土壤细菌呼吸对西双版纳热带森林恢复的响应

揭示不同恢复阶段热带森林土壤细菌呼吸季节变化及其主控因素,对于探明土壤细菌呼吸对热带森林恢复的响应机制具有重要的科学意义。以西双版纳不同恢复阶段热带森林(白背桐群落、崖豆藤群落和高檐蒲桃群落)为研究对象,运用真菌呼吸抑制法及高通量宏基因组测序技术分别测定土壤细菌呼吸速率和细菌多样性,并采用回归分析及结构方程模型揭示热带森林恢复过程中土壤细菌多样性、pH、土壤碳氮组分变化对土壤细菌呼吸速率的影响特征。结果表明：1)不同恢复阶段热带森林土壤细菌呼吸速率表现为：高檐蒲桃群落((1.51±0.62)CO₂ mg g^-1 h^-1)显著高于崖豆藤群落((1.16±0.56)CO₂ mg g^-1 h^-1)和白背桐群落((0.82±0.60)CO₂ mg g^-1 h^-1)(P<0.05)。2)不同恢复阶段土壤细菌呼吸速率呈显著的单峰型季节变化(P<0.05),最大值均出现在9月：高檐蒲桃群落((...     

细菌生物膜在农田土壤污染修复中的应用研究进展

全球农田土壤污染日趋严重。重金属、农药、微塑料作为常见的土壤污染物,已对农田生态系统与粮食安全造成严重威胁。细菌生物膜(bacterial biofilm,BF)作为分布于细菌表面的多组分聚集体,近年来已被证明在环境保护领域具有较高的应用价值。本文主要介绍了细菌生物膜的组成和功能,并对近年来细菌及其生物膜在重金属、有机物污染土壤修复中的应用及机理进行综述,展望生物膜群落结构在污染土壤中的修复潜力,以期深入理解细菌生物膜的关键作用,为挖掘更多细菌生物膜在环境保护方面的应用潜力提供理论指导。     

退耕还湿后土壤细菌群落结构和生物量变化过程

土壤细菌对湿地生态系统功能和健康维持起着重要作用。以菜子湖原始湿地、不同退耕年限湿地(3a、7a、11a和21a)和仍耕作油菜地土壤为研究对象,应用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)、高通量测序和磷脂脂肪酸(PLFAs)方法分析土壤细菌群落组成和生物量,探讨它们在退耕还湿后的变化过程及其影响因素。结果表明,退耕还湿后土壤变形菌门(α-变形菌纲、β-变形菌纲和δ-变形菌纲)、酸杆菌门(酸杆菌纲和全噬菌纲)、Nitrospinae(Nitrospinia纲)和硝化螺旋菌门(硝化螺旋菌纲)相对丰度先增高后降低;这些参与氮循环的土壤细菌对退耕后湿地生态恢复过程中土壤氮素提升起着重要作用。与农业生产活动密切关联的厚壁菌门(芽孢杆菌纲和梭菌纲)和放线菌门(放线菌纲)相对丰度逐渐降低。湿地土壤细菌多样性在退耕初期(3—7a)上升达到最大,退耕中后期逐渐降低。表层土壤各类群细菌生物量逐渐升高,亚表层土壤各类群细菌生物量则先降后升再降。水分条件和容重是与研究区土壤细菌群落结构和多样性密切相关的土壤因子,而全氮是与土壤细菌生物量密切相关的土壤因子。研究从生态过程视角解析了土壤细菌群落较详...     

Aerobic biodegradation of methyl tert-butyl ether (MTBE) by pure bacterial cultures isolated from contaminated soil

Summary Methyl tert-Butyl Ether (MTBE) has been used in gasoline as a substitute for lead-based additives, which have been demonstrated to be toxic. MTBE however, is persistent in soil and water, showing high affinity for water and low affinity for soil, and has become an important contaminant. Therefore, the aim of this work was to isolate and identify soil microorganisms capable of degrading MTBE. Two samples were taken from a gasoline-contaminated soil at a service station and 59 different bacterial strains were isolated by enrichment culture with three consecutive selective transfers. Biochemical and morphological characterization of the bacterial isolates classified them into the following groups: Bacillus, Rhodococcus, Micrococcus, Aureobacterium and Proteus. Twelve strains were selected for evaluation of MTBE biodegradation depending on visual growth and biomass production of the isolates in minimal salt broth. Six strains significantly reduced MTBE concentration (22–37%) compared to an abiotic control after 5 days of incubation. Although it has been considered that MTBE is degraded mainly by cometabolism, our results demonstrate that these microorganisms are able to reduce MTBE concentration when MTBE is the sole source of carbon.     

不同林龄杉木林土壤细菌群落结构与土壤酶活性变化研究

探究不同林龄杉木人工纯林土壤中的微生物的群落演变与结构特征与酶活性变化,为杉木人工林可持续经营管理提供依据。以福建省南平市的五片不同林龄杉木林表层土壤作为研究对象,通过16SrDNA测定细菌的群落组成,分析与土壤质量密切相关的四种土壤酶活性变化,揭示细菌群落与土壤酶活性的变化机理。结果表明,微生物的多样性指数与OTU都随着林龄的增加而增加,且幼龄林、中龄林、近熟林、成熟林土壤微生物结构差异较大;不同林龄杉木人工林中包含了29个细菌门,其中酸杆菌门与变形菌门为优势菌群,根据各种群相对丰度变化以及冗余分析,放线菌门、浮霉菌门与疣微菌门等均随林龄增长出现较大变化,且与土壤可溶性有机质以及速效养分有显著相关性（P<0.05）,说明这几种细菌群落对土壤养分变化较敏感;土壤养分变化会影响土壤酶活性,蔗糖酶与全碳呈显著正相关（P<0.05）,与速效钾呈显著负相关（P<0.05）,与放线菌门呈极显著负相关（P<0.01）。脲酶与速效氮呈显著负相关（P<0.05）,脲酶与变形菌门、绿弯菌门、放线菌门、硝化螺旋菌门以及拟杆菌门均存在较强相关性。综上,不同的土壤细菌种群与酶活性对各养分变化的响应程度不一,细菌群落结构与酶活性能反映不同林龄杉木林土壤的质量变化,适量延长杉木人工林种植年限有益于土壤质量恢复。本研究结果对指导杉木人工林优质经营有重要意义。     

Tolerance of Soil Bacterial Complexes to Salt Shock

Investigations showed that bacteria present in soil are resistant to one-day exposure to a saturated solution of ammonium nitrate and can well develop when transferred to laboratory nutrient media. The evaluated number of bacteria in NH₄NO₃-treated soil samples was nearly the same as in native soil samples, while it was 1.5–2.5 times smaller in the former than in the latter case when microbial succession in the soil samples was initiated by wetting them. Bacteria (particularly gram-negative ones) occurring at the early stages of succession were the most sensitive to salt stress. Bacteria in soil were found to be much more resistant to salt stress than the same bacteria isolated in pure cultures.     

黑龙江省表层冻土细菌群落结构组成和功能特征

冻土是气候变化的敏感区,冻土中细菌对于预测冻土和气候之间的潜在反馈机制至关重要,研究冻土区土壤中细菌的多样性和种群结构将有助于及时检测环境变化并采取有效的应对措施。以黑龙江省表层冻土为研究对象,运用Illumina MiSeq高通量测序分析技术,系统分析黑龙江表层冻土细菌群落结构组成和功能特征,探究影响因素。结果表明,得到的785640条原始序列可划分为30个门,109个纲,209个目,326个科,512个属,598个种。优势菌门主要包括Proteobacteria、Actinobacteria、Acidobacteria、Chloroflexi、Bacteroidetes、Verrucomicrobia,在P<0.01水平上,Proteobacteria与Actinobacteria、Chloroflexi极显著负相关,与Acidobacteria极显著正相关,Actinobacteria与Acidobacteria极显著负相关,与Chloroflexi极显著正相关,Acidobacteria和Chloroflexi之间则无显著相关性。属水平上,Aetherobacter属在各...     

鄂尔多斯台地盐沼滩涂湿地土壤细菌群落结构及特征

依据植被分类法将鄂尔多斯高原盐沼滩涂湿地划分为肉质耐盐草甸(B)、苔草草甸(C)、禾草草甸(D)和杂类草草甸(E)等4个植被亚型,并以盐沼裸地(A)为对照样地,共计5种盐沼滩涂湿地景观类型。运用高通量测序技术分别研究其土壤细菌群落结构特征、分布情况,以及土壤盐分与土壤细菌之间的关系。结果表明:(1) 5种盐沼滩涂湿地的土壤细菌样品共12213条OTUs,属于45个门,122个纲,365个目,663个科,1375个属,2882个种。(2)变形菌门(26.19%)是盐沼滩涂湿地平均相对丰度最高的门,其次为放线菌门(17.15%),绿弯菌门(12.62%),芽单胞菌门(Gemmatimonadetes,11.23%),拟杆菌门(Bacteroidetes,9.38%),酸杆菌门(Acidobacteria,8.83%)厚壁菌门(Firmicutes,2.96%);芽单胞菌纲中的未定细菌(norank＿c＿Gemmatimonadetes)是丰度最高的属,平均丰度为5.75%。(3)鄂尔多斯盐沼滩涂湿地自西南向东北,空间位置相近的土壤细菌群落结构具有更大的相似性,随着土壤盐分变化,变形菌门相对...     

种植阔叶树种和毛竹对土壤有机碳矿化与微生物群落结构的影响

为探明种植阔叶树种和毛竹对土壤有机碳矿化与微生物群落特征的影响,本研究通过盆栽试验和室内培养法比较分析种植香樟、木荷、青冈等阔叶树种与毛竹的土壤有机碳矿化速率和累计矿化量,并结合末端限制性片段长度多态性(T-RFLP)以及荧光定量PCR技术,分析土壤细菌、真菌群落组分与数量特征.结果表明:与种植阔叶树种的土壤相比,种植...     

杉木连栽对土壤细菌群落及其抑病能力的影响

连栽导致土壤退化是制约杉木初级生产力实现的重要障碍因素,而土壤对病原菌的抑制能力决定着植物能否有效抵御病原菌侵害,是人工林土壤地力状况的重要表现。以一代、二代、三代杉木人工林和天然次生林为对象,采用平板隔空、直接对峙的方法,分析了不同代际杉木林土壤细菌群落对尖孢镰刀菌和立枯丝核菌的抑制能力。进一步利用高通量测序技术,研究了杉木林土壤细菌群落影响土壤抑病能力的生态过程。结果表明：土壤磷元素随连栽呈显著积累趋势,而土壤pH和有机质(SOM)等含量随连栽代数的增加而下降,但这些下降指标在三代杉木林与天然林土壤间无显著差异。而杉木连栽导致土壤对病原菌的抑制能力逐代降低,天然林土壤较杉木人工林对病原菌具有显著的高抑制能力。同时杉木连栽显著改变了土壤细菌群落组成,而对群落整体α-多样性影响较小,说明土壤中一些关键类群对杉木连栽响应的敏感性高于整体细菌群落的变化。进一步利用随机森林模型预测与回归分析,揭示了杉木连栽引起的土壤一些关键细菌类群丰度的降低是土壤抑病能力下降的重要原因,这些类群主要受土壤pH、SOM、TP等土壤理化因子的调控。由此,杉木长期连栽会引起土壤微环境失衡,致使土壤抑制病原菌能力下...     

呼伦贝尔草原不同退化梯度土壤细菌多样性季节变化

为了研究草地退化程度与土壤微生物多样性的关系,在呼伦贝尔草地上选取羊草草甸草原和贝加尔针茅草甸草原两个典型放牧点,按照轻度、中度和重度划分取样点,分别于6、8月份和10月份3个不同季节采集土壤样品。应用变性梯度凝胶电泳技术(PCR-DGGE)研究两个放牧地点不同退化程度、不同季节草地的细菌群落结构变化。结果表明,呼伦贝尔草地不同退化梯度的草地土壤中细菌种类较为丰富。从丰富度和Shannon-Winner指数的变化看,两个放牧点8月份丰富度和Shannon-Winner指数最高,8月份的丰富度平均为32.4,比6月和10月份分别高11%和7.4%;8月份Shannon-Winner指数平均为4.15,比6月和10月份分别高7.7%和5.4%。DGGE图谱聚类分析结果显示,随着季节变化和草地退化程度由轻至重的变化,土壤中的细菌优势种群没有受到明显的影响。回收DGGE图谱中10个条带进行测序分析,结果显示,所有序列与GenBank数据库中的相似度在87%100%之间。基于98%的相似度,可将其中的7个鉴定为Proteobacteria(变形菌门),将其中的1个鉴定为Actinobacteria(放线菌门)。另外2个同已知序列相似性较低,可能是未知的细菌。结果表明,Proteobacteria(变形菌门)为呼伦贝尔草原土壤中的优势细菌类群,尽管所选取样点草地植被有不同程度的退化,但土壤微生物优势种群并没有发生变化。     

Bacterial Communities of Polychlorinated Biphenyls Polluted Soil Around an E-waste Recycling Workshop

Polychlorinated biphenyls (PCBs) contamination that has resulted from the recycling of electronic power capacitors and transformers could lead to deterioration in soil quality and high ecological risk. As microorganisms are generally considered to be the best indicators of soil pollution, the diversity of bacterial communities in the soil around an e-waste recycling workshop in the Taizhou e-waste recycling area of China was studied using denaturing gradient gel electrophoresis (DGGE). The results indicated that the PCBs content in the soil decreased with increasing distance from the recycling workshop. Moreover, a gradual change in soil bacteria diversity along the PCBs pollution gradient was observed. Furthermore, the predominance of Proteobacteria and Bacteroidetes was found on the basis of sequence analysis and some highly similar subsequences of microorganisms were also found to be closely related to the catabolism of PCBs and other organic compounds. In a word, our results indicated that PCBs pollutants had an evident impact on the structure of the soil microbial community and the enriched microbes might serve to decompose PCBs in soil.     

五大连池火山土壤细菌多样性及其群落结构

火山熔岩生境孕育了独特的土壤微生物群落。为了解火山生态系统土壤细菌群落多样性和群落结构及其关键影响因子,选择五大连池新、老期火山为研究样点,非火山为对照,基于高通量测序方法,分析不同采样点土壤细菌群落结构和多样性,结合土壤理化指标,进一步分析影响火山生态系统土壤细菌群落多样性的环境因子。结果表明：细菌操作分类单元（OTUs）、Ace指数、Chao1指数和Simpson指数变化趋势一致,表现为非火山 > 新期火山 > 老期火山。三个样点土壤的共有OUTs数量为713个,各自特有的OTUs数量不尽相同。三个样点土壤中检测到共有细菌16个类群,其中变形菌门、酸杆菌门、放线菌门和绿弯菌门为优势菌群,老期火山土壤中酸杆菌门、疣微菌门、Rokubacteria相对丰度最大,而Patescibacteria相对丰度最小。三个样点的土壤细菌群落具有明显的空间关系,相似性差异较大,但不符合随地理距离的增加而降低的模型。土壤理化性质测定结果标明：老期火山土壤pH、有机质、全氮、全磷、铵态氮和硝态氮显著高于新期火山和非火山,新期火山土壤含水量和速效磷显著低于老期火山和非火山。喷发时间和火成岩基质等特性会导致不同火山土壤理化性质的差异,进而影响土壤细菌多样性和群落结构。Pearson相关性分析表明：土壤pH显著影响细菌的多样性指数。冗余分析（RDA）结果表明：土壤氮含量、pH和有机质是影响火山森林生态系统土壤细菌群落结构的主要因子。     

Evaluation of Bacterial Diversity in Soil Microcosms at Different Moisture Contents

The succession analysis of bacterial diversity in the A horizons (rich in organic matter) of three contrasting types of soil—burozem, soddy gley soil, and chernozem—showed that the bacterial diversity of soil microcosms in humid regions can be adequately evaluated only if soil samples are incubated at different soil moisture contents. A complete account of actinobacteria and proteobacteria requires the levels of soil moisture corresponding to the maximum capillary–sorption moisture and capillary moisture, respectively. The bacterial diversity, whose value was maximum on the 40th day of succession, was higher in soddy gley soil than in burozem. The taxonomic structures of the bacterial communities of these two types of soil were different. After wetting chernozem samples from arid regions, the soil bacterial community changed insignificantly with time and drastically differed from that of soils from humid regions. The difference in the bacterial diversity of soils was the most distinct when it was evaluated by measuring the proportion between proteobacteria and actinobacteria.     

Seasonal fluctuations in the population of denitrifying and N2-fixing bacteria in an acid soil of a Norway spruce forest

The seasonal fluctuations in the concentration of cultured denitrifying and N₂-fixing bacteria were followed in an ammonium fertilised and a control soil of a Norway spruce forest near Villingen/Black Forest from December 1994 to August 1998. The horizontal distribution of bacteria in three layers was determined by the MPN-method and by molecular probing (colony hybridisation) using specific 0.4–0.7 kb DNA probes for denitrification steps (narG, nirS, nirK and nosZ) and for N₂-fixation (nifH). The data showed that highest bacterial counts and higher numbers of denitrifying and N₂-fixing bacteria were generally detectable in the upper (= 5 cm) soil layer and that their amount decreased with soil depth. The concentration of these cultured bacteria showed seasonal fluctuations with highest numbers in autumn/winter/early spring and with low counts in summer. Denitrifying and N₂-fixing bacteria amounted to less than 10% of the total number of cultured bacteria determined by the MPN-method. Fertilisation with ammonium did not cause a shift in the population of these bacteria. These findings were corroborated by hybridisation experiments with genomic DNA isolated from the different layers. Strongest DNA–DNA hybridisation band intensities were obtained in the upper soil layer and their intensities decreased with soil depth. Soil samples from Villingen assayed in the laboratory produced N₂O (in dependence of nitrate and C₂H₂ added to the vessels) and utilised this gas with higher activities in the assays with the fertilised soil. It is concluded that molecular techniques can successfully be applied for assessing seasonal fluctuations of bacterial populations in soil. Relative abundance of denitrifying and N₂-fixing bacteria can be determined from experiments with DNA isolated from soils. Attempts to transform these results to the total population of soil bacteria on a single cell basis are faced with many uncertainties.     

The application of the herbicide bromoxynil to a model soil-derived bacterial community: impact on degradation and community structure

AIMS: Bromoxynil degradation by soil micro-organisms has been shown to be co-oxidative in character. In this study, we investigate both the impact of the application of increasing bromoxynil concentrations on soil-derived bacterial communities and how these changes are reflected in the degradation of the compound. Our aim was to test the hypothesis that the addition of bromoxynil to a soil-derived bacterial community, and the availability of a readily utilizable carbon source would have an impact on bromoxynil degradation, and that would be reflected in the bacteria present in the soil community. METHODS AND RESULTS: Degradation of bromoxynil was observed in soil-derived communities containing 15 mg l(-1), but not 50 mg l(-1) of the compound, unless glucose was added. This suggests that the addition of carbon stimulates co-oxidative bromoxynil degradation by the members of the bacterial community. Measurable changes in the bacterial community indicated that the addition of bromoxynil led to deterministic selection on the bacterial population, i.e. the communities observed arise through the selection of specific micro-organisms that are best adapted to the conditions in the soil. The addition of bromoxynil was also shown to have a negative impact on the presence of alpha and gamma-proteobacteria in the soil community. CONCLUSION: Bromoxynil degradation is significantly inhibited in bacterial soil communities in the absence of readily accessible carbon. The application of bromoxynil appears to exert deterministic selection on the bacterial community. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the effects of increasing bromoxynil concentrations on a model bacterial population derived from soil. Soil communities show qualitative and quantitative differences to bromoxynil application depending on the availability of organic carbon. These findings might have implications for the persistence of bromoxynil in agricultural soils.     

基于高通量测序技术分析使它隆对玉米土壤细菌多样性的影响

【目的】研究使它隆对玉米土壤细菌多样性的影响。【方法】利用Illumina Miseq高通量测序技术,分别测定了玉米土壤细菌的16S rRNA基因的V4-V5可变区序列,进而对不同时期的不喷施除草剂和喷施除草剂的玉米土壤中细菌群落组成和多样性进行分析。【结果】研究共获得260940条有效序列,167191条优质序列,12656个OTUs。多样性分析结果表明,使它隆处理10 d后的土壤细菌多样性和丰度降低;使它隆处理60 d后的土壤细菌多样性和丰度提高。对土壤细菌群落组成分析发现,5个土壤样品中的优势菌门均为酸酐菌门、变形菌门、放线菌门、绿弯菌门和芽单胞菌门。使它隆处理10 d后的样品酸杆菌门的比例增加,放线菌门和绿弯菌门的比例降低;使它隆处理60 d后样品变形菌门的比例降低,绿弯菌门的比例明显增加。【结论】使它隆对玉米土壤细菌多样性产生一定影响,其影响随施药时间而异。     

Total bacterial diversity in soil and sediment communities—A review

Advances in microbial methods have demonstrated that microorganisms globally are the dominating organisms both concerning biomass and diversity. Their functional and genetic potential may exceed that of higher organisms. Studies of bacterial diversity have been hampered by their dependence on phenotypic characterization of bacterial isolates. Molecular techniques have provided the tools for analyzing the entire bacterial community including those which we are not able to grow in the laboratory. Reassociation analysis of DNA isolated directly from the bacteria in pristine soil and marine sediment samples revealed that such environments contained in the order of 10 000 bacterial types. The diversity of the total bacterial community was approximately 170 times higher than the diversity of the collection of bacterial isolates from the same soil. The culturing conditions therefore select for a small and probably skewed fraction of the organisms present in the environment. Environmental stress and agricultural management reduce the bacterial diversity. With the reassociation technique it was demonstrated that in heavily polluted fish farm sediments the diversity was reduced by a factor of 200 as compared to pristine sediments. Here we discuss some molecular mechanisms and environmental factors controlling the bacterial diversity in soil and sediments.