23 mg B kg^?1) calcareous soil of Missa series (Typic Ustochrept). Six rates of B, ranging from 0 to 8 mg B kg^?1 soil, were applied as H₃BO₃ along with adequate basal fertilization of nitrogen (N), phosphorus (P), potassium (K), and zinc (Zn). Four plants of sunflower were grown in each pot; two were harvested after 4 weeks of germination and the other two after 8 weeks. Maximum crop biomass was produced with 1.0 mg B kg ^?1, and application of ≥2.0 mg B kg^?1 proved toxic, resulting in drastic yield suppressions. Critical B concentration range for deficiency diagnosis in 4‐week‐old sunflower whole shoots appears to be 46–63 mg B kg^?1. However, critical concentration in 8‐week‐old plants was much less (i.e., 36 mg B kg^?l), presumably due to a dilution effect. As plant's internal B requirement can vary, in fact manifold, depending on the species, plant part, and plant age, only a relevant criterion can help in diagnosing the deficiency effectively.     

Inoculation with arbuscular mycorrhizal fungus Acaulospora mellea decreases Cu phytoextraction by maize from Cu-contaminated soil

A pot culture experiment was carried out to study the growth of and Cu uptake by maize (Zea mays) inoculated with or without arbuscular mycorrhizal (AM) fungus Acaulospora mellea in sterilized soil with different Cu amounts added (0, 100, 200, 400, 800 mg kg⁻¹). Root colonization rates were significantly lower with the addition of 400 and 800 mg kg⁻¹ Cu. AM inoculation increased shoot dry weights at 200 and 400 mg kg⁻¹ Cu added but showed no effects at other levels, while increased root dry weights at all Cu addition levels except 800 mg kg⁻¹. Compared with the nonmycorrhizal plants, shoot Cu concentrations in mycorrhizal plants were higher when no Cu was added but lower at other levels, while root Cu concentrations were lower at 400 and 800 mg kg⁻¹ Cu added but not affected at other levels. Thus, shoot Cu uptake in mycorrhizal plants increased with no Cu added but decreased at other levels, while mycorrhizal effects on root Cu uptake varied. Compared with nonmycorrhizal controls, Cu uptake efficiency and phytoextraction efficiency in mycorrhizal plants were higher when no Cu was added but lower at other levels, and Cu translocation efficiency was lower at all Cu addition levels. AM inoculation improved shoot and root P nutrition at all Cu addition levels. Soil pH was higher in mycorrhizal treatment than in the control when 200 mg kg⁻¹ or more Cu was added. These results indicate that A. mellea ZZ may be not suitable for Cu phytoextraction by maize, but shows a potential role in phytostabilization of soil moderately polluted by Cu.     

Promotion of plant growth by an auxin-producing isolate of the yeast Williopsis saturnus endophytic in maize (Zea mays L.) roots

A plant-growth-promoting isolate of the yeast Williopsis saturnus endophytic in maize roots was found to be capable of producing indole-3-acetic acid (IAA) and indole-3-pyruvic acid (IPYA) in vitro in a chemically defined medium. It was selected from among 24 endophytic yeasts isolated from surface-disinfested maize roots and evaluated for their potential to produce IAA and to promote maize growth under gnotobiotic and glasshouse conditions. The addition of l-tryptophan (L-TRP), as a precursor for auxins, to the medium inoculated with W. saturnus enhanced the production of IAA and IPYA severalfold compared to an L-TRP-non-amended medium. The introduction of W. saturnus to maize seedlings by the pruned-root dip method significantly (P<0.05) enhanced the growth of maize plants grown under gnotobiotic and glasshouse conditions in a soil amended with or without L-TRP. This was evident from the increases in the dry weights and lengths of roots and shoots and also in the significant (P<0.05) increases in the levels of in planta IAA and IPYA compared with control plants grown in L-TRP-amended or non-amended soil. The plant growth promotion by W. saturnus was most pronounced in the presence of L-TRP as soil amendment compared to seedlings inoculated with W. saturnus and grown in soil not amended with L-TRP. In the glasshouse test, W. saturnus was recovered from inside the root at all samplings, up to 8 weeks after inoculation, indicating that the roots of healthy maize may be a habitat for the endophytic yeast. An endophytic isolate of Rhodotorula glutinis that was incapable of producing detectable levels of IAA or IPYA in vitro failed to increase the endogenous levels of IAA and IPYA and failed to promote plant growth compared to W. saturnus, although colonization of maize root tissues by R. glutinis was similar to that of W. saturnus. Both endophytic yeasts, W. saturnus and R. glutinis, were incapable of producing in vitro detectable levels of gibberellic acid, isopentenyl adenine, isopentenyl adenoside or zeatin in their culture filtrates. This study is the first published report to demonstrate the potential of an endophytic yeast to promote plant growth. This is also the first report of the production of auxins by yeasts endophytic in plant roots.     

The millimetre-scale distribution of 2,4-D and its degraders drives the fate of 2,4-D at the soil core scale

The biodegradation of organic compounds in soil is a key process that has major implications for different ecosystem services such as soil fertility, air and water quality, and climate regulation. Due to the complexity of soil, the distributions of organic compounds and microorganisms are heterogeneous on sub-cm scales, and biodegradation is therefore partly controlled by the respective localizations of organic substrates and degraders. If they are not co-localized, transfer processes become crucial for the accessibility and availability of the substrate to degraders. This spatial interaction is still poorly understood, leading to poor predictions of organic compound dynamics in soils. The objectives of this work were to better understand how the mm-scale distribution of a model pesticide, 2,4-dichlorophenoxyacetic acid (2,4-D), and its degraders drives the fate of 2,4-D at the cm soil core scale. We constructed cm-scale soil cores combining sterilized and “natural” soil aggregates in which we controlled the initial distributions of 2,4-D and soil microorganisms with the following spatial distributions: i) a homogeneous distribution of microorganisms and 2,4-D at the core-scale, ii) a co-localized distribution of microorganisms and 2,4-D in a single spot (360 mm³) and iii) a disjoint localization of microorganisms and 2,4-D in 2 soil spots (360 mm³) separated by 2 cm. Two sets of experiments were performed: one used radiolabeled ¹⁴C-2,4-D to study the fate of 2,4-D, and the other used ¹²C-2,4-D to follow the dynamics of degraders. Microcosms were incubated at 20 °C and at field capacity (−31.6 kPa). At the core scale, we followed 2,4-D mineralization over time. On three dates, soil cores with microorganisms and 2,4-D localized in soil spots, were cut out in slices and then in 360 mm³ soil cubes. The individual soil cubes were then independently analysed for extractable and non-extractable ¹⁴C and for degraders (quantitative PCR of tfdA genes). Knowing the initial position of each soil cube allowed us to establish 3D maps of 2,4-D residues and degraders in soil. The results indicated that microorganisms and pesticide localizations in soil are major driving factors of i) pesticide biodegradation, by regulating the accessibility of 2,4-D to degrading microorganisms (by diffusion); and ii) the formation of non-extractable residues (NER). These results also emphasized the dominant role of microorganisms in the formation and localization of biogenic NER at a mm-scale. To conclude, these results demonstrate the importance of considering micro-scale processes to better understand the fate of pesticides and more generally of soil organic substrates at upper scales in soil and suggest that such spatial heterogeneity should not be neglected when predicting the fate of organic compounds in soils.