Improvement of Brassica napus growth under cadmium stress by cadmium-resistant rhizobacteria

This study focuses on the characterization of four bacterial isolates from heavy metal-polluted rhizosphere in order to examine their plant growth promoting (PGP) activity. The PGP activity on the canola (Brassica napus) of the strains which showed cadmium resistance and multiple PGP traits was assessed in the presence and in the absence of Cd²⁺. The strains, Pseudomonas tolaasii ACC23, Pseudomonas fluorescens ACC9, Alcaligenes sp. ZN4 and Mycobacterium sp. ACC14 showed 1-aminocyclopropane-1-carboxylate deaminase (ACCD) activity. They also synthesized ACCD enzyme in vitro when 0.4 mM Cd²⁺ was added to the growth medium. The presence of the metal, however, reduced the ACCD activity in Alcaligenes sp. ZN4 and Mycobacterium sp. ACC14, while it did not affect the ACCD activity of P. tolaasii ACC23 and P. fluorescens ACC9. ACC9 and ACC23 produced indole acetic acid (IAA) and siderophores, while ACC14 produced only IAA. IAA and siderophores were produced more actively under Cd-stress.Root elongation assays conducted on B. napus under gnotobiotic conditions demonstrated increases (from 34% up to 97%) in root elongation of inoculated canola seedlings compared to the control plants. Subsequently, the effect of inoculation with these strains on growth and uptake of Cd²⁺ in roots and shoots of canola was studied in pot experiments using Cd-free and Cd-treated (15 μg Cd²⁺ g^?1 dw) soil. Inoculation with P. tolaasii ACC23, P. fluorescens ACC9 and Mycobacterium sp. ACC14 promoted the growth of plants at concentrations of 0 and 15 μg Cd²⁺ g^?1 soil. The maximum growth was observed in the plants inoculated with P. tolaasii ACC23. The strains did not influence the specific accumulation of cadmium in the root and shoot systems, but all increased the plant biomass and consequently the total cadmium accumulation.The present observations showed that the bacterial strains used in this study protect the plants against the inhibitory effects of cadmium, probably due to the production of IAA, siderophores and ACCD activity.     

Mercury affects the distribution of culturable species of Pseudomonas in soil

Pseudomonas bacteria isolated during 52 days on Gould's S1 agar from soil spiked with 0, 3.5 and 15 mg Hg(II) kg soil⁻¹ were characterised to reveal whether mercury affected them differently. Isolates from the treatments with 0 and 15 mg Hg kg⁻¹ were characterised using FT-IR characterisation and subsequent 16S rDNA partial sequencing of representative isolates. To verify the selectivity of Gould's S1 agar and the FT-IR characterisation, all 450 isolates were subjected to the following tests: Gram-determination, catalase and oxidase activity, pigment production on PDA and growth at different temperatures. Furthermore, the isolates were tested for their ability to grow on agar amended with 10 mg Hg kg⁻¹ as an indication of mercury resistance. We found that up to 80% of the isolates in soil amended with 15 mg Hg kg⁻¹ were mercury-resistant, whereas only up to 20% were resistant in the treatments with 0 and 3.5 mg Hg kg⁻¹. We found two groups of Pseudomonas, which probably represent non-described species since they did not group closely with any known species of Pseudomonas in the dendrogram. Hg-enhanced isolates were closely related to P. frederiksbergensis. Furthermore, Hg resistance was almost exclusively restricted to P. frederiksbergensis and P. migulae groups. We conclude that Hg caused a shift in the dominating species of culturable Pseudomonas.     

Soil available phosphorus status determines indigenous mycorrhizal colonization of field and glasshouse-grown spring wheat from Argentina

Wheat production (Triticum aestivum L.) has increased across the world during last century with the intensification of agriculture. Phosphorus (P) fertilization is a common practice to improve wheat growth in Argentina. We investigate whether indigenous arbuscular mycorrhizal colonization (AMC) of hard red spring wheat is controlled by shoot P content (SPc) or by available soil P in an agricultural soil from the southeastern Argentine Pampas. In the field, AMC was monitored four times during two growing seasons of a conventional wheat crop. Treatments were: without P supply, annual supply of 11 and 22 kg P ha⁻¹ during the last 5 years, and 164 kg P ha⁻¹ applied once 5 years before the experiment. In the glasshouse, AMC was assessed three times in wheat growing in pots filled with the soil from unfertilized plots; treatments were: P (0 and 20 mg P pot⁻¹), and nitrogen (N) fertilization (0 and 150 mg N pot⁻¹). A range of soil P between 6 and 60 mg P kg⁻¹ was obtained and the AMC ranged from 1% to 67% of root length colonized under both field and glasshouse conditions. P supplied annually increased growth and SPc but decreased AMC. N fertilization did not affect growth or AMC. Variations in SPc did not account for AMC. Variability in AMC was best accounted for local current soil available P content (r² = 0.59). A linear-plateau relationship between soil P and indigenous AMC was established in wheat plants growing under contrasting environmental and experimental (field and glasshouse) conditions. Indigenous AMC was depressed by available soil P in the range 0–27 mg P kg⁻¹ (a decrease of 2.8% mg P⁻¹ kg⁻¹). Above 27 mg P kg soil⁻¹, AMC was stabilized at about 10%. Grain yield increased with fertilization and the highest relative shoot dry matter in field was obtained at 15.5 mg P kg soil⁻¹. The soil P range that ensures high wheat production without deterring indigenous AMC is discussed.     

Arbuscular mycorrhizal fungi induce differential Cd and P acquisition by Alfred stonecrop (Sedum alfredii Hance) and upland kangkong (Ipomoea aquatica Forsk.) in an intercropping system

A greenhouse pot experiment was conducted to study cadmium (Cd) and phosphorous (P) acquisition of upland kangkong (Ipomoea aquatica Forsk.) intercropped with Alfred stonecrop (Sedum alfredii Hance) in a Cd-contaminated soil inoculated with arbuscular mycorrhizal (AM) fungi. There were four treatments, including monoculture of kangkong (control), intercropping with stonecrop (IS), and intercropping with stonecrop plus inoculation with Glomus caledonium (IS + Gc) or Glomus versiforme (IS + Gv). Both kangkong and stonecrop plants were harvested at week 8 after seeding or cutting. Compared with the control, IS tended to decrease Cd and P acquisition by neighboring kangkong via competition for phytoavailable Cd and P. The inoculation of Gc, but not Gv, significantly elevated Cd acquisition by stonecrop, and hence resulted in significantly lower Cd acquisition by kangkong and the subsequent Cd concentrations in both roots and shoots of kangkong. Both Gc and Gv significantly increased mycorrhizal colonization rates in stonecrop roots, as well as acid phosphatase activities and available P concentrations in the soil. However, only Gc significantly elevated P acquisition and shoot biomass of the host plant (stonecrop), while Gv significantly increased P acquisition and shoot biomass of neighboring kangkong rather than of stonecrop, causing a significant dilution effect on kangkong shoot Cd concentration. In addition, both Gc and Gv inoculation significantly decreased soil DTPA-extractable (phytoavailable) Cd concentrations by elevating soil pH. The results showed that Gc and Gv played totally different roles in the intercropping system for vegetable production and phytoremediation of Cd-contaminated soils.     

Rapid estimation of microbial biomass in grassland soils by ultra-violet absorbance

A reliable and simple technique for estimating soil microbial biomass (SMB) is essential if the role of microbes in many soil processes is to be quantified. Conventional techniques are notoriously time-consuming and unreproducible. A technique was investigated that uses the UV absorbance at 280 nm of 0.5 M K₂SO₄ extracts of fumigated and unfumigated soils to estimate the concentrations of carbon, nitrogen and phosphorus in the SMB. The procedure is based on the fact that compounds released after chloroform fumigation from lysed microbial cells absorb in the near UV region. Using 29 UK permanent grassland soils, with a wide range of organic matter (2.9–8.0%) and clay contents (22–68%), it was demonstrated that the increase in UV absorbance at 280 nm after soil fumigation was strongly correlated with the SMB C (r=0.92), SMB N (r=0.90) and SMB P (r=0.89), as determined by conventional methods. The soils contained a wide range of SMB C (412–3412 μg g⁻¹ dry soil), N (57–346 μg g⁻¹ dry soil) and P (31–239 μg g⁻¹ dry soil) concentrations. It was thus confirmed that the UV absorbance technique described was a rapid, simple, precise and relatively inexpensive method of estimating soil microbial biomass.     

Impact of carbon-rich dairy factory effluent on growth of perennial ryegrass (Lolium perenne) and soil microorganisms

A pot experiment was conducted to investigate the impact of high carbon dairy factory effluent application on the growth of perennial ryegrass (Lolium perenne L.), plant nutrient uptake, soil microbial biomass carbon and nitrogen, populations of soil-microorganisms, root colonising fungi and the microbial functional diversity. The effluent was added at rates of 0, 100,000, 200,000 and 300,000 l ha^–1. These rates are equivalent to 0, × 1, × 2 and × 3 normal field application rates. The added effluent contained (g l^–1), C; 19.42, total P; 0.65; S, 0.75, K; 1.33, Na; 4.55, Mg; 0.11, NH₄; 0.073, total N; 0.073 and had a pH of 4.33. Replicate pots (incubated in a controlled-environment room at 20 °C, with 16 h light/8 h dark) were harvested at 32, 61, and 130 days after setting up of the experiment. In the first sampling, shoot dry matter levels declined significantly (P < 0.01) with increased effluent. By the third sampling the trend was reversed with treated pots having greater amounts of shoot dry matter. The initial depression of growth was possibly due to a combination of factors including excess levels of available carbon (C) for microbes leading to immobilisation of nutrients, particularly nitrogen (N) and sulphur (S). Shoot N and S concentrations were lower (P < 0.001) and the phosphorus concentrations were higher in effluent-treated samples. Soil microbial biomass-C (480 and 770 μg g⁻¹ of biomass C in untreated and treated soil, respectively) and microbial-N (81 and 123 μg g^–1 of microbial-N in untreated and treated soil, respectively) were significantly (P < 0.001) greater in effluent-treated pots at all times. Populations of total culturable bacteria were higher (P < 0.01) in the treated pots in the first sample (log₁₀ populations g^–1 were 7.3 in untreated pots compared to 8.0 averaged across three treatments) but there were no differences in the subsequent two samples. Effluent also increased yeast populations (log₁₀ numbers g^–1 were 0.6 in untreated pots and 3.1 in treated pots averaged across treatments and times P < 0.01) at all three sampling times. The Shannon-Weiner Diversity Index of root fungi decreased with increasing effluent application (P < 0.01) while the species richness decreased with effluent as well as with time (P < 0.1). Potential root pathogens Fusarium oxysporum, total Fusarium spp. and Pythium spp. significantly increased (P < 0.05) in treated samples but in the final sampling, Codinaea fertilis significantly (P < 0.05) decreased with effluent treatment. The microbial functional diversity pattern and the average well colour development (AWCD) in soil were significantly changed by the effluent application but effects were not detectable after 130 days.     

The significant contribution of mycorrhizal fungi and earthworms to maize protection and phytoremediation in Cd-polluted soils

Mycorrhizal fungi and earthworms can individually or interactively influence plant growth and heavy metal uptake. The influence of earthworms and arbuscular mycorrhizal (AM) fungi either alone or in combination on maize (Zea mays L.) growth and cadmium (Cd) uptake was investigated in a calcareous soil artificially spiked with Cd. Soils were contaminated with Cd (10 and 20 mg Cd kg⁻¹), inoculated or un-inoculated with the epigeic earthworm Lumbricus rubellus and two AM fungal species (Rhizophagus irregularis and Funneliformis mosseae) for two months of growth under greenhouse conditions. Generally, earthworms alone increased both shoot P uptake and biomass but decreased shoot Cd concentration and root Cd uptake. AM fungi individually often increased total maize P uptake, declined shoot Cd concentration, and consequently produced higher total biomass. However, R. irregularis enhanced shoot Cd uptake at low Cd level and root Cd uptake at high Cd level. In plants inoculated with F. mosseae species, earthworms increased shoot biomass and Cd uptake, decreased root biomass and Cd uptake at all Cd levels, and increased shoot Cd concentration at low Cd level. In plants colonized by R. irregularis species, however, earthworm addition decreased maize biomass only at high Cd level and root Cd concentration and total maize Cd uptake at both Cd levels. Earthworm activity decreased Cd transfer from the soil to maize roots at low Cd level, but this was counterbalanced in the presence of F. mosseae. Mycorrhizal symbiosis significantly reduced the transfer of Cd from roots to shoots, independence of earthworm effect. Overall, it is concluded that L. rubellus and AM fungi, in particular F. mosseae isolate, improved maize tolerance to Cd toxicity both individually and interactively by increasing plant growth and P nutrition, and restricting Cd transfer to the aboveground biomass. Consequently, the single and interactive effects of the two soil organisms might potentially be important not only in protecting maize plants against Cd toxicity, but also in Cd phytostabilization in soils polluted by this highly toxic metal.     

Co-inoculation of Halomonas maura and Ensifer meliloti to improve alfalfa yield in saline soils

Salinity is the major environmental factor limiting crop production. Alfalfa is a legume with high nutritional value that establishes a symbiosis relation with Ensifer meliloti. Under saline conditions the alfalfa yield decreases and this symbiosis is affected. The aim of this work is to study the effect of the co-inoculation of alfalfa plants with Halomonas maura (a moderately halophile bacterium) and E. meliloti in saline soils to improve their productivity and growth under greenhouse and field conditions. Alfalfa plants were grown in Leonard jar under greenhouse conditions, using a N-free mineral solution to mimic the conditions of an Orthic Solonchak. Then alfalfa plants were grown in the field in the same soil type. Seeds were inoculated with E. meliloti, H. maura, co-inoculated with E. meliloti and H. Maura, or non-inoculated as a control in both experiments. In greenhouse experiments the co-inoculation of alfalfa plants increased significantly the shoot dry weight (0.64 ± 0.02 vs. 0.79 ± 0.02), the leghaemoglobin content (10.17 ± 0.03 vs. 11.25 ± 0.06) and water potential (−3.12 ± 0.02 vs. −2.79 ± 0.02) compared with the single inoculation with E. meliloti. In the field experiments, biomass of co-inoculated plants clearly outyielded those of plants inoculated with any inoculant. The co-inoculation of H. maura and E. meliloti enhances alfalfa productivity in saline soils, thus contributing to the agricultural exploitation of low productive areas. H. maura and E. meliloti could be considered in formulation of bioinoculants to contribute in the reduction of the overuse of chemical fertilizers and their environmental impacts.     

Azospirillum brasilense Az39 and Bradyrhizobium japonicum E109, inoculated singly or in combination,promote seed germination and early seedling growth in corn (Zea mays L.) and soybean (Glycine max L.)

Inoculants are biological formulations that combine a stable microorganism population and various types of compounds produced and released during fermentation, such as phytohormones and plant growth regulators. Azospirillum brasilense strain Az39 and Brayrhizobium japonicum strain E109 were previously shown to produce indole 3-acetic acid (IAA), gibberellic acid (GA₃) and zeatin (Z). We tested the hypothesis that such compounds are responsible for early growth promotion in inoculated corn (Zea mays L.) and soybean (Glycine max L.) seedlings. Seeds were inoculated with Az39, E109, or both, and kept in a chamber at 20–30 °C under a controlled photoperiod to evaluate seed germination. To evaluate root and shoot length and dry weight, and number of nodules and percentage of nodulated seedlings, in soybean, seedlings were kept in a growth chamber for 14 days under similar photoperiod and temperature conditions. Az39 and E109, singly or in combination, showed the capacity to promote seed germination, nodule formation, and early development of corn and soybean seedlings. Both strains were able to excrete IAA, GA₃ and Z into the culture medium, at a concentration sufficient to produce morphological and physiological changes in young seed tissues.     

Isolation of culturable phosphobacteria with both phytate-mineralization and phosphate-solubilization activity from the rhizosphere of plants grown in a volcanic soil

Chilean volcanic soils contain large amounts of total and organic phosphorus, but P availability is low. Phosphobacteria [phytate-mineralizing bacteria (PMB) and phosphate-solubilizing bacteria (PSB)] were isolated from the rhizosphere of perennial ryegrass (Lolium perenne), white clover (Trifolium repens), wheat (Triticum aestivum), oat (Avena sativa), and yellow lupin (Lupinus luteus) growing in volcanic soil. Six phosphobacteria were selected, based on their capacity to utilize both Na-phytate and Ca-phosphate on agar media (denoted as PMPSB), and characterized. The capacity of selected PMPSB to release inorganic P (Pi) from Na-phytate in broth was also assayed. The results showed that from 300 colonies randomly chosen on Luria–Bertani agar, phosphobacteria represented from 44% to 54% in perennial ryegrass, white clover, oat, and wheat rhizospheres. In contrast, phosphobacteria represented only 17% of colonies chosen from yellow lupin rhizosphere. This study also revealed that pasture plants (perennial ryegrass and white clover) have predominantly PMB in their rhizosphere, whereas PSB dominated in the rhizosphere of crops (oat and wheat). Selected PMPSB were genetically characterized as Pseudomonas, Enterobacter, and Pantoea; all showed the production of phosphoric hydrolases (alkaline phosphatase, acid phosphatase, and naphthol phosphohydrolase). Assays with PMPSB resulted in a higher Pi liberation compared with uninoculated controls and revealed also that the addition of glucose influenced the Pi-liberation capacity of some of the PMPSB assayed.     

Influence of AM fungi on the growth and physiological status of Erythrina variegata Linn. grown under different water stress conditions

The present study investigated the effects of arbuscular mycorrhizal (AM) fungus, Glomus mosseae on the growth and physiology state of Erythrina variegata Linn, grown in sandy loam soil with four water stress levels viz. ?0.06 MPa (well watered/control), ?1.20 MPa (mild), ?2.20 MPa (moderate) and ?3.20 MPa (severe) in a completely randomized design. Plants were harvested after 90 days (60 days after stress induction) of growth. Growth parameters (root &, shoot, dry weight and, leaf area); physiological parameters (chlorophyll content, carotenoids, soluble starch, sugar, protein and proline in shoots); and microbiological parameter (percentage of mycorrhizal infection) were determined. AM fungal plants had significantly higher plant biomass, higher chlorophyll content (chlorophyll a and b), carotenoids and protein content in shoots than non-AM-plants. The AM-inoculation in stressed plants significantly declined the soluble sugar and starch in shoots. Moreover, AM-inoculation also reduced the proline accumulation in shoots and the reduction was significant when plants were severely stressed (?3.2 MPa). Mycorrhizal colonization in roots of E. variegata depressed significantly due to increased water stress. However, the AM colonization did not decline below 11% and enabled the plants to maintain osmotic adjustments and enhanced the plants tolerance against water stress.     

Effect of salinity on growth,nodulation and nitrogen assimilation in nodules of faba bean (Vicia faba L.)

A commercial cultivar (Alborea) of faba bean (Vicia faba L. var. minor) was inoculated with salt-tolerant Rhizobium leguminosarum biovar. viciae strain GRA19 in solution culture with different salt concentrations (0, 50, 75 and 100 mmoles l⁻¹ NaCl) added immediately at the time of inoculation. The results indicated that Rhizobium leguminosarum strain GRA19 formed an infective and effective symbiosis with faba bean under saline and nonsaline conditions. Salinity significantly decreased shoot and root dry weight, nodule weight and mean nodule weight. Roots were more sensitive than shoots, and N₂ fixation was more sensitive to salinity than was plant growth. Analyses of ammonium assimilating enzymes in the nodule showed that glutamine synthetase appeared to be more tolerant to salinity than glutamate synthase, and that it limits ammonium assimilation under saline stress.     

Implications of Azospirillum brasilense inoculation and nutrient addition on maize in soils of the Brazilian Cerrado under greenhouse and field conditions

The combination of nitrogen and Azospirillum can ensure greater nutrient absorption and crop yield in agricultural areas using high technology. Thus, the objective was to evaluate maize response to Azopirillum brasilense (AZ) inoculation and nutrient (macronutrients and micronutrients) application under greenhouse and field conditions in clay and sandy soils of the Brazilian Cerrado. In the greenhouse assays, the following parameters were measured: shoot dry weight (SDW), root dry weight (RDW), and root volume (RV). In the field experiments, the maize yield was determined after drying the grains at 60 °C for 48 h. In clay soil, there was a significant increase in the SDW, RDW and RV in the treatment with AZ concentrated (10¹¹ cells ml⁻¹ of inoculum) when compared with the control treatment and the treatment with AZ diluted (10⁶ cells ml⁻¹ of inoculum). In this soil, adding micronutrients did not affect the maize response under greenhouse conditions. In sandy soil, there was no difference between the AZ treatment and the control, except for treatments where nutrients and AZ were both added leading to a significant increase in the maize response. In both soils, the RV:RDW ratio was higher in the treatment with AZ concentrated compared to that in the treatment with AZ diluted, but the yield response depended on the addition of nutrients. Inoculation with A. brasilense gave comparable yield to the nitrogen treatment. The grain production was increased by 29% in the treatment with A. brasilense and nitrogen compared to nitrogen fertilization alone. In this study, the yield response was affected significantly when maize was inoculated with A. brasilense, but this response was dependent on the soil type under greenhouse conditions.     

Contribution of plant photosynthate to soil respiration and dissolved organic carbon in a naturally recolonising cutover peatland

The aim of this study was to investigate how three vascular plant species (Calluna vulgaris, Eriophorum angustifolium and Eriophorum vaginatum) colonising an abandoned cutover peatland affect fluxes of recent photosynthate to dissolved organic carbon (DOC), soil and plant respiration and shoot biomass. We used in situ ¹³CO₂ pulse labelling to trace carbon (C) throughout a 65 day pulse chase period. Between 16 and 35% of the pulse of ¹³C remained in shoot biomass after 65 days with significant differences between C. vulgaris and E. angustifolium (P = 0.009) and between C. vulgaris and E. vaginatum (P = 0.04). A maximum of 29% was detected in DOC beneath labelled plants and losses of ¹³C from peat respiration never exceeded 0.16% of the original pulse, showing that little newly fixed C was allocated to this pool. There were no significant differences between the different plant species with respect to ¹³C recovered from DOC or via peat respiration. More C was lost via shoot respiration; although amounts varied between the three plant species, with 4.94–27.33% of the ¹³C pulse respired by the end of the experiment. Significant differences in ¹³C recovered from shoot respiration were found between C. vulgaris and E. angustifolium (P = 0.001) and between E. angustifolium and E. vaginatum (P = 0.032). Analysis of δ¹³C of microbial biomass indicated that recently assimilated C was allocated to this pool within 1 day of pulse labelling but there were no significant differences in the ¹³C enrichment of the microbial biomass associated with the different plant species. The data suggest that peat respiration represents a small flux of recent assimilate compared to other fluxes and pools and that different vascular plant species show considerable variation in the quantities and dynamics of C allocated to DOC.     

Removal of benzo (a) pyrene from soil using an endogeic earthworm Pontoscolex corethrurus ()

The endogeic earthworm Pontoscolex corethrurus (Müller, 1857) was the most abundant species (75%) in soil contaminated with hydrocarbons, mostly benzo(a)pyrene (BaP), in the state of Tabasco (Mexico). The earthworm P. corethrurus was tested for its capacity to remove 100 mg BaP kg⁻¹ from an Anthrosol soil (sterilized or not) and amended with legume Mucuna pruriens (L.) DC. var. utilis (Wall. ex Wight) Baker ex Burck (3%) or the grass Brachiaria humidicola (L.) DC (3%) (recently renamed as Urochloa humidicola (Rendle) Morrone & Zuloaga) in an aerobic incubation experiment. P. corethrurus removed 26.6 mg BaP kg⁻¹ from the sterilized soil and application of B. humidicola as feed increased this to 35.7 mg BaP kg⁻¹ and M. pruriens to 34.2 mg BaP kg⁻¹ after 112 days. The autochthonous microorganisms removed 9.1 mg BaP kg⁻¹ from the unsterilized soil and application of B. humidicola increased this to 18.0 mg BaP kg⁻¹ and M. pruriens to 11.2 mg BaP kg⁻¹. Adding P. corethrurus to the unsterilized soil accelerated the removal of BaP and 36.1 mg kg⁻¹ was dissipated from soil. It was found that the autochthonous microorganisms removed BaP from soil, but addition of P. corethrurus increased the dissipation 4-fold. The endogeic earthworm P. corethrurus can thus be used to remediate hydrocarbon-contaminated soils in tropical regions.     

Physiological studies and comparative analysis of rock phosphate solubilization abilities of Actinomycetales originating from Moroccan phosphate mines and of Streptomyces lividans

Actinomycete strains originating from Moroccan phosphate mines (MPM) were selected for their ability to use the insoluble ground hydroxyapatite called rock phosphate (RP), present in their biotope, as sole phosphate (P) source. Physiological studies carried out with these strains and with the reference strains, Streptomyces lividans and Streptomyces griseus, demonstrated that all strains were able to grown in a synthetic minimal medium (SMM) containing either soluble (SP) or insoluble (RP) phosphate as sole P source. The MPM strains and S. griseus took up glucose much more actively and exhaustively than S. lividans, constituting more abundant glycogen reserves than the latter. All strains took up soluble P at comparable rates, storing it as polyphosphates. In SMM + RP, a sharp increase in the concentration of soluble P was detected in the culture broths of all MPM strains and S. griseus, at stationary phase, but not in that of S. lividans. The P peak detected in the supernatant of these strains correlated with the successive appearance of two compounds absorbing at 320 nm and 430 nm, respectively. These compounds are thought to be strong ion chelators involved in the destruction of the hydroxyapatite structure leading to soluble P release. The good growth of S. lividans in SMM + RP indicated that this strain was also able to release P from RP but consummed it as soon as it was released, unlike the other strains. Our study is expected to lead to the development of a novel type of slow release bio-phosphate fertilizer constituted by the association of the MPM strains and ground RP. This novel product would precisely supply plant needs and thus limit the pollution of the environment.     

Stimulatory effect of phosphate-solubilizing fungal strains (Aspergillus awamori and Penicillium citrinum) on the yield of chickpea (Cicer arietinum L. cv. GPF2)

The effect of six phosphate-solubilizing fungi (PSF, two strains of Aspergillus awamori, and four of Penicillium citrinum) isolated from rhizosphere of various crops, was observed on the growth and seed production of chickpea plants (Cicer arietinum L. cv. GPF2) in pot experiments. The phosphate (P) solubilizing activity of PSF in liquid varied from 38 to 760 μg ml^?1 for tricalcium phosphate (TCP) and 28–248 μg ml^?1 for mussoorie rock phosphate (MRP). All PSF isolates were biocompatible and produced growth-promoting hormone, Indole acetic acid (IAA), varying in concentration from 2.5 to 9.8 μg ml^?1. Of the various pot experiments carried out in green house, maximum stimulatory effect on chickpea plants growth was observed by inoculation of two A. awamori strains. This treatment resulted in 7–12% increase in shoot height, nearly three-fold increase in seed number and two-fold increase in seeds weight as compared to the control (un-inoculated) plants. Inoculation of four strains of P. citrinum exhibited lesser stimulatory effect. It showed 7% increase in shoot height, two-fold increase in seed number and 87% increase in seeds weight as compared to the control plants. However, a consortium of all the six fungal isolates showed no stimulatory effect on chickpea plants growth.     

Forms of phosphorus in bacteria and fungi isolated from two Australian soils

To understand the origin of organic and condensed forms of phosphorus (P) in soils, detailed information about P forms in microorganisms is required. We isolated 7 bacteria and 8 fungi from two Australian soils and analyzed the P forms in their pure cultures by extraction with NaOH-EDTA followed by ³¹P solution nuclear magnetic (NMR) spectroscopy. The bacteria belonged to the actinobacteria and the fungi to the ascomycota, as determined by rDNA sequencing. The proportions of broad forms of P were significantly different between the bacterial and fungal isolates (analysis of similarities, p = 0.001). Ortho-, pyro- and polyphosphate were present in higher proportions in fungi, while monoester and diester P were present in higher proportions in bacteria. Spectral deconvolution of the monoester region revealed 15 distinct resonances. The three major ones, which were identified by spiking experiments as glycerol 1-phosphate, glycerol 2-phosphate and adenosine-5′-monophosphate (AMP), comprised 56–74% of P in the monoester region. Ordination by principal component analysis and testing for treatment effects using analysis of similarities showed significant separation of P distribution in the monoester region between bacterial and fungal isolates (p = 0.007). However, neither group of microorganisms had a specific single P form which might be considered characteristic. As such, it may be difficult to distinguish soil P from bacterial or fungal origins, with the possible exception of a predominantly fungal origin of pyro- and polyphosphate. The identification of three major resonances in the monoester region of microorganisms is important, since the same resonances are found in ³¹P NMR spectra of soil extracts.     

Chemotaxis of deleterious rhizobacteria to birdsfoot trefoil

Intact seeds and seed and seedling root exudates of birdsfoot trefoil (Lotus corniculatus L.) were used as chemoattractants in experiments to determine the relative importance of chemotaxis in spermosphere and rhizosphere colonization by selected rhizobacteria. Results for soft-agar, capillary tube and soil chemotaxis assays indicated that selected deleterious rhizobacteria were attracted to seed and seedling root exudates. Several sugars and phenolic fractions detected in exudates were chemoattractants for these rhizobacteria. Using soil-chemotaxis assemblies, migration of rhizobacterial isolates through 2 cm distances of soil toward birdsfoot trefoil seeds was detected within 24 h. Isolates were not detected at the same site in soils without seeds until 72 h after inoculation. These results suggest that attraction of deleterious rhizobacteria toward seeds and seedling roots mediated by exudates (chemotaxis) might be the first step in the establishment and subsequent colonization of bacteria involved in soilborne disease complexes of birdsfoot trefoil.     

Climatic influences on active fractions of soil organic matter

Biologically active fractions of soil organic matter are important in understanding decomposition potential of organic materials, nutrient cycling dynamics, and biophysical manipulation of soil structure. We evaluated the quantitative relationships among potential C and net N mineralization, soil microbial biomass C (SMBC), and soil organic C (SOC) under four contrasting climatic conditions. Mean SOC values were 28±11 mg g⁻¹ (n=24) in a frigid–dry region (Alberta/British Columbia), 25±5 mg g⁻¹ (n=12) in a frigid–wet region (Maine), 11±4 mg g⁻¹ (n=117) in a thermic–dry region (Texas), and 12±5 mg g⁻¹ (n=131) in a thermic–wet region (Georgia). Higher mean annual temperature resulted in consistently greater basal soil respiration (1.7 vs 0.8 mg CO₂–C g⁻¹ SOC d⁻¹ in the thermic compared with the frigid regions, P<0.001), greater net N mineralization (2.8 vs 1.3 mg inorganic N g⁻¹ SOC 24 d⁻¹, P<0.001), and greater SMBC (53 vs 21 mg SMBC g⁻¹ SOC, P<0.001). Specific respiratory activity of SMBC was, however, consistently lower in the thermic than in the frigid regions (29 vs 34 mg CO₂–C g⁻¹ SMBC d⁻¹, P<0.01). Higher mean annual precipitation resulted in consistently lower basal soil respiration (1.1 vs 1.3 mg CO₂–C g⁻¹ SOC d⁻¹ in the wet compared with the dry regions, P<0.01) and lower SMBC (31 vs 43 mg SMBC g⁻¹ SOC, P<0.001), but had inconsistent effects on net N mineralization that depended upon temperature regime. Specific respiratory activity of SMBC was consistently greater in the wet than the dry regions (≈33 vs 29 mg CO₂–C g⁻¹ SMBC d⁻¹, P<0.01). Although the thermic regions were not able to retain as high a level of SOC as the frigid regions, due likely to high annual decomposition rates, biologically active soil fractions were as high per mass of soil and even 2–3-times greater per unit of SOC in the thermic compared with the frigid regions. These results suggest that macroclimate has a large impact on the portion of soil organic matter that is potentially active, but a relatively small impact on the specific respiratory activity of SMBC.