Effect of heavy metals on growth and survival of Macrophomina phaseolina (Tassi) Goid.

Summary The effects of Cd, Co, Ni and Zn on growth and survival of Macrophomina phaseolina were studied in vitro. Cd, Ni and Co at 500 g ml^–1 inhibited growth by 78.8%, 73.6% and 11.8%, respectively, after 4 days at 25 ± 1°C. The mycelial dry weight yield was enhanced by 2.1% at 100 g ml^–1 Zn. The population of the pathogen declined in soil amended separately with Cd, Co, Ni and Zn. Cd (4000 mg kg^–1) proved to be the most toxic, by completely inhibiting the survival of the pathogen in soil after 20 days.     

Reduction in dose of chemical fertilizers and growth enhancement of sesame (Sesamum indicum L.) with application of rhizospheric competent Pseudomonas aeruginosa LES4

Pseudomonas aeruginosa LES4, an isolate of tomato rhizosphere was found to be positive for several plant growth-promoting attributes like production of indole acetic acid, HCN and siderophore, solubilization of inorganic phosphate along with urease, chitinase and β-1-3-glucanase activity. In addition, it showed strong antagonistic effect against Macrophomina phaseolina and Fusarium oxysporum. P. aeruginosa LES4 caused halo cell formation and other morphological deformities in mycelia of M. phaseolina and F. oxysporum. Root colonization was studied with Tn5 induced streptomycin resistant transconjugants of spontaneous tetracycline-resistant LES4 (designated LES4^tetra+strep+) after different durations. The strain was significantly rhizospheric competent, as 17.4% increase in its population was recorded in sesame rhizosphere. Seed bacterization with LES4 resulted in significant increase in vegetative growth parameters and yield of sesame over non-bacterized seeds. However, application of LES4 with half dose of fertilizers resulted in growth equivalent to full dose treatment, without compromising with the growth and yield of sesame. Moreover, the oil yield increased by 33.3%, while protein yield increased by 47.5% with treatment of half dose of fertilizer along with LES 4 bacterized seeds, as compared to full dose of fertilizers.     

Effects of solar treatment on germination of sclerotia (Sclerotium rolfsii Sacc.) and on other soil mycoflora

Summary Only 10% of sclerotia germinated in a tarpcovered plot after 45 days of treatment while 50% of sclerotia germinated in the control plot. However, complete inhibition of sclerotia germination was not achieved. The maximum temperature recorded was 53°C, after 45 days of treatment in a tarped plot at 1 cm depth. The incidence of Acrophialophora fusispora, Aspergillus spp., Spicaria sp., and Trichoderma viride increased in soil after 45 days of the solar treatment.     

Plant canopy effects on litter accumulation and soil microbial biomass in two temperate forests

The objective of this study was to determine whether differences in canopy structure and litter composition affect soil characteristics and microbial activity in oak versus mixed fir-beech stands. Mean litter biomass was greater in mixed fir-beech stands (51.9t ha⁻¹) compared to oak stands (15.7t ha⁻¹). Canopy leaf area was also significantly larger in mixed stands (1.96m² m⁻²) than in oak stands (1.73m² m⁻²). Soil organic carbon (C _org) and moisture were greater in mixed fir-beech stands, probably as a result of increased cover. Soil microbial biomass carbon (C _mic), nitrogen (N _mic), and total soil nitrogen (N _tot) increased slightly in the mixed stand, although this difference was not significant. Overall, mixed stands showed a higher mean C _org/N _tot ratio (22.73) compared to oak stands (16.39), indicating relatively low rate of carbon mineralization. In addition, the percentage of organic C present as C _mic in the surface soil decreased from 3.17% in the oak stand to 2.26% in the mixed stand, suggesting that fir-beech litter may be less suitable as a microbial substrate than oak litter.     

Fate, tree growth effect and potential impact on soil microbial communities of mycorrhizal and bacterial inoculation in a forest plantation

The knowledge of the survival of inoculated beneficial fungal and bacterial strains in the field and the effects of their release on the indigenous microbial communities has been of great interest since the practical use of selected natural or genetically modified microorganisms has been developing. The aim of this study was to monitor, 4 years after plantation into the field site, the effects of Douglas fir (Pseudotsuga menziesii) co-inoculation with the mycorrhiza helper bacterial strain Pseudomonas fluorescens BBc6R8 and/or the fungal strain Laccaria bicolor S238N on seedling growth and on the indigenous bacterial and ectomycorrhizal communities using quantitative and qualitative approaches. The field persistence of the inoculated strains was also monitored. The seedling shoot volume estimate was statistically significantly higher in the fungal inoculated plots in comparison to the non-inoculated plots but no treatment-related changes in the quantitave or qualitative microbial measurements were observed and the inoculated strains could not be detected after 4 years.     

Changes in active microbial biomass by earthworms and grass amendments in agricultural soil

The influence of the earthworm Aporrectodea caliginosa on the biomass and the proportion of active and dormant soil microorganisms after the addition of cut perennial ryegrass (Lolium perenne) to upper soil from agricultural field was studied in a microcosm experiment. During a 2-month period, soil samples were taken 1, 8, 22, 36, 50, and 64 days after cut grass addition. A substrate-induced respiration (SIR) method was used to analyse the samples for total microbial biomass and the distribution of active and dormant microbial biomass. It was found that the addition of grass increased the microbial biomass (SIR) because of an increase in the active microbial biomass. After the initially high values, the active microbial biomass decreased slowly, and at day 64, it was still higher in the grass-amended soils than in the control treatment without grass addition. After 1 day, the active microbial biomass was higher in the soil with A. caliginosa than without the earthworm. At the subsequent samplings, there were no differences in microbial biomass or the proportion of dormant vs active microorganisms between the grass-amended soils. The average from all sampling occasions of SIR was higher in earthworm-treated soil.     

Agricultural factors affecting methane oxidation in arable soil

CH₄ oxidation activity in a sandy soil (Ardoyen) and agricultural practices affecting this oxidation were studied under laboratory conditions. CH₄ oxidation in the soil proved to be a biological process. The instantaneous rate of CH₄ consumption was in the order of 800 mol CH₄ kg^–1 day^–1 (13 mg CH₄ kg^–1 day^–1) provided the soil was treated with ca. 4.0 mmol CH₄ kg^–1 soil. Upon repeated supplies of a higher dose of CH₄, the oxidation was accelerated to a rate of at least 198 mg CH₄ kg^–1 day^–1. Addition of the plant-growth promoting rhizopseudomonad strains Pseudomonas aeruginosa 7NSK2 and Pseudomonas fluorescens ANP15 significantly decreased the CH₄ oxidation by 20 to 30% during a 5-day incubation. However, with further incubation this suppression was no longer detectable. Growing maize plants prevented the suppression of CH₄ oxidation. The numbers of methanotrophic bacteria and fungi increased significantly after the addition of CH₄, but there were no significant shifts in the population of total bacteria and fluorescent pseudomonads. Drying and rewetting of soil for at least 1 day significantly reduced the activity of the indigenous methanotrophs. Upon rewetting, their activity was regained after a lag phase of about 3 days. The herbicide dichlorophenoxy acetic acid (2,4-D) had a strong negative effect on CH₄ oxidation. The application of 5 ppm increased the time for CH₄ removal; at concentrations above 25 ppm 2,4-D CH₄–oxidizing activity was completely hampered. After 3 days of delay, only the treatments with below 25 ppm 2,4-D showed recovery of CH₄–oxidizing activity. This finding suggests that it can be important to include a CH₄–removal bioassay in ecotoxicology studies of the side effects of pesticides. Changes in the native soil pH also affected the CH₄–oxidizing capacity. Permanent inhibition occurred when the soil pH was altered by 2 pH units, and partial inhibition by 1 pH unit change. A rather narrow pH range (5.9–7.7) appeared to allow CH₄ oxidation. Soils pre-incubated with NH ₄ ⁺ had a lower CH₄–removal capacity. Moreover, the nitrification inhibitor 2-chloro-6-trichloromethyl pyridine (nitrapyrin) strongly inhibited CH₄ oxidation. Probably methanotrophs rather than nitrifying microorganisms are mainly responsible for CH₄ removal in the soil studied. It appears that the causal methanotrophs are remarkably sensitive to soil environmental disturbances.     

Relationship of root necrosis potential of soil to asparagus (Asparagus officinalis) decline in Switzerland

Summary The possible relationship between asparagus decline and the root necrosis potential (RNP) of soil was investigated for 11 asparagus and 4 nonasparagus soils. Asparagus seedlings were grown in each soil in climate chambers. A root necrosis index was used to determine RNP. RNP was correlated with the decline observed in the field for seven white asparagus crops, but not for three white and one green asparagus crops. Low RNP values were obtained from the soils not planted to asparagus.Fusarium oxysporum andF. oxysporum var. redolens accounted for 7070 of the fungi isolated from necrotic roots in RNP tests. BothFusarium were obtained from all soils. AllF. oxysporum andF. oxysporum var. redolens isolates tested were highly virulent on asparagus seedlings in inoculation experiments.F. solani, Penicillium verrucosum var.corymbiferum, andRhizoctonia violacea were isolated less frequently but were also highly virulent. It is concluded that soil RNP indicated a risk of decline caused by fusaria, but other soil factors were likely to be involved in the aetiology of decline. The study of these factors is necessary to develop a method for the prognosis of decline associated with fusaria.     

Leaching of genetically modified Pseudomonas fluorescens through intact soil microcosms: Influence of soil type

Summary The leaching of a genetically modified Pseudomonas fluorescens through soil was investigated using intact (undisturbed) soil microcosms over a 2-month period. The microcosms comprised large cylindrical cores of three contrasting soil types (a loamy sand, a sandy loam, and a clay loam) supporting a grass/clover sward. Late log-phase cells of Pseudomonas fluorescens containing lux genes encoding for bioluminescence were applied to the surface of the soil cores. Eighteen hours after application of the inocula, the microcosms were subjected to simulated rain events (9 mm per event) at 3-day intervals and leachates were analysed for the concentration of genetically modified bacteria. The lux-modified pseudomonads were detected immediately in leachate from the clay looam with a steady decline in the concentration of cells with time. Leaching of pseudomonads from the sandy loam and loamy sand only occurred over a few rain events and total recoveries from the leachate were lower than from the clay loam. Leaching patterns are discussed in relation to differences in structure of topsoil and subsoil, which determine the pathways of water flow, and to the matric potential at inoculation, which determines the pore-size classes into which cells were first introduced.     

Effects of Folsomia candida (Collembola) on the microbial biomass in a grassland soil

Summary The effects of the presence of Folsomia candida on substrate-induced respiration, CO₂-C evolution, bacterial count and NH ₄ ⁺ -N were investigated in a grassland soil. Differences in these parameters, with the exception of NH ₄ ⁺ , were correlated with the age of the collembolan Folsomia candida. In the presence of juvenile animals total CO₂-C evolution was enhanced, but substrate-induced respiration and the bacterial count were unchanged. In fumigated soil with imagos, substrate-induced respiration and the number of bacteria were increased, but total CO₂-C evolution was unaltered. Different food selection strategies between adults and juvenile animals may explain the results.     

Effect of N-(n-butyl) thiophosphoric triamide on hydrolysis of urea by plant,microbial, and soil urease

Summary Comparison of the effects of N-(n-butyl) thiophosphoric triamide (NBPT) and phenylphosphorodiamidate (PPD) on hydrolysis of urea by plant (jackbean), microbial (Bacillus pasteurii), and soil urease showed that whereas NBPT was considerably more effective than PPD for inhibiting hydrolysis of urea added to soil, it was much less effective than PPD for inhibiting hydrolysis of urea by plant or microbial urease. Studies to account for this observation indicated that NBPT is rapidly decomposed in soil to a compound that is much more effective than NBPT for inhibition of urease activity and that this compound is N-(n-butyl) phosphoric triamide.     

Survival and potential denitrifying activity of Azospirillum lipoferum and Bradyrhizobium japonicum inoculated into sterilized soil

Summary Potential denitrifying activity and population dynamics of Azospirillum lipoferum (137C) and Bradyrhizobium japonicum (G2sp) inoculated into a -sterilized soil were studied for a period of 3 weeks. The denitrifying enzyme potential of soil inoculated independently with each bacterial species was strongly stimulated by the presence of a plant (Zea mays L.). Simultaneous inoculation of both bacteria also produced a higher denitrifying enzyme potential than simple inoculation. Even with double inoculation, the presence of a plant did not modify the evolution of the activity. The response of the population dynamics to these treatments followed a different pattern. The population dynamics of A. lipoferum was not affected by the presence of the plant or by the presence of B. japonicum. In contrast, the presence of both a plant and of A. lipoferum seemed to promote the growth of B. japonicum.     

Diversity and Characterization of Potential H2-Dependent Fe(Ⅲ)-Reducing Bacteria in Paddy Soils

Microbial ferric iron reduction, with organic carbon or hydrogen as the electron donor, is one of the most important biogeochemical processes in anoxic paddy soils; however, the diversity and community structure of hydrogen-dependent dissimilatory iron-reducers remain unknown. Potential H2-dependent Fe(III)-reducing bacteria in paddy soils were explored using enrichment cultures with ferrihydrite or goethite as the electron acceptor and hydrogen as the electron donor. Terminal restriction fragment length polymorphism (T-RFLP) analysis and cloning/sequencing were conducted to reveal bacterial community structure. Results showed that Geobacter and Clostridium were the dominant bacteria in the enrichment cultures. Fe(III) oxide mineral phases showed a strong effect on the community structure; Geobacter and Clostridium were dominant in the ferrihydrite treatment, while Clostridium spp. were dominant in the goethite treatment. These suggested that H2-dependent Fe(III)-reducing bacteria might be widely distributed in paddy soils and that besides Geobacter, Clostridium spp. might also be an important group of H2-dependent Fe(III)-reducing microorganisms.     

The role of the earthworm Lumbricus terrestris in the transport of bacterial inocula through soil

Two laboratory experiments were used to investigate the effect of the earthworm Lumbricus terrestris on transport of genetically marked Pseudomonas fluorescens inocula through soil microcosms. The microcosms comprised cylindrical cores of repacked soil with or without earthworms. Late log-phase cells of P. fluorescens, chromosomally marked with lux genes encoding bioluminescence, were applied to the surface of soil cores as inoculated filter paper discs. In one experiment, 5 and 10 days after inoculation, cores were destructively harvested to determine concentrations of marked pseudomonads with depth relative to the initial inoculum applied. Transport of the bacteria occurred only in the presence of earthworms. In a second experiment cores were subjected to simulated rainfall events 18 h after inoculation with lux-marked bacteria at 3-day intervals over a 24-day period. Resulting leachates were analysed for the appearance of the marked bacteria, and after 28 days cores were destructively harvested. Although some marked cells (less than 0.1% of the inoculum applied) were leached through soil in percolating water, particularly in the presence of earthworms, the most important effect of earthworms on cell transport was through burial of inoculated litter rather than an increase in bypass flow due to earthworm channels.     

Plant growth and microbial activity in a tropical soil amended with faecal pellets from millipedes and woodlice

The effect of soil fauna-mediated leaf litter (faecal pellets) versus mechanically fragmented (finely ground) leaf litter on biomass production of rice (Oryza sativa, var. Primavera) was assessed in pot tests. Rice seedlings were either grown in soil samples amended with faecal pellets of diplopods and isopods fed on leaf litter of a legume cover crop (Pueraria phaseoloides (Roxb.) Benth) and a peach palm (Bactris gasipaes) or in soil amended with finely ground leaf litter. The addition of faecal pellets caused a significant and dose-related increase in plant biomass compared to pure soil. Ground leaf litter induced a significantly smaller positive effect on plant biomass development with Pueraria litter > Bactris litter > mixed primary forest litter. In contrast, soil microbial biomass development during the 4 weeks plant test was higher in the soil amended with ground litter as compared to soil amended with feacal pellets. The results show a clear positive effect of the soil fauna on soil fertility and indicate differences in the availability of nutrients from the organic substrates to higher plants and soil microorganisms.     

转基因水稻秸秆还田对土壤硝化反硝化微生物群落的影响

转基因作物可能通过根系分泌物和植株残体组成的改变及外源基因的转移释放令土壤微生物群落产生变化,影响土壤微生物的生态功能。氨氧化细菌和反硝化细菌是驱动土壤硝化和反硝化过程的关键微生物,其群落结构的变化直接关系土壤氮素的转化与利用。本研究利用荧光定量PCR和PCR-DGGE技术分析了转cry1Ac/cpti双价抗虫基因水稻‘Kf8’秸秆还田降解过程中,土壤氨氧化细菌和反硝化细菌群落丰度与组成的变化,探讨转基因水稻是否存在影响稻田土壤氮素转化与N2O排放的可能。结果显示:无论是氨氧化细菌amo A基因还是反硝化细菌nirS基因,其丰度在转基因水稻‘Kf8’与非转基因水稻‘Mh86’的秸秆还田土壤中都没有显著差异;转基因水稻‘Kf8’和非转基因水稻‘Mh86’秸秆还田降解过程中0~10 cm土层中的amo A基因丰度均显著高于10~20 cm及20~30 cm土层(P0.05);各深度土层中的nirS基因丰度均存在随秸秆还田时间延长而增加的趋势。水稻秸秆还田降解过程中,转基因水稻‘Kf8’的土壤氨氧化细菌和反硝化细菌的群落多样性指数及组成,均与非转基因水稻‘Mh86’没有显著差异。相关分析结果表明土壤氨氧化细菌和反硝化细菌群落组成均与水稻秸秆还田时间存在显著相关性(P=0.002),反硝化细菌群落组成还与土层深度显著相关(P=0.024)。本研究表明转cry1Ac/cpti抗虫基因水稻秸秆还田对稻田土壤硝化和反硝化关键微生物群落不会产生明显影响。就土壤微生物群落而言,转cry1Ac/cpti抗虫基因水稻秸秆还田不存在影响土壤氮素转化与N2O排放的可能。     

Influence of soil properties on the vertical movement of genetically-marked Pseudomonas fluorescens through large soil microcosms

Summary Vertical translocation of the introduced transposon Tn5-tagged Pseudomonas fluorescens cells was studies after irrigation of 50-cm long soil columns of loamy sand. The soil in the columns was slowly brought to saturation using groundwater, and enough water was then slowly added to permit collection of the percolated water. Introduced bacteria were transported to lower soil layers to a significantly higher degree in undisturbed soil cores than in repacked cores; water transport was hampered in both core types due to high soil bulk densities. Soil bulk density affected the degree of transport of the introduced cells; progressively more cells were translocated to deeper soil layers and into the percolation water at decreasing soil bulk densities. Repeated percolation of soil at a bulk density of 1.25 caused an increase in Tn5-tagged cell numbers in the lower soil layers and in the percolated water. Further, cells initially introduced into a dry (5.3% moisture) soil were translocated to a lesser extent than cells introduced into a wetter (13% moisture) soil. Finally, wheat roots enhanced the water-induced transport of introduced cells to the 40- and 50-cm deep soil layers and into the effluent, but not to the remaining soil layers. Large soil columns such as those used in the present study are useful in assessing the transport and survival of introduced bacterial cells in soils under a variety of simulated environmental conditions.     

Effect of certain herbicides on soil microbial populations and their influence on saprophytic growth in soil and pathogenicity of take-all fungus

Summary The application of diquat + paraquat, glyphosate and trifluralin to unsterilized field soil increased take-all caused by the fungus, Gaeumannomyces graminis var. tritici Walker by 13.0% 16.6% and 10.8% respectively, while no effect on disease was recorded in sterilized soil treated with the same herbicides. The herbicides tested had no effect on the saprophytic growth of the pathogen with the exception of glyphosate, which increased its growth in unsterilized soil. The application of diquat + paraquat and glyphosate to unsterile soil had no effect on the numbers of actinomycetes. The diquat + paraquat treatment, however, increased populations of fungi while the glyphosate decreased the numbers of bacteria. The proportion of soil fungi antagonistic to the pathogen was reduced in glyphosate-treated soil. The frequency of occurrence of Eupenicillium euglaucum (v. Beyma) Stolk & Samson (strain B), and Penicillium verruculosum Peyr. (strain B), which were strong and low level antagonists of Ggt on agar, were reduced in their occurrence in soil by 7.7% and 2.5% respectively, following glyphosate treatment. Moreover, the numbers of Aspergillus viridinutans Ducker & Thrower, which showed moderate antagonism to the pathogen, was decreased by 1.9% and 4.1% in diquat + paraquat and glyphosate treatments respectively. The proportion of antagonists rather than total numbers of fungi appears to be related to the treatment effect observed on the soil growth and pathogenicity of G. graminis var. tritici in our investigation. The increase in disease of wheat in certain herbicide-treated soils may be due to the shift in soil microbial populations away from those which are antagonistic to the pathogen.     

Soil physicochemical and microbial drivers of temperature sensitivity of soil organic matter decomposition under boreal forests

Soil organic matter(SOM)in boreal forests is an important carbon sink.The aim of this study was to assess and to detect factors controlling the temperature sensitivity of SOM decomposition.Soils were collected from Scots pine,Norway spruce,silver birch,and mixed forests(O horizon)in northern Finland,and their basal respiration rates at five different temperatures(from 4 to 28℃)were measured.The Q_(10) values,showing the respiration rate changes with a 10℃ increase,were calculated using a Gaussian function and were based on temperature-dependent changes.Several soil physicochemical parameters were measured,and the functional diversity of the soil microbial communities was assessed using the MicroResp?method.The temperature sensitivity of SOM decomposition differed under the studied forest stands.Pine forests had the highest temperature sensitivity for SOM decomposition at the low temperature range(0–12℃).Within this temperature range,the Q_(10) values were positively correlated with the microbial functional diversity index(H'_(mic))and the soil C-to-P ratio.This suggested that the metabolic abilities of the soil microbial communities and the soil nutrient content were important controls of temperature sensitivity in taiga soils.     

Relation between soil microbial activity and the effect of seed inoculation with the rhizopseudomonad strain 7NSK2 on plant growth

Summary The relationship between the microbial activity in the soil and the effect of seed inoculation with the rhizopseudomonad strain 7NSK2 was evaluated in a series of pot experiments under greenhouse conditions. The microbial activity in plain soil, as measured by the respiratory activity, was significantly increased by the growth of the plants. Both the respiration rate of the microorganisms and the density of the bacteria and fungi in the bulk soil increased with increasing duration of the plant growth. Upon repeated short-term growth of plants on the same soil, a similar stimulation was noticed.The effect of seed inoculation on the growth of the maize cultivar Beaupré and the barley cultivar Iban was most pronounced in the microbiologically more active soils. The results suggest that the increase of the plant growth by seed inoculation is probably due to the inhibition of deleterious root microorganisms.