Abundance and distribution of ammonia-oxidizing archaea in Tibetan and Yunnan plateau agricultural soils of China

As low oxygen and high ultraviolet （UV） exposure might significantly affect the microbial existence in plateau, it could lead to a specialized microbial community. To determine the abundance and distribution of ammonia-oxidizing archaea （AOA） in agricultural soil of plateau, seven soil samples were collected respectively from farmlands in Tibet and Yunnan cultivating the wheat, highland-barley, and colza, which are located at altitudes of 3200-3800 m above sea level. Quantitative PCR （q-PCR） and clone library targeting on amoA gene were used to quantify the abundances of AOA and ammonia-oxidizing bacteria （AOB）, and characterize the community structures of AOA in the samples. The number of AOA cells （9.34 × 10^7-2.32× 10^8 g^-1 soil） was 3.86-21.84 times greater than that of AOB cells （6.91 × 10^6-1.24 × 10^8 g^-1 soil） in most of the samples, except a soil sample cultivating highland- barley with an AOA/AOB ratio of 0.90. Based Kendall＇s correlation coefficient, no remarkable correlation between AOA abundance and the environmental factor was observed. Additionally, the diversities of AOA community were affected by total nitrogen and organic matter concentration in soils, suggesting that AOA was probably sensitive to several environmental factors, and could adjust its community structure to adapt to the environmental variation while maintaining its abundance.     

铜陵新桥矿区土壤中耐Cu微生物的筛选研究

选择安徽铜陵新桥矿区富Cu的污染土壤,充分利用微生物受自然环境重金属胁迫而产生耐性这一特点,进行土壤中耐Cu微生物的筛选研究。实验过程中分别配制三种不同的培养基,细菌、真菌和放线菌培养基。在水浴恒温振荡器中对土壤中耐受重金属的微生物进行驯化,将得到的对Cu2+耐受性最高的液体培养基作为菌源,在琼脂平板培养基上进行划线分离,并将得到的纯菌株在光学显微镜下进行形态观察,菌种经试管斜面富集培养后保存在4℃冰箱中以便后续使用。研究发现,土样中细菌和真菌对Cu的耐受性低,最高耐受质量浓度分别只达到500mg·L-1和1100mg·L-1。而放线菌表现出Cu高耐受性,分离得到的放线菌耐受Cu的质量浓度最高达到10000mg·L-1,初步鉴定该放线菌株为链霉菌属。该放线菌菌种可能同时对Cu有吸附降解特性,具有成为污染土壤生物治理的高效耐受吸附菌种的潜力巨大。     

近地层臭氧浓度升高对稻田土壤氨氧化与反硝化细菌活性的影响

利用中国唯一的稻麦轮作臭氧FACE（free-air O3 concentration enrichment,开放式空气臭氧浓度增高）试验平台,研究近地层臭氧浓度升高对稻田不同生育时期植株氮吸收量、土壤氮含量与脲酶活性、土壤氨氧化细菌与反硝化细菌数量以及成熟期土壤硝化与反硝化作用强度的影响。结果发现,臭氧浓度升高条件下,单株水稻（Oryza sativaL.）的氮吸收量趋于升高,土壤全氮含量趋于下降、脲酶活性趋于增强,在成熟期土壤全氮与对照相比平均下降9%,土壤脲酶活性与对照相比平均升高13%。在水稻整个生长季节,土壤氨氧化细菌和反硝化细菌的数量均呈现出先增多后减少的趋势,且均在开花期达到峰值,但臭氧熏蒸土壤与对照相比趋于升高;在水稻成熟期,土壤中单个氨氧化细菌和反硝化细菌的活性均趋于下降,土壤硝化强度和反硝化强度与对照相比也平均下降17%和24%。结果表明,近地层臭氧浓度升高条件下,稻田土壤氮素转化因为水稻氮素吸收增强而加快,土壤氨氧化细菌和反硝化细菌的数量均趋于增多,但它们的生理代谢活性均趋于下降。     

铜陵新桥矿区土壤中耐Cu微生物的筛选研究

选择安徽铜陵新桥矿区富Cu的污染土壤,充分利用微生物受自然环境重金属胁迫而产生耐性这一特点,进行土壤中耐Cu微生物的筛选研究。实验过程中分别配制三种不同的培养基,细菌、真菌和放线菌培养基。在水浴恒温振荡器中对土壤中耐受重金属的微生物进行驯化,将得到的对cu2＋耐受性最高的液体培养基作为菌源,在琼脂平板培养基上进行划线分离,并将得到的纯菌株在光学显微镜下进行形态观察,菌种经试管斜面富集培养后保存在4℃冰箱中以便后续使用。研究发现,土样中细菌和真菌对cu的耐受性低,最高耐受质量浓度分别只达到500mg·L-1和1100mg·L-1。而放线菌表现出cu高耐受性,分离得到的放线菌耐受Cu的质量浓度最高达到10000mg·L-1,初步鉴定该放线菌株为链霉菌属。该放线菌菌种可能同时对Cu有吸附降解特性,具有成为污染土壤生物治理的高效耐受吸附菌种的潜力巨大。     

铜胁迫对农田土壤酶活性、细菌和古菌数量的影响

为了研究铜矿周边农田土壤中不同Cu含量对土壤酶活性、细菌和古菌基因拷贝数的影响,在铜矿周边农田土壤中采集了8个不同Cu含量的土壤,测定土壤的酶活性、细菌和古菌基因拷贝数,结果表明,土壤总Cu含量为28.30~1 019.27mg·kg~(-1),酸可提取态Cu含量为335~415.11 mg·kg~(-1),细菌16S rRNA基因拷贝数在每克干土3.10×10~(10)~1.84×10~(11)个之间,古茵16S rRNA基因拷贝数在每克干土737×10~8~6.82×10~9个之间。土壤总Cu和酸可提取态Cu含量对土壤脲酶活性影响较大,但对土壤脱氢酶和转化酶活性影响不显著。土壤细菌基因拷贝数均与总Cu含量、各形态Cu含量呈极显著的负相关关系,同样土壤古菌基因拷贝数与总Cu含量、各形态Cu含量也均呈极显著的负相关关系,说明Cu胁迫对土壤细菌和古菌活性具有较大的抑制作用。     

Impact of anaerobically treated and untreated (raw) distillery effluent irrigation on soil microflora, growth, total chlorophyll and protein contents of Phaseolus aureus L

Impact of distillery effluent (untreated and treated) irrigation on soil microflora of the pots used for growing Phaseolus aureus L. was investigated. The growth of the P. aureus plants as affected by distillery effluent irrigation was also evaluated. The irrigation of the pots by 1-10% distillery effluent (anaerobically treated) stimulated the growth of the soil microflora (increased number of bacteria, fungi and actinomycetes) and P. aureus plants (increased shoot and root lengths, biomass, chlorophyll and protein contents). Further, 15-20% distillery effluent (anaerobically treated) had toxic effect on soil micro flora as indicated by reduced number of bacteria, fungi and actinomycetes. Reduction in shoot, root, lengths, biomass, chlorophyll, protein contents of P. aureus was also observed when irrigated by 15-20% treated distillery effluent. All the concentrations of raw distillery effluent reduced the bacterial population. However, the treated distillery effluent concentrations <10% had stimulatory effect on fungal and actinomycetes population. However, raw effluent concentrations >5% reduced the same. Raw distillery effluent was more toxic to P. aureus than treated distillery effluent as concentrations >5%, had reduced the growth (shoot, root length and biomass) of the test plant. Raw distillery effluent had adverse effect to total chlorophyll contents and all the test concentrations reduced the total chlorophyll level. However, untreated (raw) distillery effluent stimulated the protein content initially. It has been concluded from-present study that lower concentrations of the raw distillery effluent (1-5%) and treated distillery effluent (1-10%) had stimulated the growth of P. aureus and soil microflora except soil bacteria (inhibited by all the concentration of the raw effluent). However, higher concentrations (raw effluent: 10-20%; treated effluent 15-20%) had toxicity to test parameters.     

Community dynamics of ammonia oxidizing bacteria in a full-scale wastewater treatment system with nitrification stability

To determine whether the functional stability of nitrification was correlated to a stable community structure of ammonia oxidizing bacteria (AOB) in a full-scale wastewater treatment plant, the AOB community dynamics in a wastewater treatment system was monitored over one year. The community dynamics were investigated using specific PCR followed by terminal restriction fragment length polymorphism (T-RFLP) analysis of the amoA gene. The T-RFLP results indicated that during the period of nitrification stability, the AOB community structure in the full-scale wastewater treatment system was relatively stable, and the average change rate every 15 d of the system was 6.6%±5.8%. The phylogenetic analysis of the cloned amoA gene showed clearly that the dominant AOB in the system was Nitrosomonas spp. The results of this study indicated that throughout the study period, the AOB community structure was relatively stable in the full-scale wastewater treatment system with functional stability of nitrification.     

Carbon sequestration and soil microbes in purple paddy soil as affected by long-term fertilization

Soil organic carbon (SOC) sequestration impacts on food security and climate change and may be affected by soil microbes in fertilized croplands. A 12-year field experiment under the rice–wheat system was used to evaluate the effect of the long-term fertilization on the SOC accumulation, culturable soil microbes, and their interaction in purple paddy soil. Results showed that varied fertilizations resulted in a significant increase of the SOC content and stock in the plow layer, as well as rise in populations of major soil microbes, including bacteria, actinomycetes, and fungi compared with no fertilization. Soil with combined application of chemical NPK fertilizer and organic amendment (pig manure or rice straw return) on average had the highest organic carbon content and stock, amounts of bacteria, actinomycetes, and fungi, which were 7.8%, 5.8%, 75.8%, 130.5%, and 16.2% higher than the NPK fertilization alone. Fertilization differentially altered populations of the functional anaerobic bacteria in paddy soil. With the combined application of chemical NPK fertilizer and organic amendment, soil displayed higher amounts of anaerobic cellulolytic bacteria, anaerobic fermentative bacteria, hydrogen-producing acetogen, methanogenic bacteria, denitrifying bacteria, and sulphate-reducing bacteria than that with the NPK fertilization alone or no fertilization. Populations of all three major soil microbes showed significantly positive correlations with the SOC content, indicating their interaction was of mutual promotion. Data suggest that the combined application of the NPK fertilizer with organic amendment especially by the rice straw return is recommended to sustain the soil biological fertility and mitigate the emission of the greenhouse gas by the SOC sequestration in purple paddy soil.     

典型四环素类抗生素对土壤微生物及植物生长的影响

四环素类抗生素(TCs)在畜禽养殖中的大量使用甚至滥用,导致其在动物粪便中高浓度残留。随着畜禽粪便有机肥的农田施用,TCs持续进入土壤并且不断累积,由此带来的土壤生态危害和健康风险值得关注。以四环素(TC)、土霉素(OTC)为研究对象,采用室内培养试验法,考察2种典型TCs对土壤微生物、酶活性的影响及对植物生长的毒性作用。结果表明,低浓度TC和OTC作用下,土壤细菌和真菌数量即显著降低,土壤细菌较真菌对TCs的污染更为敏感。除TC对土壤酸性磷酸酶和OTC对土壤过氧化氢酶活性主要表现为激活作用外,总体上TC、OTC作用后土壤酶活性呈低浓度促进、高浓度抑制的变化趋势。80 mg·L~(-1)的TC、OTC暴露下,绿豆芽芽伸长被显著抑制,并且随着抗生素浓度的增大,绿豆芽伸长抑制率大幅升高。相同浓度、相同暴露时间条件下的TC对绿豆芽伸长的毒性大于OTC。     

磺胺类兽药对土壤微生物数量的影响

采用室内培养的方法,研究磺胺类兽药(磺胺二甲基嘧啶、磺胺甲噁唑)污染对土壤微生物数量(细菌、真菌、放线菌、固氮菌、兼气性固氮菌、硝化细菌、亚硝化细菌、反硝化细菌及氨化细菌)的影响.结果表明,磺胺类兽药对土壤细菌数量有一定的激活作用,其最大激活率在700%以上.兽药对土壤真菌数量的影响主要呈现抑制作用,其最高抑制率为92.9%.兽药对土壤放线菌数量有一定的抑制作用,对土壤固氮菌数量则有一定的激活作用.兽药对土壤兼气性固氮菌数量的影响表现为较低浓度时(10 mg.kg-1)抑制,而较高浓度时(50 mg.kg-1)则激活.兽药对土壤硝化细菌、亚硝化细菌、反硝化细菌及氨化细菌有一定的激活作用,其中对硝化细菌的最大激活率可达1000%以上.     

施肥与设施栽培措施对土壤微生物区系的影响

微生物肥料的施用能显著增加土壤中放线菌的数量，而降低细菌和霉菌的数量；施用有机肥对土壤微生物有较大的影响，其中，有机肥与无机肥，有机肥与生物肥配合施用的影响尤为明显，与对照相比，土壤中细菌数量分别增加109．8％和405．0％，放线菌数量分别增加320．3％和215．0％，而真菌数量分别降低44．0％和47．2％。大棚栽培条件下土壤微生物数量高于露地；在大棚中，种植年限长的土壤微生物数量低于种植年限短的。     

Preface

The combination of bioremediation and electrokinetics, termed bioelectrokinetics, has been studied constantly to enhance the removal of organic and inorganic contaminants from soil. The use of the bioleaching process originating from Fe- and/or S-oxidizing bacteria may be a feasible technology for the remediation of heavy metal–contaminated soils. In this study, the bioleaching process driven by injection of S-oxidizing bacteria, Acidithiobacillus thiooxidans, was evaluated as a pre-treatment step. The bioleaching process was sequentially integrated with the electrokinetic soil process, and the final removal efficiency of the combined process was compared with those of individual processes. Tailing soil, heavily contaminated with Cd, Cu, Pb, Zn, Co, and As, was collected from an abandoned mine area in Korea. The results of geochemical studies supported that this tailing soil contains the reduced forms of sulfur that can be an energy source for A. thiooxidans. From the result of the combined process, we could conclude that the bioleaching process might be a good pre-treatment step to mobilize heavy metals in tailing soil. Additionally, the electrokinetic process can be an effective technology for the removal of heavy metals from tailing soil. For the sake of generalizing the proposed bioelectrokinetic process, however, the site-specific differences in soil should be taken into account in future studies.     

西藏当雄拉屋矿区污染土壤微生物及其活性研究

通过现场采样及室内培养分析,研究了西藏当雄拉屋矿区污染土壤微生物区系组成、主要生理类群及其活性。结果表明：矿区土壤受重金属Cu、Zn、Pb、Cd不同程度污染,矿区污染土壤几种重金属质量分数比非矿区土壤有明显的增加。矿区土壤微生物区系组成和各生理类群发生了明显变化,土壤细菌、真菌、放线菌以及各生理类群数量均显著降低,且3大微生物以及各生理类群对矿区污染的敏感性大小分别表现为放线菌〉细菌〉真菌,硝化细菌〉氨化细菌〉纤维分解菌。矿区土壤酶活性较低,对照土壤的各种酶活性最高,而矿区土壤基础呼吸和代谢商则受到刺激明显提高,其中土壤脱氢酶的活性变化最大,作为矿区重金属污染的指标更灵敏。可见,土壤中微生物区系组成及参与物质转化的生理类群种类、数量及土壤酶活和微生物活性在一定程度上可反映该矿区污染生境的重金属污染特征及其生态功能的演变规律。     

Biomarkers indicate mixture toxicities of fluorene and phenanthrene with endosulfan toward earthworm (<Emphasis Type="Italic">Eisenia fetida</Emphasis>)

α-Endosulfan and some polycyclic aromatic compounds (PAHs) are persistent in the environment and can reach crop products via contaminated agricultural soils. They may even be present as mixtures in the soil and induce mixture toxicity in soil organisms such as earthworms. In this study, the combined toxicities of PAHs with α-endosulfan were determined in Eisenia fetida adults using an artificial soil system. α-Endosulfan and five PAHs were tested for their acute toxicity toward E. fetida in artificial soils. Only α-endosulfan, fluorene, and phenanthrene showed acute toxicities, with LC₅₀ values of 9.7, 133.2, and 86.2 mg kg^?1, respectively. A mixture toxicity assay was conducted using α-endosulfan at LC₁₀ and fluorene or phenanthrene at LC₅₀ in the artificial soils. Upon exposure to the mixture of fluorene and α-endosulfan, earthworms were killed in increasing numbers owing to their synergistic effects, while no other mixture showed any additional toxicity toward the earthworms. Along with the acute toxicity results, the biochemical and molecular changes in the fluorene- and phenanthrene-treated earthworms with or without α-endosulfan treatment demonstrated that enhancement of glutathione S-transferase activity was dependent on the addition of PAH chemicals, and the HSP70 gene expression increased with the addition of α-endosulfan. Taken together, these findings contribute toward understanding the adverse effects of pollutants when present separately or in combination with other types of chemicals.     

烟区烟草根际与非根际土壤微生物的生态因子作用分析

烟草根际和非根际土壤的理化性质、微生物区系和土壤酶之间的相互关系尚缺乏系统研究。对南阳烟区6种土壤包括黄褐土和黄棕壤等共13个取样点的烟草根际和非根际土壤的分析表明,根际与非根际微生物之间没有对应相关关系。根际土壤真菌数与解钾菌数、硝化细菌数显著正相关;非根际土壤荧光假单胞菌数与细菌芽孢数极显著正相关,氨化细菌数与放线菌数显著负相关。与土壤容重和空隙度显著相关的仅有根际真菌数(正相关)。与土壤全氮呈显著正相关的分别有根际荧光假单胞菌、纤维素分解菌(正相关)以及根际解磷菌和非根际真菌(负相关)。土壤全C与根际细菌总数显著负相关。土壤pH与非根际纤维素分解菌显著正相关。速效P与非根际放线菌数呈显著正相关。根际土壤蛋白酶活性与根际细菌总数极显著负相关。非根际土壤蛋白酶活性则分别与非根际放线菌和解钾菌显著正相关。根际土壤脲酶活性分别与根际真菌、解磷菌显著正相关,非根际脲酶活性则与非根际纤维分解菌显著正相关。非根际土壤过氧化氢酶活性与非根际细菌总数显著正相关。     

铜陵矿集区土壤中污染元素对微生物特性的影响研究

通过铜陵矿集区土壤中重金属污染元素含量与土壤中微生物生物量之间的对应关系,研究其环境效应。采集深度为20cm的土样,分别制备不同微生物的培养基以期对土壤中的微生物进行分离计数,细菌和放线菌采用稀释平板涂布法计数,高铁还原菌采用最大或然数法计数。研究结果表明,铜陵矿集区土壤中As、Au、Cd、Zn、Cu、Pb元素含量与土壤中细菌、放线菌、高铁还原菌数量之间的相关关系为：As、Au、Zn、Cu与细菌和放线菌,Cd、Pb与放线菌的含量呈负相关关系,指示了土壤中高浓度污染元素对微生物的抑制作用;Cd、Pb与细菌,Cd、Zn与高铁还原菌的含量呈正相关关系,反映部分微生物受污染元素的胁迫已产生了耐受性;Au、Pb、As与高铁还原菌数量的关系不明显。通过实验数据发现,微生物在重金属含量高的区域和含量低的区域的数量下降了大约2～3个数量级,有些区域微生物数量下降了约50%。矿集区土壤中三种菌种对污染元素的敏感程度不同,其敏感程度依次是：放线菌〉细菌〉高铁还原菌。矿集区部分土壤中,细菌、高铁还原菌由于其本身的耐受性或由于受胁迫而产生了耐受性,对Cd、Zn、Pb等三种重金属污染元素的抗性水平较高,具有作为土壤重金属元素污染修复微生物菌种的潜力。     

铝对土壤微生物区系组成的影响

通过大豆盆栽试验,研究了金属铝对土壤微生物区系组成的影响。结果表明,当铝质量分数<1.1 g/kg时,土壤细菌、放线菌的数量随着铝质量分数的增加而增加;当铝质量分数>1.1 g/kg时,土壤细菌、放线菌的数量随着铝质量分数的增加而减少。铝对丝状真菌的生长具有明显的促进作用。在低质量分数下,铝对土壤氨化细菌、硝化细菌、反硝化细菌具有一定的刺激作用;当质量分数>1.1 g/kg时,则表现为抑制作用,最大抑制率分别达48%、96%和95%;土壤硝化细菌、反硝化细菌可以作为铝污染土壤生态环境的指示菌。自生固氮菌的数量随铝质量分数的升高而逐渐下降。铝对土壤微生物区系产生抑制作用的临界质量分数在1.3 g/kg左右。     

不同放牧梯度下草甸草原土壤微生物和酶活性研究

通过小区控制放牧实验,研究呼伦贝尔草甸草原不同放牧强度下草地土壤微生物和酶活性的变化。结果表明：不同处理土壤微生物数量表现为细菌〉放线菌〉真菌。不同土层土壤微生物总数不放牧处理大于放牧处理,0～30 cm土层土壤微生物生物量碳、氮含量在轻牧区较高,在中牧区较低。土壤脲酶和过氧化氢酶活性轻度放牧和中度放牧高于不放牧和重度放牧。土壤微生物数量、生物量、土壤蛋白酶和过氧化氢酶活性均随土壤深度的增加呈递减趋势。相关分析表明,土壤微生物数量、微生物生物量以及土壤酶活性相互之间密切相关,土壤微生物量N与细菌达到极显著正相关（P〈0.01）,与真菌和放线菌呈显著相关（P〈0.05）。土壤微生物量C与真菌达到极显著负相关（P〈0.01）,与放线菌呈显著负相关（P〈0.05）。土壤微生物数量、土壤微生物量N与转化酶、蛋白酶、过氧化氢酶活性呈显著或极显著正相关。土壤微生物量C与转化酶、蛋白酶、过氧化氢酶呈显著负相关（P〈0.05）。     

Community diversity and distribution of ammonia-oxidizing archaea in marsh wetlands in the black soil zone in North-east China

AOA amoA genes in the soils of the two wetlands affiliated with three lineages. The main drivers of AOA community were pH and total organic carbon and ammonium. The soil characteristics rather than the vegetation control the AOA community. Since its first detection, ammonia-oxidizing archaea (AOA) have been proven to be ubiquitous in aquatic and terrestrial ecosystems. In this study, two freshwater marsh wetlands- the Honghe wetland and Qixinghe wetland – in the black soil zone in North-east China were chosen to investigate the AOA community diversity and distribution in wetland soils with different vegetation and depth. In the Honghe wetland, two sampling locations were chosen as the dominant plant transited from Deyeuxia to Carex. In the Qixinghe wetland, one sample location that was dominated by Deyeuxia was chosen. Samples of each location were collected from three different depths, and Illumina MiSeq platform was used to generate the AOA amoA gene archive. The results showed that the AOA amoA genes in the soils of the two wetlands were affiliated with three lineages: Nitrososphaera, Nitrosotalea, and Nitrosopumilus clusters. The different dominant status of these AOA lineages indicated their differences in adapting to acidic habitat, oxygenic/hypoxic alternation, organic matter, and other environmental factors, suggesting high diversity among AOA in marsh soils. The main driver of the AOA community was pH, along with organic carbon and ammonium nitrogen, which also played an important role combined with many other environmental factors. Thus, soil physiochemical characteristics, rather than vegetation, were the main cause of AOA community diversity in the wetlands in the black soil zone in China.     

长期连作及秸秆还田对棉田土壤微生物量及种群结构的影响

通过采用常规微生物培养技术分析长期不同连作年限及秸秆还田棉田土壤微生物量及种群结构变化,揭示棉花连作及秸秆还田与土壤微生物多样性关系。研究表明,随着连作年限增加,秸秆还田连作模式下真菌数量持续增加,细菌和微生物总量在连作5-20 a逐渐下降,25 a后又上升;非秸秆还田连作模式随着连作年限增加其微生物种群变化与秸秆还田连作模式变化一致,但非秸秆还田连作模式土壤细菌和微生物总量分别比秸秆还田连作模式在连作5、10和15 a降低42.9%、57.9%、70.6%和41.9%、54.7%和65.7%,而真菌数量增加28.4%、80.8%和116.7%。说明棉花秸秆还田能够增加微生物数量,改善棉田土壤微生物种群结构。连作棉田土壤Simpson指数和Shannon指数呈随连作年限增加呈增加的趋势,连作棉田土壤微生物细菌/真菌（B/F）和放线菌/真菌（A/F）比值在秸秆还田模式下先逐渐减小至连作25 a有升高,而非秸秆还田模式下持续下降。