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.     

Methanogenic archaeal communities developed in paddy fields in the Kojima Bay polder, estimated by denaturing gradient gel electrophoresis, real-time PCR and sequencing analyses

Methanogenic archaeal communities inhabiting the paddy field soils in the Kojima Bay polder were investigated using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), real-time PCR and sequencing analyses. Soil samples of the plow and subsoil layers were collected in 2006 from four paddy fields that were reclaimed between 1692 and 1954. The DGGE band patterns of the targeted 16S rRNA genes amplified from the extracted DNA from the samples were different from the patterns from the paddy field soils in diluvial and alluvial areas. The numbers of targeted 16S rRNA genes, which were involved with methanogenic archaeal and other archaeal sequences, were approximately 10⁷–10⁸ and 10⁶ g⁻¹ dry soil in the plow and subsoil layers, respectively. Sequences of methanogenic archaeal 16S rRNA genes belonging to Methanocellales (Rice cluster I), Methanosarcinales and Methanobacteriales were obtained from the major DGGE bands. Whereas sequences in Methanomicrobiales, which were predominant methanogens in the diluvial and alluvial paddy fields, were not recovered. Known halophilic and methylotrophic methanogens, which are characteristic of saline and marine environments, were not detected. These results indicate that distinctive methanogenic archaeal communities have developed in the paddy field soils in the Kojima Bay polder.     

Effects of Silver Nanoparticles on Soil Ammonia-Oxidizing Microorganisms Under Temperatures of 25 and 5℃

The excellent bactericidal performance of silver nanoparticles (Ag NPs) has led to their wide applications, resulting in increasing concerns about their potential environmental impacts. This study evaluated the influences of different concentrations of Ag NPs (0, 1, 10, and 100 μg g^-1 dry soil) on the ammonia-oxidizing microorganisms in soil at cultivation temperatures of 25 and 5℃ for 37 d. The results showed that 1 μg g^-1 dry soil of Ag NPs had no acute effects on the ammonia-oxidizing microorganisms. However, 10 and 100 μg g^-1 dry soil of Ag NPs levels were found to significantly inhibit the activities of soil nitrification, with a decrease of 69.89% and 94.55%, respectively, at 25℃ and 61.65% and 83.79%, respectively, at 5℃ compared to the control (0 μg g^-1 dry soil of Ag NPs). These levels of Ag NPs also obviously decreased soil urease activity from about 380.47 ±0.07 (at 5℃) and 529.76 ±13.44 (at 25℃) mg N g^-1 dry soil d^-1 to 61.70 ±2.97 and 68.29 ±8.22 mg N g^-1 dry soil d^-1, respectively, after 37 d of cultivation. Quantitative polymerase chain reaction showed the abundance of ammonia-oxidizing archaea and bacteria. For the same exposure time, the effects of Ag NPs on the activities of ammonia-oxidizing microorganisms and urease decreased with decreasing temperature. The threshold concentration of Ag NPs that induced negative effects on ammonia-oxidizing microorganisms was higher at 5℃ than at 25℃. Therefore, the temperature has a major impact on the toxicity of Ag NPs to ammonia-oxidizing microorganisms and on the urease activity, with toxicity being reduced with decreasing temperature.     

绿肥稻田土壤甲烷减排材料筛选及减排机制研究

绿肥在培育健康土壤和保障水稻高产优质方面具有重要作用,然而高量有机物还田可能引发稻田土壤的甲烷（CH₄）增排,须研发相应的管控路径。本研究通过在添加绿肥后的稻田土壤中施用田菁炭、过氧化钙、硫磺、2-氯乙烷磺酸钠和针铁矿5种CH₄减排材料,研究其CH₄减排效果,并针对减排效果较好的材料开展了作用机制研究。结果表明：与无绿肥（CK）相比,绿肥还田增加了稻田土壤CH₄排放,添加减排材料后CH₄排放降低。其中,与单独绿肥（GM）相比,添加田菁炭（GM+C）和硫磺（GM+S）CH₄累积排放量降幅较小,分别为19.5%和23.8%;2-氯乙烷磺酸钠（GM+CES）相较于GM的CH₄减排效果最佳,CH₄累积排放量降低了66.7%,全球增温潜势（GWP）降低了 52.5%。培养结束时（90 d）,与 GM 相比,添加 2-氯乙烷磺酸钠的土壤 pH降低了 0.4个单位,产甲烷古菌mcrA丰度降低了29.9%。随机森林模型和相关性分析发现产甲烷古菌mcrA基因丰度与CH₄排放呈显著正相关,说明产甲烷古菌是驱动绿肥稻田CH₄排放的重要因子。研究表明,绿肥稻田土壤可通过添加减排材料实现CH₄减排,本研究中以2-氯乙烷磺酸钠的效果最优,其专一、高效抑制产甲烷古菌群落,是未来值得探索的绿肥稻田土壤CH₄减排材料。     

鸡粪好氧堆肥过程中氨氧化古菌群落结构的动态变化

应用聚合酶链式反应-变性梯度凝胶电泳（PCR DGGE）技术,研究了好氧堆肥过程氨氧化古菌（ammonia oxidizing archaea, AOA）的群落结构和多样性变化。结果表明,不同堆肥时期鸡粪好氧堆肥AOA菌群的群落结构发生了明显的变化。与AOA HH 2(GU2258721)亲缘关系较近的b条带(相似性96%)和未培养泉古菌属［uncultured crenarchaeote NM 152(HQ8752251)］的m条带(相似性99%)是堆肥过程一直存在的AOA菌属。条带c、 b、 f、 i和条带m、 k、 l、 n代表的种属分属两个大类。AOA群落的Shannon Weiner指数（H）、 均匀度指数（EH）均表现为第30 d＞第5 d＞第25 d第45 d＞第3 d第12 d＞第1 d第15 d。对AOA种群数据与不同时期堆体温度、 pH、 全氮、 铵态氮、 亚硝态氮和硝态氮等环境因子的冗余分析表明,堆体温度、 全氮、 亚硝态氮和硝态氮对氨氧化古菌群落演替有着显著的影响（P0.05）,AOA的群落结构在堆肥第1 d、 第5 d、 第25 d、 第30 d、 第45 d变化较大。说明这些参数能有效控制鸡粪好氧堆肥过程中AOA的群落结构。     

种植水稻对古水稻土与现代水稻土微生物功能多样性的影响

通过温室盆栽试验,研究种植水稻对长江三角洲绰墩山遗址的古水稻土与现代水稻土的微生物功能多样性和氨氧化细菌数量的影响。在60d的试验期内观察发现,现代水稻土含有0．416～1、235MPN mg^-1的氨氧化细菌,古水稻土在温室培养后也检测到0．013—0、055MPN mg^-1的氨氧化细菌,但数量远远低于现代水稻土;在不施氮的条件下种植水稻会降低古水稻土与现代水稻土中氨氧化细菌的数量,但施加硫酸铵在种植水稻30d时增加了其数量,而施加硝酸钠对其数量没有显著影响。现代水稻土的碳底物利用能力明显弱于古水稻土,微生物群落的Shannon指数、Simpson指数和McIntosh指数也不同程度的低于古水稻土;种植水稻60d后,现代水稻土中微生物群落的整体活性显著提高,三种多样性指数也达到与古水稻土相同的水平,但施加氮肥又减缓了种植水稻对微生物整体活性的增强作用。种植水稻和施用氮肥对古水稻土微生物群落的三种多样性指数均没有显著影响,而且3320a古水稻土和6280a古水稻土的微生物功能多样性也没有显著差异。     

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...     

模拟氮沉降对贝加尔针茅草原土壤氮转化微生物的影响

草地土壤是温室气体重要的源和汇,认识草地生态系统氮转化过程有助于预测氮循环对未来氮沉降增加的响应与反馈机制。依托于2010年在内蒙古贝加尔针茅草原设置的长期模拟氮沉降增加的氮添加试验,共设置了8个氮添加水平(0、15、30、50、100、150、200、300 kg N·hm~(-2)·a~(-1))。应用荧光定量PCR方法,研究氮转化功能基因丰度对不同氮添加水平的响应。2015年8月取样分析结果表明:固氮微生物(nifH)基因丰度随着氮添加水平的升高,表现为先升高后降低的趋势。低于200 kg N·hm~(-2)·a~(-1)硝酸铵处理有利于固氮菌生长。低氮添加(N15、N30和N50)对氨氧化细菌(AOB-amoA)和氨氧化古菌(AOA-amoA)基因丰度无显著影响。高氮添加(N100、N150、N200和N300)显著提高了AOB基因丰度,降低了AOA基因丰度。高氮添加(N150、N200和N300)显著降低了nirK基因丰度。随着氮添加量的增加,高氮添加促进了AOB主导的氨氧化过程,而反硝化微生物丰度的减少提高了氨氧化产物硝酸盐的积累,继而提高了土壤硝酸盐含量。