Comparison of different microbial biomass and activity measurement methods in metal-contaminated soils

The aims of this study were: (1) to compare different microbial methods of detecting the effects of heavy metals on the functioning of the soil ecosystem; and (2) to evaluate the effect of incubation on microbial biomass and microbial activity in soils that were not pre-incubated after sampling in order to determine their suitability for measuring the effects of heavy metals on the soil microbial ecosystem. The microbial biomass methods (included: biomass C, N and ninhydrin-N by fumigation-extraction (FE); substrate-induced respiration (SIR); soil ATP content and microbial activity as evolved CO2-C and arginine ammonification. All were tested in soils from the Woburn Market Garden Experiment. Due to past sludge application the soils contained, Zn, Cu or Ni at around current European Union upper limits and Cd at up to three times the limit. The amount of microbial biomass in metal-contaminated soils was about half of that found in soils from the experiment that received uncontaminated organic manure or inorganic fertilizer. The amount of biomass measured by FE and soil ATP content in incubated soils showed little change over 20 days incubation. However, SIR measurements were statistically affected over the first few days of incubation. The rates of arginine ammonification were higher in this order: farmyard manure (FYM)>inorganic fertilizer>sewage-sludge throughout the incubation. However, the evolved CO2-C rates were not significantly different among the treatments. Discriminant analysis confirmed smaller amounts of biomass in the metal-contaminated soils than in the other treatments. Linked properties, such as relationships between biomass and soil organic matter, or biomass-specific respiration rates, may provide "internal control" which may help overcome problems of establishing suitable control, or comparative measurements, when moving from experimental to natural environments.     

Measurement of microbial biomass phosphorus in rhizosphere soil

³²P-labelled monocalcium phosphate solution was supplied by point injection to the root system of wheat plants grown in soil cores in a controlled environment. There was no detectable incorporation of³²P into organic P fractions in the soil remaining after roots were removed, confirming field observations. The techniques used to measure organic P (including biomass P) could detect an incorporation of³²P into soil microbial biomass equivalent to 0.3 μgP.g^?1 soil, compared to a total soil biomass P content estimated to be ca. 6.5 μgP.g^?1 soil. The limited incorporation of the added P into microbial biomass in the root-free soil may be due partly to a limited diffusion of³²P into the non-rhizosphere soil and partly to the removal of³²P-labelled microbial biomass adhering to or in very close association with the root surface. it is proposed that in studies of soil nutrient status, total soil biomass P (roots + soil flora + microfauna) should be measured, rather than attempting an estimate of microbial P. A sequential extraction procedure using a single soil sample, where a biocide is added to the extracting solution, is proposed as an alternative to the conventional procedure for measuring soil biomass P where two soil samples, one treated with a biocide, are extracted simultaneously.     

增温对高寒灌丛根际和非根际土壤微生物生物量碳氮的影响

本文对青藏高原东缘窄叶鲜卑花高寒灌丛生长季根际和非根际土壤微生物生物量碳和氮对增温的响应进行研究.结果表明: 窄叶鲜卑花灌丛生长季初期根际和非根际土壤微生物生物量碳和氮均显著高于生长季中期和末期.在多数时期,增温对根际土壤微生物生物量碳和氮的影响不显著.在非根际土壤中,增温对土壤微生物生物量碳和氮的影响因不同生长季节而不同: 增温使生长季初期土壤微生物生物量碳显著降低,而使土壤微生物生物量氮显著提高;生长季中期增温使土壤微生物生物量碳和氮显著提高;而在生长季末期增温对土壤微生物生物量碳和氮的影响不显著.土壤微生物生物量碳和氮的根际效应也因不同生长季节而不同: 土壤微生物生物量碳和氮在生长季初期表现为负根际效应,而在生长季中期表现为正根际效应;在生长季末期,土壤微生物生物量碳表现为负根际效应,土壤微生物生物量氮则表现为正根际效应.增温在生长季初期使土壤微生物生物量碳和氮的根际效应显著提高,而在生长季中期和末期使土壤微生物生物量碳和氮的根际效应降低.本研究初步阐明了气候变暖背景下高寒灌丛根际和非根际土壤生物学过程变化机理.     

荒漠草地土壤微生物生物量和微生物熵对沙漠化的响应

采用空间序列代替时间演替的方法,分析宁夏中北部盐池县荒漠草地不同沙漠化阶段(荒漠草地、固定沙地、半固定沙地和流动沙地)土壤微生物生物量(SMB)和微生物熵(qMB)的变化特征及其影响因子.结果表明:从荒漠草地到流动沙地,土壤微生物生物量碳、氮、磷分别降低46.1%、80.8%和30.0%.随着荒漠草地沙漠化程度的加剧,土壤微生物熵碳(qMBC)、土壤微生物熵氮(qMBN)、土壤微生物熵磷(qMBP)均表现为荒漠草地>固定沙地>半固定沙地>流动沙地,而土壤-微生物化学计量不平衡性(C∶Nimb、C∶Pimb、N∶Pimb)基本呈增加趋势.土壤微生物生物量氮与C∶Nimb呈显著正相关,与N∶Pimb呈显著负相关;土壤微生物生物量磷与C∶Pimb呈显著正相关.冗余分析(RDA)显示,土壤生态化学计量(C∶N、C∶P)对微生物熵碳的负效应最明显.荒漠草地沙漠化显著影响土壤微生物生物量和微生物熵.     

Diversity and potential biogeochemical impacts of viruses in bulk and rhizosphere soils

Viruses can affect microbial dynamics, metabolism and biogeochemical cycles in aquatic ecosystems. However, viral diversity and functions in agricultural soils are poorly known, especially in the rhizosphere. We used virome analysis of eight rhizosphere and bulk soils to study viral diversity and potential biogeochemical impacts in an agro-ecosystem. The order Caudovirales was the predominant viral type in agricultural soils, with Siphoviridae being the most abundant family. Phylogenetic analysis of the terminase large subunit of Caudovirales identified high viral diversity and three novel groups. Viral community composition differed significantly between bulk and rhizosphere soils. Soil pH was the main environmental driver of the viral community structure. Remarkably, abundant auxiliary carbohydrate-active enzyme (CAZyme) genes were detected in viromes, including glycoside hydrolases, carbohydrate esterases and carbohydrate-binding modules. These results demonstrate that virus-encoded putative auxiliary metabolic genes or metabolic genes that may change bacterial metabolism and indirectly contribute to biogeochemical cycling, especially carbon cycling, in agricultural soil.     

Predicted microbial biomass in the rhizosphere of barley in the field

Summary Laboratory data for the loss of root material by barley and field data for the growth of barley plants in Syria and in England have been combined to predict the amount of material lost by barley roots during a season, and to predict the resulting microbial biomass in the rhizosphere. The predicted microbial biomass C in the rhizosphere ranged from 10–34% of the total plant biomass C depending mainly upon the value used for rate of loss of root material. Total loss of root material predicted during a season in England constituted 7.7–25.4 percent of C fixed by photosynthesis. The major assumptions made in these calculations are considered, and the predicted values discussed in relation to reported values for soil microbial biomass, CO₂ fluxes from soil and associative nitrogen fixation.     

玉米幼苗根际土壤微生物活性对芘污染的响应

用根际袋法土培试验研究了玉米幼苗根际与非根际土壤微生物量碳、微生物熵、代谢熵和土壤酶活性对不同芘污染水平(50、200、800mg·kg-1,记为T1、T2、T3)的响应差异。结果表明,较低浓度芘可适当的刺激玉米幼苗的生长,而较高浓度芘则抑制幼苗生长,其抑制作用随芘处理浓度的提高而增强;芘对玉米根系的影响要大于对茎叶的影响。玉米幼苗能够明显促进土壤中芘的去除。根际和非根际土壤中芘的去除率分别为56.67%-76.18%和32.64%-70.44%,根际土壤中芘的平均去除率比非根际土壤高16.06%。同处理中根际土壤芘含量显著低于非根际土壤,随着芘处理浓度的提高其差异更加显著。根际土壤微生物量碳、微生物熵、多酚氧化酶活性、脱氢酶活性和磷酸酶活性均高于非根际土壤,代谢熵低于非根际土壤,且其差异随芘处理浓度的提高而增大。在不同芘污染水平下,微生物量碳、微生物熵和脱氢酶活性根际和非根际土壤为T1T2T3,代谢熵为T3T2T1;多酚氧化酶活性根际土壤为T2T1T3,非根际土壤为T1T2T3;磷酸酶活性根际土壤为T3T1T2,非根际土壤为T1T2T3。土壤中残余芘含量与土壤微生物量碳、微生物熵、多酚氧化酶、脱氢酶和磷酸酶活性呈显著负相关,与代谢熵呈显著正相关。     

Assessing short-term responses of prokaryotic communities in bulk and rhizosphere soils to tall fescue endophyte infection

In contrast to endophyte-free (E−) tall fescue, endophyte-infected (E+) tall fescue pastures appear to enhance soil carbon sequestration. A hypothetical mechanism that may account for the enhanced carbon sequestration is that the E+ tall fescue affects the soil microbial community or components of it that are involved in organic carbon turnover. A 60-week mesocosm study with a factorial arrangement of soil type, loamy sand (LS) and clay loam (CL), and E+ and E− tall fescue was conducted to determine if the soil microbial communities were affected by the presence of the endophyte. Bulk and rhizosphere soil samples were fixed in paraformaldehyde, and prepared for total direct microbial counts, and with a combination of one of a domain or subdivision fluorescent oligonucleotide probe for enumerating metabolically active Eubacteria, bacterial subdivisions, and Archaea. E+ tall fescue suppressed the archaeal and high G+C gram-positive bacterial communities of the bulk CL, the delta-proteobacterial community in the rhizosphere CL, and the Planctomycetes community of the rhizosphere LS. In the long-term, suppression of these microbial communities may be a factor in enhanced soil carbon sequestration associated with E+ tall fescue.     

Impact of soil salinity on the microbial structure of halophyte rhizosphere microbiome

The rhizosphere microbiome plays a significant role in the life of plants in promoting plant survival under adverse conditions. However, limited information is available about microbial diversity in saline environments. In the current study, we compared the composition of the rhizosphere microbiomes of the halophytes Urochloa, Kochia, Salsola, and Atriplex living in moderate and high salinity environments (Khewra salt mines; Pakistan) with that of the non-halophyte Triticum. Soil microbiomes analysis using pyrosequencing of 16S rRNA gene indicated that Actinobacteria were dominant in saline soil samples whereas Proteobacteria predominated in non-saline soil samples. Firmicutes, Acidobacteria, Bacteriodetes and Thaumarchaeota were predominant phyla in saline and non-saline soils, whereas Cyanobacteria, Verrucomicrobia, Gemmatimonadetes and the unclassified WPS-2 were less abundant. Sequences from Euryarchaeota, Ignavibacteriae, and Nanohaloarchaeota were identified only from the rhizosphere of halophytes. Dominant halophilic bacteria and archaea identified in this study included Agrococcus, Armatimonadetes gp4, Halalkalicoccus, Haloferula and Halobacterium. Our analysis showed that increases in soil salinity correlated with significant differences in the alpha and beta diversity of the microbial communities across saline and non-saline soil samples. Having a complete inventory of the soil bacteria from different saline environments in Pakistan will help in the discovery of potential inoculants for crops growing on salt-affected land.     

Extractable N and P in relation to microbial biomass in UK acid organic soils

Summary Samples of peat and mor humus were treated with CHCl₃ to kill microbial cells and the amounts of C as CO₂, N as soluble- and mineral-N, and P as inorganic-P released by the treatment were compared with estimates of the microbial biomass by the Anderson and Domsch test and ATP determination. Increased amounts of soluble-N and inorganic-P, extracted with 1M KCl and 0.01M CaCl₂ respectively, were detected immediately after the fumigation treatment. However, the subsequent rates of production of CO₂–C and mineral-N measured during a 10-day incubation period at 25°C were low and variable, resulting in anomalously low estimates of microbial biomass. The flush of inorganic-P was more consistent and, in mor humus, generally related to biomass-C as measured by the Anderson and Domsch test.     

Microscale fumigation-extraction and substrate-induced respiration methods for measuring microbial biomass in barley rhizosphere

Changes in microbial biomass in the rhizosphere of young barley seedlings was studied. A fumigation-extraction (FE) method with measurement of ninhydrin-reactive nitrogen (NR-N) and a substrate-induced respiration (SIR) method were applied on a microscale to rhizosphere soil samples of approximately 0.1 g. Rhizosphere soil was defined as the soil adhering to the roots when they were carefully separated from the bulk soil. The rhizosphere soil was gently washed off the roots with either distilled water (FE) or with glucose solution (SIR). Shaking and mild sonication was used to disperse the soil without disrupting the roots. Fumigation was carried out by direct addition of liquid chloroform to the isolated soil. These techniques were proven to give reliable results under the experimental conditions of this investigation. Rhizosphere soil was isolated from segments of the roots representing different distances to the seed different root ages. In the rhizosphere of young barley seedlings, biomass NR-N increased significantly compared to the bulk soil from day 6 after sowing (average increases of 33–97%), especially where adventitious roots had developed. From this time, SIR rates were also significantly higher in the rhizosphere than in bulk soil (average increases 72–170%). The average ratio of SIR rate to biomass NR-N was found to be approximately 50% higher in the rhizosphere than in the bulk soil, which may indicate that a larger fraction of the microbial community is potentially active in the rhizosphere as compared to the bulk soil.     

Studies on microbial antagonism in rhizosphere soils of potato varieties differentially susceptible to black-scurf and wilt

Summary The antagonistic microflora in the rhizosphere of plants of three varieties of potato, viz. Ps 194' Up to date, and a local variety of different ages were analysed. Antagonists were selected either by random isolation or by Wilska's spraying technique. The antagonists were always found in higher frequency in rhizosphere compared with the control soil. The highest percentage of actinomycetes antagonists were found in the variety Up to date; of fungal antagonists in the variety Ps 194; while the local variety was found to be the poorest in this regard. Further, the antagonistic activity of these organisms was estimated by a cross-streak method and the resistant antagonists were also selected by cross-antagonism. The most resistant antagonists found were Trichoderma viride T 56 and T 7, Chaetomium trilaterate F 75, Streptomyces alboflavus Rs 40, Streptomyces sp. Cs 73, Pseudomonas sp. Rs 1 50 and Pseudomonas sp III Ps 63.     

Factors determining soil microbial biomass and nutrient immobilization in desert soils

We examined the 10-day response of soil microbial biomass-N to additions of carbon (dextrose) and nitrogen (NH₄NO₃) to water-amended soils in a factorial experiment in four plant communities of the Chihuahuan desert of New Mexico (U.S.A.). In each site, microbial biomass-N and soil carbohydrates increased and extractable soil N decreased in response to watering alone. Fertilization with N increased microbial biomass-N in grassland soils; whereas, fertilization with C increased microbial biomass-N and decreased extractable N and P in all communities dominated by shrubs, which have invaded large areas of grassland in the Chihuahuan desert during the last 100 years. Our results support the hypothesis that the control of soil microbial biomass shifts from N to C when the ratio of C to N decreases during desertification.     

大豆连作对土体和根际微生物群落功能的影响

试验采用Biolog方法在大田试验条件下研究了不同连作年限大豆在结荚期和收获期根际和土体微生物群落功能多样性的变化.试验结果表明,在结荚期和收获期根际微生物群落功能多样性、均匀度指数和AWCD均显著高于土体.在结荚期迎茬和连作8a的土体微生物多样性、AWCD均高于正茬和连作4a土体;在收获期正茬和迎茬处理的土体微生物多样性、AWCD高于连作4a和8a.结荚期迎茬、连作4a和8a处理的根际微生物群落AWCD均显著高于收获期,说明在大豆植株生长旺盛的结荚期微生物群落的根际效应比收获期更明显.在这两个生长期降解氨基酸,糖类和羧酸类碳源的微生物可能是连作影响的主要土体微生物类群.     

Periodic carbon flushing to roots of Quercus rubra saplings affects soil respiration and rhizosphere microbial biomass

Patterns of root/shoot carbon allocation within plants have been studied at length. The extent, however, to which patterns of carbon allocation from shoots to roots affect the timing and quantity of organic carbon release from roots to soil is not known. We employed a novel approach to study how natural short-term variation in the allocation of carbon to roots may affect rhizosphere soil biology. Taking advantage of the semi-determinate phenology of young northern red oak (Quercus rubra L.), we examined how pulsed delivery of carbon from shoots to roots affected dynamics of soil respiration as well as microbial biomass and net nitrogen mineralization in the rhizosphere. Young Q. rubra exhibit (1) clear switches in the amount of carbon allocated below-ground that are non-destructively detected simply by observing pulsed shoot growth above-ground, and (2) multiple switches in internal carbon allocation during a single growing season, ensuring our ability to detect short-term effects of plant carbon allocation on rhizosphere biology separate from longer-term seasonal effects. In both potted oaks and oaks rooted in soil, soil respiration varied inversely with shoot flush stage through several oak shoot flushes. In addition, upon destructive harvest of potted oaks, microbial biomass in the rhizosphere of saplings with actively flushing shoots was lower than microbial biomass in the rhizosphere of saplings with shoots that were not flushing. Given that plants have evolved with their roots in contact with soil microbes, known species-specific carbon allocation patterns within plants may provide insight into interactions among roots, symbionts, and free-living microbes in the dynamic soil arena.     

Expediting COD removal in microbial electrolysis cells by increasing biomass concentration

Microorganisms catalyse the reaction and in this study, mainly the effect of different concentration of biomass on COD removal was investigated. Three sets of two-compartment reactors were established. The cation exchange membrane (CEM) was employed in each reactor and 0.5 V of electricity was supplied. Graphite rod employed in cathodic part and a combination of graphite rod and graphite granules were used in anodic chamber. The highest rate of COD removal (40 ± 2.0 ppm/h) was achieved in the reactor which had initial VSS at 6130 mg/l, whereas the slowest rate of 23 ± 1.2 ppm/h in the reactor started with 3365 mgVSS/l. Some ammonia removal was also noticed during the operation. Further understanding and improvement is needed to be competitive against traditional wastewater treatment processes.     

紫云英根际微生物碳源利用多样性研究

为探究绿肥根际调控效果的原因,采用碳素利用法-BIOLOG生态版,分析紫云英根际微生物种群结构和功能多样性。结果表明:紫云英根际土壤微生物总活性比非根际高,其主要贡献者是以氨基酸、糖类、酯类和醇类四大碳源为生的微生物,而与以磷酸盐、羧酸和胺类为碳源的微生物关系不大。进一步分析发现:紫云英根际产生了以α-酮丁酸、2-羟基安息香酸、4-羟基安息香酸、D-半乳糖醛酸、α-环式糊精、D-木糖、β-甲基-D-葡萄糖、D-甘露醇和L-精氨酸作为碳源的新微生物种群,完全抑制了以γ-羟基丁酸和苯乙基胺为碳源的微生物种群活性;根泌物抑制了以D-苹果酸、衣康酸和肝糖为碳源的微生物活性,而对以D-化纤二糖、N-乙酰基-D-葡萄胺、丙酮酸甲酯、吐温40、吐温80、L-苯基丙氨酸、L-天门酰胺酸和腐胺为碳源的微生物有促进作用,但对以D-氨基葡糖酸、α-D-乳糖和I-赤藻糖醇为碳源的微生物活性无显著影响;另外,即使是以同一类物质为碳源的微生物种群,在紫云英根际的表现也不完全一样,如以D-乳酸γ-内酯和L-丝氨酸为碳源的可快速培养微生物受到抑制,可慢速培养的微生物活性则有提高,而以甘氨酰基-L-谷氨酸为碳源的微生物的表现正好与之相反。     

荒漠土壤微生物量碳、氮变化对降水的响应

以腾格里沙漠东南缘的典型荒漠植被为研究对象,通过遮雨棚和滴灌系统设置5个降水梯度,即极端干旱处理、中度干旱处理、对照、增水处理I和增水处理II,研究了荒漠土壤微生物量碳(MBC)、氮(MBN)和微生物碳氮比(MBC/MBN)对季节、降水和土壤深度的响应规律,以期为极端降水事件影响干旱荒漠区土壤微生物量碳、氮及其循环规律的深入研究提供科学依据。结果表明:(1)MBC、MBN和MBC/MBN对降水处理的响应存在差异,三者的变化范围为:230.14—272.87 mg/kg,13.82—17.58 mg/kg,19.78—36.06。其中,降水处理对MBC、MBN的影响显著,对MBC/MBN的影响不显著,在极端干旱处理下,MBC、MBN均显著高于其他降水处理;(2)两年间的MBC、MBN和MBC/MBN差异显著,2017年较2016年MBC、MNB显著减少,MBC/MBN显著增加;(3)MBC、MBN和MBC/MBN变化均表现季节性差异,变化范围分别为:153.31—337.09 mg/kg,7.89—22.29 mg/kg,14.82—46.04,其中MBC、MBN为春季最高、秋季最低,M...