Comparing Metabolic Functionalities,Community Structures,and Dynamics of Herbicide-Degrading Communities Cultivated with Different Substrate Concentrations

Two 4-chloro-2-methylphenoxyacetic acid (MCPA)-degrading enrichment cultures selected from an aquifer on low (0.1 mg liter⁻¹) or high (25 mg liter⁻¹) MCPA concentrations were compared in terms of metabolic activity, community composition, population growth, and single cell physiology. Different community compositions and major shifts in community structure following exposure to different MCPA concentrations were observed using both 16S rRNA gene denaturing gradient gel electrophoresis fingerprinting and pyrosequencing. The communities also differed in their MCPA-mineralizing activities. The enrichments selected on low concentrations mineralized MCPA with shorter lag phases than those selected on high concentrations. Flow cytometry measurements revealed that mineralization led to cell growth. The presence of low-nucleic acid-content bacteria (LNA bacteria) was correlated with mineralization activity in cultures selected on low herbicide concentrations. This suggests that LNA bacteria may play a role in degradation of low herbicide concentrations in aquifers impacted by agriculture. This study shows that subpopulations of herbicide-degrading bacteria that are adapted to different pesticide concentrations can coexist in the same environment and that using a low herbicide concentration enables enrichment of apparently oligotrophic subpopulations.     

Effect of the insecticide actellic on soil microorganisms and their activity

The insecticide Actellic or its active component pirimiphosmethyl affected the growth and reproduction of some soil bacteria and fungi, fixation of atmospheric nitrogen, nitrification and overall soil metabolic activity. Both bacterial growth and the activities under study were negatively influenced by high concentrations of the agent (as compared with actual pirimiphosmethyl concentrations routinely used in agriculture) whereas lower concentrations stimulated the overall metabolic soil activity.     

Nitrogen Fixation by Photosynthetic Bacteria in Lowland Rice Culture

Propanil (3′,4′-dichloropropionanilide) was a potent inhibitor of the nitrogenase activity of blue-green algae (cyanobacteria) in flooded soil, but the herbicide at comparable concentrations was not toxic to rice, protozoa, and nitrogen-fixing bacteria. Ethanol-amended flooded soils treated with propanil exhibited higher rates of nitrogenase activity than those not treated with the herbicide. The enhanced nitrogenase activity in propanil-treated soils was associated with a rise in the population of purple sulfur bacteria, especially of cells resembling Chromatium and Thiospirillum. By employing propanil and a means of excluding light from the floodwater to prevent the development of phototrophs during rice growth under lowland conditions, the relative activities of blue-green algae, photosynthetic bacteria, and the rhizosphere microflora were determined. The results suggest that the potential contribution of photosynthetic bacteria may be quite high.     

Comparative effects of four herbicides on physiological aspects in Triticum sativum L.

Four herbicide (topik, puma super, granstar and derby) treatments at the recommended field dose (RFD) were applied for 5 weeks on wheat (Triticum sativum L. cv. Giza 168) crop. These selected herbicides are recommended to control two types (narrow and broad) of weeds in this crop. The growth parameter, oxidative stress defence system, nitrate reduction, proline level and protein contents were used to evaluate the toxic effect of these herbicides on wheat crop. The toxicity of herbicides used on this crop was as follows: puma super > topik > derby > granstar. The correlation between the herbicide concentrations and the activities of the antioxidant enzymes was monitored.     

除草剂二氯喹啉酸对水稻田土壤中微生物种群的影响

对好氧微生物采用平板稀释法,厌氧微生物采用最大或然计数法和滚管法研究土壤中施入0．33、0．67、1．00、1．33、2．00μg·g^-1干土除草剂二氯喹啉酸后对土壤可培养微生物种群数量的影响。结果表明,各种微生物对二氯喹啉酸的反应随其施加浓度的不同而有差异．二氯喹啉酸对水稻田土壤中好氧性细菌、水解发酵细菌、反硝化细菌数量的影响都是短暂的,第33d时均能恢复至接近对照水平,浓度在1．33μg·g^-1干土以下时二氯喹啉酸促进真菌数量增加,高于该浓度时则具有抑制作用．施用各浓度二氯喹啉酸初期,对土壤中放线菌和产甲烷菌有一定程度的抑制效应,但低浓度时抑制效应在培养后期消失．正常土壤施用量的二氯喹啉酸(即0．67μg·g^-1干土)对水田土壤微生物各种群无实质危害级农药。     

Effect of Zytron and Its Degradation Products on Soil Microorganisms

There was no adverse effect of Zytron, o-2,4-dichlorophenyl o-methyl isopropyl phosphoramidothioate, a herbicide, upon molds, actinomycetes, and soil bacteria in field plots, or upon selected soil microorganisms in model systems. 2,4-Dichlorophenol, a degradation product, was found to be toxic at levels above 10 ppm to molds, but levels this high were not found in soil from treated plots. Aspergillus clavatus degraded both Zytron and 2,4-dichlorophenol. Sodium o-methyl isopropyl phosphoramidothioate, another degradation product of Zytron, stimulated the growth of a species of Penicillium.     

Plant growth promoting bacteria from Crocus sativus rhizosphere

Present study deals with the isolation of rhizobacteria and selection of plant growth promoting bacteria from Crocus sativus (Saffron) rhizosphere during its flowering period (October–November). Bacterial load was compared between rhizosphere and bulk soil by counting CFU/gm of roots and soil respectively, and was found to be ~40 times more in rhizosphere. In total 100 bacterial isolates were selected randomly from rhizosphere and bulk soil (50 each) and screened for in-vitro and in vivo plant growth promoting properties. The randomly isolated bacteria were identified by microscopy, biochemical tests and sequence homology of V1–V3 region of 16S rRNA gene. Polyphasic identification categorized Saffron rhizobacteria and bulk soil bacteria into sixteen different bacterial species with Bacillus aryabhattai (WRF5-rhizosphere; WBF3, WBF4A and WBF4B-bulk soil) common to both rhizosphere as well as bulk soil. Pseudomonas sp. in rhizosphere and Bacillus and Brevibacterium sp. in the bulk soil were the predominant genera respectively. The isolated rhizobacteria were screened for plant growth promotion activity like phosphate solubilization, siderophore and indole acetic acid production. 50 % produced siderophore and 33 % were able to solubilize phosphate whereas all the rhizobacterial isolates produced indole acetic acid. The six potential PGPR showing in vitro activities were used in pot trial to check their efficacy in vivo. These bacteria consortia demonstrated in vivo PGP activity and can be used as PGPR in Saffron as biofertilizers.This is the first report on the isolation of rhizobacteria from the Saffron rhizosphere, screening for plant growth promoting bacteria and their effect on the growth of Saffron plant.     

“使它隆”对玉米根部不同微生环境细菌群落多样性的影响

【目的】研究除草剂"使它隆"施用后对玉米根部不同微生环境细菌群落结构和多样性的影响。【方法】利用Illumina Miseq高通量测序技术,对玉米根系内生菌、根际和非根际土壤细菌16S rRNA的V4–V5可变区序列进行测定,分析不同生长期喷施除草剂使它隆对玉米土壤细菌及根系内生菌群落结构和多样性的影响。【结果】本研究15个样品共得到544393条有效序列,333565条优质序列。多样本共有OTU分析表明,非根际和根际土壤的群落结构更为相似,在一定程度上说明玉米根部相关细菌的定殖具有选择性并且是从根际到根系逐步专一化。丰度等级曲线和Alpha多样性结果显示非根际和根际土壤群落的丰富度和均匀度较高,而玉米根系内生菌群落的丰富度和均匀度都比较低,且成熟期玉米根系内生菌群落的丰富度在施用除草剂使它隆后下降比较剧烈。群落组成分析发现,使它隆除草后,玉米根部相关细菌各时期在门及属水平上的分布都发生了较大变化。菌群代谢功能预测结果表明玉米生长从苗期到成熟期,微生物的生长压力逐渐加大,需要消耗更多的能量用于新陈代谢和环境适应。【结论】施用除草剂使它隆后会降低玉米根部土壤细菌群落的多样性,使它隆对成熟期玉米根系内生菌群落影响最为显著。     

Contribution to nitrogen fixation and soil fertility by photosynthetic bacteria

Summary The nitrogen fixation ability ofRhodopseudomonas capsulatus (a member of the photosynthetic bacteria) has been investigated. This organism can fix N₂ most effectively under illuminated anaerobic conditions. However, in mixed culture in symbiotic association with heterotrophic bacteria, this microorganism using pyruvic acid excreted by the heterotrophs is capable of fixing nitrogen even under an apparent aerobic environment. It has been demonstrated that some correlation exists between the growth of photosynthetic bacteria and the reproduction of the rice plant. Compared to the mineral fertilizer, application of photosynthetic bacteria at the reproductive stage of rice plants increased the yield of grain. This was confirmed by the fact that the root system of rice is capable of absorbing amino acids and nucleic acids excreted by photosynthetic bacteria. Uracil and proline have the most influence on rice reproduction. This is also true for tomato plants. Many toxic molecules such as hydrogen sulfide, amines, etc. are found in soil. They are metabolized by photosynthetic bacteria, which contributes to the detoxication of soil. Such findings were extended to the purification of polluted waste waters from industry and domestic sources,etc. It can be concluded that photosynthetic bacteria contribute very significantly to soil fertility and improvement of the plant growth condition.     

Successive soil treatment with captan or oxytetracycline affects non-target microorganisms

Changes in microbial biomass and activity were determined in a sandy-loam soil treated with successive dosages of oxytetracycline (a bactericide) or captan (a fungicide) throughout 98 days of incubation under laboratory conditions. The numbers of culturable bacteria and fungi, total bacterial and fungal biomass (as amounts of phospholipid fatty acids, PLFA), the fungal/bacterial ratio, activities of acid and alkaline phosphatases and urease as well as concentrations of N-NH₄ ⁺ and N-NO₃ ⁻ were assessed. Both oxytetracycline and captan significantly decreased numbers of culturable bacteria whereas total bacterial biomass (bactPLFA) was not affected. Oxytetracycline did not effect on the fungal biomass, however their numbers were reduced after the first and second time of soil amendment with the bactericide. Conversely, fungal numbers and biomass (PLFA 18:2ω6,9) significantly decreased in response to soil treatment with the fungicide. Compared to oxytetracycline, captan significantly decreased activities of acid and alkaline phosphatases. For urease activity, the decreased activity was only observed in the soil after the third dosage of captan. Both biocides significantly increased concentrations of N-NH₄ ⁺ and decreased concentrations of N-NO₃ ⁻ after the soil treatments. The results of this study indicate that successive soil treatment with oxytetracycline or captan dosages may negatively affect non-target soil microorganisms and their activities.     

The effects of Fusarium oxysporum f.sp. lycopersici (Sacc.) Snyder & Hansen on tomato in diphenamid-treated soil

Pre-emergence soil application of the herbicide diphenamid in concentrations exceeding the normal field rate increased the resistance of tomato plants towards infection by the wilt fungus Fusarium oxysporum f.sp. lycopersici. This was detected as significant increases in the percentage emergence of seedlings although growth parameters of the raised seedlings were reduced. Treated plants exhibited no wilt symptoms, although the pathogen maintained its population at detectable levels in the rhizosphere of tomato plants. However, the growth inhibition caused by diphenamid alone was much less than that reported for the combined application of pathogen and herbicide. Growth activities of F. oxysporum f.sp. lycopersici were inhibited by high concentrations of diphenamid in vitro. It is possible that the biodegradation of this herbicide by species such as Aspergillus candidus (present in substantial counts in treated rhizospheres) was one of the causes of increased tolerence of the pathogen to the herbicide in situ.     

Silicon increases boron tolerance and reduces oxidative damage of wheat grown in soil with excess boron

The effect of silicon on the growth, boron concentrations, malondialdehyde (MDA) content, lipoxygenase (LOX) activity, proline (PRO) and H₂O₂ accumulation, and the activities of major antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX)] and non-enzymatic antioxidants (AA) of wheat grown in soil originally with toxic B concentrations were investigated. Applied of 5.0 and 10.0 mM Si to the B toxic soil significantly increased Si concentration of the wheat and counteracted the deleterious effects of B on shoot growth. The contents of PRO, H₂O₂, MDA, and LOX activity of wheat grown in B toxic soil were significantly reduced by Si treatments. Compared with control plants, the activities of SOD, CAT, APX and content of AA were decreased by applied Si. Based on the present work, it can be concluded that Si alleviates B toxicity of wheat by preventing oxidative membrane damage and also translocation of B from root to shoot and/or soil to plant.     

Responses of soil nitrogen-fixing, ammonia-oxidizing, and denitrifying bacterial communities to long-term chlorimuron-ethyl stress in a continuously cropped soybean field in Northeast China

Chlorimuron-ethyl is a type of long-residual herbicide applied widely to soybean fields in China, but little information is available about the long-term impact of this herbicide on soil nitrogen-transforming microbial communities. Soil samples (0–20 cm) were collected from three treatments (no, 5-year and 10-year application of chlorimuron-ethyl) in a continuously cropped soybean field. Plate count (CFU), most probable number (MPN) count, and clone library analyses were conducted to investigate the abundance and composition of nitrogen-fixing, ammonia-oxidizing, and denitrifying bacterial communities, and a chlorate inhibition method was adopted to measure the soil nitrification potential. Long-term chlorimuron-ethyl application reduced the abundance of soil culturable nitrogen-fixing, ammonia-oxidizing, and denitrifying bacteria. Moreover, chlorimuron-ethyl decreased the diversity of nitrogen-fixing and ammonia-oxidizing bacteria but promoted that of denitrifying bacteria. Chlorimuron-ethyl restrained some uncultured nitrogen-fixing bacteria, ammonia-oxidizing bacteria Nitrosospira sp. cluster 3a and 3d, and some novel or putative denitrifying bacteria. The nitrogen-fixing bacteria were closely related to Bradyrhizobium sp., ammonia-oxidizing bacteria Nitrosospira sp. cluster 3b and 3c, and most denitrifying bacteria were resistant to chlorimuron-ethyl. There was a negative correlation between the nitrification potential and the residual amount of soil chlorimuron-ethyl (R²?=?0.88, n?=?3, P?<?0.05). Therefore, long-term application of chlorimuron-ethyl in the continuously cropped soybean field could seriously disturb soil N-transforming communities, and might impact soybean soil biological quality and soybean growth. Further studies should address rational amendment models of this herbicide to reduce the possible ecological risks of long-term application of this herbicide to soybean fields.     

Responses of the Anaerobic Bacterial Community to Addition of Organic C in Chromium(VI)- and Iron(III)-Amended Microcosms

Chromium (VI) is toxic to microorganisms and can inhibit the biodegradation of organic pollutants in contaminated soils. We used microcosms amended with either glucose or protein (to drive bacterial community change) and Fe(III) (to stimulate iron-reducing bacteria) to study the effect of various concentrations of Cr(VI) on anaerobic bacterial communities. Microcosms were destructively sampled based on microbial activity (measured as evolution of CO₂) and analyzed for the following: (i) dominant bacterial community by PCR-denaturing gradient gel electrophoresis (DGGE) of the 16S rRNA gene; (ii) culturable Cr-resistant bacteria; and (iii) enrichment of iron-reducing bacteria of the Geobacteraceae family by real-time PCR. The addition of organic C stimulated the activities of anaerobic communities. Cr(VI) amendment resulted in lower rates of CO₂ production in glucose microcosms and a slow mineralization phase in protein-amended microcosms. Glucose and protein amendments selected for different bacterial communities. This selection was modified by the addition of Cr(VI), since some DGGE bands were intensified and new bands appeared in Cr(VI)-amended microcosms. A second dose of Cr(VI), added after the onset of activity, had a strong inhibitory effect when higher levels of Cr were added, indicating that the developing Cr-resistant communities had a relatively low tolerance threshold. Most of the isolated Cr-resistant bacteria were closely related to previously studied Cr-resistant anaerobes, such as Pantoea, Pseudomonas, and Enterobacter species. Geobacteraceae were not enriched during the incubation. The studied Cr(VI)-contaminated soil contained a viable anaerobic bacterial community; however, Cr(VI) altered its composition, which could affect the soil biodegradation potential.     

Effects of sulfate on cadmium uptake by Swiss chard: II. Effects due to sulfate addition to soil

Sulfate complexation of Cd in nutrient solution has been shown to have little impact on Cd uptake by plants. This study examined the effect of sulfate added to soil on Cd concentrations in soil solution and Cd uptake by Swiss chard (Beta vulgaris L. cv. Fordhook Giant). Swiss chard was grown in soil which was wetted with complete nutrient solution containing equivalent salt concentrations of NaNO₃ or Na2SO₄. Plant growth was reduced by increasing both NO₃ and SO₄ concentrations in soil solution, with growth reductions similar for both salts. The Cd concentration in soil solution increased P< 0.05) more consistently with increasing concentrations of SO₄ compared to NO₃ in soil solution. Solution speciation, calculated with GEOCHEM-PC, showed significant increases of Cd2+ activities with increasing salt rates. Shoot Cd content in 19-day-old Swiss chard plants was marginally but significantly P <0.05) increased with increasing SO₄ concentration but no effect was observed with increasing NO₃ concentration. These results are compared with earlier work on the marked effect of Cl- salinity on Cd availability in Swiss chard. Possible mechanisms explaining the smaller effect of SO₄ compared to Cl on Cd availability are proposed.     

Using Soil Available P and Activities of Soil Dehydrogenase and Phosphatase as Indicators for Biodegradation of Organophosphorus Pesticide Methamidophos and Glyphosate

The heavy use of organophosphorus pesticides in northeastern China strongly affects the ecological functions and the quality of the soil environment. In this work, a 30-day soil incubation experiment was conducted to evaluate the potential of using soil available P and the activities of soil dehydrogenase and acid phosphatase as indicators of the application of methamidophos and glyphosate. Two kinds of unpolluted soils, phaiozem and burozem, were selected as the test soils. The higher application rate of organophosphorus pesticide to the two soils caused more release of PO₄ ^3? which finally entered the soil available P pool, suggesting that soil available P is one of the effective chemical markers for biodegradation of organophosphorus pesticides. Methamidophos exhibited a significant inhibitory effect on the activity of soil dehydrogenase. The extent of enzyme inhibition was almost positively correlated with the insecticide concentration, and the enzyme activity was gradually restored after day 15. However, its effect on soil acid phosphatase activity (stimulation or inhibition) seemed to be indefinite, and varied with the application rate, soil type, and incubation time. In the case of glyphosate, soil acid phosphatase activity was depressed significantly and the depressing extent could be a function of herbicide concentration and incubation time, but soil dehydrogenase activity showed an irregular variation with the herbicide application rate and soil type. In general, dehydrogenase activity was a good biochemical indicator for the biodegradation of methamidophos, but for glyphosate biodegradation the indicator was acid phosphatase activity.     

Soil microbial community indices as predictors of soil solution chemistry and N leaching in Picea abies (L.) Karst. forests in S. Sweden

High rates of inorganic nitrogen (N) deposition or internal N turnover increases the risks of N loss from forests with negative effects on stream water quality. We hypothesized that soil fungi may be more important N sinks than bacteria, and thus examined the impact of soil microbial community composition on N leaching from forests. We studied 19 spruce stands to examine relationships between microbial community composition, stem growth, soil-, and lysimeter-collected soil solution characteristics, and N leaching. We used nitrate concentration in the soil solution below the rooting zone as an N leaching index and phospholipid fatty acid (PLFA) analysis for characterisation of microbial communities. Microbial community composition in the organic horizon and soil solution chemistry below the rooting zone was highly correlated. Stands with low concentrations of nitrate (NO₃ ^?) and aluminium (Al) had higher fungi: bacteria ratio compared with stands with higher concentrations of NO₃ ^? and Al. Stem growth and fungi: bacteria ratio explained 70 % of the variation in N and Al leaching. We identified three microbial predictors of variation in soil solution chemistry, of which the fungi: bacteria was the strongest. The other two were putative indicators of microbial C limitation, a condition known to stimulate N mineralisation and nitrification.     

Impact of an Herbicide Combination of Bromoxynil and Prosulfuron on Soil Microorganisms

Soil microcosm experiments were used to investigate the effects on growth and activity of soil microorganisms of an herbicide combination (60% bromoxynil + 3% prosulfuron) frequently used to provide a broad spectrum control of weed species. Culturable aerobic bacteria, fungi, and actinomycetes, the fundamental groups of heterotrophic microorganisms, and nitrifiers, considered a very sensitive group to these compounds, were evaluated. Since herbicides have been found to inhibit decomposition of cellulose in soil, the effects on cellulolytic bacteria and fungi were determined. Dehydrogenase activity as a measure of microbial activity was another parameter considered. The results emphasized a tendency of reversible stimulatory/inhibitory effects of the tested compounds on soil microorganisms, with fungi as an exception. A long-lasting negative action on the activity of the dehydrogenase (DHA), commonly used as an index of the overall microbial activity in soil, was found. The magnitude of these effects were dependent on the assayed concentrations of the herbicides mixture. We concluded that the presence of bromoxynil + prosulfuron could induce significant changes in the microbial populations of the soil, concerning the activity and balance of microbial community. Possible environmental risks must be considered. Dehydrogenase activity was shown to be an important indicator of side-effects attributed to these herbicides.     

Ammonia concentration determines differential growth of ammonia-oxidising archaea and bacteria in soil microcosms

The first step of nitrification, oxidation of ammonia to nitrite, is performed by both ammonia-oxidising archaea (AOA) and ammonia-oxidising bacteria (AOB) in soil, but their relative contributions to ammonia oxidation and existence in distinct ecological niches remain to be determined. To determine whether available ammonia concentration has a differential effect on AOA and AOB growth, soil microcosms were incubated for 28 days with ammonium at three concentrations: native (control), intermediate (20 μg NH₄⁺-N per gram of soil) and high (200 μg NH₄⁺-N per gram of soil). Quantitative PCR demonstrated growth of AOA at all concentrations, whereas AOB growth was prominent only at the highest concentration. Similarly, denaturing gradient gel electrophoresis (DGGE) analysis revealed changes in AOA communities at all ammonium concentrations, whereas AOB communities changed significantly only at the highest ammonium concentration. These results provide evidence that ammonia concentration contributes to the definition of distinct ecological niches of AOA and AOB in soil.