黄河三角洲湿地土壤微生物群落结构分析

采用末端限制性片段长度多态性分析(T-RFLP)技术和16S rDNA克隆文库的方法,分析了黄河三角洲滨海湿地土壤不同深度细菌和古菌的群落结构.研究表明,随着深度的增加,细菌群落的多样性下降,而古菌群落多样性则有上升的趋势,且土壤的细菌和古菌群落结构都呈现出规律的层状分布.该土壤包括各种硫酸盐还原菌、产甲烷古菌、光合细菌等丰富的细菌和古菌资源.图5参27     

景观人工湿地微生物群落结构的季节变化

采用PCR-DGGE技术对梦清园人工芦苇湿地不同季节的细菌群落变化进行了研究。结果表明,随着水体流动和季节更替,人工湿地中优势细菌一直在变化。测序结果显示芽孢杆菌在系统中较占优势,4个季节里都可以检测出来。恶臭假单胞菌在春夏秋3季比较有优势,而冬天枯草芽孢杆菌比较适合在该芦苇湿地中生存。经湿地处理后,水体细菌群落的多样性下降,相似性升高,部分细菌被淘汰出水体,但适应的细菌生长较快,整体细菌数量上升。对底泥的研究中,随运行时间的增加,进水口与出水口的细菌相似性分别下降,且进水口的相似性下降要明显快于出水口。     

分子技术在湿地微生物群落解析中的应用

人工湿地的研究和应用近年来受到了广泛重视。微生物是人工湿地系统的重要组成部分,其群落结构对于湿地的净化功能的发挥具有重要影响。与传统的微生物分析技术相比,分子生物学技术在解析人工湿地微生物群落结构时无需纯培养,具有高效、快速、简便的特点,使其广泛应用于环境微生物的研究。文章综述了近年来在聚合酶链反应技术（Polymerase Chain Reaction,PCR）基础上发展起来的几种新的分子生物技术,包括PCR-DGGE、LH-PCR、T-RFLP、PCR-SSCP和ARDRA,以及其在人工湿地微生物研究中的应用现状。通过这些分子技术,可以分析湿地处理特定废水过程中微生物的数量、丰度、多样性及优势种;鉴定湿地中特定功能菌群（如氨氧化细菌、反硝化细菌、除硫菌等）的数量、活动分布、空间变更及与污染物去除的关系;判断各种系统条件（如不同基质、植物、水力负荷等）的设置对微生物多样性和稳定性的影响。最后,对分子技术在湿地领域的未来发展进行了展望。     

恩诺沙星残留对土壤微生物数量及群落功能多样性的影响

对不同浓度恩诺沙星作用于土壤后三大类微生物数量及群落功能多样性进行了研究. 结果表明,恩诺沙星残留对它们影响强弱顺序为:细菌>放线菌>真菌,其影响作用随浓度从每克土 0. 01μg至 10μg的增加而加大,药物作用活性维持期为 6~8d,但对真菌作用不明显. 相对较低浓度的恩诺沙星残留(每克土中 0. 1μg, 1μg)不影响土壤微生物群落功能多样性,而相对较高浓度的恩诺沙星残留 (每克土中 10μg)则降低了其微生物群落功能多样性.图 2表 2参 9     

黄河口湿地典型盐碱植被群落土壤氮素的季节动态及根际效应

于5、8和10月对黄河口湿地典型断面盐碱植被群落进行3个航次的野外生态调查,研究了不同植被覆盖的湿地土壤氮素的时空分布特征及典型盐碱植物碱蓬和芦苇群落土壤氮素的根际效应,并基于线性模型的冗余分析(RDA)探讨土壤理化参数对氮素分布特征的影响.结果表明,黄河口湿地5月和10月土壤氮素含量高于8月,土壤氮素以有机氮为主,无机氮以铵态氮为主;总体上,表层土壤氮素含量芦苇碱蓬光滩,光滩土壤氮素含量低于碱蓬和芦苇群落土壤的30%—50%;光滩、碱蓬和芦苇群落剖面土壤总氮、有机氮和铵态氮在0—10 cm表层含量最高,随土壤深度其含量下降,硝态氮在土壤中呈现显著淋溶作用,其峰值出现在地下土壤层30 cm左右;碱蓬和芦苇群落根际土壤氮素含量均高于非根际土壤,且碱蓬根际土壤氮素含量显著高于芦苇根际土壤(P0.05).RDA分析结果表明,影响土壤氮素分布特征的主要理化参数是土壤有机碳含量和pH.植被种类影响黄河口湿地土壤氮素的累积,一般情况下土壤氮素的累积量呈现芦苇碱蓬光滩的特征,并具有明显的季节差异,相对芦苇群落,碱蓬在生长季节8月份根际土壤中积累更多的氮素.     

多氯联苯复合污染土壤的微生物群落结构多样性变化

土壤微生物群落结构多样性是指示土壤生态系统稳定性及其功能的重要传感器。采用磷脂脂肪酸(Phospholipid fatty acids,PLFAs)方法,对长江三角洲地区某POPs高风险区PCBs长期复合污染土壤的微生物群落结构多样性进行了初步研究。结果表明,PCBs重度污染土壤中格兰氏阴性菌(16:1w9、cy17:0等)和厌氧微生物(18:1w7)的PLFAs组分含量较多,而格兰氏阳性菌(如i15:0、i17:0等)、放线菌(16:0(10Me))及真菌(18:2ω6,9)和好氧性微生物的PLFAs含量较低,表明PCBs污染土壤中微生物群落结构与组成发生了明显变化。这一结果为PCBs降解微生物资源的定向筛选提供了科学依据。     

农田土壤抗生素抗性基因与微生物群落的关系

抗生素抗性基因(Antibiotic resistance genes,ARGs)作为一种新型环境污染物,近20年来在农田土壤中广泛富集,促进了耐药性的传播.土壤微生物群落在农业种植下也发生了极大的变化,由于ARGs的传播扩散与微生物息息相关,因此关于两者之间关系及相互作用的研究急速增加.本文通过综述近几年的国内外研究成果,概述了农田土壤ARGs的分布现状和优势微生物群落的变化情况,从农业种植模式、有机肥施用情况、施用有机肥的种类、土壤理化性质和土壤污染影响等角度总结ARGs富集与优势微生物群落、微生物多样性变化的关系,指出耐药菌的变化情况,基于当下研究的不足展望了继续深入探索的方向,为今后进一步深入探索两者间的科学规律提出了设想.     

盐碱地环境下芨芨草土壤微生物群落的初步分析

新疆的盐碱化土壤分布广泛,它与荒漠化过程相伴而生,不但造成了资源的破坏、农业生产的巨大损失,而且还对生物圈和生态环境构成威胁。芨芨草(Achnatherumsplendens(Trin)Nevski)有着耐寒、耐旱、耐盐、根系发达、适应性广,能够涵养水分、改良土壤等特性,因此芨芨草成为在新疆种植的改良盐碱地土壤的可选植物之一。目前,关于芨芨草在改良盐碱土地的过程中,它对改变盐碱地土壤微生物群落方面的研究还很少有人报道。作者采用稀释平板分离计数法测定了芨芨草不同地块垂直分布的各个土层中细菌、放线菌、真菌的数量,初步研究分析了种植芨芨草的盐碱地土壤微生物群落分布变化的特点,讨论了芨芨草的种植对盐碱地土壤环境的影响。试验表明,种植芨芨草对盐碱地土质的改良可能起到了一定的作用,随着芨芨草种植年限的增加,不但使细菌和放线菌的总数升高,而且细菌和放线菌数量的垂直分布表现出向上层移动的趋势,同时出现了真菌。此研究结果为种植芨芨草改良盐碱地土壤环境的进一步研究奠定基础。     

城市化对土壤微生物群落结构的影响

随着城镇化的推进,中原农业区大量的农业用地转变为城市用地,受城市化过程中人为活动的强烈影响,与城市化有关的土壤质量问题日益突出。而土壤质量与土壤微生物密切相关,后者对环境变化比较敏感,它能够较早地指示生态系统功能的变化,并反映出土壤的质量和健康状况。因此,研究土地利用方式的变化对土壤微生物的影响是评价土壤环境质量的重要指标。以"中部崛起"六省中城镇化率增长最快的地区之———河南省商丘市为例,采用以空间梯度代替时间顺序的方法,以城市建成区、城市外围的郊区及农田为研究对象,分别测定不同城市化水平下土壤微生物的数量和组成,研究土壤由农业用地转变为城市用地过程中,土壤微生物群落结构的变化。以城市中心为起点,分别在东、南、西、北四个方向上取样,在同一方向上沿城区—郊区—农田梯度上进行取样。采用平板涂抹计数法测定土壤中细菌、真菌、放线菌的数量。结果表明,农田、郊区和城市土壤微生物总数分别为14.68×106 cfu·g-1、8.19×106 cfu·g-1和10.99×106 cfu·g-1,即城市化使土壤中微生物总数量减少;同时,土壤微生物的组成和比例结构也发生变化,其中,城市建成区土壤中细菌和真菌所占微生物总数的比例高于郊区和农村,而放线菌在农村占的比例高于城市建成区和郊区。城市建成区不同用地类型下土壤微生物总数的变化趋势为公园校园居住区道路,其中,细菌占微生物总数的比例在公园土壤中最大,而放线菌和真菌则相反,二者占微生物总数的比例在道路土壤中最大,在公园土壤中比例最小。土壤微生物数量和组成的这些变化反映出土地由农业用地转变为城市用地过程中,土壤质量和健康状况及生态系统功能的变化,可以为评价土壤环境质量及土壤管理提供参考依据。     

南极中山站排污口土壤中微生物群落的DGGE分析

以南极中山站排污口土壤为研究对象,提取样品中微生物宏基因组DNA,用变性梯度凝胶电泳(DGGE)对宏基因组DNA中的16S rRNA基因V3可变区进行分析,测定了31个不同条带所代表的序列.比较结果显示,排污口土壤样品与相同生境对照样品中的微生物类群组成均以CFB (Cytophaga-Flexibacteria-Bacteroides)类群和γ-变形菌纲(γ-Proteobacteria)为主,另外还有少量的α-, β-, δ-变形菌纲,和一些原核的藻类和革兰氏阳性细菌(G ). 但在排污口土壤样品中检出了传染性微生物毛球菌属(Trichococcus)和狡诈菌属(Dolosigranulum),以及丰富的小球藻(Chlorella),说明污水的排放已经对该地微生物群落造成明显的影响.     

Removal of tetrachlorobisphenol A and the effects on bacterial communities in a hybrid sequencing biofilm batch reactor-constructed wetland system

SBBR-CW system was proposed to effectively treat wastewater containing TCBPA. CW unit contributed more than SBBR to the removal of TCBPA. TCBPA changed the composition and structure of bacterial community in the system. GAOs massively grew in SBBR, but did not deteriorate TP removal efficiency. Tetrachlorobisphenol A (TCBPA) released into the sewage may cause environmental pollution and health risk to human beings. The objective of this study was to investigate the removal of TCBPA and bacterial community structures in a laboratory-scale hybrid sequencing biofilm batch reactor (SBBR)-constructed wetland (CW) system. The results showed that the removal efficiency of chemical oxidation demand (COD), ammonia, total nitrogen and total phosphorus in the SBBR-CW system was 96.7%, 97.3%, 94.4%, and 88.6%, respectively. At the stable operation stage, the system obtained a 71.7%±1.8% of TCBPA removal efficiency with the influent concentration at 200 mg/L. Illumina MiSeq sequencing of 16S rRNA gene revealed that the presence of TCBPA not only reduced the bacterial diversity in the SBBR-CW system, but also altered the composition and structure of bacterial community. After the addition of TCBPA, Proteobacteria increased from 31.3% to 38.7%, while Acidobacteria and Parcubacteria decreased greatly in the SBBR. In contrast, Acidobacteria replaced Proteobacteria as the dominant phylum in the upper soils of CW. The results indicated that TCBPA stimulated the growth of GAOs in the SBBR without deteriorating the phosphorus removal due to the presence of sufficient carbon sources. The ammonia oxidizing bacteria, Nitrosomonas, and denitrification bacteria, Hyphomicrobium and Pseudomonas, were inhibited by TCBPA, resulting in a decreasing the removal efficiency of TN and ammonia.     

Impact of heavy metals on bacterial communities from mangrove soils of the Mahanadi Delta (India)

This study aimed to assess soil nutrient status and heavy metal content and their impact on the predominant soil bacterial communities of mangroves of the Mahanadi Delta. Mangrove soil of the Mahanadi Delta is slightly acidic and the levels of soil nutrients such as carbon, nitrogen, phosphorous and potash vary with season and site. The seasonal average concentrations (μg/g) of various heavy metals were in the range: 14 810–63 370 (Fe), 2.8–32.6 (Cu), 13.4–55.7 (Ni), 1.8–7.9 (Cd), 16.6–54.7 (Pb), 24.4–132.5 (Zn) and 13.3–48.2 (Co). Among the different heavy metals analysed, Co, Cu and Cd were above their permissible limits, as prescribed by Indian Standards (Co=17 μg/g, Cu=30 μ g/g, Cd=3–6 μ g/g), indicating pollution in the mangrove soil. A viable plate count revealed the presence of different groups of bacteria in the mangrove soil, i.e. heterotrophs, free-living N₂ fixers, nitrifyers, denitrifyers, phosphate solubilisers, cellulose degraders and sulfur oxidisers. Principal component analysis performed using multivariate statistical methods showed a positive relationship between soil nutrients and microbial load. Whereas metal content such as Cu, Co and Ni showed a negative impact on some of the studied soil bacteria.     

铬、铜和镍对人工湿地硝化细菌与反硝化细菌群落结构分布的影响

在铬、铜和镍复合污染的3段波形潜流人工湿地中采集0~15 cm土壤样品,分别以单氨加氧酶的编码基因amoA和亚硝酸还原酶的编码基因nirK的部分序列作为硝化细菌和反硝化细菌的指示基因,采用实时聚合酶链式反应（实时PCR）与PCR-变性梯度凝胶电泳（PCR-DGGE）结合克隆和测序的方法,研究了硝化细菌与反硝化细菌群落结构的空间分布。结果表明,在人工湿地中,amoA和nirK基因丰度均在第3段最高,分别为（8.60×2.75）×106拷贝·g-1和（137.76×42.16）×10^6拷贝·g-1。回收的15个amoA基因序列中有7个属于簇3a,而8个nirK基因序列中有3个属于簇III。相关性分析表明,amoA基因拷贝数与全量铜、全量镍和铬的活化率呈显著负相关（r分别为-0.870、-0.884和-0.898）,但有效铜与amoA香农-维纳指数和辛普森指数之间均呈显著正相关（r分别为0.892和0.940）,铜的活化率与amoA基因均匀度呈显著正相关（r=0.887）;nirK基因拷贝数与全量铜、全量镍和铬的活化率也均呈显著负相关（r分别为-0.818、-0.820和-0.860）,但有效铜与nirK基因均匀度呈显著正相关（r=0.917）。以上结果表明,硝化细菌和反硝化细菌在人工湿地各个阶段的变化规律基本相同,这二者的丰度与多样性指数对重金属污染的反应总体上呈相反的变化。     

Effects of salinity on soil bacterial and archaeal community in estuarine wetlands and its implications for carbon sequestration: verification in the Yellow River Delta

Soils from two typical tidal salt marshes with varied salinity in the Yellow River Delta wetland were analysed to determine possible effects of salinity on soil carbon sequestration through changes in soil microbiology. The mean soil respiration (SR) of the salt water–fresh water mixing zone (MZ) was 2.89 times higher than that of the coastal zone (CZ) (4.73 and 1.63?μmol?m^?2?s^?1, respectively, p?Pseudomonas sp. and Limnobacter sp. that might have led to its higher dehydrogenase activity and respiratory rates. Additionally, the CZ possessed more Halobacteria and Thaumarchaeota with the ability to fix CO₂ than the MZ. Significantly lower soil salinity in MZ (4.25?g?kg^?1) was suitable for β-Proteobacteria, but detrimental for Halobacteria compared with CZ (7.09?g?kg^?1, p?相似文献   

滨海盐沼及其植物群落的分布与多样性

滨海盐沼是广泛存在于世界中、高纬度地区的一种湿地生态系统,具有抵御风暴潮灾害、净化污染物和为珍稀濒危生物提供适宜生境等重要的生态和经济价值。滨海盐沼因随高程变化而急剧变化的环境梯度和植物带状分布现象而为生态学者阐释自然界物种的分布机制提供了理想系统。主要概述了滨海盐沼的定义、特点、类型、全球分布以及潮汐作用、土壤盐度等环境因子特征;阐述了不同尺度下滨海盐沼的植物群落分布和多样性特征。在滨海盐沼植物群落的分布特征上,重点阐述了中尺度下的植物带状分布,即植物群落往往在白海向陆渐高的不同高程梯度上表现出显著的分带分布,不同植物各自占据该梯度上的一定区域。通常认为,带状分布是植物竞争和物理性胁迫共同调控的结果,但其在不同地理区域的普适性仍存争议。滨海植物群落多样性往往较低,在中、小尺度上盐沼植物多样性受控于盐度、潮汐等物理性胁迫、植物间相互作用等因子的作用;在大尺度上盐沼植物多样性可能随纬度增大而增加。系统深入地认识滨海盐沼植物群落生态格局和过程,将为气候变化、生物入侵等人类影响下的滨海盐沼生态系统的管理和恢复提供有益经验。     

A comparative study of microbial communities in soils amended by manures from pigs fed with organic versus synthetic feeds

We applied pyrosequencing to determine bacterial communities in soil samples taken from test fields amended by manures of pigs fed with green feeds (GPMS), synthetic feeds (APMS), and the reference field without swine manure amendment (NPMS), respectively. For each sample, 7050 effective sequences were selected and utilized to do the bacterial diversity and abundance analysis, respectively. In total, 2986, 2873, and 1602 operational taxonomic units were obtained at a 3% distance cut-off in GPMS, APMS, and NPMS, respectively. Bacterial phylotype richness in GPMS was higher than the other samples. NPMS had the least richness. The most dominant class in both GPMS and APMS is Betaproteobacteria, whereas Alphaproteobacteria is dominant in NPMS. Circa 7.5% sequences in GPMS were found to be affiliated with Burkholderiales order. Bacterial diversity and abundance in the soil varied with swine manure amendment, due to nutrient elements as well as toxic metals in swine manures. Compared with the soil amendment by manures from pigs bred with synthetic feeds, more bacterial diversity but less potentially pathogenic bacterial genera in the soil amended by manures of pigs fed with green feeds were found, which indicated that these manures were better land-applied organic fertilizers.     

Spatial and seasonal variations in bacterial communities of the Yellow Sea by T-RFLP analysis

Four typical coastal sites (rocky shore, sandy shore, mud flat shore, and artificial harbor) at the Yellow Sea were chosen to investigate the spatial and seasonal variations in bacterial communities. This was accomplished by using terminal restriction fragment length polymorphism (T-RFLP) analysis of PCR amplified 16S rDNA fragments. Two kinds of tetrameric restriction enzymes, HhaI and MspI, were used in the experiment to depict the bacterial community diversity in different marine environments. It was found that the community compositions digested by the two enzymes separately were different. However, the results of bacterial community diversity derived from them were similar. The MDA analysis results of T-RFLP profiles coming from HhaI and MspI both exhibited a significant seasonal community shift for bacteria and a relatively low spatial variation among the four locations. With HhaI as the sample, the pair wise T-tests also revealed that variations were minor between each pair of marine environments, with R ranging from 0.198 to 0.349. However, the bacterial community structure in the mud flat site depicted a larger difference than each of the other three sites (R ranging from 0.282 to 0.349).     

宿根甘蔗根际土壤细菌多样性分析中培养法与非培养法比较研究

采用培养法和非培养法提取甘蔗根际土壤微生物的总DNA,基于通用引物PCR扩增构建两种方法的细菌16SrDNA文库,并进行核糖体DNA扩增片段限制性内切酶分析(ARDRA),通过部分克隆序列测定构建细菌克隆文库的系统发育树,进而对基于传统的细菌平板培养法和直接提取总细菌DNA的非培养法进行比较分析.结果显示,非培养法文库的香侬–威纳指数、辛普森指数、丰富度分别为4.94、0.998、28.91,均高于培养法文库中相应的多样性参数(分别为4.27、0.996、16.79),均一度数值都>0.95.结合统计学数据和序列测定信息,反映出甘蔗根际土壤存在着丰富的细菌多样性,但平板培养法所展现的多样性低于直接提取法,表明前者存在着很大的局限性,而非培养法文库中主要是未培养微生物的16S rDNA序列,这从一个侧面反映了土壤样品中未培养微生物占很大比例.因此,在土壤微生物群落研究中,必须结合新的分子生态学技术手段,如采用直接提取总DNA的方法进行研究,才能比较全面地认识土壤微生物多样性.图3表3参18     

三江平原湿地典型植物群落氮的积累与分配

采用野外采样和室内分析结合的方法,研究了三江平原湿地典型植物群落氮的积累与分配特征。结果表明,小叶章（Calamagrostics angustifolia）、乌拉苔草（Carex meyeriana）和毛苔草（Carex lasiocarpa）群落地上器官氮含量不存在显著差异,根的氮含量存在显著差异,枯落物的氮含量表现为乌拉苔草群落〉小叶章群落〉毛苔草群落（p〈0.01）。小叶章、乌拉苔草和毛苔草群落不同部分的氮积累量和积累速率（VN）季节变化明显,三者地上器官、枯落物的氮积累量和VN表现为乌拉苔草群落〉小叶章群落〉毛苔草群落;三者根的氮积累量表现为小叶章群落〉毛苔草群落〉乌拉苔草群落,VN表现为毛苔草群落〉小叶章群落〉乌拉苔草群落。小叶章、乌拉苔草和毛苔草群落不同部分的氮分配比在各时期差异明显,根是重要的氮储库,其分配比高达（87.76±2.55）%、（79.84±7.53）%和（89.25±5.49）%;地上部分的氮分配比均以叶最高,茎较低;地上与地下的氮分配比呈相反变化规律,反映了其在氮供给方面的密切联系。     

三江平原湿地土壤磷的吸附与解吸研究

采用恒温培养法研究了三江平原典型沼泽湿地表层土壤中磷的吸附与解吸行为及其动力学过程。结果表明:湿地土壤对磷的自然释放量有限(1.89～13.25mg·kg-1),对磷酸盐具有较强的吸附能力,吸附量与土壤本身理化性质及水中磷酸盐初始质量浓度有关,其吸附/解吸平衡浓度(EPC0)在0.034~0.161mg·L-1之间。研究区域湿地表层土壤磷的吸附解吸特征趋于一致,吸附过程表现为快慢两种反应。六种动力学方程都能很好的拟合土壤磷吸附动力学过程,其中以langmuir型方程的拟合性最好,Evolich方程次之。