奶牛肾小管上皮细胞的原代培养和鉴定

为建立奶牛肾小管上皮细胞的原代培养、传代以及鉴定方法,本研究将肾脏皮质剪碎,并网筛过滤选取肾小管后分别进行直接培养、胶原酶1消化培养、胰蛋白酶消化培养和胰蛋白酶和0．02％EDTA混合消化培养,分别应用上述四种方法进行原代培养和传代培养,然后采用CK-18、PCK和Vimentin三种蛋白进行免疫组化鉴定奶牛肾小管上皮细胞。结果表明,采用消化培养方法都能够得到奶牛肾小管上皮细胞,但是胶原酶1消化得到的活细胞更多,能更快地成功培养奶牛肾小管上皮细胞。胶原酶1消化法是培养奶牛肾小管上皮细胞的理想培养法。     

3种兔球虫18S rDNA部分序列测定与系统发育分析

采用单卵囊分离法从河北某兔场分离大型艾美耳球虫、黄艾美耳球虫及肠艾美耳球虫,接种无球虫兔后获得大量纯种卵囊,CTAB法提取孢子化卵囊基因组DNA。利用艾美耳属球虫18S rDNA保守引物,PCR扩增3种兔球虫18S rDNA片段,产物纯化后测序。将3种球虫18S rDNA测序结果与GenBank中发布的兔球虫18S rDNA序列用DNAStar软件进行比对。使用MEGA4.0软件对兔球虫18S rDNA进行同源性比较,并绘制遗传进化树。结果表明,大型艾美耳球虫扩增出大小为1 521bp的18S rDNA片段;黄艾美耳球虫及肠艾美耳球虫均扩增出大小为1 520bp的18S rDNA片段。序列比对结果显示,3种河北株兔球虫与GenBank中相应的3种兔球虫18S rD-NA(EF694016、EF694011、EF694012)相似性分别为99.6%、99.6%和100%。3种河北株兔球虫序列和GenBank中兔球虫18S rDNA序列(EF694007-EF694017)位于一个单系集群。     

三系辣椒新品种沈研18号的选育

沈研18号是以辣椒细胞质雄性不育系A074—3为母本,恢复系0952—7为父本配制的一代杂交种。果实长灯笼形,果纵径15．0cm果横径9．7cm,果色绿,果面略皱,微辣,单果质量195．0g,可食率85％以上。Vc含量61．7mg／kg（鲜质量）,可容性总糖4．08％。适于辽宁、河北、山东、吉林、黑龙江、内蒙古、新疆、...     

下辽河平原潮棕壤撂荒地和人工林N2O、CH4排放研究

采用静态箱(暗箱)气相色谱法对下辽河平原潮棕壤撂荒地和人工林N2O、CH4排放进行了为期1年的原位测量,同时测量了土壤温湿度、气温,土壤NO3-N、NH4 -N含量等相关因子.结果表明,在观测期间内人工林地土壤的N2O排放通量明显高于撂荒地.撂荒地是大气CH4的源,而人工林则是CH4的汇.对人工林的观测结果表明,N2哦O排放与CH4排放之间存在负相关关系(R2=-0.351,p＜0.05,n=36);而撂荒地两种气体排放之间无明显相关关系.N2O排放通量和温度变化有很好的相关性.当土壤含水量在200 g/kg以上时,土壤含水量与CH4气体排放具有较好的正相关关系.     

稻季施肥管理措施对后续麦季N2O排放的影响

2003─2004年选用江苏省宜兴市稻-麦轮作试验田,研究了水稻生长季秸杆(0和3.75×103kg/hm2两个水平)和N肥施用(N0、200和270kg/hm23个水平)对后续麦季N2O排放的影响。结果表明:稻季秸杆施用显著减少了后续麦季N2O的排放,这些减少量主要体现在小麦播种-返青期,方差分析达显著水平(P<0.05)。稻季施用N肥,后续麦季N2O排放减少,但N200和270kg/hm2N肥施用水平的处理间无显著差异。麦季土壤水分情况与N2O排放通量存在显著正相关(P<0.05)。     

The N2O product ratio of nitrification and its dependence on long-term changes in soil pH

The contribution of nitrification to the emission of nitrous oxide (N₂O) from soils may be large, but its regulation is not well understood. The soil pH appears to play a central role for controlling N₂O emissions from soil, partly by affecting the N₂O product ratios of both denitrification (N₂O/(N₂+N₂O)) and nitrification (N₂O/(NO₂⁻+NO₃⁻). Mechanisms responsible for apparently high N₂O product ratios of nitrification in acid soils are uncertain. We have investigated the pH regulation of the N₂O product ratio of nitrification in a series of experiments with slurries of soils from long-term liming experiments, spanning a pH range from 4.1 to 7.8. ¹⁵N labelled nitrate (NO₃⁻) was added to assess nitrification rates by pool dilution and to distinguish between N₂O from NO₃⁻ reduction and NH₃ oxidation. Sterilized soil slurries were used to determine the rates of chemodenitrification (i.e. the production of nitric oxide (NO) and N₂O from the chemical decomposition of nitrite (NO₂⁻)) as a function of NO₂⁻ concentrations. Additions of NO₂⁻ to aerobic soil slurries (with ¹⁵N labelled NO₃⁻ added) were used to assess its potential for inducing denitrification at aerobic conditions. For soils with pH?5, we found that the N₂O product ratios for nitrification were low (0.2-0.9‰) and comparable to values found in pure cultures of ammonia-oxidizing bacteria. In mineral soils we found only a minor increase in the N₂O product ratio with increasing soil pH, but the effect was so weak that it justifies a constant N₂O product ratio of nitrification for N₂O emission models. For the soils with pH 4.1 and 4.2, the apparent N₂O product ratio of nitrification was 2 orders of magnitude higher than above pH 5 (76‰ and 14‰). This could partly be accounted for by the rates of chemodenitrification of NO₂⁻. We further found convincing evidence for NO₂⁻-induction of aerobic denitrification in acid soils. The study underlines the role of NO₂⁻, both for regulating denitrification and for the apparent nitrifier-derived N₂O emission.     

Stable carbon isotope ratios of water-extractable organic carbon affected by application of rice straw and rice straw compost during a long-term rice experiment in Yamagata,Japan

ABSTRACT

Hot-water- and water-extractable organic matter were obtained from soil samples collected from a rice paddy 31 years after the start of a long-term rice experiment in Yamagata, Japan. Specifically, hot-water-extractable organic carbon and nitrogen (HWEOC and HWEON) were obtained by extraction at 80°C for 16 h, and water-extractable organic carbon and nitrogen (WEOC and WEON) were obtained by extraction at room temperature. The soil samples were collected from surface (0–15 cm) and subsurface (15–25 cm) layers of five plots that had been treated with inorganic fertilizers alone or with inorganic fertilizers plus organic matter, as follows: PK, NPK, NPK plus rice straw (RS), NPK plus rice straw compost (CM1), and NPK plus a high dose of rice straw compost (CM3). The soil/water ratio was 1:10 for both extraction temperatures. We found that the organic carbon and total nitrogen contents of the bulk soils were highly correlated with the extractable organic carbon and nitrogen contents regardless of extraction temperature, and the extractable organic carbon and nitrogen contents were higher in the plots that were treated with inorganic fertilizers plus organic matter than in the PK and NPK plots. The HWEOC and WEOC δ¹³C values ranged from ?28.2% to ?26.4% and were similar to the values for the applied rice straw and rice straw compost. There were no correlations between the HWEOC or WEOC δ¹³C values and the amounts of HWEOC or WEOC. The δ¹³C values of the bulk soils ranged from ?25.7% to ?23.2% and were lower for the RS and CM plots than for the PK and NPK plots. These results indicate that HWEOC and WEOC originated mainly from rice plants and the applied organic matter rather than from the indigenous soil organic matter. The significant positive correlations between the amounts of HWEOC and HWEON and the amount of available nitrogen (P < 0.001) imply that extractable organic matter can be used as an index for soil fertility in this long-term experiment. We concluded that the applied organic matter decomposed more rapidly than the indigenous soil organic matter and affected WEOC δ¹³C values and amounts.     

地膜覆盖对土壤中N2O释放的影响

研究地膜覆盖下土壤中N₂O释放可为进一步探明膜下土壤中N₂O的传输、消耗和排放到大气的动力学过程提供理论依据。在2001年3月至6月和2001年10月至2002年6月连续两个冬小麦生长季，采用静态漏斗法和揭膜—封闭箱法测定了地膜覆盖下耕层5、10、20 cm土层处和地表处N₂O的释放特征及相应土壤性质。结果表明：地膜覆盖下，地表和耕层10、20 cm土层处N₂O释放通量显著增加；0～5 cm土层土壤水分和10～20 cm土层土壤硝态氮的浓度的变化分别解释了休闲地和冬小麦地土壤中N₂O释放通量85.23%和92.11%的变异，它们是膜下休闲地和冬小麦地土壤中N₂O释放通量增加的主要原因。该结论对地膜覆盖下科学地控制农田水分、养分以及地膜覆盖在中国西北地区的科学使用和推行具有实际意义。     

Effect of intermittent drainage in reduction of methane emission from paddy soils in Hokkaido,northern Japan

ABSTRACT

Emission of methane (CH₄), a major greenhouse gas, from submerged paddy soils is generally reduced by introducing intermittent drainage in summer, which is a common water management in Japan. However, such a practice is not widely conducted in Hokkaido, a northern region in Japan, to prevent a possible reduction in rice grain yield caused by cold weather. Therefore, the effects of intermittent drainage on CH₄ emission and rice grain yield have not been investigated comprehensively in Hokkaido. In this study, we conducted a three-year field experiment in Hokkaido and measured CH₄ and nitrous oxide (N₂O) fluxes and rice grain yield to elucidate whether the reduction in CH₄ emission can be achieved in Hokkaido as well as other regions in Japan. Four experimental treatments, namely, two soil types [soils of light clay (LiC) and heavy clay (HC) textures] and two water management [continuous flood irrigation (CF), and intermittent drainage (ID)], were used, and CH₄ and N₂O fluxes were measured throughout the rice cultivation periods from 2016 to 2018. Cumulative CH₄ emissions in 2016 were markedly low, suggesting an initially low population of methanogens in the soils presumably due to no soil submergence or crop cultivation in the preceding years, which indicates a possible reduction in CH₄ emission by introducing paddy-upland crop rotation. Cumulative CH₄ emissions in the ID-LiC and ID-HC plots were 21–91% lower than those in the CF-LiC and CF-HC plots, respectively, whereas the cumulative N₂O emissions did not significantly differ between the different water managements. The amount of CH₄ emission reduction by the intermittent drainage was largest in 2018, with a comparatively long period of the first drainage for 12 days in summer. Rice grain yields did not significantly differ between the different water managements for the entire 3 years, although the percentage of well-formed rice grains was reduced by the intermittent drainage in 2018. These results suggest that CH₄ emission from paddy fields can be reduced with no decrease in rice grain yield by the intermittent drainage in Hokkaido. In particular, the first drainage for a long period in summer is expected to reduce CH₄ emission markedly.     

N2O emissions from different cropping systems and from aerated,nitrifying and denitrifying tanks of a municipal waste water treatment plant

Nitrous oxide emissions, nitrate, water-soluble carbon and biological O₂ demand (BOD₅) were quantified in different cropping systems fertilized with varying amounts of nitrogen (clayey loam, October 1991 to May 1992), in an aerated tank (March 1993 to March 1994), and in the nitrification-denitrification unit (March to July 1994) of a municipal waste water treatment plant. In addition, the N₂O present in the soil body at different depths was determined (February to July 1994). N₂O was emitted by all cropping systems (mean releases 0.13–0.35 mg N₂O m^-2 h^-1), and all the units of the domestic waste water treatment plant (aerated tank 0–6.2 mg N₂O m^-2 h^-1, nitrification tank 0–204,3 mg N₂O m^-2, h^-1, denitrifying unit 0–2.2 mg N₂O m^-2 h^-1). During the N₂O-sampling periods estimated amounts of 0.9, 1.5, 2.4 and 1.4 kg N₂O–N ha^-1, respectively, were released by the cropping systems. The aerated, nitrifying and denitrifying tanks of the municipal waste water treatment plant released mean amounts of 9.1, 71.6 and 1.8 g N₂O–N m^-2, respectively, during the sampling periods.The N₂O emission were significantly positively correlated with nitrate concentrations in the field plots which received no N fertilizer and with the nitrogen content of the aerated sludge tank that received almost exclusively N in the form of NH ₄ ⁺ . Available carbon, in contrast, was significantly negatively correlated with the N₂O emitted in the soil fertilized with 80 kg N ha^-1 year. The significant negative correlation between the emitted N₂O and the carbon to nitrate ratio indicates that the lower the carbon to nitrate ratio the higher the amount of N₂O released. Increasing N₂O emissions seem to occur at electron donorto-acceptor ratios (C_H2_O or BOD₅-to-nitrate ratios) below 50 in the cropping systems and below 1200–1400 in the waste water treatment plant. The trapped N₂O in the soil body down to a depth of 90 cm demonstrates that agricultural production systems seem to contain a considerable pool of N₂O which may be reduced to N₂ on its way to the atmosphere, which may be transported to other environments or which may be released at sometime in the future.Dedicated to Professor J.C.G. Ottow on the occasion of his 60th birthday