Linking soil bacterial biodiversity and soil carbon stability

Native soil carbon (C) can be lost in response to fresh C inputs, a phenomenon observed for decades yet still not understood. Using dual-stable isotope probing, we show that changes in the diversity and composition of two functional bacterial groups occur with this ‘priming'' effect. A single-substrate pulse suppressed native soil C loss and reduced bacterial diversity, whereas repeated substrate pulses stimulated native soil C loss and increased diversity. Increased diversity after repeated C amendments contrasts with resource competition theory, and may be explained by increased predation as evidenced by a decrease in bacterial 16S rRNA gene copies. Our results suggest that biodiversity and composition of the soil microbial community change in concert with its functioning, with consequences for native soil C stability.Substrate inputs can stimulate decomposition of native soil organic carbon (SOC; Kuzyakov et al., 2000), a phenomenon known as the ‘priming effect'' (Kuzyakov, 2010), and is considered large enough to influence ecosystem C balance (Wieder et al., 2013). Two functionally distinct groups of microorganisms are postulated to mediate priming: one that grows rapidly utilizing labile C, and one that grows slowly, breaking down recalcitrant SOC (Fontaine et al., 2003; Blagodatskaya et al., 2007). However, distinguishing these groups is technically challenging. Here, we used dual-stable isotope probing with ¹³C-glucose and ¹⁸O-water to identify bacteria in these two groups growing in response to single and repeated pulses of glucose. Organisms that utilize labile C for growth assimilate both ¹³C-glucose and ¹⁸O-water into their DNA, whereas organisms that grow using SOC incorporate only ¹⁸O-water. Differential isotope incorporation leads to a range of DNA densities separable through isopycnic centrifugation, which can then be characterized by sequencing (Radajewski et al., 2000).We sequenced fragments of bacterial 16S rRNA genes following single and repeated glucose pulses. We hypothesized that the single pulse of labile C would stimulate growth of opportunistic organisms, thus immobilizing nutrients and suppressing growth and diversity of the SOC-utilizing community, decreasing SOC decomposition (negative priming), a response analogous to that observed in plant communities in response to chronic N additions (Tilman, 1987; Clark and Tilman, 2008). We hypothesized that multiple glucose additions would stimulate growth of a more diverse bacterial community, including more native SOC-utilizing organisms that possess enzymes to decompose recalcitrant compounds, causing positive priming (Fontaine et al., 2003; Kuzyakov, 2010).Soil from a ponderosa pine ecosystem was amended weekly for 7 weeks with 500 μg C-glucose per gram soil (2.65 atom % ¹³C) in 100 μl deionized water or with 100 μl deionized water (n=5). Measurements of δ¹³C–CO₂ and [CO₂] enabled the partitioning of CO₂ into that derived from added glucose or from native SOC (C_SOC):where C_total is CO₂–C from glucose-amended samples, δ_total is the δ¹³C–CO₂ from glucose-amended samples, δ_glucose is the δ¹³C of the added glucose and δ_SOC is the δ¹³C–CO₂ evolved from the non-amended samples. Priming was calculated as the difference between SOC oxidation of the amended and non-amended samples. With this approach, any evolved CO₂ carrying the ¹³C signature of the added glucose is considered respiration of glucose, including ¹³C-labeled biomass and metabolites derived from prior glucose additions. Thus, this approach quantifies priming as the oxidation of SOC present at the beginning of the experiment, consistent with many other studies of priming (Cheng et al., 2003; De Graaff et al., 2010).In a parallel incubation for dual-stable isotope probing, the repeated-pulse samples received unlabeled glucose (500 μg C-glucose per gram soil) for 6 weeks while the non-amended and single-pulse samples received sterile deionized water. In week 7, samples received one of four isotope treatments (n=3): 97 atom % H₂ ¹⁸O (non-amended soil), 99 atom % ¹³C-glucose and 97 atom % H₂ ¹⁸O (single- and repeated-pulse soil), ¹²C-glucose and 97 atom % H₂ ¹⁸O (repeated-pulse soil) or ¹²C-glucose and H₂ ¹⁶O (repeated-pulse soil). After incubating for 7 days, soil was frozen at −40 °C. DNA was extracted, separated through isopycnic centrifugation, and two density ranges were sequenced for the bacterial 16S rRNA gene (Supplementary Figure 1): 1.731–1.746 g ml⁻¹ (hereafter called the SOC-utilizing community) and 1.759–1.774 g ml⁻¹ (hereafter called the glucose-utilizing community).Amplicons of the V3–V6 16S rRNA region were bar coded with broad-coverage fusion PCR primers and pooled before sequencing on a Genome Sequencer FLX instrument. These sequence data have been submitted to the GenBank database under accession number SRP043371. Data were checked for chimeras (Edgar et al., 2011), demultiplexed and quality checked (Caporaso et al., 2010). Taxonomy was assigned to genus at the ⩾80% bootstrap confidence level (Cole et al., 2009).We used the Shannon''s diversity index (H′), commonly used in microbial systems (Fierer and Jackson, 2006), to assess changes in microbial diversity. Analysis of variance was used to compare the amount of DNA within densities between isotope treatments (Supplementary Figure 2) and to test the effects of the treatments on the Shannon''s diversity (Figure 2) and Pielou''s evenness (Supplementary Figure 3) of the active bacterial communities, with post hoc Student''s t-tests, α=0.05. PRIMER 6 and PERMANOVA were used to create the nonmetric multidimensional scaling ordination and to compare bacterial communities between glucose treatments and the two sequenced density ranges.The single pulse of glucose suppressed SOC oxidation, whereas repeated pulses increased SOC oxidation (Figure 1). Few experiments to date have examined priming in response to repeated substrate amendments (Hamer and Marschner, 2005; Qiao et al., 2014), even though in nature soil receives repeated substrate pulses from litterfall and rhizodeposition. Our results demonstrate the dynamic response of SOC decomposition to repeated labile C inputs.Open in a separate windowFigure 1Weekly priming rates calculated as the difference in SOC respired between glucose-amended and non-amended soil (n=5).Dual-stable isotope probing was able to separate the growing bacteria into two groups with distinct DNA densities (P<0.001, PERMANOVA; Figure 3a), indicating differential uptake of ¹³C-glucose and ¹⁸O-water. In response to the initial glucose addition, the diversity of the growing glucose- and SOC-utilizing bacterial communities declined compared with the non-amended community (P<0.001, t-tests; Figure 2), driven by a strong decrease in evenness (Supplementary Figure 3). In the SOC-utilizing community, where DNA was labeled with ¹⁸O only, the relative abundance of Bacillus increased 4.9-fold compared with the non-amended control to constitute 31.6% of the community (Figure 3b). Bacillus survives well under low-nutrient conditions (Panikov, 1995), and is able to synthesize a suite of extracellular enzymes capable of degrading complex substrates (Priest, 1977), traits that are conducive for using SOC for growth. In the glucose-utilizing community, where DNA was labeled with both ¹³C and ¹⁸O, Arthrobacter increased 67.7-fold relative to the non-amended control to constitute 75.5% of the growing bacteria (Figure 3b). In culture experiments, Arthrobacter can rapidly take up and store glucose for later use (Panikov, 1995) and here we find it dominating the high-density DNA fractions, signifying that it is using the labeled glucose to grow. The increased biomass of Arthrobacter may have resulted in greater resource competition, thus reducing the diversity of the growing community, as is frequently found in plant communities (Bakelaar and Odum, 1978; Clark and Tilman, 2008).Open in a separate windowFigure 2Shannon''s diversity index (H′) of the non-amended, single-pulse, and repeated-pulse treatments (n=3) in the SOC- (mid-density) and glucose-utilizing (high-density) communities. Treatments with the same letter are not significantly different from each other (Student''s t, α=0.05).Open in a separate windowFigure 3(a) Nonmetric multidimensional scaling ordination showing differences in growing bacterial communities at the genus taxonomic level in the SOC-utilizing (mid-density; open symbols) and glucose-utilizing (high-density; closed symbols) groups of non-amended (Δ), single-pulse (○) and repeated-pulse (□) treatments (n=3). (b) Pie charts of genera in the SOC- and glucose-utilizing communities of the single- and repeated-pulse treatments (n=3). Genera with relative abundances >5% are listed in the figure legend.After repeated glucose amendments, the diversity of the growing community recovered to non-amendment levels (Figure 2) without strongly dominant organisms (Figure 3b and Supplementary Figure 3). The higher diversity found after repeated glucose pulses may be explained by trophic interactions where predators graze on prey populations that have been enlarged by resource addition, suppressing competition between prey species and causing secondary mobilization of nutrients (Clarholm, 1985). The decrease in total bacterial 16S rRNA gene copies in the repeated-pulse—compared with the single-pulse—treatment (Supplementary Figure 4) supports predation as a potential mechanism explaining the observed diversity increase after repeated glucose pulses.The recovery of diversity after repeated glucose pulses contrasts with resource competition theory (Tilman, 1987). When chronic additions of a limiting resource are applied, species diversity and evenness typically decrease (Bakelaar and Odum, 1978; Clark and Tilman, 2008) because competitive organisms become dominant. We observed this after the single glucose pulse, but not after repeated pulses. This diversity response may be the result of community shifts facilitated by short bacterial life cycles and the tens to hundreds of generations expected during the 7-week incubation (Behera and Wagner, 1974). In contrast, systems on which most ecological theory is based (for example, plants) might achieve perhaps 20 generations in a multi-decadal field experiment (Bakelaar and Odum, 1978; Clark and Tilman, 2008). With more generations, more community dynamics can occur, including increased resource cascades in which extracellular enzymes, metabolites or lysed cells of one functional group increase substrates for another (Blagodatskaya and Kuzyakov, 2008). Our results highlight the opportunity to test ecological theories in microbial ecosystems (Prosser et al., 2007), particularly as the short life cycles of microbes makes them well suited for pursuing ecological questions in an evolutionary framework (Jessup et al., 2004).The priming effect is ubiquitous, yet its drivers remain elusive. Our results suggest that changes in the diversity and composition of the growing bacterial community contribute to priming, and thus that ecosystem properties such as soil C storage may be sensitive to soil microbial biodiversity.     

植物、土壤及土壤管理对土壤微生物群落结构的影响

土壤微生物是土壤生态系统的重要组成部分,对土壤微生物群落结构多样性的研究是近年来土壤生态学研究的热点。本文综述了有关植物、土壤类型以及土壤管理措施对土壤微生物群落结构影响的最新研究结果,指出植物的作用因植物群落结构多样性、植物种类、同种植物不同的基因型,甚至同一植物不同根的区域而异;而土壤的作用与土壤质地和有机质含量等因素有关;植物和土壤类型在对土壤微生物群落结构影响上的作用存在互作关系。不同的土壤管理措施对土壤微生物群落结构影响较大,长期连作、大量的外援化学物质的应用降低了土壤微生物的多样性;而施用有机肥、免耕可以增加土壤微生物群落结构多样性,有利于维持土壤生态系统的功能。     

Distinct impacts of reductive soil disinfestation and chemical soil disinfestation on soil fungal communities and memberships

Applied Microbiology and Biotechnology - Soil disinfestation is an important agricultural practice to conquer soil-borne diseases and thereby ensure crop productivity. Reductive soil disinfestation...     

Available soil nitrogen in relation to fractions of soil nitrogen and other soil properties

The available N of 27 soils from England and Wales was assessed from the amounts of N taken up over a 6-month period by perennial ryegrass grown in pots under uniform environmental conditions. Relationships between availability and the distribution of soil N amongst various fractions were then examined using multiple regression. The relationship: available soil N (mg kg^–1 dry soil)=(N_min×0.672)+(N_inc×0.840)+(N_mom×0.227)–5.12 was found to account for 91% of the variance in available soil N, where N_min=mineral N, N_inc=N mineralized on incubation and N_mom=N in macro-organic matter. The N mineralized on incubation appeared to be derived largely from sources other than the macro-organic matter because these two fractions were poorly correlated. When availability was expressed in terms of available organic N as % of soil organic N (N_ao) the closest relationship with other soil characteristics was: N_ao=[N_inc×(1.395–0.0347×CN_mom]+[N_mom×0.1416], where CN_mom=CN ratio of the macro-organic matter. This relationship accounted for 81% of the variance in the availability of the soil organic N.The conclusion that the macro-organic matter may contribute substantially to the available N was confirmed by a subsidiary experiment in which the macro-organic fraction was separated from about 20 kg of a grassland soil. The uptake of N by ryegrass was then assessed on two subsamples of this soil, one without the macro-organic matter and the other with this fraction returned: uptake was appreciably increased by the macro-organic matter.     

The porosity of soil aggregates from bulk soil and from soil adhering to roots

Summary Total porosity and pore-size distribution (p.s.d.) were determined in soil aggregates taken in plots planted with maize and treated with farmyard manure and three rates of compost. Soil aggregates were collected from the soil adherent to the maize roots (root soil aggregates) and from bulk soil (bulk soil aggregates). Mercury intrusion porosimetry was used to evaluate the total porosity and the p.s.d. Treatments did not affect the total porosity of the bulk soil aggregates. The same was observed for the root soil aggregates. However the total porosity of the root soil aggregates was always lower than that of the bulk soil aggregates. The loss of total porosity was found to be due to a decrease in the percentage of larger pores with respect to the total.     

Estimating respiration of roots in soil: Interactions with soil CO2, soil temperature and soil water content

Little information is available on the variability of the dynamics of the actual and observed root respiration rate in relation to abiotic factors. In this study, we describe I) interactions between soil CO₂ concentration, temperature, soil water content and root respiration, and II) the effect of short-term fluctuations of these three environmental factors on the relation between actual and observed root respiration rates. We designed an automated, open, gas-exchange system that allows continuous measurements on 12 chambers with intact roots in soil. By using three distinct chamber designs with each a different path for the air flow, we were able to measure root respiration over a 50-fold range of soil CO₂ concentrations (400 to 25000 ppm) and to separate the effect of irrigation on observed vs. actual root respiration rate. All respiration measurements were made on one-year-old citrus seedlings in sterilized sandy soil with minimal organic material.Root respiration was strongly affected by diurnal fluctuations in temperature (Q₁₀ = 2), which agrees well with the literature. In contrast to earlier findings for Douglas-fir (Qi et al., 1994), root respiration rates of citrus were not affected by soil CO₂ concentrations (400 to 25000 ppm CO₂; pH around 6). Soil CO₂ was strongly affected by soil water content but not by respiration measurements, unless the air flow for root respiration measurements was directed through the soil. The latter method of measuring root respiration reduced soil CO₂ concentration to that of incoming air. Irrigation caused a temporary reduction in CO₂ diffusion, decreasing the observed respiration rates obtained by techniques that depended on diffusion. This apparent drop in respiration rate did not occur if the air flow was directed through the soil. Our dynamic data are used to indicate the optimal method of measuring root respiration in soil, in relation to the objectives and limitations of the experimental conditions.     

The breakdown of malathion in soil and soil components

The disappearance of the organophosphorus insecticide, malathion, from a silt loam soil and from its organic and inorganic components was examined. Half-lives and the time taken for 90% decomposition in nonsterile, sodium azide-treated, and 2.5 Mrad-irradiated soils were similar (3/4–1 1/2 days and 4–6 days, respectively) but breakdown in autoclaved soils was negligible. Decay in nonsterile sand, silt, and clay minus organic matter fractions was 3–6 times slower than that recorded in the original soil. Breakdown of malathion in the clay plus organic matter fraction (organo-mineral complex) was rapid (half-life, 1 day), as was the case in the separated organic matter (half-life, 1 3/4 days). Filter-sterilized organic matter was not as effective in catalyzing the breakdown of malathion (half-life, 4 days), and no loss occurred from any of the autoclaved components. Irradiation doses of 2.5 and 5.0 Mrad had little influence on the ability of soil to degrade malathion. Thereafter, increases up to 20 Mrad had a more drastic, though far from totally inhibitory, effect. Our results suggest that either the colloidal organic matter itself, or a fraction associated with it, is the most important single factor concerned with the rapid breakdown of malathion in the soil studied. Direct microbial metabolism is a slower process and may have a significant role in malathion disappearance in coarsetextured soils low in colloidal organic matter. The catalytic component of the organic matter is suggested to be a stable exoenzyme and is supportive of reports by other workers. The quantitative effect of organo-mineral complex (containing the active degradative ingredient) additions to sand and silt fractions on the rate of subsequent malathion decay is also described.     

Crop diversification reinforces soil microbiome functions and soil health

Plant and Soil - Intensive agriculture with continuous monocropping and massive chemical inputs has adversely affected belowground microbial composition and functions, resulting in soil sickness...     

Harvester ant nests, soil biota and soil chemistry

Many ant species accumulate organic debris in the vicinity of their nests. These organic materials should provide a rich resource base for the soil biota. We examined the effect of harvester ant nests (Pogonomyrmex barbatus) on the soil community and soil chemistry. Ant nest soils supported 30-fold higher densities of microarthropods and 5-fold higher densities of protozoa than surrounding, control soils. The relative abundances of the major groups of protozoa differed as well: amoebae and ciliates were relatively overrepresented, and flagellates underrepresented, in ant nest versus control soils. Densities of bacteria and fungi were similar in the two soil types. Concentrations of nitrate, ammonium, phosphorus, and potassium were significantly higher in ant nest soils, while concentrations of magnesium, calcium, and water were similar in nest and control soils. Ant nest soils were marginally more acidic than controls. The results demonstrate that P. barbatus nests constitute a significant source of spatial heterogeneity in soil biota and soil chemistry in arid grasslands. Received: 17 March 1997 / Accepted: 10 June 1997     

土壤活性有机质及其与土壤质量的关系

活性有机质是土壤的重要组成部分 ,主要包括溶解性有机碳、微生物生物量、轻组有机质。它在土壤中具有重要作用 :(1)可以表征土壤物质循环特征、评价土壤质量 ,可以作为土壤潜在生产力以及由土壤管理措施引起土壤有机质变化的早期指标 ;(2 )在养分周转中起重要作用 ,是植物的养分库 ,可以提供植物所需要的养分如氮、磷、硫等 ;(3)能稳定土壤结构 ,对维持团粒结构稳定性有重要作用。从土壤养分、土壤物理、化学性质方面讨论了活性有机质与土壤质量的关系。土壤中的溶解性有机碳、微生物生物量碳氮含量与土壤有机碳、全氮和碱解氮等物质的含量呈正相关。活性有机质受土壤质地、含水量、温度等因素影响 ,与土壤酸碱度、阳离子交换量等也有关。土壤微生物生物量碳和微生物量 C/有机碳比与土壤粘粒、粉粒含量呈正相关、与砂粒含量呈负相关     

水土保持林土壤抗蚀性能评价研究

运用土壤有机质含量、水稳性团聚体含量、水稳性团粒平均重量直径、团聚度和分散系数等各项指标,对不同树种组成、不同林龄水土保持林的土壤抗蚀性能进行分析、评价.结果表明,水土保持林对于提高土壤抗蚀性能具有重要作用,这种作用主要针对表层土壤而言;与油松纯林相比,油松阔叶树混交林土壤有机质含量较高,水稳性团聚体含量增加了1.71%～38.53%;且随着林龄的增长,水土保持林土壤抗蚀性能不断增强.     

土壤侵蚀对土壤肥力及土地生物生产力的影响

通过对红壤坡地不同土地利用方式土壤肥力及土地生物生产力的空间分异研究,揭示了土壤侵蚀对土壤肥力和土地生产力的负面影响．即侵蚀导致Ｎ、Ｐ、Ｋ等土壤速效养分含量减少及其在坡面上部的相对贫乏和下部的相对富集;土壤有机质含量降低;土壤机械组成中砂、粉、粘粒比率发生变化,表现为土壤沙化,土地生物生产力下降．     

原油污染对黄绵土和风沙土水分入渗的影响

原油进入土壤后会堵塞土壤孔隙,影响土壤斥水性,改变土壤水分运动状况。本研究利用土柱模拟的方法,研究了不同原油污染程度(0、0.5%、1%、2%、4%)对黄绵土和风沙土水分入渗过程的影响。结果表明: 随着原油含量的增加,两种土壤湿润锋的推进速度和入渗速率均减小,土壤原油污染程度为4%时湿润锋运移到土柱底部的所需时间最长,污染程度为0时湿润锋运移到土柱底部的所需时间最短,黄绵土湿润锋达到土柱底部所需最长时间是最短时间的5倍,风沙土最长时间是最短时间的48倍;当湿润锋运移到土柱底部时,黄绵土的累积入渗量随原油含量的增加而减小,而风沙土的累积入渗量先增大后减小;在高浓度(2%、4%)原油处理下,风沙土的累积入渗量曲线出现“翘尾”现象。Kostiakov入渗模型和Philip入渗模型比Green-Ampt模型能更好地模拟不同原油处理下的黄绵土土壤水分入渗过程,但对风沙土而言,两种模型对低浓度(0、0.5%、1%)原油处理的土壤水分入渗过程拟合较好。原油污染能够显著影响土壤水分入渗过程,且对风沙土的影响更大。     

Acid rain and soil

A summary of important chemical properties of soil is given and the way in which acid rain may affect these properties is discussed. Acid rain may suppress microbiological decomposition and nitrification processes, thus influencing the nutrient status of soils. It has also been found that soil organic matter is less soluble in more acid solutions. Changed nutrient availability patterns are predicted in a low pH environment and enhanced leaching of essential elements from the soil exchange complex has been observed. Increased solubility of potentially toxic elements such as aluminium may also occur from soils which have been exposed to acidified rainfall.     

Mycorrhizas and soil structure

In addition to their well-recognized roles in plant nutrition and communities, mycorrhizas can influence the key ecosystem process of soil aggregation. Here we review the contribution of mycorrhizas, mostly focused on arbuscular mycorrhizal fungi (AMF), to soil structure at various hierarchical levels: plant community; individual root; and the soil mycelium. There are a suite of mechanisms by which mycorrhizal fungi can influence soil aggregation at each of these various scales. By extension of these mechanisms to the question of fungal diversity, it is recognized that different species or communities of fungi can promote soil aggregation to different degrees. We argue that soil aggregation should be included in a more complete 'multifunctional' perspective of mycorrhizal ecology, and that in-depth understanding of mycorrhizas/soil process relationships will require analyses emphasizing feedbacks between soil structure and mycorrhizas, rather than a uni-directional approach simply addressing mycorrhizal effects on soils. We finish the discussion by highlighting new tools, developments and foci that will probably be crucial in further understanding mycorrhizal contributions to soil structure.     

Earthworms and soil fertility

Summary Earthworms redistribute organic materials within the soil, increase soil penetrability and, und certain conditions, influence ion transport in soils. Root distribution may be modified and microbial activity increased by their burrowing and feeding activities. Earthworms influence the supply of nutrients in several ways. Not only is earthworm tissue and cast material enriched in certain nutrients, relative to the soil matrix, but ingestion of organic material increases the rate of cycling. Certain farm-management practices, such as cultuvation and the use of acidic fertilizers, reduce the ability of earthworm to improve plant growth. Where other inorganic fertilizers increase the growth of plants, an increase in earthworm numbers can be expected because of the increased food supply. Lime, in particular, and possibly drainage also increase earthworm activity. Further research is required on the physical and biological effects of earthworms on nutrient supply, so that suitable management practices can be developed to optimise the beneficial effects of earthworms on soil fertility.Introductory lecture