四年O3熏气对小麦根际土壤氮素微生物转化的影响

采用开顶箱（open top chambers,OTC） 法设置3个臭氧浓度梯度,连续4a对小麦生长季土壤进行臭氧增加试验。测定小麦拔节、孕穗、抽穗、灌浆和成熟期的根际土壤微生物量氮、氨氧化细菌、反硝化细菌数量以及硝化和反硝化强度。结果显示微生物量氮、氨氧化细菌数量和硝化强度随小麦生育进程的推进,呈先升高后降低的趋势,4a变化趋势相同。O₃胁迫下,小麦根际微生物量氮、氨氧化细菌数量、硝化强度亦降低,与对照比较,均达显著水平（P < 0.05）;随O₃作用年限增加,抑制效应越强,第4年 O₃对其的抑制率显著高于第1年的抑制率。反硝化细菌数量在前期没有显著变化,成熟期则增加两个数量级,O₃显著促进成熟期反硝化细菌数量增加。4a试验有相同的变化趋势。在较短时间里O₃对土壤反硝化强度没有显著影响,而作用3个生长季后,反硝化强度显著升高。结果说明,O₃浓度升高降低了麦田土壤微生物量氮、氨氧化细菌数量和硝化强度,并有一定的积累效应。O₃剂量和作用时间的累积量达到一定阈值,显著增加土壤反硝化细菌数量和反硝化强度,增加麦田土壤N₂O排放的风险。土壤氮素微生物转化受小麦生长发育状况和O₃剂量、作用时间的共同影响,不同形态的氮素对O₃的敏感阈值不同,响应的时间和变化的范围有差异。     

大气CO2浓度增高对麦田土壤硝化和反硝化细菌的影响

硝化和反硝化细菌是土壤中与氮转化有关的微生物菌群 ,大气CO2 浓度升高可能对它们的数量产生影响。位于中国无锡的稻 麦轮作农田生态系统FACE平台 2 0 0 1年 6月开始运行。本试验在 2 0 0 3年小麦生长季研究了土壤 (0～ 5cm和 5～ 10cm土层 )中硝化和反硝化细菌在大气CO2 浓度升高条件下的变化。试验采用最大可能法 (MPN)计这两种微生物菌群的数量。结果表明 ,0～ 5cm土层硝化菌数拔节期和成熟期FACE低于对照 ,而孕穗期FACE高于对照 ,5～ 10cm土层硝化菌数越冬期与成熟期FACE低于对照 ,大气CO2 浓度升高使得麦田土壤硝化细菌数目减少。 0～ 5cm土层各个生长期反硝化菌数FACE与对照均没有明显差异 ,5～ 10cm土层反硝化菌数拔节期FACE低于对照 ,大气CO2 浓度升高对麦田土壤反硝化菌的影响不大。     

设施菜田不同碳氮管理对反硝化菌结构和功能的影响

【目的】通过6年长期定位试验,比较设施菜田不同碳氮管理下反硝化菌结构和功能的差异。【方法】采用末端限制性片段多态性(T-RFLP)和变性梯度凝胶电泳(DGGE)方法分别分析nir K/nir S和nos Z型反硝化菌群结构特征,利用自动连续在线培养监测体系(Robot系统)测定分析NO/(NO3-+NO2-)和N2O/(N2O+N2)产物比,并通过乙炔抑制法测定反硝化酶活性。【结果】传统施肥处理(CN)显著改变了nir K和nos Z型反硝化菌的结构,增加了NO/(NO3-+NO2-)和N2O/(N2O+N2)产物比。nir S型菌受碳氮管理影响较小。减氮(RN)和添加秸秆处理(RN+S)的nir K和nos Z型反硝化菌结构与CN处理的差异性显著,且会显著降低NO/(NO3-+NO2-)和N2O/(N2O+N2)产物比;与CN和RN相比,RN+S显著增加反硝化酶活性。【结论】设施菜田长期传统施肥措施改变了反硝化菌的结构和功能,增加土壤自身的NO产生能力并减弱了N2O还原N2的能力。减氮和添加秸秆管理能形成自身的反硝化菌群结构,并降低NO和N2O排放风险;秸秆的添加会促进反硝化潜在速率,降低菜田NO3-淋洗风险。     

O3浓度升高和太阳辐射减弱对小麦根际土壤微生物功能多样性的影响

采用开顶箱(OTC)法和遮光网技术,设置100 n L/L臭氧熏气与3个辐射减弱梯度结合,模拟臭氧浓度升高和太阳辐射减弱的复合大气背景。用BIOLOG生态测试板,采用孔平均颜色变化率法(AWCD)测定冬小麦根际土壤微生物利用不同碳源的能力,计算微生物群落多样性指数,对不同碳源的利用率进行了主成分分析。两年试验结果显示,臭氧熏气与太阳辐射减弱复合作用,降低了土壤微生物对碳源的利用速度和利用总量;除了聚合物以外其它碳源利用率显著降低;对土壤微生物多样性没有直接的影响;对碳源降解的抑制效应大于增强的O3与减弱的太阳辐射两因素各自的单独作用。太阳辐射减弱20%,一定程度上增加了对聚合物类的分解。O3熏气条件下太阳辐射减弱,糖类、胺类代谢变异度较高,受环境影响较大。     

地表太阳辐射减弱对小麦根际土壤微生物碳代谢指纹的影响

采用遮荫网设置4个太阳辐射通量梯度(20%、40%、60%、100%),对小麦生长季麦田土壤进行太阳辐射减弱的试验,用BIOLOG生态测试板(ECO MicroPlate),采用孔平均颜色变化率法(average well-color development,AWCD)测定不同太阳辐射通量处理下的冬小麦根际土壤微生物利用单一碳源的能力,计算微生物群落多样性指数,对不同碳源的利用率进行主成分分析,重复2年的试验,研究太阳辐射减弱对麦田土壤微生物多样性的影响。结果表明:1)太阳辐射减弱,土壤微生物对单一碳源的利用速度降低,碳源分解的总量降低;2)土壤微生物优势种群、群落丰富度和个体数量在群落中的分布状况未受到太阳辐射减弱的直接影响;3)微生物对大多数碳源的代谢能力减弱,糖类代谢受辐射减弱的影响最大,变异度最高。结果说明,小麦根际土壤微生物碳代谢受地表太阳辐射减弱的影响,这为了解大气环境变化对土壤微生态的影响提供了参考。     

分离于河口区芦苇湿地1株好氧反硝化菌的鉴定及其反硝化特性

【目的】筛选高效脱氮且N_2O释放量少的好氧反硝化细菌,并对菌株的反硝化特性进行研究,可为河口湿地富营养化水体的生物修复提供技术支撑。【方法】经BTB培养基初筛和反硝化能力测定,从辽河河口区芦苇湿地土壤中分离得到1株具有较高反硝化能力的好氧反硝化菌C3。经形态观察、生理生化鉴定和16S rRNA序列分析,对菌株进行鉴定。研究温度、碳源、pH及C/N对其生长量、反硝化能力及N2O释放的影响。【结果】筛选得到的高效好氧反硝化细菌C3,经鉴定属于假单胞菌属(Pseudomonas sp.)。反硝化特性研究结果表明,该菌最适碳源为柠檬酸三钠,在温度为30°C、pH为7.0、C/N为10时生长速率和脱氮效率最高且N_2O释放量较少。在此条件下,该菌在36 h内使NO_3~–由179.55 mg/L降至5.08 mg/L,脱氮率高达97.17%。该菌株在整个反硝化过程中中间产物N_2O的最大累积量较低,为0.22 mg/L。【结论】从湿地土壤中分离所得好氧反硝化菌C3为假单胞菌属的1个种(Pseudomonas sp.),该菌株在高效除氮和低N_2O累积方面均具有明显优势,对后续河口湿地富营养化水体治理具有重要意义。     

施肥制度对水稻土壤nosZ型反硝化细菌群落的影响

由含氧化亚氮还原酶(NOS)的反硝化细菌驱动的氧化亚氮(N2O)还原成氮气(N2)的过程是N2O排放的重要调控途径。为探明施肥对稻田土壤nosZ型反硝化细菌群落的影响,采用荧光定量PCR和高通量测序等方法,研究了湖南省宁乡县长达30年的定位试验条件下4种施肥制度[不施肥(CK)、化肥(CF)、70%化肥+30%有机肥(CFM1)和40%化肥+60%有机肥(CFM2)]对水稻土壤nosZ型反硝化细菌数量和群落结构的影响。结果表明:不同施肥处理nosZ基因丰度为2.14×10~8～6.09×10~8copies·g~(-1)干土,施肥处理nosZ基因拷贝数比对照低47.3%～64.8%(P0.05),但不同有机肥配施比例处理间nosZ基因拷贝数差异不显著;变形菌门是优势门水平类群,占总序列的60.2%～77.5%; Bacteria_unclassified、Proteobacteria_unclassified、Betaproteobacteria_unclassified和根瘤菌目为优势目水平类群,占总序列的93.6%～95.9%。施肥显著降低了Proteobacteria_unclassified的相对丰度(P0.01),但显著提高了根瘤菌目、environmental_samples和红环菌目的相对丰度(P0.05);施肥显著改变nosZ型反硝化细菌的群落结构,但有机肥配施比例对其影响较弱;除碳氮比外,其他土壤理化性质均显著影响nosZ型反硝化细菌的数量和群落结构,其中,硝态氮和土壤p H是驱动nosZ型反硝化细菌群落变化的主要因子;施肥显著影响nosZ型反硝化细菌数量和群落结构,有机肥配施比例对nosZ型反硝化细菌群落的影响较弱,研究结果为进一步阐述施肥制度对土壤反硝化微生物的影响提供依据。     

中亚热带水热条件对农田置换土壤硝化强度的影响

水热条件是影响土壤硝化过程的主要环境因素.选择我国东部地区3种主要农田土壤(黑土、潮土、红壤),在江西鹰潭设置野外土壤置换试验,模拟研究中亚热带水热条件对不同土壤硝化强度的影响.2006年的试验结果表明:玉米生育期中,水热条件的变化影响土壤硝化强度的变化,从玉米种植前(2006年4月17日)到生长旺盛期(2006年7月10日),月平均气温由19.4℃上升到30.2℃,月降水量由335.6mm减少到59.2mm,3种土壤硝化强度均下降,黑土、潮土和红壤分别下降63 0%～84.8%、42.9%～66.7%和43.7%～46.2%;到玉米成熟期(月平均气温29.25℃,月降水175.6mm),土壤硝化强度又有所升高.土壤类型显著影响了土壤硝化细菌数量和硝化强度,硝化细菌数和硝化强度大小顺序为:潮土>黑土>红壤.土壤pH显著影响土壤硝化强度,相关系数r=0.778(P<0.01).此外,在玉米生育期,施用N、P、K化肥后,3种土壤的硝化细菌数以及硝化强度均升高.总体上,区域水热状况、土壤性质和施肥均影响土壤硝化强度,而且土壤与气温以及施肥处理之间存在显著的交互作用.     

黄土性土壤剖面不同层次N2O浓度的原位监测

用土壤探头法对黄土性土壤玉米-小麦轮作体系下不同剖面层次N2O浓度变化进行了3a的田间原位监测.结果证实了黄土性土壤深层反硝化作用的存在,且N2O浓度有着明显的时间和空间变异.表现为N2O浓度受土壤气候条件(温度和降水)和生产管理措施的影响,丰水年明显高于亏水年;在降水或灌溉后出现瞬时N2O浓度峰.由于小麦和玉米生长特点和作物生长季气侯特点的差异,玉米生长季土壤剖面各层N2O浓度显著高于小麦生长期土壤剖面各层的浓度.统计分析结果表明:土壤剖面中不同土层N2O浓度的变化对照处理为60cm ≈ 90cm ≈150cm＞ 30cm＞ 10cm,而施肥处理为60cm > 90cm ≈150cm＞ 30cm＞ 10cm.深层土壤N2O的主要来源是土壤的反硝化作用,施肥显著地增加各土壤层次N2O的产生量.     

不同水分管理下稻田土壤CH4和N2O排放与微生物菌群的关系

采用MPN计数法对黑土(海伦)和草甸棕壤(沈阳)稻田生长季4种微生物菌群数量进行了测定,同时采用封闭式箱法对CH4和N2O通量进行观测,以深入了解稻田生物源温室气体排放的微生物学过程,两地试验田均采用长期淹灌与间歇灌溉两种不同水分管理,对实验结果多元回归分析,结果表明,海伦与沈阳两地稻田两种水分管理条件下CH4通量季节变化与产甲烷菌数季节变化存在极显著正相关关系沈阳稻田生长季CH4通量季节变化与甲烷氧化菌数季节变化具有显著正相关性,间歇灌溉条件下黑土稻田N2O通量与反硝化菌数呈显著性正相关关系,两种水分管理条件下沈阳稻田N2O通量与硝化菌数具有显著正相关关系,间歇灌溉条件下沈阳稻田N2O通量与反硝化菌数呈显著性正相关关系。     

近地层臭氧对小麦抗氧化酶活性变化动态的影响

摘要:利用开顶式气室（TOCs）模拟了地表O3浓度升高的大田试验条件,测定了小麦各生长期(返青期、拔节期、孕穗期、抽穗期、灌浆期、成熟期)叶片抗氧化酶的活性。通过两年的重复试验,得到相似结果。试验表明,过氧化氢酶（CAT）和过氧化物酶（POD）活性在小麦的生长进程中,先升高后降低。O3浓度升高使CAT酶和POD酶活性的峰值期提前,POD酶峰值期明显提前所需的O3浓度比CAT酶高,O3诱导小麦生长前期的CAT、POD酶活性升高,后期则起抑制作用。POD酶对低浓度O3的胁迫响应时间较晚。当O3浓度提高到150 nL?L-1时,超氧化物歧化酶（SOD）活性随小麦生长期的变化,由原来的双峰曲线变为单峰曲线,高浓度O3使SOD酶活性在抽穗期后持续显著降低。O3不明显改变抗坏血酸过氧化物酶（APX）对胁迫的响应时间,但一定浓度范围内O3能诱导APX酶活性升高。不同浓度的O3对SOD酶和APX酶的作用效应和强度显著不同, CAT酶和POD酶对O3比较敏感。 因此,近地层O3浓度增加,改变了CAT、POD和SOD酶活性变化的时序特征,抗氧化酶对O3胁迫的响应特征取决于小麦的生育期、O3的浓度和薰气时间。     

The Effect of Elevated Ozone Concentrations with Varying Shading on Dry Matter Loss in a Winter Wheat-Producing Region in China

Surface-level ozone pollution causes crop production loss by directly reducing healthy green leaf area available for carbon fixation. Ozone and its precursors also affect crop photosynthesis indirectly by decreasing solar irradiance. Pollutants are reported to have become even more severe in Eastern China over the last ten years. In this study, we investigated the effect of a combination of elevated ozone concentrations and reduced solar irradiance on a popular winter wheat Yangmai13 (Triticum aestivum L.) at field and regional levels in China. Winter wheat was grown in artificial shading and open-top-chamber environments. Treatment 1 (T1, i.e., 60% shading with an enhanced ozone of 100±9 ppb), Treatment 2 (T2, i.e., 20% shading with an enhanced ozone of 100±9 ppb), and Control Check Treatment (CK, i.e., no shading with an enhanced ozone of 100±9 ppb), with two plots under each, were established to investigate the response of winter wheat under elevated ozone concentrations and varying solar irradiance. At the field level, linear temporal relationships between dry matter loss and cumulative stomatal ozone uptake were first established through a parameterized stomatal-flux model. At the regional level, ozone concentrations and meteorological variables, including solar irradiance, were simulated using the WRF-CMAQ model (i.e., a meteorology and air quality modeling system). These variables were then used to estimate cumulative stomatal ozone uptake for the four major winter wheat-growing provinces. The regional-level cumulative ozone uptake was then used as the independent variable in field data-based regression models to predict dry matter loss over space and time. Field-level results showed that over 85% (T1: R² = 0.85 & T2: R² = 0.89) of variation in dry matter loss was explained by cumulative ozone uptake. Dry matter was reduced by 3.8% in T1 and 2.2% in T2 for each mmol O₃·m^-2 of cumulative ozone uptake. At the regional level, dry matter loss in winter wheat would reach 50% under elevated ozone concentrations and reduced solar irradiance as determined in T1, and 30% under conditions as determined in T2. Results from this study suggest that a combination of elevated ozone concentrations and reduced solar irradiance could result in substantial dry matter loss in the Chinese wheat-growing regions.     

臭氧胁迫对水稻生长以及C、N、S元素分配的影响

采用开顶式气室(Open-top Chamber, OTC),对水稻"3694繁"(Oryza sativa L., 3694 Fan)在浙江嘉兴进行田间原位臭氧(O₃)熏气实验,研究不同臭氧浓度熏气对水稻生长以及C、N,S元素分配的影响。实验设置分4个水平:过滤大气组(CF,10 nL/L)、自然大气组(NF,40 nL/L)和两个不同浓度的臭氧处理组(O₃-1:100 nL/L; O₃-2:150 nL/L)。主要结果表明:(1)开始臭氧熏气时,各个处理组单茎水稻各组分生物量没有差异. 在熏气后期(水稻成熟期),臭氧处理使单茎水稻根、茎和穗生物量显著下降,根冠比降低,株高显著降低,表明臭氧胁迫增加水稻地上部分的干物质分配,且对株高的影响可能大于对地上生物量的影响;(2)臭氧处理使水稻根和茎C元素含量下降,叶C元素含量上升,表明臭氧胁迫提高了叶片中碳分配,而降低了根和茎的碳分配;(3)各个组分N元素含量上升和碳氮比下降;(4)茎、叶和穗S元素含量上升,可能会增强水稻抗氧化系统的作用,从而抵抗臭氧胁迫。所有实验结果表明臭氧浓度升高会对水稻生长产生严重不利影响,从而导致水稻各个组分的C、N、S元素分配格局发生改变。     

Effects of increased UV-B radiation and elevated levels of tropospheric ozone on physiological processes in European beech (Fagus sylvatica)

As a consequence of the ongoing reduction of the stratospheric ozone layer, the vegetation is exposed to increasing levels of UV-B radiation (280–320 nm). In addition ozone in the troposphere is a pollutant and also capable of affecting the photosynthetic machinery. In this study, 5-year-old European beech trees were exposed from 1 July to October 1993 to two levels of UV-B radiation and two levels of ozone, alone and in combination, in open-top chambers equipped with lamps. The simulated UV-B levels corresponded to either clear sky ambient level or a 14% decrease in the stratospheric ozone column over eastern Denmark, resulting in a 23% difference in biologically effective UV-B (UV-B_BE) irradiance. The maximum UV-Bbe given was 8.61 kJ m⁻² day⁻¹. The ozone levels were either the ambient (average 32 nl l⁻¹) or ambient with ozone addition (average resulting concentration 71 nl l⁻¹). Compared to the control treatment (ambient UV-B, ambient O₃) the elevated levels of UV-B and O₃ affected the trees negatively, expressed as declines in net photosynthesis (P_n), stomatal conductance (g_s), chlorophyll fluorescence (F_v/F_m) and acceleration of senescence, measured as yellowing of the leaves. The UV-B treatment induced stomatal closure before the other treatments did. The magnitude of the decreases in P_n and F_v/F_m occurred in the order: control 3 3. Compared to the control, the combination treatment with high levels accelerated the visual senescence processes by ca 27 days, while for high UV-B and O₃ alone, there was an acceleration by 14 and 21 days, respectively. UV-B and O₃ in combination enhanced the negative effects compared with UV-B and O₃ alone. The P_n and F_v/F_m results could be related to this acceleration process. The chamber effect was investigated by comparing the control plots with a plot without open-top chamber. The trees in the chambers showed a higher P_n and F_v/F_m and a 14-day delayed senescence compared to the trees outside, probably caused by higher temperatures, a more protected environment and altered conditions inside the chambers.     

Comparison of crop yield sensitivity to ozone between open‐top chamber and free‐air experiments

Assessments of the impacts of ozone (O₃) on regional and global food production are currently based on results from experiments using open‐top chambers (OTCs). However, there are concerns that these impact estimates might be biased due to the environmental artifacts imposed by this enclosure system. In this study, we collated O₃ exposure and yield data for three major crop species—wheat, rice, and soybean—for which O₃ experiments have been conducted with OTCs as well as the ecologically more realistic free‐air O₃ elevation (O₃‐FACE) exposure system; both within the same cultivation region and country. For all three crops, we found that the sensitivity of crop yield to the O₃ metric AOT40 (accumulated hourly O₃ exposure above a cut‐off threshold concentration of 40 ppb) significantly differed between OTC and O₃‐FACE experiments. In wheat and rice, O₃ sensitivity was higher in O₃‐FACE than OTC experiments, while the opposite was the case for soybean. In all three crops, these differences could be linked to factors influencing stomatal conductance (manipulation of water inputs, passive chamber warming, and cultivar differences in gas exchange). Our study thus highlights the importance of accounting for factors that control stomatal O₃ flux when applying experimental data to assess O₃ impacts on crops at large spatial scales.     

Response of gas exchange and yield components of field-grown Triticum aestivum L. to elevated ozone in China

To assess photosynthesis and yield components’ response of field-grown wheat to increasing ozone (O₃) concentration (based on diurnal pattern of ambient O₃) in China, winter wheat (Triticum aestivum L.) cv. Jia 403 was planted in open top chambers and exposed to three different O₃ concentrations: O₃-free air (CF), ambient air (NF), and O₃-free air with additional O₃ (CF+O₃). Diurnal changes of gas exchange and net photosynthetic rate (P _N) in response to photosynthetic photon flux density (PPFD) of flag leaves were measured at the filling grain stage, and yield components were investigated at harvest. High O₃ concentration altered diurnal course of gas exchange [P _N, stomatal conductance (g _s), and intercellular CO₂ concentration (C _i)] and decreased significantly their values except for C _i. Apparent quantum yield (AQY), compensation irradiance (CI), and saturation irradiance (SI) were significantly decreased, suggesting photosynthetic capacity was also altered, characterized as reduced photon-saturated photosynthetic rate (P _Nmax). The limit of photosynthetic activity was probably dominated by non-stomatal factors in combination with stomatal closure. The significant reduction in yield was observed in CF+O₃ treatment as a result of a marked decrease in the ear length and the number of grains per ear, and a significant increase in the number of infertile florets per ear. Even though similar responses were also observed in plants exposed to ambient O₃ concentration, no statistical difference was observed at current ambient O₃ concentration in China.     

Interactive effects of ozone and drought stress on pigments and activities of antioxidative enzymes in Pinus halepensis

The aim of this study was to investigate the interactive effects of ozone (O₃) and drought on pigments and antioxidant enzymes of Aleppo pine (Pinus halepensis). Two‐year‐old seedlings were exposed in open‐top chambers to charcoal‐filtered air or non‐filtered air plus an additional 40 nL L^?1 of ozone. After 20 months of O₃ exposure, a subset of plants was subjected to drought stress by withholding water supply for 11 d. Ozone induced higher guaiacol peroxidase, catalase and KCN‐resistant superoxide dismutase (SOD) activities in young needles, while drought stress increased glutathione reductase and CuZnSOD. One‐year‐old needles showed lower capacity to activate these enzymes in response to stress. Both ozone and drought activated the xanthophyll cycle pool and reduced chlorophyll contents in both current and 1‐year‐old needles. The combined effects of ozone and drought decreased antioxidant enzyme activities and the capacity of recovering after re‐watering. Similarly, interactive effects of O₃ and drought reduced xanthophyll‐mediated photoprotection capacity in 1‐year‐old needles but induced a higher conversion of the cycle in current‐year needles. These results showed that ozone modified the Aleppo pine response to drought stress, suggesting that this pollutant might be reducing the ability of this species to withstand other environmental stresses.