Bacteria-grazing amoebae in situ in the rhizosphere

Summary An ultrastructural investigation of amoebae in situ in the rhizosphere showed that the protozoa are closely associated with soil aggregates and produce long pseudopodia that penetrate micropores. This could partly explain why bacteria are generally confined to the interiors of microaggregates. The presence of cytologically intact and partly digested bacteria in the food vacuoles indicates that rhizosphere bacteria are both ingested and digested by the amoebae. This digestion could lead to the recycling of P and N immobilized in rhizosphere microorganisms.     

Earthworms change the abundance and community structure of nematodes and protozoa in a maize residue amended rice-wheat rotation agro-ecosystem

The influence of earthworms on nematodes and protozoan communities was determined during the wheat phase of a six year rice-wheat rotation agro-ecosystem. Experimental plots in the rotation had five treatments, i.e. incorporation or mulching of maize residues with or without added earthworms and a control. The addition of maize residues to soil strongly affected the abundance and community structure of nematodes and protozoa in the absence of earthworms. The presence of earthworms gave significantly lower total nematode numbers at all soil depths following maize residue incorporation than the same treatment without earthworms, and also gave lower (although not significantly) total nematode numbers in the upper soil layer following maize residue mulching than the same treatment without earthworms. This was mainly due to a significant decrease in bacterial-feeding nematode numbers. Earthworms also strongly affected the distribution of the number of total nematodes and two trophic groups (bacterial and plant feeders) with soil depth. In the presence of earthworms, total protozoan and flagellate numbers significantly increased at all soil depths following both incorporation and mulching of maize residues, while numbers of amoebae increased only when maize residues were mulched. Additionally, in earthworm casts total nematode numbers (mainly bacterial and fungal feeders) were significantly higher, whereas total protozoa numbers (mainly flagellates and amoebae) were significantly lower than that in soil from 0 to 5 cm layer.These results indicated that earthworm activity could affect the abundance and community structure of microfauna, and change their distribution between soil layers and cast material, depending on the mode of application of organic residues.     

Migration of bacterial-feeding nematodes,but not protozoa,to decomposing grass residues

Summary Populations of bacterial-feeding nematodes and protozoa developing in soil amended with dried grass powder or a nutrient solution were monitored in experimental systems designed to prevent migration from surrounding unamended soil. The addition of nutrient solution stimulated both microbial activity, as determined by dehydrogenase activity, and protozoa, but brought about no increase in nematode numbers. Amendment of soil with grass, however, caused an increase in both types of grazer, with the maximum biomass of protozoa (180 g g^-1) exceeding that of bacterial-feeding nematodes (42 g^-1). The decomposing grass was rapidly colonised by rhaditid nematodes, mainly Caenorhabditis sp. Incubating grass-amended soil in the absence of any surrounding soil, to prevent migration, changed the microflora from predominantly bacterial to predominantly fungal, and so could not be used to compare treatments with and without migration. Surrounding the amended soil with sterilised soil prevented migration and caused no detectable change in the microflora. This treatment demonstrated that migration plays an important part in the colonisation of decomposing substrates by nematodes, but that protozoa do not migrate in soil. The nematodes migrated from a volume of unamended soil that was equivalent to eight times the volume of amended soil. The potential effects of the large grazing pressure on the subsequent decomposition of the grass residue are discussed.     

Synergists and antagonists in the rhizosphere modulate microbial communities and growth of Quercus robur L.

The synergistic and antagonistic interactions among biotic components in the rhizosphere play a crucial role in plant defence against soil-borne pathogens. We investigated if the rhizosphere helper bacterium Streptomyces sp. AcH 505 (HB) indirectly protects the plant from the parasitic nematode Pratylenchus penetrans by modifying the rhizosphere microbial community structure and whether these interactions are dependent on the growth stage of oaks. Changes in the abundance of Streptomyces sp. AcH 505 and the phospholipid fatty acid (PLFA) composition of the rhizosphere soil as well as oak shoot and root biomass were assessed. Investigated were the bud resting stage A and the bud swelling stage B with maximal root elongation of oak microcuttings at two successive harvest times. The deleterious effect of P. penetrans on oak biomass was dependent on plant development, being limited to oak microcuttings growing at the stage B. In comparison to control and HB inoculated soils, shoot biomass decreased by about 33% and 41%, and root biomass by about 33 and 48%, respectively. The antagonistic effect of Streptomyces against the nematode was linked to shifts in the rhizosphere microbial community. The Streptomyces AcH505 strain promoted growth of oak microcuttings at bud swelling stage B during maximal root elongation and enhanced the abundance of saprophytic and ectomycorrhizal fungi in the rhizosphere by 158% with respect to controls. Our results highlight the importance of Streptomyces for counteracting the damage of nematodes and promoting plant growth in natural ecosystems such as forests.     

The mycorrhizal fungus (Glomus intraradices) affects microbial activity in the rhizosphere of pea plants (Pisum sativum)

Pea plants were grown in γ-irradiated soil in pots with and without addition of the AM fungus Glomus intraradices at sufficient N and limiting P. Depending on the growth phase of the plant presence of AM had negative or positive effect on rhizosphere activity. Before flowering during nutrient acquisition AM decreased rhizosphere respiration and number of protozoa but did not affect bacterial number suggesting top-down regulation of bacterial number by protozoan grazing. In contrast, during flowering and pod formation AM stimulated rhizosphere respiration and the negative effect on protozoa decreased. AM also affected the composition of the rhizosphere bacterial community as revealed from DNA analysis (DGGE). With or without mycorrhiza, rhizosphere respiration was P-limited on very young roots, not nutrient limited at more mature roots and C-limited at withering. This suggests changes in the rhizosphere community during plant growth also supported by changes in the bacteria (DGGE).     

Trophic interactions between rhizosphere bacteria and bacterial feeders influenced by phosphate and aphids in barley

The aim was to study the effects of P fertilization and leaf aphid attack on the trophic interactions of bacteria and bacterial feeders in the rhizospheres of barley plants. The density of protozoa peaked in the rhizospheres of plants fertilized with N and P, whereas nematodes peaked in the rhizospheres of plants to which only N had been added. Fingerprinting of bacterial communities by length heterogeneity polymerase chain reaction revealed differences in community structure between NP rhizospheres and N rhizospheres as well as aphid-related differences within N rhizospheres. Specifically, α-proteobacteria increased with P addition. To evaluate if differences in bacteria in terms of their quality as food could partly explain the observed differences in protozoan and nematode abundances, growth of the flagellate Cercomonas sp. was assessed with 935 bacteria isolated from the different treatments. This assay indicated that bacterial isolates were of higher food quality to Cercomonas sp. in NP than in N rhizospheres when plants were subjected to aphid attack. Bacteria of high and low food quality for Cercomonas sp., respectively, were fed to the nematode Caenorhabditis elegans and larval production examined. α-Proteobacteria supported the growth of Cercomonas sp. well, whereas Actinobacteria did not. In contrast, C. elegans reproduced poorly on most α-proteobacteria but were able to reproduce well on some Actinobacteria. These results suggest that the different response of protozoa and nematodes to P addition could be mediated through a food quality-related change in community composition of bacteria and that leaf aphid attack may interfere with nutrient effects on bacterial assemblages of rhizospheres.     

A comparison of soil food webs beneath C<Subscript>3</Subscript>- and C<Subscript>4</Subscript>-dominated grasslands

Soil food webs influence organic matter mineralization and plant nutrient availability, but the potential for plants to capitalize on these processes by altering soil food webs has received little attention. We compared soil food webs beneath C₃- and C₄-grass plantings by measuring bacterial and fungal biomass and protozoan and nematode abundance repeatedly over 2 years. We tested published expectations that C₃ detritus and root chemistry (low lignin/N) favor bacterial-based food webs and root-feeding nematodes, whereas C₄ detritus (high lignin/N) and greater production favor fungal decomposers and predatory nematodes. We also hypothesized that seasonal differences in plant growth between the two grassland types would generate season-specific differences in soil food webs. In contrast to our expectations, bacterial biomass and ciliate abundance were greater beneath C₄ grasses, and we found no differences in fungi, amoebae, flagellates, or nematodes. Soil food webs varied significantly among sample dates, but differences were unrelated to aboveground plant growth. Our findings, in combination with previous work, suggest that preexisting soil properties moderate the effect of plant inputs on soil food webs. We hypothesize that high levels of soil organic matter provide a stable environment and energy source for soil organisms and thus buffer soil food webs from short-term dynamics of plant communities.     

Approaches to measuring the contribution of nematodes and protozoa to nitrogen mineralization in the rhizosphere

Abstract. Nematodes and protozoa assist nitrogen mineralization by consuming bacteria, and this may increase the amount of nitrogen taken up by plants so that it exceeds the amount lost by root exudation. One of the methods used to determine bacterial consumption by protozoan grazers in aquatic ecosystems, namely progressive dilution of the grazing pressure, was tested in a sandy soil. The result suggests that a basic assumption of the technique, that grazing is directly proportional to the dilution factor, is untrue for soils, possibly because the increase in moisture content with increasing dilution enhances grazer activity.     

不同茬口设施番茄栽培的根圈基质中酶活性与养分效应

研究了番茄设施有机基质栽培的正茬、迎茬、连茬根圈生物性状和理化性状的变化, 分析了不同茬口对番茄栽培基质中酶活性、有机质含量和主要养分含量的影响。结果表明, 番茄连茬栽培基质的蛋白酶、磷酸酶、脲酶和过氧化氢酶活性下降, 有机质、全氮、铵态氮、碱解氮、全磷、速效磷、全钾及速效钾等主要养分含量降低, 而正茬栽培对基质酶活性、有机质和主要养分含量的影响较小。基质酶活性与理化性质的相关性分析表明, 番茄有机基质栽培中, 连茬、迎茬和正茬条件下基质的化学性质与生物学性质密切相关, 从而导致了番茄连茬基质微生态环境的改变。所有指标的相关性中, 脲酶与有机质和主要养分含量的相关系数最大, 说明脲酶活性是影响基质有机质含量和主要养分的重要因子。各茬口中, 速效钾、速效磷、全钾和过氧化氢酶活性的可塑性指数均最大, 不同茬口基质各指标的可塑性指数表现为新基质的最大、正茬次之、连茬最小, 说明连茬基质不适宜于番茄生长, 而正茬基质的生物和理化性质与新基质差异不大, 适宜于番茄生长发育。     

Grassland plants affect dissolved organic carbon and nitrogen dynamics in soil

Dissolved organic carbon (DOC) and nitrogen (DON) are central in many nutrient cycles within soil and they play an important role in many pedogenic processes. Plants provide a primary input of DOC and DON into soil via root turnover and exudation. Under controlled conditions we investigated the influence of 11 grass species alongside an unplanted control on the amount and nature of DOC and DON in soil. Our results showed that while the presence of plants significantly increases the size of a number of dissolved nutrient pools in comparison to the unplanted soil (e.g. DOC, total phenolics in solution) it has little affect on other pools (e.g. free amino acids). Grass species, however, had little effect on the composition of the DOC, DON or inorganic N pools. While the concentration of free amino acids was the same in the planted and unplanted soil, the flux through this pool was significantly faster in the presence of plants. The presence of plants also affected the biodegradability of the DOC pool. We conclude that while the presence of plants significantly affects the quantity and cycling of DOC and DON in soil, comparatively, individual grass species exerts less influence.     

Localisation of nitrate in the rhizosphere of biochar-amended soils

A wheat seedling rhizobox approach was used to differentiate between the rhizosphere and non-rhizosphere (bulk) soil amended with low and high rates of biochar (20 and 60 t ha⁻¹ vs. control). Nitrate (NO₃⁻) was added as the main nitrogen (N) source because emerging biochar research points to reduced NO₃⁻ loss through leaching and gaseous loss as nitrous oxide. The rhizosphere under the different treatments were distinct (P = 0.021), with greater soil-NO₃⁻ and biochar-NO₃⁻ contents in the high biochar treatment. Biochar addition increased wheat root length ratio (P = 0.053) and lowered root N uptake (P = 0.017), yet plant biomass and N content were similar between treatments. The results indicate localisation of NO₃⁻ within the rhizosphere of biochar-amended soils which has implications for NO₃⁻ loss and improved nitrogen use efficiency.     

Protease and deaminase activities in wheat rhizosphere and their relation to bacterial and protozoan populations

Protease and deaminase activities and population dynamics of bacteria and protozoa were measured in the rhizosphere of wheat to study their interactions with the mineralization of nitrogen. The experimental design allowed the separation of roots and soil material by means of a gauze. The most pronounced rhizosphere effect was detected for all the measured variables in the soil closest to the gauze. The number of bacteria was significantly higher in the presence than in the absence of plants up to 4 mm away from the soil-root interface and the closer to this interface the higher the number. Protozoan and bacterial population dynamics were positively correlated; generally, populations of flagellates and amoebae were comparable and their sum accounted for the population of total protozoa. For both enzyme activities the rhizosphere effect extended up to 2 mm away from the soil-root interface. The histidinase activity was of bacterial origin, while it is likely that bacteria, protozoa and root hair all contributed to the overall caseinase activity. Decomposition of root exudates and native organic matter in the rhizosphere, reflected by a growing microbial population, is associated with nitrogen mineralization through increases in caseinhydrolysing and L-histidine-deaminating activities. The adopted soil-plant microcosm is suitable for the study of the rhizosphere effect over time of incubation and distance gradient from the soil-root interface.     

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     

Magnitude,direction, and drivers of rhizosphere effect on soil nitrogen and phosphorus in global agroecosystem

The rhizosphere is the most active soil area for material transformation and energy flow of soil, root, and microorganism, which plays an important role in soil biochemical cycling. Although the rhizospheric nitrogen (N) and phosphorous (P) were easily disturbed in the agroecosystem, the effects of rhizosphere on the dynamics of soil N and P cycling have not yet been systematically quantified globally. We summarized the magnitude, direction, and driving forces of rhizosphere effects on agroecosystem's N and P dynamics by 1063 observations and 15 variables from 122 literature. Rhizosphere effects increased available N (AN, 9%), available P (AP, 11%), and total P (TP, 5%), and decreased nitrate N (NO₃–N, 18%) and ammonia N (NH₄–N, 16%). The effect of rhizosphere on total N (TN) was not significant. These effects improved AN in tropical (12%) and subtropical (14%) regions. The effect of rhizosphere on TP was greater under subtropical conditions than in other climates. The most substantial effects of the rhizosphere on TP and AP were observed under humid conditions. Rhizosphere effects increased AN and AP in vegetables more than in other crop systems. Application of N > 300 kg ha⁻¹ had the most significant and positive rhizosphere effects on TN and AN. P application of 100–150 kg ha⁻¹ had the greatest rhizosphere effects on TP and AP. These effects also improved the microbial (biomass N and P) and enzymatic aspects (urease, acid phosphatase, and alkaline phosphatase) of soil P and N cycling. Structural equation modeling suggested that aridity indices, fertilizer application rate, soil pH, microbial biomass, and soil enzymes strongly influence the magnitude and direction of the rhizosphere's effect on the P and N cycles. Overall, these findings are critical for improving soil nutrient utilization efficiency and modeling nutrient cycling in the rhizosphere for agricultural systems.     

Denitrification in the rhizosphere of plants with inherently different aerenchyma formation: Wheat (Triticum aestivum) and rice (Oryza sativa)

Summary Denitrification in the rhizosphere of wheat and rice was studied in relation to aerenchyma formation. Seedlings were grown in quartz silt amended with mineral nutrients at given bulk densities and water tensions. In adventitious wheat roots the formation of cortical lacunae was strongly dependent on soil aeration. Growing the wheat plants in dry (–20 kPa) and moist substrate (–2 kPa) established aerenchyma contents of 3% and 15%, respectively. Denitrification was measured after the introduction of equal moisture levels in the substrates of both treatments. The higher aerenchyma content of roots pregrown in the wetter substrate did not counteract denitrification in the rhizosphere which had doubled in this treatment. In contrast to the unspecific lysis of cortical cell walls, the well organized formation of aerenchyma in rice roots was independent of soil aeration. Root porosity averaged 14%. As in wheat, it was not related to denitrification. However, the level of denitrification per mg of root dry matter was about four times lower than that of wheat. The addition of decomposable organic matter (cellulose) to the substrate stimulated aerenchyma formation in rice and considerably increased denitrification. The results suggest that denitrification in the rhizosphere is independent of aerenchyma formation.     

Carbon and nitrogen budgets of nematodes in arable soil

Summary The amounts of C and N that pass through the nematode biomass in four cropping systems, barley without and with N fertilization, grass ley and lucerne, has been estimated. The nematodes were sampled at the field site of a Swedish integrated research project Ecology of Arable Land: The Role of Organisms in Nitrogen Cycling. The nematode biomass was lower (200 mg dry weight m^–2) in the annual (barley) than in the perennial (grass and lucerne, 350 mg dry weight m^–2) crops. For respiration, the nematodes used 4–71 O₂m^–2 year^–1 corresponding to C liberation of 1.3%–2.0% of the carbon input to the soil. A higher relative contribution by bacterial-feeding nematodes to the C and N fluxes and a higher turnover rate of the nematode biomass is an indication of more rapid nutrient circulation in the annual than in the perennial cropping systems.     

Soil protozoa and forest tree growth: non-nutritional effects and interaction with mycorrhizae

Mycorrhizal (Lactarius rufus Fr.) and non-mycorrhizal Norway spruce seedlings (Picea abies Karst.) were grown in a sand culture and inoculated with protozoa (naked amoebae and flagellates) extracted from native forest soil or with protozoa grown on agar cultures. A soil suspension from which the protozoa were eliminated by filtration or chloroform fumigation was used as a control. After 19 weeks of growth in a climate chamber at 20–22°C, the seedlings were harvested. Protozoa reduced the number of bacterial colony-forming units extracted from the rhizoplane of both non-mycorrhizal and mycorrhizal seedlings and significantly increased seedling growth. However, concentrations of mineral nutrients in needles were not increased in seedlings with protozoan treatment. It is concluded that the increased growth of seedling was not caused by nutrients released during amoebal grazing on rhizosphere micro-organisms. The protozoa presumably affected plant physiological processes, either directly, via production of phytohormones, or indirectly, via modification of the structure and performance of the rhizosphere microflora and their impact on plant growth. Mycorrhizal colonization significantly increased the abundance of naked amoebae at the rhizoplane. Our observations indicate that protozoa in the rhizosphere interact significantly with mycorrhizae.     

Grazing of protozoa on rhizosphere bacteria alters growth and reproduction of Arabidopsis thaliana

Plant roots are densely colonized by bacteria which form the basis of the rhizosphere bacterial food web with protozoa as most effective predators. We established a well defined laboratory system with Arabidopsis thaliana as model plant allowing to investigate in detail the effect of rhizosphere interactions on plant performance. We used this system to analyse separate and combined effects of natural rhizobacteria and the protozoa Acanthamoeba castellanii on plants.Protozoa and bacteria increased plant growth with the effect of protozoa markedly exceeding that of bacteria only. Arabidopsis immediately responded to the presence of protozoa by increasing carbon but not nitrogen uptake. Later protozoa enhanced plant uptake of nitrogen from organic material and prolonged vegetative growth of Arabidopsis resulting in strongly increased seed production. It is concluded that the immediate plant response was based on changes in rhizosphere signalling inducing increased plant carbon fixation rather than on protozoa-mediated increase in nitrogen availability. The subsequently increased plant nitrogen uptake presumably originated from nitrogen fixed in bacterial biomass made available by protozoan grazing, i.e. the microbial loop in soil. The results suggest that Arabidopsis prepared for the upcoming mobilization of nitrogen by increasing carbon fixation and root carbon allocation which paid-off later by increased nutrient capture and strongly increased plant reproduction.     

化感植物根际生物学特性研究现状与展望

植物的根际是一个复杂的微生态系统,植物的根必须与入侵的邻近植物根及大量以有机物质为营养的细菌、真菌、土存害虫相互竞争空间、水分、矿质营养等。在土壤中,根与根际生物体的相互作用相当复杂且受到许多土壤因素的影响,地下根际生物体以根分泌物为媒介相互作用的机制比发生在地表的生物体的相互作用复杂的多。越来越多的试验表明,根分泌物在根与根、根与根际微生物间起着重要作用,并以其为媒介在植物与环境的相互作用中起着传递信息的作用。本文在简要综述前人研究的基础上,深入探讨了化感植物根际生物学问题及攻克途径,以期为深入研究植物化感作用提供依据。