低氮胁迫下苦荞根际土壤纤维素酶活性的响应机制：荧光光谱法测定

土壤纤维素酶参与土壤中养分的循环过程,分解纤维素为小分子糖分微生物生命活动提供能源物质,并且对生态系统有重要的指示作用。如何快速、精确的测定纤维素酶活性对土壤生态系统诊断、恢复等有重要指示意义。自然界中不同的氮肥处理及种植不同的苦荞品种均会对土壤纤维素酶活性产生影响。利用德国进口全自动进样酶标仪(InfiniteR 200 PRO),以4-Methylumbelliferyl β-D-cellobiopyranoside (Cel) 纤维素荧光物质为底物,将不同耐瘠性苦荞(迪庆苦荞-耐低氮;黑丰1号-不耐低氮)根际土壤以悬液形式用排枪吸取微量和荧光底物混合加入到96孔微孔板进行充分反映、培养测定土壤中纤维素酶活性,其中将对照板和样品放在同一个板块上,减少培养空间占用和操作时间。结果表明,除了苦荞成熟期的常氮处理,苗期和开花期的低氮处理条件下,纤维素酶活性均表现为迪庆苦荞根际土壤显著高于黑丰1号,迪庆根际纤维素酶活性在苗期低氮处理下高于黑丰39.50%;纤维素酶活性在迪庆苦荞的开花期和成熟期氮处理间有显著性差异,表现为常氮高于其他处理,而黑丰1号仅在开花期时常氮处理和其他处理之间有显著差异;灭菌处理酶活性显著低于常氮处理的54.29%,说明迪庆苦荞根际纤维素酶一部分来源于微生物的贡献,养分充足时微生物的活性更高,贡献越大。荧光光谱法分析出的纤维素酶活性表明耐瘠性品种迪庆苦荞可以通过增加根际土壤酶活性的方法来抵抗外界环境的胁迫,面对黄土高原养分贫瘠的土壤,可以考虑选种耐贫瘠的品种来降低成本增加收益。试验过程中,荧光标记的酶标仪测定方法与传统的测定方法相比更加快捷、准确和节约,同样适合大批量样品测定,可以为未来精准农业生产,测土配方施肥提供及时的数据支持。     

外源植酸酶添加对土壤磷酸酶活性影响的荧光光谱法研究

外源植酸酶加入土壤中,能直接或者间接提高有机磷的生物有效性。但外源植酸酶添加后,对土壤本身的磷酸酶(单酯酶、二酯酶)活性会产生何种影响尚不明确。采用荧光物质为底物,将96微孔板和荧光检测法相结合,利用多功能酶标仪测定外源植酸酶(1 g/50 g干土)添加条件下红壤、棕壤以及褐土中磷酸单酯酶(酸性和碱性)和二酯酶的活性响应。结果表明,外源植酸酶添加后,酸性磷酸单酯酶活性在红壤中极显著增加(p≤0.01),而在褐土中显著降低;碱性磷酸单酯酶活性在棕壤和褐土中增加,其中在褐土中增加极显著(p≤0.01),而在红壤中显著降低;磷酸二酯酶活性在三种土壤中均不同程度地增加,其中在棕壤中增加极显著(p≤0.01);不同土壤中,培养到8天时不同酶活性亦保持增加,即红壤中的酸性磷酸单酯酶,棕壤中的磷酸二酯酶以及褐土中的碱性磷酸单酯酶分别保持活性增加,说明外源植酸酶的添加能够有效地增强土壤磷酸酶活性,这种作用不仅与土壤性质如pH和磷形态相关,并且可能与刺激微生物分泌酶量有关。运用荧光光谱法研究外源植酸酶对土壤磷酸酶活性的影响在国内外尚属首次。与传统的分光光度法相比, 微孔板荧光法是一种灵敏、准确、快速、简便的土壤磷酸酶活性关系的测定方法。     

壳聚糖固定化脲酶活性的X射线微区分析

利用X射线微区分析方法,对壳聚糖固定化脲酶活性进行了定位分析。筛选出了能捕捉固定化脲酶活性部位的捕捉剂BaCl₂,以尿素作为底物,在Tris-HCl缓冲溶液中,底物经固定化脲酶催化产生NH₃和CO₂,后者和捕捉剂反应生成BaCO₃沉淀, 沉积在固定化脲酶的催化活性部位,借此确定其催化活性部位。结果表明BaCl₂可用做固定化脲酶活性定位的捕捉剂。同时得到了壳聚糖固定化脲酶活性定位的最佳实验条件。     

热重红外光谱法考察木质生物质综纤维素热转化特性

热重红外光谱联用考察了木质生物质纤维素、半纤维素以及综纤维素的热转化特性,并与微晶纤维素和木聚糖等模型化合物进行了对比分析。应用三维扩散模型计算了活化能、指前因子等热转化动力学参数,拟合效果良好。通过分析气相产物三维IR图谱,在最大失重速率附近,观察到了H₂O,CO,CO₂,CH₄和含氧化合物的明显特征峰。讨论了主要气体产物可能的生成途径,发现其产量顺序为CO₂>H₂O>CO≈CH₄。综合分析得出,综纤维素的热转化过程是纤维素主导下、纤维素和半纤维素综合作用的结果。     

外源植酸酶添加对土壤磷酸酶活性影响的荧光光谱法研究（英文）

外源植酸酶加入土壤中,能直接或者间接提高有机磷的生物有效性。但外源植酸酶添加后,对土壤本身的磷酸酶(单酯酶、二酯酶)活性会产生何种影响尚不明确。采用荧光物质为底物,将96微孔板和荧光检测法相结合,利用多功能酶标仪测定外源植酸酶(1g/50g干土)添加条件下红壤、棕壤以及褐土中磷酸单酯酶(酸性和碱性)和二酯酶的活性响应。结果表明,外源植酸酶添加后,酸性磷酸单酯酶活性在红壤中极显著增加(p≤0.01),而在褐土中显著降低;碱性磷酸单酯酶活性在棕壤和褐土中增加,其中在褐土中增加极显著(p≤0.01),而在红壤中显著降低;磷酸二酯酶活性在三种土壤中均不同程度地增加,其中在棕壤中增加极显著(p≤0.01);不同土壤中,培养到8天时不同酶活性亦保持增加,即红壤中的酸性磷酸单酯酶,棕壤中的磷酸二酯酶以及褐土中的碱性磷酸单酯酶分别保持活性增加,说明外源植酸酶的添加能够有效地增强土壤磷酸酶活性,这种作用不仅与土壤性质如pH和磷形态相关,并且可能与刺激微生物分泌酶量有关。运用荧光光谱法研究外源植酸酶对土壤磷酸酶活性的影响在国内外尚属首次。与传统的分光光度法相比,微孔板荧光法是一种灵敏、准确、快速、简便的土壤磷酸酶活性关系的测定方法。     

基于非分散红外(NDIR)技术的土壤剖面二氧化碳浓度的测定

为了探索土壤剖面CO₂浓度以及不同土壤层(腐殖质H层、A层、B层、C层)土壤呼吸的变化规律,应用非分散红外(NDIR)技术的新方法,持续不间断的测量土壤剖面二氧化碳浓度。实验所用的主要仪器为硅基非分散红外测量仪,能在高湿、高粉尘、污垢及其他恶劣环境中进行光谱数据采集。通过2013年全年光谱测定值的采集,并应用梯度法模型计算不同深度土壤碳通量,同时利用LI-8100碳通量自动监测系统持续监测的土壤碳通量值进行回归分析。结果显示：土壤剖面CO₂浓度呈现明显的梯度变化,即随着土壤深度的增加,土壤CO₂浓度增大;梯度法模型得出的不同土壤层的土壤呼吸模拟值与实测土壤呼吸值之间具有较好的线性相关,H,A,B,C层的模型预测的决定系数(R2)分别为0.906 9,0.718 5,0.838 2,0.903 0,均方根误差(RMSE)分别为0.206 7,0.104 1,0.015 6,0.009 6。均达到了较好的预测结果,表明该方法对定量分析不同土壤层碳通量是可行的。该方法具有清晰揭示土壤CO₂在不同土壤层之间的传输规律,以及有助于分析不同土壤层土壤呼吸特性的优点,能为全球土壤剖面碳通量计算提供基础数据,是一种具有发展前途的传感器。     

地下储存CO2 泄漏胁迫下地表植被光谱变化特征及识别研究

随着全球气候变暖,减少温室气体排放成为全世界所关注的问题,而碳捕捉与储存(carbon capture and storage,CCS)技术可以减少温室气体CO₂排放量,但储存在地下的CO₂有泄漏的风险。本工作的目的是通过野外模拟实验,研究地表植被(甜菜)在CO₂轻微泄漏胁迫下其叶片叶绿素含量、水分含量及光谱变化特征,结果表明CO₂泄漏胁迫的甜菜叶绿素与叶片含水量明显降低,叶片反射率在550 nm减小,而在680 nm增大。设计了比值指数R₅₅₀/R₆₈₀进行识别CO₂泄漏胁迫的甜菜,发现该指数能够在胁迫发生7天后识别出胁迫的甜菜,且该指数具有较强的敏感性、稳健性及识别能力。研究结果对于未来CCS项目选址、地表生态监测评估、遥感监测CO₂泄漏点等都具有重要的现实意义与应用价值。     

2 μm附近CO2谱线参数测量及应用

吸收光谱技术用于痕量气体浓度监测,特别是在气体分子稳定同位素丰度探测中,吸收谱线参数的准确性非常重要,目前普遍使用的HITRAN数据库中给出的各项参数具有一定的不确定性。为利用2.0 μm激光波段进行CO₂浓度及其同位素丰度探测,需要对该波段的CO₂吸收谱线参数进行校准,采用窄线宽分布反馈式二极管激光器作为光源,结合自行搭建的谱线参数测量系统,采集了2.0 μm波段10条CO₂吸收谱线,获得了各谱线的位置、强度、自加宽系数和N₂加宽系数,并与HITRAN2012数据库中相应的数据进行对比发现两者之间吻合较好,CO₂谱线强度和自加宽系数相对偏差均小于2%。实测实验室大气的CO₂浓度为440 ppm,13CO₂的丰度值δ为-9‰。测量结果为该波段应用于CO₂浓度及13CO₂同位素丰度的实时在线探测提供了重要参考依据。     

荧光光谱法测定生物炭/秸秆输入土壤后酶活性的变化

土壤α-葡萄糖苷酶、β-葡萄糖苷酶直接参与土壤有机物质的矿化过程,对维持生态系统碳和养分的循环起重要作用。秸秆或者秸秆炭化物还田是提高碳储量的一种有效措施,输入到土壤后对生物学活性产生一定影响,对碳库转化具有直接或者间接作用。采用荧光物质作为底物,将96微孔板和荧光检测法结合,利用多功能酶标仪测定生物炭/秸秆(2.5 g/50 g干土)添加条件下黑土中α(β)葡萄糖苷酶活性。结果表明,秸秆添加到土壤后,土壤α(β)葡萄糖苷酶活性增强,水稻秸秆处理β-葡萄糖苷酶活性高于玉米秸秆处理,40天后仍然保持较强活性,说明秸秆的输入有助于土壤碳素转化。可能是因为秸秆炭添加提高土壤中养分含量,促进微生物活性,使得土壤酶活性提高,水稻秸秆添加增强土壤酶活性,而玉米秸秆没有促进作用,可能与材料来源不同有关,材料来源与添加效应的关系有待进一步研究。而生物炭添加对黑土中α(β)葡萄糖苷酶活性影响不大,这与生物炭含速效养分少、自身难降解特性有关。与传统的分光光度法相比,微孔板荧光法可以灵敏的检测到土壤悬液中的酶活性,可批量检测样品,是一种快速、准确、简便的土壤酶活性测定方法。     

同位素质谱法对土壤反硝化酶活性的测定研究

在评价硝化抑制剂的作用效果时, 需要确认此硝化抑制剂对反硝化过程有无影响。而对此过程的确认是通过反硝化酶活性(denitrifying enzyme activity, 简写为DEA)的测定来实现的。采用加入同位素标记底物结合同位素质谱仪来测定新型硝化抑制剂3,4-二甲基吡唑磷酸盐作用下的反硝化酶活性。结果表明, 此新型方法能够较准确的测定培养体系中的N₂O的浓度,与气相色谱法的测定结果具有良好的相关性(MS_N2O=-0.45+1.03GC_N2O,R2=0.995)。通过测定15N₂O和15N₂的丰度能够较好的区分2种反硝化酶活性(硝酸还原酶和N₂O还原酶), 且克服了传统测定中需要加入乙炔作为酶抑制剂的弊端。对DMPP作用下土壤反硝化酶的测定表明,DMPP对反硝化酶无显著影响, 说明DMPP在使用中不会影响土壤中的反硝化过程。     

Isotopic ((13)C) fractionation during plant residue decomposition and its implications for soil organic matter studies

Carbon isotopic fractionations in plant materials and those occurring during decomposition have direct implications in studies of short-and longer-term soil organic matter dynamics. Thus the products of decomposition, the evolved CO(2) and the newly formed soil organic matter, may vary in their (13)C signature from that of the original plant material. To evaluate the importance of such fractionation processes, the variations in (13)C signatures between and within plant parts of a tropical grass (Brachiaria humidicola) and tropical legume (Desmodium ovalifolium) were measured and the changes in (13)C content (signatures) during decomposition were monitored over a period of four months. As expected the grass materials were less depleted in (13)C (-11.4 to -11.9 per thousand) than those of the legume (-27.3 to -25.8 per thousand). Root materials of the legume were less (1.5 per thousand) depleted in (13)C compared with the leaves. Plant lignin-C was strongly depleted in (13)C compared with the bulk material by up to 2.5 per thousand in the legume and up to 4.7 per thousand in the grass. Plant materials were subsequently incubated in a sand/nutrient-solution/microbial inoculum mixture. The respiration product CO(2) was trapped in NaOH and precipitated as CaCO(3), suitable for analysis using an automated C/N analyser coupled to an isotope ratio mass spectrometer. Significant depletion in (13)C of the evolved CO(2) was observed during the initial stages of decomposition probably as a result of microbial fractionation as it was not associated with the (13)C signatures of the measured more decomposable fractions (non-acid detergent fibre and cellulose). While the cumulative CO(2)-(13)C signatures of legume materials became slightly enriched with ongoing decomposition, the CO(2)-C of the grass materials remained depleted in (13)C. Associated isotopic fractionation correction factors for source identification of CO(2-)C varied with time and suggested errors of 2-19% in the estimation of the plant-derived C at 119 days of incubation in a soil of an intermediate (-20.0 per thousand) (13)C signature. Analysis of the residual material after 119 days of incubation showed little or no change in the (13)C signature partly due to the incomplete decomposition at the time of harvesting. Copyright 1999 John Wiley & Sons, Ltd.     

Sampling soil-derived CO2 for analysis of isotopic composition: a comparison of different techniques

A new system for soil respiration measurement [P. Rochette, L.B. Flanagan, E.G. Gregorich. Separating soil respiration into plant and soil components using analyses of the natural abundance of carbon-13. Soil Sci. Soc. Am. J., 63, 1207-1213 (1999).] was modified in order to collect soil-derived CO2 for stable isotope analysis. The aim of this study was to assess the suitability of this modified soil respiration system to determine the isotopic composition (delta13C) of soil CO2 efflux and to measure, at the same time, the soil CO2 efflux rate, with the further advantage of collecting only one air sample. A comparison between different methods of air collection from the soil was carried out in a laboratory experiment. Our system, as well as the other dynamic chamber approach tested, appeared to sample the soil CO2, which is enriched with respect to the soil CO2 efflux, probably because of a mass dependent fractionation during diffusion and because of the atmospheric contribution in the upper soil layer. On the contrary, the static accumulation of CO2 into the chamber headspace allows sampling of delta13C-CO2 of soil CO2 efflux.     

冬小麦不同生育时期叶面积指数反演方法

针对当前作物叶面积指数遥感反演过程中,在不同生育时期采用相同的植被指数进行反演存在叶面积指数反演精度较低的问题。以冬小麦为研究对象,选取了对冬小麦覆盖度响应程度不同的六种宽带和四种窄带共10种植被指数,分析比较了在冬小麦整个生育期选用当前广泛使用的归一化植被指数(NDVI)反演冬小麦的LAI和在冬小麦不同生长阶段选用不同的植被指数反演冬小麦LAI的结果差异。在冬小麦整个生育期内使用NDVI反演小麦LAI得到的LAI反演值和真实值之间的R2=0.558 5,RMSE=0.320 9。改进的比值植被指数(mSR)适合于反演冬小麦生长前期(拔节期之前)的LAI,得到的LAI反演值和真实值之间的相关系数r=0.728 7,均方根误差RMSE=0.297 1;比值植被指数(SR)适于反演冬小麦生长中期(拔节到抽穗前),得到的LAI反演值和真实值之间的R2=0.654 6,RMSE=0.306 1;NDVI适于反演冬小麦生长后期(抽穗到成熟期)的LAI,得到的LAI反演值和真实值之间的R2=0.679 4,均方根误差RMSE=0.316 4。 研究表明：在冬小麦的不同生育时期,根据地表作物覆盖度的变化和反射率的变化,选择不同的植被指数建立冬小麦LAI的反演模型获得的反演精度均高于在冬小麦整个生育期使用NDVI获得的反演结果。说明在冬小麦的不同生育时期选择不同的植被指数构建LAI的分段反演模型可以改善冬小麦LAI的反演精度。     

Soil matrix tracer contamination and canopy recycling did not impair ¹³CO₂ plant-soil pulse labelling experiments

When conducting (13)CO(2) plant-soil pulse labelling experiments, tracer material might cause unwanted side effects which potentially affect δ(13)C measurements of soil respiration (δ(13)C(SR)) and the subsequent data interpretation. First, when the soil matrix is not isolated from the atmosphere, contamination of the soil matrix with tracer material occurs leading to a physical back-diffusion from soil pores. Second, when using canopy chambers continuously, (13)CO(2) is permanently re-introduced into the atmosphere due to leaf respiration which then aids re-assimilation of tracer material by the canopy. Accordingly, two climate chamber experiments on European beech saplings (Fagus sylvatica L.) were conducted to evaluate the influence of soil matrix (13)CO(2) contamination and canopy recycling on soil (13)CO(2) efflux during (13)CO(2) plant-soil pulse labelling experiments. For this purpose, a combined soil/canopy chamber system was developed which separates soil and canopy compartments in order to (a) prevent diffusion of (13)C tracer into the soil chamber during a (13)CO(2) canopy pulse labelling and (b) study stable isotope processes in soil and canopy individually and independently. In combination with laser spectrometry measuring CO(2) isotopologue mixing ratios at a rate of 1 Hz, we were able to measure δ(13)C in canopy and soil at very high temporal resolution. For the soil matrix contamination experiment, (13)CO(2) was applied to bare soil, canopy only or, simultaneously, to soil and canopy of the beech trees. The obtained δ(13)C(SR) fluxes from the different treatments were then compared with respect to label re-appearance, first peak time and magnitude. By determining the δ(13)C(SR) decay of physical (13)CO(2) back-diffusion from bare soils (contamination), it was possible to separate biological and physical components in δ(13)C(SR) of a combined flux of both. A second pulse labelling experiment, with chambers permanently enclosing the canopy, revealed that (13)CO(2) recycling at canopy level had no effect on δ(13)C(SR) dynamics.     

Soil respiration rates and δ13C(CO2) in natural beech forest (Fagus sylvatica L.) in relation to stand structure

Soil respiration rates were studied as a function of soil type, texture and light intensity at five selected natural beech forest stands with contrasting geology: stands on silicate bedrock at Kladje and Bricka in Pohorje, a stand on quartz sandstone at Vrhovo and two stands on a carbonate bedrock in the Karstic-Dinaric area in Kocevski Rog, Snezna jama and Rajhenav, Slovenia, during the growing season in 2005-2006. Soil respiration exhibited pronounced seasonal and spatial variations in the studied forest ecosystem plots. The CO(2) flux rates ranged from minimum averages of 2.3?μmol CO(2) m(-2) s(-1) (winter) to maximum averages of about 7?μmol CO(2) m(-2) s(-1) (summer) at all the investigated locations. An empirical model describing the relationship between soil respiration and soil temperature predicted seasonal variations in soil respiration reasonably well during 2006. Nevertheless, there were also some indications that soil moisture in relation to soil texture could influence the soil CO(2) efflux rates in both sampling seasons. It was shown that spatial variability of mean soil respiration at the investigated sites was high and strongly related to root biomass. Based on the [image omitted] data, it was shown that new photoassimilates could account for a major part of the total soil respiration under canopy conditions in forest ecosystems where no carbonate rocks are present, indicating that microbial respiration could not always dominate bulk soil CO(2) fluxes. At Snezna jama and Rajhenav, the abiotic CO(2) derived from carbonate dissolution had a pronounced influence on CO(2) efflux accounting, on average, to ～17%. Further spatial heterogeneity of soil respiration was clearly affected by management practice. Higher respiration rates as well as higher variability in respiration rates were observed in the virgin forest (Rajhenav) than in the management forest (Snezna jama) and could be related to a higher amount of detritus and consequently to less pronounced influence of inorganic pool to CO(2) efflux, lower mixing with atmospheric CO(2) and higher sensitivity to environmental changes. Major differences in soil carbon dynamics among these five forest ecosystems can be explained by the influence of bedrock geology (particularly, the presence or absence of carbonate minerals) and soil texture (affecting gas exchange with overlying air and soil moisture).     

Mechanistic study of the CO oxidation reaction on the CuO (111) surface during chemical looping combustion

Cu-based oxides oxygen carriers and catalysts are found to exhibit attractive activity for CO oxidation, but the dispute with respect to the reaction mechanism of CO and O₂ on the CuO surface still remains. This work reports the kinetic study of CO oxidation on the CuO (111) surface by considering the adsorption, reaction and desorption processes based on density functional theory calculations with dispersion correction (DFT-D). The Eley–Rideal (ER) CO oxidation mechanism was found to be more feasible than the Mars-van-Krevelen (MvK) and Langmuir–Hinshelwood (LH) mechanisms, which is quite different from previous knowledge. The energy barrier of ER, LH, and MvK mechanisms are 0.557, 0.965, and 0.999 eV respectively at 0 K. The energy barrier of CO reaction with the adsorbed O species on the surface is as low as 0.106 eV, which is much more active in reacting with CO molecules than the lattice O of CuO (111) surface (0.999 eV). A comparison with the catalytic activity of the perfect Cu₂O (111) surface shows that the ER mechanism dictates both the perfect Cu₂O (111) and the CuO (111) surface activity for CO oxidation. The activity of the perfect Cu₂O (111) surface is higher than that of the perfect CuO (111) surface at elevated temperatures. A micro-kinetic model of CO oxidation on the perfect CuO (111) surface is established by providing the rate constants of elementary reaction steps in the Arrhenius form, which could be helpful for the modeling work of CO catalytic oxidation.     

洪涝胁迫的水稻叶面积指数变化及其光谱响应研究

通过水稻淹水实验模拟洪涝胁迫状态,分析不同生育期、不同洪涝胁迫强度下的水稻叶面积指数变化及其冠层高光谱响应规律,建立洪涝胁迫下水稻叶面积指数(LAI)的估测模型。结果表明,分蘖期、拔节期、抽穗期水稻LAI均随淹水深度的增加而降低;水稻冠层680~760 nm光谱的一阶微分具有“双峰”现象,主峰位于724~737 nm,701和718 nm有次峰出现,并随淹水深度的增加出现“三峰”;冠层红边位置在各个生育期出现“蓝移”,红边幅值、红边面积在分蘖期、拔节期、灌浆期均呈现“蓝移”,在抽穗期出现“红移”;生育前期水稻LAI与光谱特征参量存在显著相关,生育后期相关性不显著;最后以D_λ737/D_λ718光谱参量建立了水稻生育前期的LAI估测模型。表明在洪涝胁迫下,水稻LAI的变化幅度直接反映胁迫强度的大小,估测模型LAI=3.138(D_λ737/D_λ718)-0.806可用于估测洪涝胁迫不同强度下的水稻叶面积指数。     

Anisotropic Nucleation,Growth and Ripening under Stirring—A Phenomenological Model

The anisotropic formation of elongated metal-oxide aggregates in water under intensive stirring is analyzed. It is treated in terms of anisotropic ballistically mediated aggregation kinetics in open systems. The basic kinetic equations describing the stages of homogeneous nucleation, independent growth, and ripening of the aggregates are formulated for the open system under the external influence with the stirring intensity as the main parameter governing the process. The most significant elongation of the aggregates is shown to evolve at the ripening stage.