小麦苗期根系分泌物对根际反硝化细菌的影响

本文通过无菌水培试验，研究了两种不同基因型小麦菌期根系分泌物对反硝化细菌优势种生长量和反硝化活性的影响。结果表明：两种不同基因型的小麦根系对反硝化细菌均有明显的根际效应，其菌数和反硝化活性又以郑引１号大于宝丰７２２８；根分泌物中氨基酸组分两者相似，但其总量也以郑引１号大于宝丰７２２８，对根际反硝化细菌产生直接影响。而反硝化活性不受菌株耐药标记的影响。     

好氧反硝化细菌的筛选及反硝化效率测定

从土壤中分离得到21株能以硝酸盐为唯一氮源提供的细菌,其中,能进行反硝化生成亚硝酸盐氮的细菌共12株,从中选择效果较好的2株作为研究对象,分别标记为DM-8和DM-13。研究其生长曲线、最适生长pH值范围,得到DM-8最适生长的pH值范围是6.0~8.0,DM-13最适生长的pH值范围是6.0~7.5。测定反硝化过程中硝酸盐氮和亚硝酸盐氮的浓度变化,DM-8和DM-13在培养48 h后,硝酸盐氮的去除率分别为33%和97%。     

Nitrification activity in tropical rain forest soils of the Coastal Lowlands and Atherton Tablelands,Queensland, Australia

Intact soil cores from a montane tropical rain forest site in the Atherton Tablelands (Kauri Creek) and from a lowland tropical rain forest site in the Coastal Lowlands (Bellenden Ker), Queensland, Australia were investigated during different hygric seasons for the magnitude of gross nitrification rates using the Barometric Process Separation technique (BaPS). Pronounced seasonal variations of gross nitrification rates were found at both sites with highest values during the transition period between dry and wet season (montane site: 24.0 mg N (kg SDW)^—1 d^—1; lowland site: 13.1 mg N (kg SDW)^—1 d^—1) and significantly lower rates of gross nitrification during the dry and wet season. Rates of gross nitrification were always higher at the montane site than at the lowland site, but the opposite was found for N₂O emissions. The results indicated that the high losses of N₂O at the lowland tropical rain forest site may be contributed largely by high denitrification activity due to its wetter and warmer climate as compared to the dryer and colder climate at the montane tropical rain forest site. This conclusion was supported by analysis of cell numbers of microbes involved in N‐cycling. Higher numbers of denitrifiers were present at the lowland site, whereas higher numbers of nitrifiers were found at the montane site.     

End-of-pipe single-sludge denitrification in pilot-scale recirculating aquaculture systems

A step toward environmental sustainability of recirculat aquaculture systems (RAS) is implementation of single-sludge denitrification, a process eliminating nitrate from the aqueous environment while reducing the organic matter discharge simultaneously. Two 1700 L pilot-scale RAS systems each with a 85 L denitrification (DN) reactor treating discharged water and hydrolyzed solid waste were setup to test the kinetics of nitrate and COD removal. Nitrate removal and COD reduction efficiency was measured at two different DN-reactor sludge ages (high θ_X: 33–42 days and low θ_X: 17–23 days). Nitrate and total N (NO₃⁻ + NO₂⁻ + NH₄⁺) removal of the treated effluent water ranged from 73–99% and 60–95% during the periods, respectively, corresponding to an overall maximum RAS nitrate removal of approximately 75%. The specific nitrate removal rate increased from 17 to 23 mg NO₃⁻-N (g TVS d)⁻¹ and the maximal potential DN rate (measured at laboratory ideal conditions) increased correspondingly from 64–68 mg NO₃⁻-N (g TVS d)⁻¹ to 247–294 mg NO₃⁻-N (g TVS d)⁻¹ at high and low θ_X, respectively. Quantification of denitrifiers in the DN-reactors by qPCR showed only minor differences upon the altered sludge removal practice. The hydrolysis unit improved the biodegradability of the solid waste by increasing volatile fatty acid COD content 74–76%. COD reductions in the DN-reactors were 64–70%. In conclusion, this study showed that single-sludge denitrification was a feasible way to reduce nitrate discharge from RAS, and higher DN rates were induced at lower sludge age/increased sludge removal regime. Improved control and optimization of reactor DN-activity may be achieved by further modifying reactor design and management scheme as indicated by the variation in and between the two DN-reactors.     

Effect of urease and nitrification inhibitors on ammonia volatilization and abundance of N‐cycling genes in an agricultural soil

The effect of the combined application of urease and nitrification inhibitors on ammonia volatilization and the abundance of nitrifier and denitrifier communities is largely unknown. Here, in a mesocosm experiment, ammonia volatilization was monitored in an agricultural soil treated with urea and either or both of the urease inhibitor N‐(n‐butyl) thiophosphoric triamide (NBPT) and the nitrification inhibitor 3,4‐dimethylpyrazole phosphate (DMPP), with 50% and 80% water‐filled pore space (WFPS). The effect of the treatments on the abundance of bacteria and archaea was estimated by quantitative PCR (qPCR) amplification of their respective 16S rRNA gene, that of nitrifiers using amoA genes, and that of denitrifiers by qPCR of the norB and nosZI denitrification genes. After application of urea, N losses due to NH₃ volatilization accounted for 23.0% and 9.2% at 50% and 80% WFPS, respectively. NBPT reduced NH₃ volatilization to 2.0% and 2.4%, whereas DMPP increased N losses by up to 36.8% and 26.0% at 50% and 80% WFPS, respectively. The combined application of NBPT and DMPP also increased NH₃ emissions, albeit to a lesser extent than DMPP alone. As compared with unfertilized control soil, both at 50% and 80% WFPS, NBPT strongly affected the abundance of bacteria and archaea, but not that of nitrifiers, and decreased that of denitrifiers at 80% WFPS. Regardless of moisture conditions, treatment with DMPP increased the abundance of denitrifiers. DMPP, both in single and in combined application with NBPT, increased the abundance of nitrification and denitrification genes.     

模拟水位下降与刈割对高寒湿地土壤氨氧化与反硝化微生物的影响

湿地土壤是温室气体重要的源和汇,认识湿地生态系统氮循环过程有助于预测氮循环对未来气候变化的响应与反馈机制。为探讨硝化作用和反硝化作用对土壤水位变化和刈割的响应机制,依托于2013年在青藏高原东部若尔盖泥炭地南部湿地设置的野外实验,通过在样地周围挖掘不同深度的排水沟模拟水位下降,结合刈割处理,研究水位下降和刈割对泥炭地土壤氨氧化古菌(Ammonia-oxidizing archaea,AOA)、氨氧化细菌(Ammonia-oxidizing bacteria,AOB)和反硝化细菌(Denitrifying bacteria)丰度的影响。2014年7月取样分析结果表明:水位下降显著降低土壤含水量,水位下降与刈割均显著降低土壤呼吸;氨氧化及反硝化微生物功能基因丰度在各处理间无显著差异,但刈割及其与水位下降的交互作用显著影响AOA-amo A与AOB-amo A基因丰度比。刈割处理显著增加AOB-amo A基因相对丰度,但对AOA-amo A基因丰度无显著影响,揭示AOB可能在湿地土壤硝化过程中占主导地位。土壤nir S基因丰度显著高于nir K基因,表明nir S基因对水位下降及刈割的响应更为敏感。随着土壤水位的下降,刈割促进了由AOB主导的氨氧化过程,而反硝化微生物丰度的增加削减了氨氧化产物硝酸盐的积累,继而降低了土壤硝酸盐含量。     

Simulated animal trampling of a free‐draining stony soil stimulated denitrifier growth and increased nitrous oxide emissions

Winter forage grazing systems in New Zealand cause compaction of soil by grazing animals, especially when the soil is wet. However, there is little information on the effects of animal trampling on denitrifiers in soil, despite their importance for N₂O production. Here, we report a field study of the abundance of the denitrifying genes nirS, nirK, and nosZ and N₂O emissions following the application of dairy cow urine in a free‐draining stony soil. Importantly, we found that simulated animal trampling altered some of the denitrifying microbial communities, thus leading to increased N₂O emissions. Over the 111 day measurement period, the abundance of nitrite (NO₂^?)‐reducing nirS gene copy numbers increased significantly by 87% in the trampled soil with urine (P < 0.01) and increased by 40% in the trampled soil without urine (P < 0.05), but the nirS gene abundance did not change significantly in the nontrampled soil. The abundance of NO₂^? reducing nirK gene copy numbers was not affected by trampling, but increased significantly following urine application. The abundance of N₂O‐reducing nosZ clade I and nosZ clade II gene copy numbers increased significantly in the trampled soil, but did not change significantly in the nontrampled soil. N₂O emissions from the trampled soil were about twice that from the nontrampled soil without urine (1.20 and 0.62 kg N₂O‐N per ha, respectively) and about eight times greater (6.24 kg N₂O‐N per ha) than from nontrampled soil (0.80 kg N₂O‐N per ha) when urine was applied. These results strongly suggest that animal trampling during winter forage grazing can have a major impact on denitrifying communities in soil, which in turn stimulate greater denitrification with increased N₂O emissions.     

土壤中“接力反硝化”机制的部分证据

设想土壤中存在不同类型的不完全反硝化细菌 ;这些细菌可以彼此配合 ,此菌产物作为彼菌的底物 ,共同完成完整的反硝化过程。该机制称之为“接力反硝化”机制 ,有别于传统的反硝化机制。本文为“接力反硝化”机制的存在提供部分证据。以土壤浸提液为培养基、N2 O为电子受体富集土壤微生物 ,获得了 1株仅完成NO-3 →NO-2 反应的细菌 (原始编号 2 1 6 9 2 )、1株仅完成NO-2 →N2 O反应的细菌 (原始编号 1 9 5 3)、1株仅完成NO-2 →N2 O→N2 反应的细菌 (原始编号 2 1 6 3 6 )。把菌株 2 1 6 9 2和 1 9 5 3两菌株以适当的数量比例混合于灭菌的土壤中 ,不添外来碳源 ,仅添加NO-3 ,厌气培养 1周后 ,测得土壤中剩余的NO-3 仅为原添加量的 39 4 %～ 5 3 0 % ,与此同时有 5 2 %～ 13 9%的NO-3 被还原成NO-2 ,有 2 8 6 %～ 30 8%以N2 O形态被回收 ,总回收率为 75 4 %～ 95 5 % ,说明两者可以相互配合 ,菌株 2 1 6 9 2的硝酸根还原产物可以被菌株 1 9 5 3用作底物 ,共同完成反硝化过程 ,从而支持我们设想的“接力反硝化”机制。     

种植模式影响施肥导致的土壤反硝化势变化及其微生物机制

水田土壤反硝化势(soil denitrification potential,SDP)往往高于旱地土壤,但施肥对水田和旱地SDP的影响差异往往基于不同气候条件下的不同土壤类型获取,其准确性可能受外界条件干扰.以发育自同一母质的相邻水田和旱地长期试验为平台,比较不同施肥模式下水田和旱地SDP的变化及其与功能基因(nar...     

温度和氮添加对旱地红壤反硝化功能基因丰度的影响

研究旱地红壤反硝化微生物功能基因nirS、nirK、nosZ I和nosZ Ⅱ的丰度对温度和氮添加的响应，可为农田红壤养分管理和生态环境保护提供指导建议。本研究以长期常规氮磷钾施肥的旱地红壤为研究对象，设置0 mg N/kg、25 mg N/kg、50 mg N/kg三个氮添加处理，15 ℃、25 ℃、35 ℃三个温度处理，进行微宇宙培养实验。在培养的第7和30天破坏性采集土样，进行DNA提取，测定反硝化微生物功能基因丰度。结果表明：培养7天后，nirS、nirK、nosZ I和nosZ Ⅱ基因丰度都在25 ℃时最高。培养30天后，nirS、nirK、nosZ I和nosZ Ⅱ基因丰度在15 ℃时最高，且随着温度升高而下降。氮添加对反硝化微生物功能基因丰度无显著影响。三因素方差分析表明，温度、氮添加和培养时间的交互作用显著影响反硝化微生物功能基因丰度。综上，旱地农田反硝化功能基因丰度受氮添加影响较小，但受温度显著影响，其丰度可能会呈现出日变化和季节变化，在土壤采样和氧化亚氮动态监测时应特别注意。