Involvement of Bradyrhizobium japonicum denitrification in symbiotic nitrogen fixation by soybean plants subjected to flooding

Denitrification by Bradyrhizobium japonicum bacteroids contributes to nitric oxide (NO) production within soybean nodules in response to flooding conditions. However, the physiological relevance of NO production by denitrification in B. japonicum-Glycine max symbiosis is still unclear. In this work, soybean plants were inoculated with B. japonicum strains lacking the nirK or norC genes which encode the copper-containing nitrite reductase and the c-type nitric oxide reductase enzymes, respectively. 14 days flooding increased nodule number of plants inoculated with the WT and norC strains, but not of plants inoculated with the nirK mutant. However, nodule dry weight was not affected by 14 days flooding regardless of the strain used for inoculation. Supporting this observation, individual nodule growth was significantly higher in plants inoculated with nirK than those inoculated with WT or norC after 14 days flooding. Nodule functioning was strongly inhibited by flooding since leghemoglobin content of the nodules induced by any of the strains was significantly decreased after 7 or 14 days flooding compared to control plants. However, this effect was more relevant in nodules of plants inoculated with the WT or norC mutant than in those inoculated with the nirK mutant. Nitrogen fixation was also estimated by analyzing nitrogen content derived from biological nitrogen fixation in shoots, using the ¹⁵N isotope dilution technique. By using this approach, we observed that the negative effect of 14 days flooding on nitrogen fixation was more pronounced in plants inoculated with the norC mutant. However, nitrogen fixation of plants inoculated with nirK showed the highest tolerance to 14 days flooding. These findings allowed us to demonstrate the previously proposed hypothesis which suggests that NO formed by copper-containing nitrite reductase in soybean nodules, in response to flooding, has a negative effect on nitrogenase activity. We propose that inoculation of soybeans with a B. japonicum nirK mutant, which does not produce NO from nitrate, increases the tolerance of symbiotic nitrogen fixation to flooding.     

Changes in forest soil organic matter pools after a decade of elevated CO2 and O3

The impact of rising atmospheric carbon dioxide (CO₂) may be mitigated, in part, by enhanced rates of net primary production and greater C storage in plant biomass and soil organic matter (SOM). However, C sequestration in forest soils may be offset by other environmental changes such as increasing tropospheric ozone (O₃) or vary based on species-specific growth responses to elevated CO₂. To understand how projected increases in atmospheric CO₂ and O₃ alter SOM formation, we used physical fractionation to characterize soil C and N at the Rhinelander Free Air CO₂-O₃ Enrichment (FACE) experiment. Tracer amounts of ¹⁵NH₄⁺ were applied to the forest floor of Populus tremuloides, P. tremuloides-Betula papyrifera and P. tremuloides-Acer saccharum communities exposed to factorial CO₂ and O₃ treatments. The ¹⁵N tracer and strongly depleted ¹³C-CO₂ were traced into SOM fractions over four years. Over time, C and N increased in coarse particulate organic matter (cPOM) and decreased in mineral-associated organic matter (MAOM) under elevated CO₂ relative to ambient CO₂. As main effects, neither CO₂ nor O₃ significantly altered ¹⁵N recovery in SOM. Elevated CO₂ significantly increased new C in all SOM fractions, and significantly decreased old C in fine POM (fPOM) and MAOM over the duration of our study. Overall, our observations indicate that elevated CO₂ has altered SOM cycling at this site to favor C and N accumulation in less stable pools, with more rapid turnover. Elevated O₃ had the opposite effect, significantly reducing cPOM N by 15% and significantly increasing the C:N ratio by 7%. Our results demonstrate that CO₂ can enhance SOM turnover, potentially limiting long-term C sequestration in terrestrial ecosystems; plant community composition is an important determinant of the magnitude of this response.     

Compartmentalization of the soil animal food web as indicated by dual analysis of stable isotope ratios (N/N and C/C)

The soil animal food web has become a focus of recent ecological research but trophic relationships still remain enigmatic for many taxa. Analysis of stable isotope ratios of N and C provides a powerful tool for disentangling food web structure. In this study, animals, roots, soil and litter material from a temperate deciduous forest were analysed. The combined measurement of δ¹⁵N and δ¹³C provided insights into the compartmentalization of the soil animal food web. Leaf litter feeders were separated from animals relying mainly on recent belowground carbon resources and from animals feeding on older carbon. The trophic pathway of leaf litter-feeding species appears to be a dead end, presumably because leaf litter feeders (mainly diplopods and oribatid mites) are unavailable to predators due to large size and/or strong sclerotization. Endogeic earthworms that rely on older carbon also appear to exist in predator-free space. The data suggest that the largest trophic compartment constitutes of ectomycorrhizal feeders and their predators. Additionally, there is a smaller trophic compartment consisting of predators likely feeding on enchytraeids and potentially nematodes.     

采用15N同位素稀释法研究不同层次土壤氮素总转化速率

采用15N同位素稀释方法,开展短期(7天)室内培养实验,估算了一水稻土0~20、20~60和60~90 cm土层土壤主要N素转化过程的总转化速率,结果表明,标记N溶液加入后2 h内各土层土壤的总矿化、硝化、固定速率显著高于其他时间段(p<0.01)。2 h后,矿化速率在小范围内起伏。0~20 cm土层土壤N素的硝化速率随培养时间延长而降低,另外两层土壤则基本保持稳定,硝化速率的变化与硝化作用底物NH4+-N浓度的变化呈显著正相关。值得注意的是,外源无机N溶液加入后2 h内,大量NH4+-N和NO3--N被固定,并认为N素的非生物固定起主导作用。2 h后,出现了N素在固定与再矿化间反复转换的现象。实验结果表明,与净转化速率相比总转化速率能更好地描述单个N素转化过程,但由于外源N加入对N素转化的影响、再矿化作用以及忽略了N素转化过程中的气体损失、DNRA(硝态氮异化还原为铵)过程等,本研究结果与真实值间存在一定差异。     

施氮水平对甘蔗氮素吸收与利用的影响

以新台糖22号(ROC22)为试材,设15N标记尿素2.5、5.0和7.5g/盆(相当于225、450和675kg/hm2)3个处理,采用网室盆栽试验方式,研究施氮水平对甘蔗氮素吸收与利用的影响。结果表明:甘蔗吸收的氮素17.27%～27.28%来自施用的氮肥,72.72%～82.73%来自土壤和种茎;甘蔗氮肥利用率为34.21%～42.46%。随施氮水平提高,甘蔗干物质积累、氮素积累及来源于肥料氮素的比率显著增加,同时蔗叶对氮素的吸收利用呈上升趋势,但蔗茎对氮素的吸收利用呈下降趋势,氮肥利用率也显著下降。土壤碱解氮和硝态氮在各土层中的含量随施氮水平的提高而增加,且两者在0～20cm土层的积累明显大于20～40cm土层。本试验条件下,甘蔗氮肥施用为尿素5.0g/盆(相当于450kg/hm2)及土层深度为20cm较为适宜。     

高氮和NO2-对中亚热带森林土壤N2O和NO产生的影响

利用15N同位素标记方法,研究在两种水分条件即60％和90％ WHC下,添加硝酸盐(NH4NO3,N 300 mg kg-1)和亚硝酸盐(NaNO2,N 1 mg kg-1)对中亚热带天然森林土壤N2O和NO产生过程及途径的影响.结果表明,在含水量为60％ WHC的情况下,高氮输入显著抑制了N2O和NO的产生(p＜0.01);但当含水量增为90％ WHC后,实验9h内抑制N2O产生,之后转为促进.所有未灭菌处理在添加NO2-后高氮抑制均立即解除并大量产生N2O和NO,与对照成显著差异(p＜0.01),在60％ WHC条件下,这种情况维持时间较短(21 h),但如果含水量高(90％ WHC)这种情况会持续很长时间(2周以上),说明水分有效性的提高和外源NO2-在高氮抑制解除中起到重要作用.本实验中N2O主要来源于土壤反硝化过程,而且加入未标记NO2-后导致杂合的N2O(14N15NO)分子在实验21 h内迅速增加,表明这种森林土壤的反硝化过程可能主要是通过真菌的“共脱氮”来实现,其贡献率可多达80％以上.Spearman秩相关分析表明未灭菌土壤NO的产生速率与N2O产生速率成显著正相关性(p＜0.05),土壤含水量越低二者相关性越高.灭菌土壤添加NO2-能较未灭菌土壤产生更多的NO,但却几乎不产生N2O,表明酸性土壤的化学反硝化对NO的贡献要大于N2O.     

根际注射施肥对黄土高原苹果氮素吸收利用及产量和品质的影响

【目的】在西北黄土高原地区,春季干旱少雨和肥料利用率低限制着生产优质苹果。国外通常应用水肥一体化技术来克服水肥利用率低的问题,但由于其硬件设施要求高,投资大,短期内在我国难以推广。近年来我国采用的根际注射施肥可利用施肥枪将肥料溶液直接注入根际土壤中,施肥成本低且技术简单。本研究利用同位素15N示踪技术,研究根际注射施肥对苹果氮素吸收利用及产量品质的影响,可为黄土高原果园水肥高效利用提供依据。【方法】以9年生富士/M26/新疆野苹果为试验材料,利用15N尿素标记肥料去向,最后通过MAT-251质谱计测15N丰度,得出果树各器官和土壤的肥料利用率。同时利用叶绿素仪(SPAD-502)测定标记叶片的SPAD值,用浸以磷酸甘油溶液的海绵进行田间原位测定,得到土壤氨挥发的量,用静态箱—气相色谱法测定土壤的N2O逸失量。综合对比分析黄土高原传统环状开沟撒施肥与根际注射施肥对苹果吸收利用氮素、肥料氮在土壤中残留及果实产量和品质的影响。【结果】黄土高原苹果园根际注射施肥的优越性体现在:1)施肥后一个月内,果园土壤的气态氮素损失发生变化,根际注射施肥比传统环状开沟施肥的氨挥发总量低54.9%,同时N2O的排放通量低5.0%。2)根际注射施肥后,促进了肥料在土壤中的扩散范围,扩大了根系肥水吸收容积,叶片和果实吸收的肥料氮比例(Ndff%)在整个生长季始终处于较高水平。生长季末期,根际注射施肥的整株氮素当季吸收率为53.04%,比环状开沟施肥提高12.25个百分点,表明根际注射施肥有利于氮素更快地被吸收利用,显著提高苹果树的氮素当季利用率。3)生长季末,在0—60 cm土层内,根际注射施肥的土壤氮素残留率为36.55%,而环状开沟施肥为43.13%,前者显著低于后者。4)在整个生长季内,根际注射施肥处理下的树体新梢叶片内叶绿素含量(SPAD)值一直高于环状开沟施肥。根际注射施肥能提高苹果单株产量和单果重,其单果重和单株产量分别比环状开沟施肥处理提高了3.8%和19.7%。【结论】黄土高原地区推广的果树根际注射施肥技术可以有效提高苹果树体氮素的利用率,降低了土壤中的氮素残留。此外注射施肥的深度、注射量、密度和时间均可根据不同时期的养分需要随时调整,使水肥在土壤中均匀分布,达到节水节肥的目的。同时可避免传统施肥时挖坑作业对浅土层吸收根的损伤,降低劳动力成本。综合来看,根际注射施肥是提高黄土高原区旱地苹果树肥水利用率、产量和品质的有效方式之一。     

肥料~(15)N对烟叶氮和烟碱含量的影响

利用15N示踪技术在黄壤烟区研究了烤烟不同部位烟叶氮和烟碱含量的变化及肥料氮在烟碱中的分配。结果表明,不施氮肥的情况下,烟株第10片以上烟叶烟碱含量达到3.54%;施氮量为97.5kghm-2时,10片以上烟叶烟碱含量达到4.13%以上。烟碱积累在整个生育期是呈不断增加的趋势,于移栽后7周内积累较少,烟碱积累主要集中在9～15周,到烟叶采收结束17周为122.55kghm-2。烟叶进入成熟期后,烟碱含量与各部位烟叶来自土壤氮素比例呈正相关,其中与上部烟叶相关系数达0.88。烟碱总氮中来自肥料氮于下部、中部和上部烟叶分别占28.31%,26.63%和25.45%;相应构成烟碱的氮素中,下、中、上部烟叶中有来自土壤氮分别为71.69%、73.37%和74.55%。追肥氮占施氮总量30%条件下,烟碱中氮来自追肥氮在下、中和上部烟叶分别占14.25%、15.30%和14.94%,追肥氮是烟叶烟碱总氮主要氮源。     

“小麦/玉米/大豆”套作体系中不同作物间的相互作用及氮素的转移、吸收

在根系分隔盆栽条件下,采用15N土壤稀释标记方法,研究了“小麦/玉米/大豆”三熟套作体系不同作物间的相互作用及氮素的转移、吸收利用特性。结果表明,“小麦/玉米/大豆”套作体系促进小麦对肥料氮和土壤氮的吸收,不分隔处理的生物产量、15N总吸收量和总回收率得到显著提高,土壤残留15N丰度及总氮含量明显降低;玉米表现出套作优势（Awc<0,NCRwc<0）,不分隔处理的籽粒产量、籽粒15N吸收量、15N总回收率、土壤残留15N丰度及总氮含量较分隔处理提高17.17%、24.52%、17.63%、13.9%和10.1%;大豆表现出套作劣势,不分隔处理的15N总吸收量、籽粒15N吸收量、15N总回收率和土壤残留15N丰度降低,土壤总氮含量提高6.06%。“小麦/玉米/大豆”套作体系存在氮素的双向转移,以玉米向小麦、大豆向玉米转移为主。     

不同施氮量对水稻氮素吸收与分配的影响

运用15N示踪法研究了不同施氮量对两个品种水稻（4007和武运粳15）干物质积累量与其对15N吸收及分配的影响。结果表明,当施氮量超过N 150 kg/hm2时, 两个品种水稻子粒产量均不再显著增加。4007在4个施氮量下(N 100,150,200和 250 kg/hm2)分别比无氮区增产22.3%,36.9%,43.2%和38.1%;武运粳15分别增产10.6%,18.8%,27.1%和21.5%。同一施氮量下,4007子粒中15N累积量显著高于武运粳15,但茎叶和根中没有差异。增加施氮量降低了15N在水稻子粒中的分配比例,但提高了茎叶中15N的分配比例。15N在根中的分配比例不受施氮量和品种的影响。研究结果还表明,同一施氮量下,4007对肥料氮的总体利用率要比武运粳15高3～6个百分点。