Optimization of an oxygen-based approach for community-level physiological profiling of soils

Current approaches for rapid assessment of carbon source utilization by whole soil communities (i.e., community-level physiological profiling or CLPP) provide a limited, biased view of microbial communities with little connection to in situ activities. We developed an alternative CLPP approach based upon fluorometric detection of dissolved oxygen consumption in a microtiter platform which offers flexible manipulation of experimental factors. In the attempt to reduce oxygen re-dissolution, the wells were filled with liquid to very near the top and sealed. We found that filling the wells with 240 vs. 150 μl of sample improved the sensitivity of the system to discern both the response to a substrate amendment as low as 10 mg l⁻¹ and un-amended, endogenous respiration. The preparation of a soil slurry facilitates inoculation into the microplate. Disruption of soil samples had a limited effect on the endogenous respiration in comparison to intact soil microbags in a 24-well microplate. Storage time (up to 33 days) reduced the level of activity in intact soil microbags but not in disrupted samples. A microcosm fertilization experiment was set to study the effects of N availability on the respiratory response in the plates. The use of soil organic carbon (SOC) and amended C-substrates (50 mg l⁻¹) was increased by the addition of nitrogen (N) in the plate, and appeared N-limited shortly after microcosm fertilization. The addition of the eukaryotic inhibitor cycloheximide delayed the initial increase in fluorescence (time to minimum response) of several C sources (casein, acetate, asparagine, coumaric acid), varying among soils, which could be explained by the fungal use of these compounds. However, the extent of the inhibition caused by cycloheximide did not increase at higher fungal to bacteria ratios as estimated by PLFA analysis, indicating that the direct estimation of the fungal biomass from cycloheximide addition is not feasible. This paper provides an optimized, standardized protocol for soil analysis, and sets the basis for further validation studies that will continue to define the underlying capabilities/biases of this approach.     

Gaseous and leaching nitrogen losses from no-tillage and conventional tillage systems following surface application of cattle manure

Previous studies have demonstrated inconsistent results on the impact of tillage systems on nitrogen (N) losses from field-applied manure. This study assessed the impact of no-tillage (NT) and conventional tillage (CT) systems on gaseous N losses, N₂O:N₂O + N₂ ratios and NO₃⁻-N leaching following surface application of cattle manure. The study was undertaken during the 2003/2004 and 2004/2005 seasons at two field sites in Nova Scotia namely, Streets Ridge (SR) in Cumberland County and the Bio-environmental Engineering Centre (BEEC) in Truro. Results showed that the NT system had higher (p < 0.05) NH₃ losses than CT. Over the two seasons, manure incorporation in CT reduced NH₃ losses on average by 86% at SR and 78% at BEEC relative to NT. At both sites and during both seasons, denitrification rates and N₂O fluxes in NT were generally higher than in CT plots, presumably due to higher soil water and organic matter content in NT. Over the two seasons, mean denitrification rates at SR were 239 and 119 g N ha⁻¹ d⁻¹, while N₂O fluxes were 120 and 64 g N ha⁻¹ d⁻¹ under NT and CT, respectively. At BEEC mean denitrification rates were 114 and 71 g N ha⁻¹ d⁻¹, while N₂O fluxes were 52 and 27 g N ha⁻¹ d⁻¹ under NT and CT, respectively. Conversely, N₂O:N₂O + N₂ ratios were lower in NT than CT suggesting more complete reduction of N₂O to N₂ under NT. When averaged across all soil depths, NO₃⁻-N was higher (p < 0.05) in CT than NT. Nitrate-N decreased with depth at both sites regardless of tillage. In most cases, NO₃⁻-N was higher under CT than NT at all soil depths. Similarly, flow-weighted average NO₃⁻-N concentrations in drainage water were generally higher under CT. This may be partly attributed to higher denitrification rates under NT. Therefore, NT may be a viable strategy to remove NO₃⁻-N from the soil, and thus, reduce NO₃⁻-N contamination of groundwater. However, it should be noted that while the use of NT reduces NO₃⁻-N leaching it may come with unintended environmental tradeoffs, including increased NH₃ and N₂O emissions.     

Soil chemical and microbiological properties in hay production systems: residual effects of contrasting N fertilization of swine lagoon effluent versus ammonium nitrate

This study characterized soil chemical and microbiological properties in hay production systems that received from 0 to 600 kg plant-available N (PAN) ha⁻¹ year⁻¹ from either swine lagoon effluent (SLE) or ammonium nitrate (AN) from 1999 to 2001. The forage systems contained plots planted with bermudagrass (Cynodon dactylon L.) or endophyte-free tall fescue (Festuca arundinaceae Schreb.). In March 2004, the plots were sampled for measurements of a suite of soil chemical and microbiological properties. Nitrogen fertilization rates were significantly correlated with soil pH and K₂SO₄-extractable soil C but not with total soil C, soil C/N ratio, electrical conductivity, or Mehlich-3-extractable nutrients. Soil supplied with SLE had significantly lower Mehlich-3-extractable nutrients than the soil supplied with AN. Two indicators of soil N-supplying capacity (potentially mineralizable N and amino sugar N) varied with plant species and the type of N fertilizer. However, they generally peaked at an application rate of 200 or 400 kg PAN ha⁻¹ year⁻¹. Soil microbial biomass C also peaked at an application rate of 200 or 400 kg PAN ha⁻¹ year⁻¹. Nitrification potential was significantly higher in soil supplied with AN than in the unfertilized control but was similar between SLE-fertilized and unfertilized soils. Our results indicated that an application rate as high as 600 kg PAN ha⁻¹ year⁻¹ did not benefit soil microbial biomass, microbial activity, and N transformation processes in these forage systems.     

Cutting regime determines allocation of fixed nitrogen in white clover

Leguminous leys are important sources of nitrogen (N), especially in forage-based animal production and organic cropping. Models for estimating total N₂ fixation of leys—including below-ground plant-derived N (BGN)—are based on grazed or harvested leys. However, green manure leys can have different proportions of above-ground plant-derived N (AGN) and BGN when subjected to different cutting regimes. To investigate the effects of cutting on N distribution in white clover, a pot experiment was carried out using ¹⁵N techniques to determine N₂ fixation, N rhizodeposition and root C and N content of cut and uncut white clover (Trifolium repens L. cv. Ramona) plants. Percentage N derived from air (%Ndfa) was lower in uncut (63%) than in cut (72%) plants, but total Ndfa was not significantly affected by cutting. The higher reliance on N₂ fixation in cut plants was thus counterbalanced by lower biomass and total N content. With BGN taken into account, total plant-derived N increased by approximately 50% compared with AGN only. Cutting did not affect the proportion of BGN to standing shoot biomass N after regrowth, but decreased the proportion of BGN to total shoot biomass production during the entire growth period. Thus, estimates of N fixation in green manure leys should consider management practices such as cutting regime, as this can result in differences in above- and below-ground proportions of plant-derived N.     

陕西关中地区肉牛产地同位素溯源技术初探

为应用牛组织中C、N同位素组成的区域分布和同位素指标溯源肉牛产地,本文利用同位素比率质谱仪(IRMS)对陕西关中不同区县来源的牛尾毛样品的1δ3C和1δ5N值进行检测,通过聚类分析,研究陕西关中地区肉牛组织中同位素组成的区域分布情况。结果表明,杨凌区、眉县牛尾毛样品聚为一类;乾县、永寿县样品聚为一类;麟游县样品聚为一类;扶风县、岐山县和凤翔县分类不十分明显,分散在上述3类中。说明陕西关中不同地区肉牛组织中同位素组成存在差异,利用它们可进行肉牛地域的小范围溯源。     

氮素形态、用量及施用时期对小青菜产量和硝酸盐含量的影响

采用田间试验研究了氮素形态、用量及施用时期对小青菜产量和硝酸盐含量的影响。结果表明,等氮量施肥下,产量最高的是硝酸钙,尿素+微量元素处理;尿素,有机无机复混肥和碳酸氢铵处理间无显著差异,DMPP处理产量最低;追施微量元素能提高小青菜的产量。硝酸盐含量随着收获时期的延后而降低,有机无机复混肥处理则略有升高,但未达到显著水平;收获期取样时,不同氮肥对硝酸盐的积累是:硝酸钙＞DMPP＞尿素＞有机无机复混肥＞尿素+微量元素＞碳酸氢铵。配施微量元素及氮磷钾的协同吸收均能降低硝酸盐含量。综合考虑产量和品质指标,以有机无机复混肥处理效果较好。小青菜产量和Vc含量随着施氮量的提高而提高,但硝酸盐含量也随之提高;小青菜产量和Vc含量随着施氮时期的延后而降低,硝酸盐含量高峰出现在追肥后20d左右。     

稻草易地还土对丘陵红壤团聚体碳氮分布的影响

利用新垦坡地、熟化旱地两个定位试验的典型处理(不施肥、化肥、稻草 NP),研究稻草易地还土对丘陵红壤水稳定性大团聚体(>0.25 mm)内碳、氮分布的影响。结果表明,稻草易地还土提高了土壤大团聚体含量;团聚体粒径越大,有机碳和全氮的含量越高;与对照相比,稻草易地还土提高大团聚体内的有机碳含量10.1%~78.5%、全氮含量9.4%~63.6%(p<0.05),同时也显著提高二者在较大粒径(1~2 mm,2~5 mm)团聚体内的分配比例;与化肥处理相比,稻草易地还土提高>0.5 mm各粒级大团聚体内的有机碳含量6.5%~46.1%,也提高了0.5~1 mm,1~2 mm,2~5 mm团聚体内的全氮含量8.7%~16.1%(p<0.05)。水稳定性大团聚体对土壤碳、氮具有强富集和物理保护作用;稻草易地还土提高水稳定性大团聚体内碳、氮的含量和分配比例,是改良丘陵红壤结构、提高并协调土壤肥力的有效保育措施。     

N,N- 二甲基甲酰胺对炔雌醚不育作用和适口性的影响

【目的】明确添加高效、安全的助剂 N,N- 二甲基甲酰胺 (N,N-Dimethylformamide,DMF) 对提高不 育剂炔雌醚（Quinestrol）药饵的不育效果,以及改善炔雌醚药饵对鼠类适口性的作用。【方法】试验设 3 个处理组, 分别投喂三组小鼠空白饵料（C 组）、炔雌醚药饵（T1 组）和 DMF- 炔雌醚药饵（T2 组）,计算饵料消耗量。 将 3 个处理组的雄、雌鼠合笼 7 d,记录雌鼠怀孕及产仔情况。高效液相色谱法检测不同配制的药饵中炔雌醚的 分布浓度。【结果】T1 和 T2 组对雄、雌鼠体重均有显著影响,且 T1 和 T2 组雄鼠肾脏器官鲜重分别低于 C 组 16.4% 和 19.7%。炔雌醚对雌性小鼠繁殖抑制作用明显,T1 和 T2 组雌鼠产仔率降低至 C 组的 37.5% 和 25.0%, 产仔数仅为 C 组的 35.0% 和 11.1%。DMF 对炔雌醚药饵适口性改善作用不明显,高效液相色谱法检测药饵中药 物的分布浓度结果表明,T2 组的糙米中炔雌醚的含量为 T1 组的 1.5 倍;DMF 提高了炔雌醚在糙米中的渗透, 从而增加了小鼠的药物摄入量。【结论】DMF 可快速有效溶解炔雌醚,并增强其在糙米中的渗透力,提高炔雌 醚药饵的抗生育作用,有望开发为具有良好应用前景的不育剂配方。     

不同氮效率油菜种质苗期氮吸收转运与利用差异研究

氮效率研究是油菜营养性状遗传改良的前提,为探究不同氮效率油菜种质苗期氮吸收、转运和利用的异同,以2个氮效率差异油菜种质H6（氮高效）和L18（氮低效）为供试材料,利用水培营养液设置正常氮（CK）和低氮（LN）2个氮浓度处理,培养14d后检测植株氮含量,计算氮累积量、氮转运系数和氮利用效率。结果显示,不同氮浓度处理对油菜苗期氮的吸收、转运和利用效率影响的差异达到极显著水平（P＜0.01）。与CK相比,LN处理的油菜生物量、氮含量和氮累积量均显著降低,而氮利用效率和根冠比显著提高;氮高效油菜种质H6的生物量、氮累积量、氮转运系数和氮生理利用效率均显著大于氮低效种质L18,分别为L18的2.07、1.42、3.23和1.56倍。从氮的吸收、转运和利用3个方面探讨了低氮胁迫处理下油菜种质苗期氮效率差异的原因,对深入开展油菜氮高效机理研究及油菜品种氮效率改良具有一定的指导意义。     

土壤有机碳对沼渣或牛粪施用后温室气体排放潜力的影响—盆栽试验结果

A change in the European Union energy policy has markedly promoted the expansion of biogas production.Consequently,large amounts of nutrient-rich residues are being used as organic fertilizers.In this study,a pot experiment was conducted to simulate the high-risk situation of enhanced greenhouse gas (GHG) emissions following organic fertilizer application in energy maize cultivation.We hypothesized that cattle slurry application enhanced CO2 and N2O fluxes compared to biogas digestate because of the overall higher carbon (C) and nitrogen (N) input,and that higher levels of CO2 and N2O emissions could be expected by increasing soil organic C (SOC) and N contents.Biogas digestate and cattle slurry,at a rate of 150 kg NH4+-N ha-1,were incorporated into 3 soil types with low,medium,and high SOC contents (Cambisol,Mollic Gleysol,and Sapric Histosol,termed Clow,Cmedium,and Chigh,respectively).The GHG exchange (CO2,CH4,and N2O) was measured on 5 replicates over a period of 22 d using the closed chamber technique.The application of cattle slurry resulted in significantly higher CO2 and N2O fluxes compared to the application of biogas digestate.No differences were observed in CH4 exchange,which was close to zero for all treatments.Significantly higher CO2 emissions were observed in Chigh compared to the other two soil types,whereas the highest N2O emissions were observed in Cmedium.Thus,the results demonstrate the importance of soil type-adapted fertilization with respect to changing soil physical and environmental conditions.