Contrasting development of soil microbial community structure under no-tilled perennial and tilled cropping during early pedogenesis of a Mollisol

A range of agricultural practices influence soil microbial communities, such as tillage and organic C inputs, however such effects are largely unknown at the initial stage of soil formation. Using an eight-year field experiment established on exposed parent material (PM) of a Mollisol, our objectives were to: (1) to determine the effects of field management and soil depth on soil microbial community structure; (2) to elucidate shifts in microbial community structure in relation to PM, compared to an arable Mollisol (MO) without organic amendment; and (3) to identify the controlling factors of such changes in microbial community structure. The treatments included two no-tilled soils supporting perennial crops, and four tilled soils under the same cropping system, with or without chemical fertilization and crop residue amendment. Principal component (PC) analysis of phospholipid fatty acid (PLFA) profiles demonstrated that microbial community structures were affected by tillage and/or organic and inorganic inputs via PC1 and by land use and/or soil depth via PC2. All the field treatments were separated by PM into two groups via PC1, the tilled and the no-tilled soils, with the tilled soils more developed towards MO. The tilled soils were separated with respect to MO via PC1 associated with the differences in mineral fertilization and the quality of organic amendments, with the soils without organic amendment being more similar to MO. The separations via PC1 were principally driven by bacteria and associated with soil pH and soil C, N and P. The separations via PC2 were driven by fungi, actinomycetes and Gram (−) bacteria, and associated with soil bulk density. The separations via both PC1 and PC2 were associated with soil aggregate stability and exchangeable K, indicating the effects of weathering and soil aggregation. The results suggest that in spite of the importance of mineral fertilization and organic amendments, tillage and land-use type play a significant role in determining the nature of the development of associated soil microbial community structures at the initial stages of soil formation.     

The role of lignin and cellulose in the carbon-cycling of degraded soils under semiarid climate and their relation to microbial biomass

A high level of biological degradation is usually observed in soils under semiarid climate where the low inputs of vegetal debris constraint the development of microbiota. Among vegetal inputs, cellulose and lignin are dominant substrates but their assimilation by the microbial community of semiarid soils is yet not understood. In the present study, ¹³C-labeled cellulose and ¹³C-labeled lignin (75 μg ¹³C g⁻¹ soil) were added to two semiarid soils with different properties and degradation level. Abanilla soil is a bare, highly degraded soil without plant cover growing on it and a total organic C content of 5.0 g kg⁻¹; Santomera soil is covered by plants (20% coverage) based on xerophytic shrubs and has a total organic C content of 12.0 g kg⁻¹. The fate of added carbon was evaluated by analysis of the carbon isotope signature of bulk soil-derived carbon and extractable carbon fractions (water and sodium-pyrophosphate extracts). At long-term (120 days), we observed that the stability of cellulose- and lignin-derived carbon was dependent on their chemical nature. The contribution of lignin-derived carbon to the pool of humic substances was higher than that of cellulose. However, at short-term (30 days), the mineralization of the added substrates was more related to the degradation level of soils (i.e. microbial biomass). Stable isotope probing (SIP) of phospholipid fatty acids (PLFA-SIP) analysis revealed that just a minor part of the microbial community assimilated the carbon derived from cellulose and lignin. Moreover, the relative contribution of each microbial group to the assimilation of lignin-derived carbon was different in each soil.     

Comparison of the microbial communities of alpacas and sheep fed diets with three different ratios of corn stalk to concentrate

The objective of this study was to investigate the characteristics of ruminal microbial communities of alpacas (Lama pacos) and sheep (Ovis aries) fed three diets with varying ratios of roughage (corn stalk) to concentrate, 3:7 (LS), 5:5 (MS) and 7:3 (HS). Six alpacas (one-year-old and weighing 29.5 ± 7.1 kg) and six sheep (one-year-old and weighing 27.9 ± 2.7 kg) were used in this study, in a replicated 3 × 3 Latin square experiment. Total protozoa concentration was determined under the microscope; total fungi and methanogens were assessed using quantitative polymerase chain reaction and expressed as a percentage of total bacterial 16S rRNA gene copies; bacterial communities were investigated by targeted 16S rRNA gene (V3–V4 region) sequencing. The percentage of fungi was significantly higher in alpacas than in sheep under the LS diet, while the concentration of protozoa was significantly lower in alpacas under HS, MS and LS diets. The alpha diversity including Shannon, Chao l and ACE indices of bacterial communities was higher in alpacas than in sheep, under the LS diet. A total of 299 genera belonging to 22 phyla were observed in the forestomach of alpaca and sheep, with Bacteroidetes and Firmicutes dominating both animal species. Phyla Armatimonadetes and Fusobacteria, as well as 64 genera, were detected only in alpacas, whereas phyla Acidobacteria and Nitrospira, as well as 44 genera, were found only in sheep. The abundance of cellulolytic bacteria, including Butyrivibrio and Pseudobutyrivibrio, was higher in alpacas than in sheep under all three diets. These differences in the forestomach microbial communities partly explained why alpacas displayed a higher poor-quality roughage digestibility, and a lower methane production. Results also revealed that the adverse effects of high-concentrate diets (70%) were lesser in alpacas than in sheep.     

高含固率木质纤维素厌氧发酵物总DNA的4种提取方法比较

不同的样本特性和提取方法对获得微生物总DNA的质量有重要影响。文章基于高含固率木质纤维素厌氧发酵物腐殖酸、酚类物质含量高、质地均一性差、微生物浓度低的特点,研究了4种方法提取不同高含固率粪秸厌氧发酵物中微生物总DNA的效果。结果表明,常规的十二烷基磺酸钠法(sodium dodecyl sulfate,SDS)、十二烷基磺酸钠和溴化十六烷基三甲铵结合法(sodium dodecyl sulfate and cetyltrimethyl ammonium bromide,SDS-CTAB)和商业的粪便试剂盒法提取的DNA质量均较差,SDS法和试剂盒法未能获得聚合酶链式反应(polymerase chain reaction,PCR)扩增目的条带,SDS-CTAB法得到的条带较模糊;改进SDS-CTAB法获得的DNA杂质少、纯度高,具有较好的稳定性,A260/A280和A260/A230值分别为1.74~1.86和1.65~1.86,每克样品的DNA浓度在50 ng·μL^-1以上,电泳条带单一齐整、清晰明亮,PCR扩增的目的条带清晰度高,适宜后续分子生物学技术的分析。林格氏液洗脱、聚乙烯吡咯烷酮-40(Polyvinyl Pyrrolidone-40,PVP-40)洗涤液除杂以及裂解液和多种酶联合破壁是改进SDS-CTAB法获得该类专一性样本高质量微生物总DNA的关键步骤。     

复合潜流人工湿地对农村生活污水的净化效果 及其微生物群落结构特征

试验探究了复合潜流人工湿地对农村生活污水的净化效果及其微生物群落结构特征。结果表明，当水力负荷为0.35 m³·(m²·d）^-1时，系统对TN、NH₄⁺-N、COD和TP去除效率分别为（89.84±7.64）%、（98.67±1.31）%、（61.63±16.01）%和（70.21±8.00）%。湿地中一共发现11个主要菌门、18个主要菌纲和33个主要菌属，变形菌门（Proteobacteria）、蓝藻门（Cyanobacteria）、拟杆菌门（Bacteroidetes）、厚壁菌门（Firmicutes）、绿弯菌门（Chloroflexi）、酸杆菌门（Acidobacteria）、放线菌门（Actinobacteria）是其中的优势菌门，它们在各级湿地中表层和底层的相对丰度分别是57.26%、61.37%、91.60%、93.22%、88.78%、88.02%。蓝藻菌纲（Cyanobacteria）、α-变形菌纲(Alphaproteobacteria) 、芽孢杆菌纲(Bacilli)和γ-变形菌纲(Gammaproteobacteria)、β-变形菌纲(Betaproteobacteria)是其中优势菌纲，约占主要菌纲的70%。共有17个优势菌属，在各级湿地中的分布情况不同，其中芽孢杆菌属（Bacillus）相对丰度最高，集中分布于第三级湿地，其次是Leptolyngbya、席藻属（Phormidium），在第一、二级湿地中广泛分布。复合潜流人工湿地具有良好污水净化效果，系统中有机物和氮素的去除主要依靠微生物作用，磷素的去除主要依靠基质吸附沉淀作用。     

秸秆还田配施氮肥对稻田土壤活性碳氮动态变化的影响

【目的】土壤微生物量碳氮和水溶性有机碳氮是土壤中最活跃的碳氮组分,是衡量土壤碳氮周转与养分有效性的重要指标。探讨秸秆配施氮肥、氮肥用量及基追比例对稻田土壤微生物量碳氮、水溶性有机碳氮、易氧化有机碳和速效氮的影响,明确秸秆还田条件下水稻生长季不同氮肥用量与基追比的土壤活性碳氮变化特征,为稻麦轮作区秸秆还田的氮肥管理提供理论依据。【方法】2012—2015年在湖北省荆门市田间试验中设置施氮量、秸秆配施氮肥和施氮时期3个大田试验。施氮量：不施氮（N0）,推荐施氮（165 kg·hm ^-2,N165）,习惯施氮（195 kg·hm ^-2,N195）;秸秆配施氮肥：秸秆移除（CK）,秸秆还田（移栽前将上季小麦秸秆全部还田,S）,秸秆还田+习惯施氮量（SN）,秸秆还田+推荐施氮量（SF）,秸秆还田+推荐施氮量+腐解菌剂（SM）;施氮时期：基施﹕拔节期﹕抽穗期氮肥施用比例为7﹕3﹕0（R1）,5﹕3﹕2（R2）,10﹕0﹕0（R3）。【结果】秸秆还田+习惯施氮量（SN）显著提高了水稻拔节期土壤微生物量碳（SMBC）含量,但是其成熟期水溶性有机碳含量（DOC）显著降低。秸秆还田+推荐施氮量（SF）显著提高了水稻拔节期土壤水溶性有机氮含量（DON）。腐解菌剂的施用显著降低了水稻成熟期DON含量,拔节期易氧化有机碳含量（ROC）也显著降低。秸秆还田下增加氮肥用量显著提高了水稻抽穗期和灌浆期土壤速效氮含量（AN）;推荐施氮处理（165 kg N·hm ^-2）的DON和AN含量显著升高;农民习惯施氮处理（195 kg N·hm ^-2）降低了DON和AN含量;增加追施氮肥比例对土壤SMBC和DOC含量无明显影响,但提高了水稻拔节期SMBN和ROC含量。【结论】施氮量及其基追比是影响秸秆还田下稻田土壤活性碳氮含量的主要因素,合理配施氮肥能提高土壤微生物量碳、速效氮及水溶性有机氮等活性碳氮组分含量,增加追肥比例也能提高水稻生育期内土壤活性碳氮含量。     

生物炭连续施用对农田土壤氮转化微生物及N2O排放的影响

【目的】研究连续添加生物炭6年后对农田土壤氮转化相关微生物功能基因的影响,揭示生物炭影响作物产量和N₂O排放的微生物学机制,并为生物炭的推广使用提供理论依据。【方法】通过在潮土农田设置0（BC0,对照)、2.25（BCL,低量）、6.75（BCM,中量）和11.25 t·hm^-2（BCH,高量）4个秸秆生物炭量处理的田间定位试验,采用田间观测、化学分析、荧光定量PCR（qPCR）技术,系统研究施用生物炭对氧化亚氮（N₂O）排放、氨单加氧酶（amoA）、亚硝酸还原酶（nirK、nirS）、氧化亚氮还原酶（nosZ）基因丰度及夏玉米产量的影响。【结果】与对照BC0处理相比,施用生物炭可显著提高夏玉米籽粒产量,且BCM处理籽粒产量达到最大值10 811 kg·hm^-2,显著降低夏玉米生育期N₂O累积排放量,并以BCM处理减少N₂O排放效果最优。研究还发现,在夏玉米多个生育时期,与对照比较,生物炭施用可以显著提高耕层土壤无机氮储量和土壤含水量。此外,随着生物炭施用量增加,土壤氨氧化古菌（AOA）基因拷贝数在夏玉米大喇叭口期和成熟期均表现为先上升后下降趋势,且两个时期均以BCM处理最高,而氨氧化细菌（AOB）基因拷贝数在夏玉米大喇叭口期和成熟期分别为BCH处理和BCM处理最高。与对照相比,中、高量生物炭施用（BCM、BCH处理）可显著提高夏玉米大喇叭口期和成熟期土壤反硝化作用功能相关基因（nirK、nirS、nosZ）拷贝数。相关性分析表明,夏玉米成熟期土壤N₂O排放通量与土壤硝态氮、土壤含水量、AOA、AOB、nirK、nirS、nosZ呈显著负相关关系。【结论】施用生物炭通过增加土壤微生物氮转化功能基因丰度进而降低土壤N₂O排放,通过增加土壤耕层无机氮储量和土壤水分含量进而提高作物产量,并以中等用量（6.75 t·hm^-2）施用效果最优。