Tolerance of soil bacterial complexes to salt shock

Investigations showed that bacteria present in soil are resistant to one-day exposure to a saturated solution of ammonium nitrate and can well develop when transferred to laboratory nutrient media. The evaluated number of bacteria in NH4NO3-treated soil samples was nearly the same as in native soil samples, while was 1.5-2.5 times smaller in the former than in the latter case when microbial succession in the soil samples was initiated by wetting them. Bacteria (particularly gram-negative ones) occurring at the early stages of succession were the most sensitive to salt stress. Bacteria in soil were found to be much more resistant to salt stress than the same bacteria isolated in pure cultures.     

Variation in the incidence of H2-oxidizing chemolithotrophic bacteria in rice grown under different cultivation conditions

Summary The incidence of H₂-oxidizing chemolithotrophic bacteria associated with rice grown under continuous wetland, upland, and rainfed wetland conditions was studied by¹⁴C-autoradiographic technique in a neutral soil at IRRI (Maahas) and an acid rainfed wetland soil (Luisiana).In Maahas soil, H₂-oxidizing chemolithotrophic bacteria were not detected in the endorhizosphere, rhizosphere, and nonrhizosphere soil of rice grown under dryland conditions. Under continuously flooded conditions a very large population of these bacteria were found in the endorhizosphere but not in the oxidized and reduced soil.A very low population of these bacteria were found in the endorhizosphere and basal culm of rice grown under rainfed wetland conditions at Luisiana. Bacteria isolated from Maahas wetland rice and inoculated to rice seedling planted in Luisiana soil failed to establish.Both Maahas and Luisiana soils consumed externally supplied H₂ and produced H₂ and CH₄ almost at the same rate when they were amended with rice straw or sucrose. This paper discusses possible causes of variation in the number of these bacteria and their distribution in rice grown under different cultural and soil conditions.     

Soil Drying As a Model for the Action of Stress Factors on Natural Bacterial Populations

The drying of soil samples reduced the abundance (especially of predominant species) and the diversity of bacteria isolated from these samples, making easier the isolation of rare bacterial species. Some bacterial species that were minor before soil drying became dominant in dried soil samples. In general, soil drying allowed the diversity of soil bacteria to be determined more adequately. The bacteria that were isolated from dried soil samples turned out to be resistant to gamma radiation (with LD₉₀ = 2.8–4.6 kGy) and desiccation. It is concluded that soil drying may serve as a model for the action of stress factors on natural bacterial populations. The hypothesis that periodic desiccation was the primary cause of formation of bacterial radioresistance in nature is discussed.     

鄂尔多斯台地盐沼滩涂湿地土壤细菌群落结构及特征

依据植被分类法将鄂尔多斯高原盐沼滩涂湿地划分为肉质耐盐草甸(B)、苔草草甸(C)、禾草草甸(D)和杂类草草甸(E)等4个植被亚型,并以盐沼裸地(A)为对照样地,共计5种盐沼滩涂湿地景观类型。运用高通量测序技术分别研究其土壤细菌群落结构特征、分布情况,以及土壤盐分与土壤细菌之间的关系。结果表明:(1) 5种盐沼滩涂湿地的土壤细菌样品共12213条OTUs,属于45个门,122个纲,365个目,663个科,1375个属,2882个种。(2)变形菌门(26.19%)是盐沼滩涂湿地平均相对丰度最高的门,其次为放线菌门(17.15%),绿弯菌门(12.62%),芽单胞菌门(Gemmatimonadetes,11.23%),拟杆菌门(Bacteroidetes,9.38%),酸杆菌门(Acidobacteria,8.83%)厚壁菌门(Firmicutes,2.96%);芽单胞菌纲中的未定细菌(norank＿c＿Gemmatimonadetes)是丰度最高的属,平均丰度为5.75%。(3)鄂尔多斯盐沼滩涂湿地自西南向东北,空间位置相近的土壤细菌群落结构具有更大的相似性,随着土壤盐分变化,变形菌门相对...     

鄂尔多斯台地盐沼滩地微生物群落与土壤条件分析

[背景]我国北方内陆区与平原区土地盐碱化问题严重,针对微生物如何在极端盐碱地植被演替过程中发挥作用的研究鲜有报道.[目的]研究鄂尔多斯台地5种不同植被类型的盐沼湿地微生物群落与土壤条件的关系,筛选出耐盐碱菌群及影响耐盐碱菌群的土壤环境因子.[方法]采用高通量测序技术,对其微生物细菌群落组成进行了比对分析.[结果]鄂尔多...     

The Culturable Soil Antibiotic Resistome: A Community of Multi-Drug Resistant Bacteria

Understanding the soil bacterial resistome is essential to understanding the evolution and development of antibiotic resistance, and its spread between species and biomes. We have identified and characterized multi-drug resistance (MDR) mechanisms in the culturable soil antibiotic resistome and linked the resistance profiles to bacterial species. We isolated 412 antibiotic resistant bacteria from agricultural, urban and pristine soils. All isolates were multi-drug resistant, of which greater than 80% were resistant to 16–23 antibiotics, comprising almost all classes of antibiotic. The mobile resistance genes investigated, (ESBL, bla _NDM-1, and plasmid mediated quinolone resistance (PMQR) resistance genes) were not responsible for the respective resistance phenotypes nor were they present in the extracted soil DNA. Efflux was demonstrated to play an important role in MDR and many resistance phenotypes. Clinically relevant Burkholderia species are intrinsically resistant to ciprofloxacin but the soil Burkholderia species were not intrinsically resistant to ciprofloxacin. Using a phenotypic enzyme assay we identified the antibiotic specific inactivation of trimethoprim in 21 bacteria from different soils. The results of this study identified the importance of the efflux mechanism in the soil resistome and variations between the intrinsic resistance profiles of clinical and soil bacteria of the same family.     

Colonization of wheat by Azospirillum brasilense Cd is impaired by saline stress

The effect of saline stress on the colonization of wheat was analyzed by using Azospirillum brasilense Cd carrying the fusion of the reporter gene lacZ (β-galactosidase) with the N₂ fixation gene promoter nifA. Colonization was also studied by inducing para-nodules on wheat roots using 2,4-D, establishing that these structures acted as bacterium protected niches. Bacteria grown under standard conditions were distributed along the whole root system, except the elongation zone, and colonized the para-nodules. Bacteria experiencing saline stress were mainly localized at the root tips and the lateral roots. In 2,4-D treated plants, most of the bacteria were present around the basal surface of the modified lateral root structures. Using the MPN method, there were not statistical differences between the numbers of control and stressed bacteria. As this method estimates endophytic colonization in contrast with the one using X-gal, which emphasizes colonization on the root surface, both procedures demonstrated to be necessary, concluding that salt treatment reduced surface colonization (X-gal) but not colonization inside the root. The bacterial counts made on inoculated wheat roots indicated higher numbers of both control and stressed bacteria in roots treated with 2,4-D compared with untreated roots. This revised version was published online in June 2006 with corrections to the Cover Date.     

蕾期土壤盐度降低后棉花叶片的生理功能恢复

试验于2011—2012年在江苏南京江苏省农业科学院经济作物研究所试验田进行,采用盆栽方法,以鲁棉研37号和苏棉22号为供试材料,设置土壤盐度降低试验(初始土壤含盐量为0.2%,棉花进入二叶期后每7d加入混合盐1次,每次增加0.1%,使土壤含盐量逐渐达到0.5%,蕾期进行盐度降低处理,使土壤含盐量降低到0.2%左右),研究蕾期土壤盐度降低后棉花叶片的生理代谢动态特征。结果表明:土壤盐度降低后,棉花叶片叶绿素(Chl)、类胡萝卜素(Car)含量和Chl/Car升高;净光合速率和气孔导度升高,且分别在土壤盐度降低后第14天和7天接近于低盐对照;土壤盐度降低后棉花叶片超氧化物歧化酶(SOD)和过氧化物酶(POD)活性升高,过氧化氢酶(CAT)活性和丙二醛(MDA)含量降低,MDA含量在土壤盐度降低后第14天接近于低盐对照;土壤盐度降低后棉花叶片中可溶性糖、游离氨基酸和脯氨酸含量降低,且接近于低盐对照。上述结果表明土壤盐度降低后,棉花叶片生理功能逐渐恢复,进而实现棉花生长发育的恢复补偿。棉花叶片生理功能在土壤盐度降低后的恢复能力存在品种间差异,鲁棉研37号较苏棉22号叶片生理功能表现出更强的恢复能力。     

Symbiotic formation of alginate lyase in mixed culture of bacteria isolated from soil

Bacteria having alginate lyase activity were screened by growing cells from various sources in a medium containing alginate as a carbon source. Among the various samples tested, the culture enriched with paddy-field bacteria showed the highest alginate lyase activity, and contained three kinds of bacteria. They were purified and identified to be Flavobacterium, Alcaligenes and Bacillus species. The alginate lyase activity in these strains was low when they were grown separately. The highest alginate lyase activity was obtained when these strains were cultured all together in the same medium.     

Epidemiology of Antimicrobial Resistance in Escherichia coli Isolates from Raccoons (Procyon lotor) and the Environment on Swine Farms and Conservation Areas in Southern Ontario

Antimicrobial resistance is a global threat to livestock, human and environmental health. Although resistant bacteria have been detected in wildlife, their role in the epidemiology of antimicrobial resistance is not clear. Our objective was to investigate demographic, temporal and climatic factors associated with carriage of antimicrobial resistant Escherichia coli in raccoons and the environment. We collected samples from raccoon paws and feces and from soil, manure pit and dumpsters on five swine farms and five conservation areas in Ontario, Canada once every five weeks from May to November, 2011–2013 and tested them for E. coli and susceptibility to 15 antimicrobials. Of samples testing positive for E. coli, resistance to ≥ 1 antimicrobials was detected in 7.4% (77/1044; 95% CI, 5.9–9.1) of raccoon fecal samples, 6.3% (23/365; 95% CI, 4.0–9.3) of paw samples, 9.6% (121/1260; 8.0–11.4) of soil samples, 57.4% (31/54; 95% CI, 43.2–70.8) of manure pit samples, and 13.8% (4/29; 95% CI, 3.9–31.7) of dumpster samples. Using univariable logistic regression, there was no significant difference in the occurrence of resistant E. coli in raccoon feces on conservation areas versus farms; however, E. coli isolates resistant to ≥ 1 antimicrobials were significantly less likely to be detected from raccoon paw samples on swine farms than conservation areas and significantly more likely to be detected in soil samples from swine farms than conservation areas. Resistant phenotypes and genotypes that were absent from the swine farm environment were detected in raccoons from conservation areas, suggesting that conservation areas and swine farms may have different exposures to resistant bacteria. However, the similar resistance patterns and genes in E. coli from raccoon fecal and environmental samples from the same location types suggest that resistant bacteria may be exchanged between raccoons and their environment.     

Comparative Toxicities of Salts on Microbial Processes in Soil

Soil salinization is a growing threat to global agriculture and carbon sequestration, but to date it remains unclear how microbial processes will respond. We studied the acute response to salt exposure of a range of anabolic and catabolic microbial processes, including bacterial (leucine incorporation) and fungal (acetate incorporation into ergosterol) growth rates, respiration, and gross N mineralization and nitrification rates. To distinguish effects of specific ions from those of overall ionic strength, we compared the addition of four salts frequently associated with soil salinization (NaCl, KCl, Na₂SO₄, and K₂SO₄) to a nonsaline soil. To compare the tolerance of different microbial processes to salt and to interrelate the toxicity of different salts, concentration-response relationships were established. Growth-based measurements revealed that fungi were more resistant to salt exposure than bacteria. Effects by salt on C and N mineralization were indistinguishable, and in contrast to previous studies, nitrification was not found to be more sensitive to salt exposure than other microbial processes. The ion-specific toxicity of certain salts could be observed only for respiration, which was less inhibited by salts containing SO₄²⁻ than Cl⁻ salts, in contrast to the microbial growth assessments. This suggested that the inhibition of microbial growth was explained solely by total ionic strength, while ion-specific toxicity also should be considered for effects on microbial decomposition. This difference resulted in an apparent reduction of microbial growth efficiency in response to exposure to SO₄²⁻ salts but not to Cl⁻ salts; no evidence was found to distinguish K⁺ and Na⁺ salts.     

Taxonomic composition of denitrifying bacteria in soddy podzolic soil

The taxonomic composition of denitrifying bacteria in soddy podzolic soil was studied by the succession analysis method. This method revealed a significant variation in the taxonomic composition of denitrifying microorganisms in the course of succession. In contrast to succession analysis, the single microbiological analysis of soil samples reflected only the late stage of succession and thus led to an underestimation of the major members of succession. Myxobacteria were found to be the most active denitrifiers at the early stages of succession, whereas bacilli dominated at its late stages. The bacilli were represented by three facultatively anaerobic species:Bacillus cereus, Bac. circulons, andBac. polymyxa.     

山东地区盐碱土花生种子际土壤微生物群落结构的研究

【目的】以不同含盐量的滨海盐土、内陆盐碱土和中等肥力非盐碱土壤为实验对象,探讨花生种子在吸水膨胀与萌发过程中,不同类型盐碱土对种子际土壤微生物多样性变化的影响。【方法】采集不同含盐量的滨海盐土、内陆盐碱土和中等肥力非盐碱土壤,通过对各样品中细菌的16S r RNA基因的V3-V4区进行PCR扩增,利用Illumina Hiseq高通量测序技术对12份V3-V4高变区PCR产物进行测序,并对测序数据进行生物信息学分析。【结果】(1)盐碱土壤的种子际细菌群落多样性高于非盐碱土壤,且以东营青坨滨海盐土种子际土壤细菌群落多样性较高。(2)不同类型土壤样本微生物群落结构在纲水平存在明显差异。4种土壤类型种子际土壤细菌共分属于6个菌纲,分别为Proteobacteria、Actinobacteria、Actinobacteria、Bacteroidetes、Acidobacteria和Firmicutes菌纲,并均以Proteobacteria和Actinobacteria菌纲为主要菌纲。全样本菌落结构分析结果表明,4种类型土壤中不同吸胀时间内种子际微生物菌落在门、属水平上的类型和丰度差异最为显著(P0.05)。(3)beta多样性分析和各样本遗传距离(phylogenetic distances)聚类树图分析表明,4个土壤类型的12个土壤样本种子际土壤中微生物群落均可聚为2大类。【结论】土壤含盐量越高其种子际土壤细菌群落多样性较高。不同类型土壤样本微生物群落结构在纲水平存在明显差异,以Proteobacteria和Actinobacteria菌纲为主要菌纲。种子吸胀萌发时间影响种子际微生物菌落在门、属水平上的类型和丰度,但对相同土壤类型样本间遗传距离无影响。     

盐碱胁迫对枸杞幼苗生物量积累和光合作用的影响

以内陆高寒区盐碱地重要的经济树种枸杞2年生幼苗为研究对象,采用盆栽控制试验方法,设置50、100、200、300mmol·L~(-1)共4个盐和碱(NaCl和NaHCO3)胁迫浓度,研究盐、碱胁迫对枸杞苗木生长和光合的影响,以明确枸杞幼苗生长的耐盐、碱浓度范围,探讨土壤盐碱含量与土壤水分含量的关系,为不同类型盐碱条件下枸杞的种植和水分管理提供理论依据。结果表明:(1)随着盐碱胁迫浓度的增大,枸杞幼苗根茎叶生物量及叶绿素含量(SPAD)、净光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)和胞间二氧化碳浓度(Ci)等光合作用参数逐渐受到显著抑制,且碱胁迫的抑制作用更强烈;但低浓度(50mmol/L)NaCl胁迫下,枸杞幼苗叶绿素含量和净光合速率并未受到显著影响。(2)在盐碱胁迫条件下,枸杞幼苗的最大净光合速率(Pnmax)、暗呼吸速率(RD)、初始量子效率(AQY)、光饱和点(LSP)均低于对照,而光补偿点(LCP)高于对照,且随着胁迫浓度的增加,碱胁迫处理下的变幅大于盐胁迫。(3)随着胁迫浓度的增大,影响净光合速率的因素由气孔限制转向非气孔限制的临界值,在盐胁迫下的临界浓度约为200mmol/L,在碱胁迫下的临界浓度约为100mmol/L。(4)按照指标值超过对照组50%标准,经回归分析确定,枸杞耐盐和耐碱阈值分别为(246.3±2.1)mmol/L和(126.7±2.7)mmol/L;在此阈值的基础上,得到土壤含水量与土壤含盐量回归曲线方程。研究认为,枸杞幼苗具有一定的耐盐能力,但过高浓度的盐碱胁迫会损坏其光合结构,降低光环境适应能力和光合作用效率,从而影响其正常生长。     

Partial identification and copper tolerance of soil bacteria from copper-impregnated bog and from a salt marsh

Abstract

Bacteria from two areas in Wales have been extracted and partially identified. The areas are a copper impregnated bog in the Coed-Y-Brenin Forest with low pH and high carbon and copper content, and the Morfa Harlech salt marshes which have neutral pH and lower copper and carbon. Bacillus spp. was the predominant bacterium in both soils, but there was increased diversity of bacteria in the salt marsh soils. Bacteria from the Cu-rich, Coed-Y-Brenin soils showed higher growth densities in nutrient agar plates containing 10, 30 and 100 μg cm^?3 Cu than bacteria from the salt marsh soils, indicating that bacteria from the Coed-Y-Brenin soils could tolerate higher copper concentrations. Pseudomonas spp. from both study areas grew better in media with added Cu at low levels. The possibility is discussed that the bacteria play a part in the bioavailability of Cu particularly in the copper-rich Coed-Y-Brenin area.     

Thymidine incorporation of bacteria sequentially extracted from soil using repeated homogenization-centrifugation

Bacteria were sequentially extracted from soil into a water suspension after shaking soil with water or mixing it in a blender followed by a low-speed centrifugation. Bacteria, which were released only after several cycles of homogenization-centrifugation, had higher growth rates as judged from thymidine and leucine incorporation, whereas bacteria that were more readily released by a gentle shaking procedure had the lowest growth rate. This indicated that bacteria more tightly bound to soil particles were growing faster than those that were more easily released into the water suspension. The same pattern was found both in an agricultural and a forest soil, with contrasting pH and organic matter content, and irrespective of whether the bacteria were labeled before or after the centrifugation steps. The different growth rates of the bacteria could not be explained by different partitioning of label between different macromolecules, different cell size, different viability of the bacteria, or different dilution of the added radioactive substrate in the different homogenization-centrifugation fractions. The total amount of phospholipid fatty acids per bacterial cell was also similar in the different fractions. Different composition of the bacterial communities in the different homogenization-centrifugation fractions was indicated by a gradually altered phospholipid fatty acid pattern of the extracted bacteria, and an increased hydrophobicity of the bacteria released only after several homogenization-centrifugation treatments. Correspondence to: E. Bååth     

The dynamics of the bacterial population associated with a salt marsh

The distribution and temporal fluctuations in the density of bacteria in the water covering a high-salinity marsh were investigated employing epifluorescence microscopy for quantification as well as by transmission and scanning electron microscopy. The observed densities ranged from about 1 to 19 × 10⁶ bacteria/ml during the course of the study. High-marsh sampling sites had an average population level of 7.8 × 10⁶ bacteria/ml which was more than double the mean density recovered from large creeks near the mouth of the marsh system. Bacteria associated with seston varied tidally and seasonally, whereas the population of free planktonic bacteria varied only seasonally. Very small fluorescing bodies were commonly observed during epifluorescent observation of samples. These small bodies were observed at densities two orders of magnitude higher than easily recognized bacteria. In a salt marsh, the relative density of epibacteria was influenced by short-term tidal effects, and the population of planktobacteria was apparently controlled by long-term seasonal phenomena.     

盐渍化土壤根际微生物群落及土壤因子对AM真菌的影响

为探明盐渍化土壤影响下AM真菌与根际土壤间的关系,试验选取宁夏碱化龟裂土、草甸盐土、盐化灌淤土3种类型4个样地上典型植被群落,测定了植物根际土壤养分含量、微生物群落结构、AM真菌侵染率以及孢子密度。结果显示:盐渍化土壤根际微生物碳源利用类型显著不同,对芳香类化合物的代谢能力整体较弱;红寺堡草甸盐土上微生物优势群落为氨基酸代谢类群,惠农盐化灌淤土为多聚化合物代谢群,西大滩碱化龟裂土为碳水化合物代谢群。AM真菌孢子密度与微生物碳源代谢群间的关系比较复杂。其中,惠农样点根际土壤孢子密度与多聚化合物微生物代谢群呈显著正相关,西大滩地区孢子密度与碳水化合物微生物代谢群呈显著正相关。土壤有机质、全盐、全氮、碱解氮等土壤肥力因子及土壤中的HCO-3、Na+、Cl-等盐基离子含量能解释AM真菌孢子密度与土壤环境因子之间相互关系的大部分信息。较高的HCO-3浓度促进了AM真菌侵染率的提高,但高盐浓度下Na+和Cl-降低了菌根侵染率。土壤对AM真菌孢子密度、侵染率的影响因土壤盐分组成类型的不同而异。研究结果为深入了解AM真菌多样性,促进宁夏盐碱地的合理开发与利用提供了理论依据。     

石灰土演替过程中颗粒态有机质和矿物结合态有机质的微生物群落特征

碳酸盐岩经风化作用并在地形、植被、气候、时间及生物等因素的影响下逐渐演替出黑色石灰土、棕色石灰土、黄色石灰土和红色石灰土。【目的】研究不同演替阶段石灰土颗粒态有机质(particulate organic matter, POM)和矿物结合态有机质(mineral-associated organic matter, MAOM)的微生物群落特征,为岩溶土壤有机质稳定机制研究提供理论依据。【方法】以广西弄岗国家级自然保护区的黑色石灰土、棕色石灰土、黄色石灰土和红色石灰土为研究对象,运用湿筛法将土壤有机质(soil organic matter, SOM)分为POM和MAOM,分析其理化性质以及微生物群落特征。【结果】石灰土演替过程中POM和MAOM的有机碳、总氮、交换性钙含量均呈下降趋势,且MAOM的C/N均大于POM,POM的C/P均大于MAOM。细菌α多样性在黑色石灰土POM和MAOM中最高,且四类石灰土MAOM的真菌多样性比POM要高。Acidobacteria、Proteobacteria、Ascomycota均为石灰土演替过程中POM和MAOM的优势菌门。总磷是影响石灰土演替过...     

Treatment of nitric oxide supplemented with nitrogen and sulfur regulates photosynthetic performance and stomatal behavior in mustard under salt stress

Nitric oxide (NO) is a hormone that connects numerous reactions in plant cells under normal and environmental stress conditions. The application of 100 µM NO as sodium nitroprusside (SNP; NO donor) applied individually or in combination with N or S in different combinations (i.e. 100 mg N or S kg⁻¹ soil applied at the time of sowing [100 N + 100S]_0d or with split, 50 mg N or S kg⁻¹ soil at the time of sowing and similar dose at 20 d after sowing [50 N + 50S]_0d + [50 N + 50S]_20d) was tested to alleviate salt stress in mustard (Brassica juncea L.). Application of 100 µM NO plus split application of N and S more significantly promoted stomatal behavior, photosynthetic and growth performance in the absence of salt stress and maximally alleviated effects of salt stress through increased N- and S-use efficiency, proline and antioxidant system. The combined application of N and S at the time of sowing was lesser effective in promoting photosynthesis and growth under salt or no salt stress conditions in presence or absence of NO. The study suggests that salt stress effects on the photosynthetic performance are mitigated more efficiently when NO was applied together with the split application of N and S given at two stages, and the photosynthetic activity was promoted under salt stress through increased N and S assimilation and antioxidant system. This strategy may be adopted in agricultural system for overcoming salt stress effects on performance of mustard.