磷矿粉、骨炭和油菜秸秆对重金属复合污染土壤细菌和古菌数量的影响

微生物的活动在土壤生态系统中发挥着重要的作用,作为检测土壤质量的敏感指标,其基因拷贝数的变化是响应外界环境的一种方式.通过土壤盆栽试验,结合Real-time PCR技术研究不同改良剂添加对重金属复合污染土壤细菌和古菌16S rRNA基因拷贝数的影响.结果表明:各处理中土壤细菌16S rRNA基因拷贝数在每克干土6.02×109～1.82×10m个之间,古菌在每克干土7.99×106～2.97×107个之间.污染土壤中添加不同改良剂对土壤中古菌和细菌16S rRNA基因拷贝数的影响有差异,种植玉米与否也影响这两种微生物在土壤的基因拷贝数.不种植玉米时,与对照相比,添加磷矿粉、骨炭和油菜秸秆处理导致土壤细菌基因拷贝数提高116.8％～163.2％;磷矿粉和骨炭处理导致土壤古菌基因拷贝数分别提高171.1％和224.8％.种植玉米时,添加骨炭和油菜秸秆处理导致土壤细菌的基因拷贝数分别比对照处理提高104.2％和92.0％;土壤中古菌的基因拷贝数除了添加骨炭处理显著比对照处理提高231.3％外,其他处理对土壤古菌基因拷贝数均没有影响.     

牛场肥水灌溉对土壤氨氧化微生物的影响

以不同牛场肥水灌溉制度下河北省徐水县梁家营长期定位施肥试验田为研究对象,通过构建氨氧化细菌和氨氧化古菌的amoA 基因克隆文库,利用T-RFLP方法研究了5种施肥处理(CK:不施肥;CF:300 kg N·hm^-2,120 kg P₂O·hm^-2,75 kg K₂O·hm^-2;T4:105 kg N·hm^-2,39 kg P₂O₅·hm^-2;T5:210 kg N·hm^-2,78 kg P₂O₅·hm^-2; T11:317 kg N·hm^-2,117 kg P₂O₅·hm^-2)下土壤中氨氧化细菌及氨氧化古菌的多样性及其群落结构演变。氨氧化细菌T-RFLP结果显示,T5 处理土壤中氨氧化细菌的 Shannon-Wiener指数(H')和 Pielou指数(E)均最高, T11处理最低,表明T5处理增加了土壤中氨氧化细菌的多样性和丰富度,T11处理一定程度上降低了氨氧化细菌的多样性和丰富度。基于amoA基因建立的系统进化树显示,土壤中氨氧化细菌主要属于Nitrosospira 和Nitrosomanas。氨氧化古菌T-RFLP结果显示,T11处理氨氧化古菌Shannon-Wiener指数(H')、Simpson 指数(Ds)和Pielou指数(E)均最高,表明T11处理增加了土壤中氨氧化古菌的多样性和丰富度。氨氧化古菌的系统发育树显示大部分氨氧化古菌amoA序列与 Cluster S 聚为一类,且占主导地位,少数序列为Unclassified-Archaea(未分类古菌)。最大优势菌与来自中国荷斯坦奶牛瘤胃样品的氨氧化古菌Clone属于同种,推测牛场肥水灌溉增加了土壤的氨氧化古菌的群落多样性。     

Ammonia oxidizer abundance in paddy soil profile with different fertilizer regimes

Studies about ammonia-oxidizing bacteria (AOB) and archaea (AOA) are often focused on topsoil, but little is known about their activity and distribution in subsoil. A long-term fertilizer experiment was conducted to assess the effects of different fertilizer treatments on AOB and AOA in vertical soil profiles of paddy soil plots that received no nitrogen fertilizer control (CK), NPK chemical fertilizers (CF), organic–inorganic mixed fertilizer (OIMF) and organic fertilizer (OF). Soil properties, potential nitrification rate (PNR) and amoA gene abundance of AOB and AOA were measured and analyzed by two-way ANOVA and correlation analysis. Quantitative PCR analysis of amoA genes showed that AOA were more abundant than AOB in all the soil samples. AOB declined sharply with soil depth. Compared with CK and OF treatments, CF and OIMF treatments had higher abundance of AOB throughout the soil profiles. However, AOA tend less responsive to soil depth and fertilizers compared to AOB. This caused the AOA/AOB ratios in subsoil higher than in topsoil, and in CK and OF higher than in CF and OIMF treatments. These results suggest that AOA are more abundant and can be better adapted to nutrient-poor subsoils than AOB, and autotrophic nitrification could likely be determined by a complex suite of environmental factors in vertical profiles of the paddy soil tested.     

水稻根际和周围土壤中微生物功能基因丰度与碳、氮状况及其通量之间的关系

Rapid nitrogen（N） transformations and losses occur in the rice rhizosphere through root uptake and microbial activities. However,the relationships between rice roots and rhizosphere microbes for N utilization are still unclear. We analyzed different N forms（NH＋4,NO-3, and dissolved organic N）, microbial biomass N and C, dissolved organic C, CH4 and N2O emissions, and abundance of microbial functional genes in both rhizosphere and bulk soils after 37-d rice growth in a greenhouse pot experiment. Results showed that the dissolved organic C was significantly higher in the rhizosphere soil than in the non-rhizosphere bulk soil, but microbial biomass C showed no significant difference. The concentrations of NH＋4, dissolved organic N, and microbial biomass N in the rhizosphere soil were significantly lower than those of the bulk soil, whereas NO-3in the rhizosphere soil was comparable to that in the bulk soil. The CH4 and N2O fluxes from the rhizosphere soil were much higher than those from the bulk soil. Real-time polymerase chain reaction analysis showed that the abundance of seven selected genes, bacterial and archaeal 16 S rRNA genes, amoA genes of ammonia-oxidizing archaea and ammonia-oxidizing bacteria, nosZ gene, mcrA gene, and pmoA gene, was lower in the rhizosphere soil than in the bulk soil, which is contrary to the results of previous studies. The lower concentration of N in the rhizosphere soil indicated that the competition for N in the rhizosphere soil was very strong, thus having a negative effect on the numbers of microbes. We concluded that when N was limiting, the growth of rhizosphere microorganisms depended on their competitive abilities with rice roots for N.     

基于宏基因组学的酸性森林土壤氨氧化微生物群落特征研究

全球30％以上陆地面积是酸性土壤(pH<5.5),而酸性土壤中氨氧化微生物群落特征研究是破译其硝化过程微生物学机理的基础.尤其随着完全硝化微生物(Complete ammonia oxidizer,comammox)的发现,亟需重新认知酸性土壤中氨氧化微生物类群.以酸性马尾松林为研究对象,综合利用荧光定量PCR(qPC...     

高通量测序分析人工养殖成年林麝粪便古菌菌群多样性

本试验的目的是采用高通量测序技术研究人工养殖成年林麝粪便中古菌的结构和组成,并对比不同性别之间的差异。选取12只3岁健康的林麝依据性别分为雄性组(M组)和雌性组(F组),每组各6只。采集其新鲜粪便,提取DNA,用古菌通用引物PCR扩增古菌16S rRNA的V4~V5区,用M i Seq 300 PE测序平台对扩增产物进行高通量测序,用QIIM E等软件对测序序列进行分析统计。结果表明:在从门到属的各级分类水平上,F组与M组古菌的相对丰度差异均不显著(P0.05)。M组和F组的组内遗传距离分别为0.16±0.03和0.27±0.06,组间为0.24±0.07,样品的相似度很高。林麝粪便中的古菌可以分为3个门,优势门为阔古细菌门(Euryarchaeota);在属水平上可以分为7个已知属,优势属为甲烷短杆菌属(Methanobrevibacter),其次为热裸单胞菌属(Thermogymnomonas)。结果提示,雄性和雌性林麝粪便中古菌的结构和组成都没有显著差异,Methanobrevibacter是林麝粪便中的优势古菌属     

A lysimeter study of nitrate leaching from grazed grassland as affected by a nitrification inhibitor,dicyandiamide, and relationships with ammonia oxidizing bacteria and archaea

Nitrate (NO₃^?) can contribute to surface water eutrophication and is deemed harmful to human health if present at high concentrations in the drinking water. In grazed grassland, most of the NO₃^?‐N leaching occurs from animal urine‐N returns. The objective of this study was to determine the effectiveness of a nitrification inhibitor, dicyandiamide (DCD), in decreasing NO₃^? leaching in three different soils from different regions of New Zealand under two different rainfall conditions (1260 mm and 2145 mm p.a.), and explore the relationships between NO₃^?‐N leaching loss and ammonia oxidizing bacteria (AOB) and ammonia oxidizing archaea (AOA). The DCD nitrification inhibitor was found to be highly effective in decreasing NO₃^?‐N leaching losses from all three soils under both rainfall conditions. Total NO₃^?‐N leaching losses from the urine patch areas were decreased from 67.7–457.0 kg NO₃^?‐N/ha to 29.7–257.4 kg NO₃^?‐N/ha by the DCD treatment, giving an average decrease of 59%. The total NO₃^?‐N leaching losses were not significantly affected by the two different rainfall treatments. The total NO₃^?‐N leaching loss was significantly related to the amoA gene copy numbers of the AOB DNA and to nitrification rate in the soil but not to that of the AOA. These results suggest that the DCD nitrification inhibitor is highly effective in decreasing NO₃^? leaching under these different soil and rainfall conditions and that the amount of NO₃^?‐N leached is mainly related to the growth of the AOB population in the nitrogen rich urine patch soils of grazed grassland.     

长期施肥对水稻土剖面氨氧化古菌和细菌丰度及组成的影响

长期施肥影响稻田土壤理化性质和硝化微生物群落,但长期施肥对稻田不同土层氨氧化古菌(AOA)和氨氧化细菌(AOB)群落结构的影响尚不明确.以湖南宁乡稻田不同施肥制度长期定位试验为平台,选取不施肥(CK)、施秸秆有机肥(ST)、有机-无机肥配施(OM)和施全量化肥(NPK)4个处理,采用实时荧光定量PCR和Illumina...     

红壤稻田不同生育期土壤氨氧化微生物群落结构

以福建省红壤稻田土壤为对象,通过提取土壤总DNA,利用特异引物进行PCR(聚合酶链反应)扩增和DGGE(变性梯度凝胶电泳)并结合DNA克隆测序,研究了水稻生长过程中稻田土壤氨氧化细菌和氨氧化古菌群落结构的变化。结果显示:稻田土壤具有丰富的氨氧化细菌和氨氧化古菌资源。水稻生长过程中土壤氨氧化细菌群落组成较为稳定,只表现出水稻生长前期(苗期、分蘖期)和中后期(孕穗期、成熟期)间存在一定差异。而土壤氨氧化古菌群落组成变化较大,在水稻生长的苗期、分蘖期、孕穗期和成熟期4个时期间均存在一定差异。在水稻生长过程中,土壤氨氧化细菌群落多样性指数无显著性变化,但氨氧化古菌群落多样性指数随水稻生长明显提高,孕穗期后才达到平稳。水稻生长前期土壤硝化势也具有显著上升趋势,孕穗期时达到最高,而后有所下降。土壤硝化势与氨氧化古菌群落多样性指数具有显著正相关性,与氨氧化细菌没有相关性。研究表明,氨氧化古菌对红壤稻田土壤硝化作用的影响程度较大,证实了氨氧化微生物尤其是氨氧化古菌在稻田土壤微生物组成及其生态系统功能中的重要性。     

藏系绵羊瘤胃古菌季节动态分析

在不同季节分别随机选取3只藏系绵羊,采用PCR-DGGE技术对12只藏系绵羊瘤胃内容物中的细菌区系进行分子生态学分析.对古菌16SrDNA的V2～V4可变区的扩增产物进行DGGE图谱分析、相似性分析和Shannon多样指数分析.结果表明:瘤胃内容物中古菌群落多样性指数冬季与春季间差异不显著(P>0.05),但冬季显著高于夏季(P<0.05),并极显著高于秋季(P<0.01);春季与夏季差异不显著(P>0.05),春季显著高于秋季(P<0.05);夏、秋季之间差异不显著(P>0.05).各季间瘤胃古菌区系组成结构相似性高于94%.冬、春季节,由于牧草中的纤维性物质含量增高,导致瘤胃古菌群落多样性增加,不同季节牧草中粗蛋白和酸性洗涤纤维含量的变化可以引起特异性的古菌群落发育,可能导致不同的优势种群.