Efficient mineral weathering is a distinctive functional trait of the bacterial genus Collimonas

The mineral weathering ability of 45 bacterial strains belonging to the genus Collimonas and coming from various terrestrial environments was compared to that of 5 representatives from the closely related genera Herbaspirillum and Janthinobacterium. Using glucose as the sole carbon source in a microplate assay for quantifying the release of iron and protons from biotite, all Collimonas strains proved to be very efficient weathering agents, in contrast to the Herbaspirillum and Janthinobacterium strains. The weathering phenotype was also evident during growth of collimonads on mannitol and trehalose, but not on gluconic acid. All Collimonas strains were able to solubilize inorganic phosphorus and produce gluconic acid from glucose, suggesting that acidification is one of the main mechanisms used by these bacteria for mineral weathering. The production of siderophores may also be involved, but this trait, measured as the ability of collimonads to mobilize iron, was shared with Herbaspirillum and Janthinobacterium strains. These findings are discussed in an ecological context that recognizes collimonads as mycophagous (fungal-eating) and efficient mineral weathering bacteria and suggests that this ability has evolved as an adaptation to nutrient-poor conditions, possibly as part of a mutualistic relationship with mycorrhizal fungi.     

Distribution with depth of protozoa,bacteria and fungi in soil profiles from three Danish forest sites

The numbers and biomass of protozoa, bacteria and fungi were measured at various depths (1.5–122.5 cm) in the unsaturated zone of three contrasting pristine Danish forest site profiles: a dry beech (Fagus silvatica) forest on mor, a wet peaty spruce (Picea abies)/birch (Betula pubescens) forest and a dry spruce (P. abies) forest on mor. All sites were situated on a Weichel moraine. Except for a bacterial peak at 42.5 cm in the peat profile, the general tendency was a decrease in biomass with increasing depth for all groups examined. Protozoa decreased more rapidly with increasing depth than the other two groups of organisms examined. An evaluation of the bacterial–protozoan relationship by a simple mathematical model indicated that the subsurface protozoan populations are active and not accidental percolated cysts. The low protozoan numbers found in shallow subsurface sites contrast markedly with the results from contaminated sites where much larger protozoan populations have been reported even at considerable depths. Consequently, the results suggest that protozoa are good indicators of organic pollution in subsurface soils; however, more work involving the comparison of polluted and unpolluted soils is needed to confirm this suggestion.     

The “soil microbial loop” is not always needed to explain protozoan stimulation of plants

Protozoa stimulate plant growth, but we do not completely understand the underlying mechanisms, and different hypotheses seek to explain this phenomenon. To test these hypotheses, we grew the grass Yorkshire fog (Holcus lanatus) in pots with soil, which contained either (1) no organisms but bacteria – or (2) bacteria and protozoa. Half of the pots received a glucose treatment so as to mimic an additional root exudation. We measured plant growth and plant nitrogen uptake, along with various microbial pools and processes that support plant growth. Protozoan presence significantly enhanced soil nitrogen mineralization, plant nitrogen uptake from organic nitrogen sources, plant nitrogen content, and plant growth. By contrast, we found no evidence that glucose addition, mimicking root exudation, increased soil nitrogen availability and plant nitrogen uptake. Moreover, although protozoan presence affected bacterial community structure, it did not affect the proportion of IAA-producing bacteria in the community or plant root morphology. These results refute the “soil microbial loop” hypotheses, which suggest that protozoan stimulation of plant growth results from complex interactions between plant roots, bacteria and protozoa. Our experiment thus favours the simple explanation that increased nitrogen availability is the key factor behind the positive protozoan effect on plant growth. To exploit natural resources in an efficient and environmentally friendly way, we need to understand in detail the functioning of ecosystems. This study stresses that to achieve this, it is still urgent, besides investigating intricate food-web and signal compound interactions, also to focus on the basic stoichiometric and energetic aspects of organisms.     

Prey-predator dynamics in communities of culturable soil bacteria and protozoa: differential effects of mercury

We investigated whether the prey-predator dynamics of bacteria and protozoa were affected by inorganic mercury at concentrations of 0, 3.5 and 15 mg Hg(II) kg soil⁻¹. The amount of bioavailable Hg was estimated using a biosensor-assay based on the mer-lux gene fusion. The numbers of bacterial CFUs on the general medium 1/100 tryptic soy agar (TSA) were significantly decreased when the soil had been amended with Hg. In contrast, no effect was seen on the number of CFUs on the Pseudomonas-specific medium Gould's S1 agar. Protozoan numbers estimated by the most probable number (MPN) method with 1/100 TSB as growth medium were also negatively affected by Hg. The different fractions of protozoa were affected to different degrees suggesting that amoebae were less sensitive than slow-growing flagellates, which again were less sensitive than the fast-growing flagellates. In contrast, Hg did not induce any detectable changes in the diversity of flagellate morphotypes. In the treatment with 15 mg Hg kg⁻¹ a transiently increased number of bacteria was seen at day 6 probably concomitant with a decrease in the numbers of protozoa. This might indicate that Hg affected the prey-predator dynamics in communities of culturable bacteria and protozoa in soil. Furthermore, we showed that the number of Pseudomonas spp. was not affected by Hg whereas the number of bacteria growing on a general medium was.     

Solubilization of zinc phosphate by a strain of Pseudomonas fluorescens isolated from a forest soil

 A strain of Pseudomonas fluorescens, able to solubilize zinc phosphate, was isolated from a forest soil. Colonies of the microorganism produced clear haloes on solid medium incorporating zinc phosphate, but only when glucose was provided as the carbon source. Solubilization of zinc phosphate occurred by both an increase in the H⁺ concentration of the medium, probably a consequence of ammonia assimilation, and the production of gluconic acid. High concentrations of gluconic acid were produced when P. fluorescens 3a was cultured in the presence of zinc phosphate. Although under some conditions gluconic acid is purportedly able to solubilize metals by the formation of chelates, no evidence of zinc chelation was obtained in our experiments. Furthermore, the increased Zn²⁺ concentration caused by the solubilization process resulted in the manifestation of toxic effects on the culture. A sample of the culture, sonicated to disrupt cells, still possessed the ability to produce gluconic acid from glucose, in the presence and absence of zinc phosphate. The lack of gluconic acid overproduction in cultures of P. fluorescens 3a which were not amended with zinc phosphate suggests that at least some of the glucose oxidation required for the zinc solubilization occurred as a result of the toxic stress caused by the high Zn²⁺ concentration. Received: 16 December 1997     

Organic-acid-producing,phytate-mineralizing rhizobacteria and their effect on growth of pigeon pea (Cajanus cajan)

Phytates represent a significant pool of organic phosphorus (Po) that is largely unavailable to plants. This study deals with phytate-mineralizing (PM), organic-acid-producing (OAP) rhizobacterial isolates, their characterization and their effect on plant growth. Their genetic diversity was assessed by 16S rRNA amplified ribosomal DNA restriction analysis (ARDRA) and selected isolates were identified by partial sequencing of 16S rRNA gene. Na-phytate and Po rich poultry farm manure (PFM) used as sources of phosphorus in semi-solid-agar (SSA) medium and soil respectively, for plant inoculation studies, where Cajanus cajan (pigeon pea) used as plant. Of thirty-nine rhizobacterial isolates, nineteen were proficient at releasing phosphate (Pi) (up to 85 μg/ml) from sparingly soluble calcium (Ca)-phytate and concomitantly decreasing the pH of minimal medium with 100 mM glucose from 8.0 to below 5. When the medium contained glycerol in place of glucose, Ca-phytate remained undissolved with no significant Pi released and no decline in pH. Genetic diversity of phytate-mineralizing (PM) rhizobacterial isolates suggests that the isolates mainly fall in two populations: acid-producing (AP) population (mainly represented by members of Enterobacteriaceae) and non-AP population. OAP-PM rhizobacterial isolates were identified as Citrobacter, Pantoea, Klebsiella and Enterobacter species. Organic acids (OAs) secreted by PM isolates were detected by HPLC, showed secretion of gluconic and acetic acids. Importance of OAs in Ca-phytate dephosphorylation was demonstrated in vitro using A. ficuum phytase. Gluconate and acetate additions enhanced phytase catalyzed dephosphorylation of Ca-phytate in vitro. Sonicated cell lysates of isolates showed significant Pi release from Ca-phytate compared to whole cells, indicating inaccessibility of Ca-phytate due to poor solubility. Selected isolates showed that they possess cell-associated acid phytase and modulators of phytase activity suggested that the enzymes are histidine acid phosphatase (HAP) type of phytase. OAP-PM isolates PP1 and DHRSS showed significant increase in dry shoot/root ratio and P content of shoot in Na-phytate containing semi-solid agar (SSA) medium, but isolate DHRSS did not increase dry shoot/root ratio in soil experiments containing poultry farm manure as source of P, although it significantly increased shoot P content of plants. The inoculation of isolates enhanced the shoot P content and dry shoot/root ratio, but did not increase the dry weight in SSA medium. It may be concluded that some OAP-PM rhizobacterial isolates that release P from Ca-phytate show increase in shoot P content in phytate containing SSA medium and in soils.     

Effect of some rare earth elements on the growth and lanthanide accumulation in different Trichoderma strains

Accumulation of rare earth elements (REE) in the soil may be due to the use of REE enriched fertilizers and to contamination by REE containing wastes. Although widely used in China for soil and foliar dressing of crops, little is known about the effect of REE applications on the soil microbial community. The effect of REE on the growth of biological control strains of Trichoderma atroviride and Trichoderma harzianum was investigated in vitro using either a mix of different REE containing different amounts of lanthanum, cerium, praseodymium, neodymium, gadolinium nitrate and lanthanum nitrate alone in comparison to treatments with potassium nitrate and water. In plate tests applied concentrations ranged from 0.1 mM to 300 mM for lanthanum and REE mix and from 0.1 mM to 900 mM for the potassium solution. In liquid culture tests applied concentrations ranged from 0.001 mM to 100 mM for lanthanum and REE mix and from 0.003 mM to 900 mM for the potassium solution. ICP-MS, TEM and TEM X-ray microanalysis were used to study the accumulation of REE in fungal biomass. All the Trichoderma strains showed a good tolerance to the presence of REE in the culture media. Some growth enhancing effects were observed in liquid cultures of T. harzianum strains but not in T. atroviride. Accumulation of REE in fungal biomass, both at intracellular level and in the extracellular matrix, was observed.     

Predation by protozoa on Escherichia coli K12 in soil and transfer of resistance plasmid RP4 to indigenous bacteria in soil

Escherichia coli K12 strain (J5-3/RP4) persisted in sandy loam for more than 70 days when incubated at 10°C or 4°C. It decreased to below the level of detection within 20 days when incubated at 25°C. No loss of multi-resistance plasmid RP4 from the E. coli cells was detected during incubation in soil. There was a positive relation between the bacterial inoculum size and the following increase of the protozoan numbers in the soil. When soil microcosms were amended with an eukaryotic inhibitor, the period of survival was increased. These observations indicate a direct involvement of protozoa in the decline of E. coli in soil. Transfer of plasmid RP4 from E. coli donor bacteria to indigenous bacteria in soil was detected already 24 h after addition of the E. coli K12 donor strain. The efficiency of transfer during the first 48 h was approximately 10⁻⁶ transconjugants per donor. Inhibition of protozoan predation increased the number of transconjugants appearing in the soil, but the transfer efficiency per donor was not affected by the decreased predation. No transfer could be detected when the donor strain was washed and resuspended in saline before addition to the soil, but transconjugants were detected in this experiment when nutrients (LB) were supplemented after two days of incubation. Plasmid RP4 was maintained in the transconjugant soil bacteria throughout the experiment. The data presented here indicate that the indigenous bacteria in soil may serve as a sink for plasmidborne traits.     

Influence of texture on habitable pore space and bacterial-protozoan populations in soil

Summary Soil texture affects pore space, and bacterial and protozoan populations in soil. In the present study we tested the hypothesis that bacteria are more protected from protozoan predation in fine-textured soils than in coarse-textured soils because they have a larger volume of protected pore space available to them. The experiment consisted of three sterilized Orthic Black Chernozemic soils (silty clay, clay loam, and sandy loam) inoculated with bacteria, two treatments (with and without protozoa), and five sampling dates. The soils were amended with glucose and mineral N on day 0. On day 4 bacterial numbers in all three soils were approximately 3×10⁹ g^–1 soil. The greatest reduction in bacteria due to protozoan grazing occurred between day 4 and day 7. Compared to the treatment without protozoa, bacteria in the treatment with protozoa were reduced by 68, 50, and 75% in the silty clay, clay loam, and sandy loam, respectively. On day 4, 2 days after the protozoan inoculation, all protozoa were active. The numbers were 10330, 4760, and 15 380 g^–1 soil for the silty clay, clay loam, and sandy loam, respectively. Between day 4 and day 7, the period of greatest bacterial decline, total protozoa increased greatly to 150480, 96160, and 192100 g^–1 soil for the three soils, respectively. Most protozoa encysted by day 7. In all soils the addition of protozoa significantly increased CO₂–C evolution per g soil relative to the treatment without protozoa. Our results support the hypothesis that bacteria are more protected from protozoan predation in fine-textured soils than in coarse-textured soils.     

A modified method based on p-nitrophenol assay to quantify hydrolysis activities of lipases in litters

Lipases are glycerol ester hydrolases (EC 3.1.1.3) produced by a wide range of microorganisms. They catalyse the hydrolysis of different esters depending on the water content of the reaction medium. Here, we developed a simple methodology to quantify lipase hydrolysis activities using two different litters: a litter of Quercus pubescens (QP) and a litter of both Q. pubescens and Q. ilex. Different p-nitrophenyl esters were used to test hydrolysis in a reaction medium with an organic solvent (heptane). We showed that these activities depended on the amount of litter, the incubation time and the substrate concentration and that they increased with temperature. Furthermore, the lipases from the studied litters were still active after 2 h at 70 °C. These activities showed common properties of lipases: the highest activities were obtained with a medium-acyl chain substrate, p-nitrophenyl laurate. Moreover abiotic hydrolysis with short-chain acyl substrates was observable. The following parameters are recommended to quantify hydrolysis activities of lipases in litters: 10 mM of p-nitrophenyl laurate in 2 ml of heptane, 1 g of litter, 2 ml of water incubated at 30 °C for 2 h.     

The impact of protozoa on the availability of bacterial nitrogen to plants

Summary Microbial N from ¹⁵N-labelled bacterial biomass was investigated in a microcosm experiment, in order to determine its availability to wheat plants. Sterilized soil was inoculated with either bacteria (Pseudomonas aeruginosa alone or with a suspension of a natural bacterial population from the soil) or bacteria and protozoa to examine the impact of protozoa. Plant biomass, plant N, soil inorganic N and bacterial and protozoan numbers were determined after 14 and 35 days of incubation. The protozoa reduced bacterial numbers in soil by a factor of 8, and higher contents of soil inorganic N were found in their presence. Plant uptake of N increased by 20010 in the presence of protozoa. Even though the total plant biomass production was not affected, the shoot: root ratios increased in the presence of protozoa, which is considered to indicate an improved plant nutrient supply. The presence of protozoa resulted in a 65010 increase in mineralization and uptake of bacterial ¹⁵N by plants. This effect was more pronounced than the protozoan effect on N derived from soil organic matter. It is concluded that grazing by protozoa strongly stimulates the mineralization and turnover of bacterial N. The mineralization of soil organic N was also shown to be promoted by protozoa.Communication No. 9 of the Dutch Programme on Soil Ecology of Arable Farming Systems     

Suppression of root-knot disease by Pseudomonas fluorescens CHA0 in tomato: importance of bacterial secondary metabolite, 2,4-diacetylpholoroglucinol

The antimicrobial metabolites 2,4-diacetylphloroglucinol (2,4-DAPG) and pyoluteorin contribute to the ability of Pseudomonas fluorescens strain CHA0 to control plant diseases caused by soil-borne pathogens. P. fluorescens strain CHA0 and its derivatives CHA89 (antibiotics-deficient) and CHA0/pME3424 (antibiotics overproducing) were investigated as potential biocontrol agents against Meloidogyne javanica the root-knot nematode. Exposure of root-knot nematode to culture filtrates of P. fluorescens under in vitro conditions significantly reduced egg hatch and caused substantial mortality of M. javanica juveniles. Nutrient broth yeast extract (NBY) medium amended with 2% (w/v) glucose or 1 mM EDTA markedly repressed hatch inhibition activity of the strain CHA0 but not that of CHA0/pME3424 or CHA89. On the other hand, NBY medium amended with glucose significantly enhanced nematicidal activity of the strain CHA0/pME3424. Neither glucose nor EDTA had an influence on the nematicidal activity of the strains CHA0 and CHA89. Under in vitro conditions, antibiotic overproducing strain CHA0/pME3424 and CHA0 expressed phl‘-’lacZ reporter gene but strain CHA89 did not. Expression of the reporter gene reflects actual production of DAPG. In general, CHA0/pME3424 expressed reporter gene to a greater extent compared to its wild type counterpart CHA0. Regardless of the bacterial strains, reporter gene expression was markedly enhanced when NBY medium was amended with glucose but EDTA had no such effect. A positive correlation between the degree of juvenile mortality and extent of phl‘-’lacZ reporter gene expression was also observed in vitro. Strain CHA0 produced zones of 4-6 mm on MM medium containing gelatin while strain CHA0/pME3424 and CHA89 did not. When MM medium containing gelatin was amended with 2% glucose of 1 mM EDTA size of haloes produced by the strain CHA0 reduced to 2 mm. Under glasshouse conditions aqueous cell suspension of the strains CHA0 or CHA0/pME3424 at various inoculum levels (10⁷, 10⁸ or 10⁹ cfu ml⁻¹) significantly reduced root-knot development. CHA89 caused significant reduction in galling when applied at 10⁹ cfu ml⁻¹. To better understand the mechanism of nematode suppression, split root bioassay was performed. Split-root experiments, that guarantee a spatial separation of inducing agent and a challenging pathogen, showed that soil treatment of one half of the root system with cell suspension of CHA0 or CHA0/pME3424 resulted in a significant systemic induced resistance leading to reduction of M. javanica infection of tomato roots in the non-baterized nematode treated half. The results clearly suggest that the antibiotic 2,4-DAPG from P. fluorescens CHA0 act as the inducing agents of systemic resistance in tomato roots. Populations of CHA0 and its derivatives declined progressively by 10-fold between first and fourth harvests (0-21 days after inoculation). However, bacterial populations increased at final harvest (28 days after application).     

Grazing of a common species of soil protozoa (Acanthamoeba castellanii) affects rhizosphere bacterial community composition and root architecture of rice (Oryza sativa L.)

We performed a controlled experiment with rice seedlings (Oryza sativa L.) growing in Petri dishes on homogeneous nutrient agar containing a simple rhizosphere food web consisting of a diverse bacterial community and a common soil protozoa, Acanthamoeba castellanii, as bacterial grazer. Presence of amoebae increased bacterial activity and significantly changed the community composition and spatial distribution of bacteria in the rhizosphere. In particular, Betaproteobacteria did benefit from protozoan grazing. We hypothesize that the changes in bacterial community composition affected the root architecture of rice plants. These effects on root architecture affect a fundamental aspect of plant productivity. Root systems in presence of protozoa were characterized by high numbers of elongated (L-type) laterals, those laterals that are a prerequisite for the construction of branched root systems. This was in sharp contrast to root system development in absence of protozoa, where high numbers of lateral root primordia and short (S-type) laterals occurred which did not grow out of the rhizosphere region of the axile root. As a consequence of nutrient release from grazed bacteria and changes in root architecture, the nitrogen content of rice shoots increased by 45% in presence of protozoa. Our study illustrates that interactions over three trophic levels, i.e. between plants, bacteria and protozoa significantly modify root architecture and nutrient uptake by plants.     

Effect of Bacillus subtilis on granite weathering: A laboratory experiment

We performed a comparative experiment to investigate: (1) how the ubiquitous soil bacterium Bacillus subtilis weathers granite; and (2) which granite-forming minerals weather more rapidly via biological processes. Batch system experiments (granite specimen in a 500 ml solution including NaCl, glucose, yeast extract and bacteria B. subtilis at 27 °C) were carried out for 30 days. Granite surfaces were observed by SEM before and after the experiment. B. subtilis had a strong influence on granite weathering by forming pits. There were 2.4 times as many pits and micropores were 2.3 times wider in granite exposed to B. subtilis when compared with bacteria-free samples. B. subtilis appear to preferentially select an optimum place to adhere to the mineral and dissolve essential elements from the mineral to live. Plagioclase was more vulnerable to bacterial weathering than biotite among the granite composing minerals.     

The mitigation potential of hippuric acid on N2O emissions from urine patches: An in situ determination of its effect

Previous laboratory studies have demonstrated that hippuric acid, a ruminant urine constituent, can mitigate nitrous oxide (N₂O) emissions from simulated urine patches. Hippuric acid has the potential to be a N₂O mitigation tool because animal diets can be manipulated to adjust its concentration in the urine. This study was conducted to determine if the effect observed in previous laboratory studies would also occur in situ under field conditions. In our field study, plots were treated with unadulterated bovine urine (56 mM hippuric acid), the same bovine urine amended with either benzoic acid (34 mM), dicyandiamide (DCD) or varying rates of hippuric acid (up to 90 mM). Soil inorganic-N, N₂O fluxes, and plant N responses were monitored over a 78 d period. Effects on microbial communities were monitored by determining the size and structure of nitrite oxidizer (nxrA) and nitrite reducer (nirS) bacterial populations using real-time PCR and denaturing gradient gel electrophoresis (DGGE), respectively. Decreases in N₂O emissions, with increasing hippuric and benzoic acid concentrations, were only seen on Day two of the trial. With the exception of the DCD treatment (0.60% of N applied) the amended urine treatments did not significantly affect emissions of N₂O as a percentage of N applied (1.28-1.65%). Soil inorganic-N and plant response were not affected by urinary amendment, except in the DCD treatment where nitrification inhibition occurred. Nitrite oxidizer community structures shifted and increased approximately 5.4-fold in size over 48 d in response to urine, although no specific response to elevated hippuric acid or benzoic acid was observed. No treatment effects were observed on community structure of the nitrite reducing bacteria but averaged over time the highest rate of hippuric acid significantly decreased nirS gene copy numbers g⁻¹ soil. We concluded that under the conditions of this field study, increasing hippuric or benzoic acid concentrations in bovine urine had no effect on N₂O mitigation in situ. We argue that the discrepancy with previous laboratory studies may be related to differences in soil pH, microbial communities and the presence of vegetation. Further research is needed to determine the potential for hippuric acid as a tool to mitigate N₂O emissions, and its effect(s) on resident N cycling microorganisms.     

Changes in the succession and diversity of protozoan and microbial populations in soil spiked with a range of copper concentrations

We studied microbial and protozoan activity, diversity and abundance as affected by Cu²⁺ amendments ranging from 0 to 1000 μg g⁻¹ over a 70-day period. At the end of the experiment the microbial population size, as indicated by substrate-induced respiration, had normalized for all Cu²⁺ concentrations, but 1000 μg g⁻¹. Protozoan abundance was negatively affected by Cu²⁺, although, only in the first few weeks. A more detailed analysis of the individual components that make up the microbial and micro-faunal populations (phospholipid fatty acid (PLFA) profile and protozoan morphotypes), however, yielded a somewhat more complex picture. For the three highest Cu²⁺ amendments (160, 400 and 1000 μg g⁻¹), there still was a significant reduction in number of differentiable protozoan morphotypes. The bacterial PLFA pattern suggested a shift from Gram-negative towards Gram-positive bacteria for the high amendments, a process where protozoan grazing most likely played a significant role. The ratio of the trans/cis isomers of the 16:1ω7 fatty acid indicated that Cu²⁺, even at low and medium concentrations, induced physiological changes in the microbial population. The relatively slight changes in total microbial and micro-faunal abundance and activity, also at the highest Cu²⁺ concentrations, probably reflected the ability of the community to compensate for loss of taxa by functional substitution.     

Isolation and characterization of phosphate-solubilizing bacteria from betel nut (Areca catechu) and their effects on plant growth and phosphorus mobilization in tropical soils

Phosphate-solubilizing bacteria (PSB) were isolated and characterized from the rhizosphere and bulk soils of Areca catechu plants. A long history of phosphate fertilizer use has elicited a direct effect on the incidence of soil PSB. Their abundance and ability to solubilize insoluble phosphate were significantly greater (P?<?0.0001) in soils with low available phosphorus (P) content than in other soil types. Three efficient PSB strains, namely, ASL12, ASG34, and ADH302, were identified as Acinetobacter pittii, Escherichia coli, and Enterobacter cloacae by characterizing 16S rRNA sequences and biochemical characteristics; they produced gluconic acid at concentrations of 7862.4, 4306.5, and 2663.8 mg L^?1, respectively. The highest amount of solubilized P was determined in Pikovskaya (PVK) medium for the bacterial strain ASL12. The secretion of gluconic acid was related to the available P of rhizosphere soils and P solubilization. Under shaded conditions, the application of these three strains significantly improved plant height, shoot and root dry weight, and nutrient uptake of A. catechu seedlings. A further increase in P solubilization was observed by co-inoculating the three strains and also applying tricalcium phosphate (TCP) or aluminum phosphate (AP). A significant (P?<?0.05) correlation was also observed between P-solubilization activity and A. catechu plant growth in pot experiments. Thus, the three strains can be potentially applied as inoculants in tropical and aluminum-rich soils.     

Phosphate solubilization potentials of soil Acinetobacter strains

Many phosphate solubilizing microorganisms (PSM) require external pyrroloquinoline quinone (PQQ) for strong phosphorus (P) solubilization in vitro. The objective of this study was to isolate efficient and PQQ-independent PSM. A total of 21 PSM were isolated from the rhizosphere soil of wheat and maize grown in the pots. Acinetobacter strains were the only PQQ-independent and most effective solubilizers of tricalcium phosphate containing agar. The mean P dissolved in liquid cultures of Acinetobacter strains in a 5-day incubation ranged from 167 to 888 μg/ml P. The pH dropped to below 4.7 from 7.8 in six isolates, which produced gluconic acid in concentrations ranging between 27.5 and 37.5 mM. There was a linear regression between soluble P and gluconic acid concentrations in the bacterial cultures (P < 0.05; R ² = 0.59). Inoculation with Acinetobacter sp. WR922 significantly (P < 0.05) increased wheat (Triticum aestivum L.) P content by 27% at 15 days after emergence (DAE) and dry matter by 15% at 30 DAE compared to the control. The plant P content in inoculated plants at 30 DAE was linearly correlated with soluble P of the bacterial cultures (P < 0.05; R ² = 0.69). Gluconic acid production directly affected phosphate solubilization in vitro, which in turn influenced plant P content of inoculated plants in PQQ-independent P-solubilizing Acinetobacter strains.     

Influence of reduced tillage on earthworm and microbial communities under organic arable farming

Although reduced tillage is an agricultural practice reported to decrease soil erosion and external inputs while enhancing soil fertility, it has still rarely been adopted by European organic farmers. The objective of this study was to assess the long-term interactive effects of tillage (conventional (CT) vs. reduced (RT)) and fertilization (slurry (S) vs. composted manure/slurry (MCS)) on earthworms and microbial communities in a clay soil under spelt in an organic 6-year crop rotation. Earthworm populations (species, density and biomass, cocoons) were investigated by handsorting the soil nine years after initial implementation of the treatments. Soil microbial carbon (C_mic) and nitrogen (N_mic) were measured by chloroform-fumigation extraction and a simplified phospholipid fatty acid (PLFA) analysis was used to separate for populations of bacteria, fungi and protozoa. Significantly increased total earthworm density in RT plots was mainly attributed to increased numbers of juveniles. Moreover, we found five times more cocoons with RT. Species richness was not affected by the treatments, but tillage treatments had differentially affected populations at the species-level. In addition, cluster analysis at the community level revealed two distinct groups of plots in relation to tillage treatments. In RT plots C_mic increased in the 0–10 cm and 10–20 cm soil layers, while PLFA concentrations indicative of Gram-negative bacteria, fungi and protozoa only increased in the topsoil. Lower bacteria-to-fungi ratios in the upper soil layer of RT plots indicated a shift to fungal-based decomposition of organic matter whereas a higher C_mic-to-C_org ratio pointed towards enhanced substrate availability. Slurry application decreased microbial biomass and enhanced density of juvenile anecic earthworms but overall fertilization effect was weak and no interactions with tillage were found. In conclusion, tillage is a major driver in altering communities of earthworms and microorganisms in arable soils. The use of reduced tillage provides an approach for eco-intensification by enhancing inherent soil biota functions under organic arable farming.     

Spatial distribution of the nematode biocontrol agent Pasteuria penetrans as influenced by its soil habitat

Root-knot nematodes belonging to the Meloidogyne genus are ubiquitous plant-parasitic pests, especially on vegetables. The Pasteuria penetrans bacterium is an obligate parasite of nematodes, parasitizing most of the Meloidogyne species. Spatial distributions of Meloidogyne javanica populations infested or not by P. penetrans and of bacterial populations were studied in a vegetable plot naturally infested by these organisms. It was observed that distributions of M. javanica populations, of populations infested by P. penetrans, and of free bacteria populations were not overlapped. Soil factors involved were investigated. Soil texture and water flow in porosity are concerned, as they directly influence the level of the pool of bacteria and then the chances of both organisms to meet. The soil solution has a direct effect on the attachment of the bacterium on the nematode cuticle.