高密度聚乙烯微塑料与氯嘧磺隆对大豆生长和根际细菌群落的复合胁迫效应

随着我国农业的大力发展，塑料地膜和农药被广泛投入到农业生产中，而塑料地膜降解形成的微塑料和农药在土壤中累积也带来诸多环境问题.目前微塑料与农药单一作用的环境生物学效应已有报道，但两者复合胁迫对作物生长和根际土壤细菌群落的影响研究较少.因此，设计高密度聚乙烯微塑料（HDPE，500目）与磺酰脲类除草剂代表品种氯嘧磺隆共处理，研究其对大豆生长的影响，并通过高通量测序技术、互作网络和PICRUSt2功能分析，探究HDPE和氯嘧磺隆复合胁迫对大豆根际土壤细菌群落多样性、结构组成、菌群网络和土壤功能的影响，阐明HDPE和氯嘧磺隆对大豆的复合毒性.结果表明1% HDPE处理延长氯嘧磺隆在土壤中的半衰期（由11.5 d升至14.3 d），并且HDPE和氯嘧磺隆复合胁迫较单一污染物对大豆生长的影响更为明显.HiSeq 2500测序表明复合胁迫下的大豆根际细菌群落由20个门、312个属组成，门和属的组成数量显著少于对照和单一处理，并降低具有潜在生物防治特性、植物促生特性等功能菌属的相对丰度（如Nocardioides和Sphingomonas等）.Alpha多样性表明复合胁迫显著降低大豆根际细菌群落的丰富度与多样性，Beta多样性则表明复合胁迫显著改变大豆根际细菌群落结构.组间样品LEfSe和PICRUSt2功能分析表明复合胁迫调控根际细菌群落的优势菌群，并减弱土壤氨基酸代谢、能量代谢和脂质代谢等二级功能层的丰度占比.由属水平网络分析推测复合胁迫降低土壤细菌间的总连接数和网络密度，使网络结构简单化，维持网络稳定的重要菌群种类也发生变化.研究结果表明HDPE和氯嘧磺隆复合胁迫显著影响大豆生长，并改变大豆根际细菌群落结构、土壤功能和网络结构，相较于单一处理，复合胁迫的潜在危害更大.研究结果可为评价聚乙烯微塑料和氯嘧磺隆生态风险，以及污染土壤修复提供指导.     

甲胺磷、乙草胺和铜单一与复合污染对黑土环境安全的胁迫研究

以黑土为环境介质,通过急性毒性试验法研究了东北黑土区普遍存在的2种农用化学品甲胺磷和乙草胺与重金属Cu对赤子爱胜蚓(Eiseniafoetida)的单一与复合毒性效应.单一毒性试验结果表明,3者对蚯蚓均有毒性,顺序为甲胺磷>乙草胺>Cu.复合毒性实验结果表明,2种有机农药与重金属Cu复合毒性效应十分复杂,与不同的浓度组合及染毒历时有关,一般随时间的延长,毒性加剧.2种有机农药通过不同途径毒害蚯蚓,复合毒性效应表现为协同作用.可见,3者对土壤生态系统环境安全性和土壤健康质量存在潜在危害,同时这几种污染物的共存进一步加大了潜在危害性,且复合毒性效应与各组浓度组合及污染暴露时间密切相关.     

乙草胺、铜污染共存对土壤甲胺磷蚯蚓降解过程的影响研究

为揭示利用蚯蚓活动强化甲胺磷降解的可行性,采用了微宇宙培养方法,通过有蚯蚓和无蚯蚓的对比实验,考察了乙草胺和铜分别与甲胺磷共存条件下污染黑土中甲胺磷降解过程的动态变化.结果表明,无论是否加入蚯蚓,甲胺磷单一污染的土壤中甲胺磷的降解符合一级反应动力学规律;蚯蚓活动能促进甲胺磷降解过程的进行.土壤中乙草胺或铜与甲胺磷共存时,均明显地干扰了甲胺磷降解过程随时间的动态变化;根据化学结构分析,推测两者对甲胺磷降解规律的影响机制可能类似.铜与甲胺磷复合污染的土壤中甲胺磷含量明显高于乙草胺与甲胺磷复合组,说明铜较乙草胺对土壤甲胺磷降解过程的延缓作用更强.     

乙草胺与硫酸铜对黑土微生物的复合生态影响

为了探究2种不同类型农药联合施用对土壤微生物的复合生态影响,以除草剂乙草胺和杀菌剂硫酸铜为例,采用传统毒理学方法和BIOLOG法对其进行评价.结果表明,乙草胺和硫酸铜的联合施用对细菌、放线菌和真菌活菌量以及土壤脱氢酶活性基本呈现急性抑制效应,但随时间延长其作用逐渐减弱甚至发生逆转,而对土壤底物诱导呼吸强度表现为明显促进.利用Shannon、Simpson和McIntosh多样性指数模型和主成分分析法对BIOLOG数据进行分析,表明2种农药的联合施用显著破坏了黑土微生物群落物种多样性的丰富度和均一性,主成分分析法也表明土壤微生物群落碳源利用多样性发生了改变.     

微塑料和镉复合污染对狼尾草根际土壤微生物群落结构和功能的影响

微塑料和重金属复合污染对植物生长和根际微生物群落和功能的影响目前尚未清晰.以狼尾草为实验材料进行盆栽模拟试验,研究重金属镉（Cd）与不同种类MPs （PE、PS）、粒径（13 μm和550 μm）、质量分数（0.1%和1%）的微塑料复合污染对狼尾草生长、重金属积累和根际微生物群落功能的影响.结果表明,MPs和Cd复合污染条件下整体呈现对植物生长胁迫增加、Cd含量和积累量降低的趋势.MPs和Cd复合污染能改变细菌群落组成,降低细菌多样性,其中550 μm 0.1% PE+Cd处理组ACE指数和Chao1指数降低最显著.与单一Cd污染相比,不同MPs种类、质量分数和粒径的MPs添加能改变新陈代谢、氨基酸的转运和代谢、能量生成和转换等功能组的基因丰度,显著影响狼尾草根际土壤细菌的功能.研究采用宏基因组学的方法分析了MPs和Cd复合污染对狼尾草根际细菌群落和功能的影响,可为MPs重金属复合污染的生态毒理效应及其生物修复提供基础数据和科学依据.     

能源植物修复镉污染土壤对根际细菌网络结构的影响

选取油脂类能源植物大豆和碳水化合物类能源植物玉米,采用高通量测序方法研究大豆、玉米修复Cd污染土壤过程中根际土壤细菌群落组成,基于高通量测序数据采用分子生态网络分析细菌相互作用.结果表明,50 mg·kg-1Cd污染土壤中两种植物根部Cd浓度和积累量最高,转移系数TF分别为玉米0.78和大豆0.35.基于细菌16S r RNA基因的群落分析表明,大豆、玉米根际土壤细菌主要包括Proteobacteria(变形菌门)、Acidobacteria(酸杆菌门)31个门细菌组成,大豆、玉米种植均能影响土壤细菌群落组成,能影响Candidate division TM7 norank、Acidimicrobiales norank、Sphingomonas等丰度.分子生态网络分析表明种植大豆和玉米增加了细菌之间的相互作用,导致其网络结构更为复杂,关键细菌从不种植物处理的1个增加到种植大豆的6个和种植玉米的10个.     

重金属在土壤-空心菜系统中的迁移分配

通过盆栽试验,研究了Cu,Zn,Pb和Cd单一与复合污染条件下在土壤-空心菜系统中的迁移分配情况.结果表明:单一和复合污染情况下,空心菜地上部重金属的富集系数大体表现为Cd>Zn>Cu>Pb,重金属转运系数则表现为Cd>Zn>Pb>Cu.与单一污染相比较,复合污染条件下,Cu,Zn,Pb和Cd富集系数下降,Cu的转运系数升高,而Zn,Pb和Cd的转运系数降低.单一污染条件下,随着土壤中重金属质量分数的增大,Cu的富集系数和转运系数都相应减小;Zn和Pb的富集系数增大而转运系数减少;Cd的富集系数增大,但转运系数变化很小.整体上看,复合污染条件下,土壤w(Cu)的增加使Zn,Pb和Cd在空心菜中的富集系数明显减小;土壤w(Zn)的增加使空心菜中Cd的富集系数明显减小;土壤w(Pb)的增加使空心菜中Cu的富集系数明显增大;土壤w(Cd)的增加使空心菜中Pb的富集系数明显增大.      

不同粒度CuO及与乙基黄原酸钾复合污染对土壤脲酶和微生物多样性的影响

纳米氧化铜不同于常规的氧化铜,纳米氧化铜的尺寸小,比表面积大,对生态环境存在潜在危害.本次通过向土壤中加入不同比例的纳米氧化铜(N CuO)和微米氧化铜(M Cu0)(0,50,200,800,1600mg/kg干土)观察它们对土壤脲酶活性和細菌群落的影响.同吋,进一步研究了N CuO和M CuO分別与乙基黄原酸钾(PEX)复合污染对土壤微生物的影响.N CuO较M CuO对脲酶活性有更强的抑制作用,只有M CuO浓度为1600mg/kg对脲酶的影响较明显.N CuO对脲酶活性的抑制可能与破坏细菌的細胞膜或刺激细菌产生活性氧,造成細菌細胞的衰退有关,N CuO对细菌群落结构的影响也较大.N CuO与PEX复合污染加剧了N CuO对土壤微生物的毒性,使脲酶的活性大幅度降低,细菌的群落结构也发生了较大的变化.N CuO及其复合污染对土壤微生物的作用机制需进一歩研究.通过比较研究N CuO和M CuO的生态毒性,纳米颗粒的生态毒性更需要引起足够的重视.     

乙草胺降解菌A-3的筛选及其降解特性

采用富集培养的方法从农药厂污泥和长期受乙草胺污染的土壤中分离到1株能以乙草胺为氮源生长的细菌命名为A-3,经16S rRNA鉴定,菌株A-3属于粘着剑菌属(Ensifer adhaerens).研究结果表明,菌株A-3能以乙草胺作为唯一氮源生长并高效降解乙草胺.在含10 mg/L乙草胺的无机盐培养基中培养10 d后,菌...     

酸性矿山废水对稻田土壤微生物菌群结构的影响

为探究酸性矿山废水(AMD)对稻田土壤微生物群落结构的影响,通过取自矿区受AMD污染和未受污染的稻田土壤进行微宇宙灌溉模拟实验,研究了AMD污染过程中土壤理化性质和微生物群落的变化,同时建立环境条件变化引起土壤微生物群落结构改变的相关性关系.结果表明,受AMD污染的土壤中SO_4~(2-)、Cd、Zn含量显著上升,土壤酸化且土壤中细菌群落的多样性下降;而恢复清洁水灌溉可提高土壤细菌群落的多样性,有利于修复AMD的污染.采用高通量测序技术分析了不同处理稻田土壤中微生物群落在门和属分类水平上的相对丰度分布变化,冗余分析(RDA)表明,土壤pH和重金属(Pb、Cu)含量是影响稻田土壤微生物群落结构的主要环境因子.研究结果不仅有助于进一步揭示AMD污染、土壤因子与土壤微生物群落的相互关系,同时可为恢复AMD污染农业土壤提供理论依据.     

Changes in functional structure of soil bacterial communities due to fungicide and insecticide applications in canola

The fungicide vinclozolin and insecticide λ-cyhalothrin are widely used to control canola (Brassica spp.) diseases and insect pests, respectively, in Canada. We investigated non-target effects of these pesticides, applied at recommended rates, on soil microbial biomass, functional bacterial diversity and functional community structure of soil bacteria (by evaluating patterns of C substrate utilization) in canola rhizosphere and bulk soil at three locations in Alberta from 2002 to 2004. Experimental treatments were (a) untreated control, (b) vinclozolin fungicide foliar application, (c) λ-cyhalothrin insecticide foliar application, and (d) vinclozolin and λ-cyhalothrin applications. No significant pesticide effects on soil microbial biomass or functional bacterial diversity were observed, but the functional structures of soil bacteria were altered. In 1 of 12 cases, the control treatment had a different soil bacterial community structure from the 3 pesticide treatments. The fungicide treatment had different bacterial community structures from the control or insecticide treatments in 3 of 12 cases, the insecticide treatment had different community structures from the control or fungicide treatments in 4 of 12 cases, and the combined fungicide and insecticide treatment had different community structures from the other treatments in 3 of 12 cases. Therefore, evaluating soil bacterial functional structures revealed pesticide effects that were not detected when bacterial diversity or microbial biomass were measured in canola rhizosphere or bulk soil.     

乙草胺、甲胺磷及组合对农田黑土细菌种群生长及多样性的毒性效应

应用传统及分子微生物生态学16S rDNA-PCR DGGE(Denaturing Gradient Gel Electrophoresis)等研究方法,分别对不同浓度乙草胺、甲胺磷及其乙草胺-甲胺磷二元组合胁迫下黑土中细菌活细胞数量(CFU Colony Forming Units)、种群丰富度(Richness)及其结构变化规律进行了研究.结果表明:单因子乙草胺及甲胺磷对细菌CFU产生急性毒性效应,但是长期毒性效应不显著;高浓度乙草胺-甲胺磷的二元组合对细菌CFU产生复合毒性效应,其对细菌CFU的复合抑制率明显高于相应组合中各单因子.乙草胺对可培养自生固氮菌表现出强烈刺激的作用,甲胺磷表现为严重抑制作用;而所有乙草胺-甲胺磷二元组合却对自生固氮菌CFU表现为复合毒性效应,其复合毒性明显高于相应组合中各单因子对自生固氮菌的毒性效应.应用16S rDNA-PCR DGGE技术研究结果显示:乙草胺、甲胺磷及乙草胺-甲胺磷组合不同程度地降低了黑土中细菌种群在分子水平上的丰富度,并使其种群组成和结构均发生了显著变化.     

Adsorption and desorption of Cu~(2+) on paddy soil aggregates pretreated with different levels of phosphate

Interactions between anions and cations are important for understanding the behaviors of chemical pollutants and their potential risks in the environment.Here we prepared soil aggregates of a yellow paddy soil from the Taihu Lake region,and investigated the effects of phosphate(P) pretreatment on adsorption-desorption of Cu~(2+) of soil aggregates,free iron oxyhydrates-removed soil aggregates,goethite,and kaolinite with batch adsorption method.The results showed that Cu~(2+) adsorption was reduced on the aggregates pretreated with low concentrations of P,and promoted with high concentrations of P,showing a V-shaped change.Compared with the untreated aggregates,the adsorption capacity of Cu~(2+) was reduced when P application rates were lower than 260,220,130 and110 mg/kg for coarse,clay,silt and fine sand fractions,respectively.On the contrary,the adsorption capacity of Cu~(2+) was higher on P-pretreated soil aggregates than on the control ones when P application rates were greater than those values.However,the desorption of Cu~(2+) was enhanced at low levels of P,but suppressed at high levels of P,displaying an inverted V-shaped change over P adsorption.The Cu~(2+) adsorption by the aggregate particles with and without P pretreatments was well described by the Freundlich equation.Similar results were obtained on P-pretreated goethite.However,such P effects on Cu~(2+)adsorption-desorption were not observed on kaolinite and free iron oxyhydrates-removed soil aggregates.The present results indicate that goethite is one of the main soil substances responsible for the P-induced promotion and inhibition of Cu~(2+) adsorption.     

Single and joint stress of acetochlor and Pb on three agricultural crops in northeast China

In order to evaluate ecological risk of agrochemicals in agricultural environment, single and joint toxic effects of an important herbicide and a typical heavy metal on root elongation of crops were investigated. Seeds of the three crops including wheat （Triticum aestivum）, Chinese cabbage （Brassica pekimensis） and soybean （Glycine max） as the main crops in northeast China were exposed to acetochior as a herbicide and lead （Pb） as a heavy metal using the pot-culture method, and meadow brown soil as one of the main soils distributed in northeast China was applied in the investigation. The results indicated that the interactive effects of the two pollutants on root elongation of the three crops were very complicated although they had markedly significant （P〈0.01） linear interrelationships based on the regression analyses. When the concentration of added Pb^2＋ reached 200 mg/kg, acetochlor and Pb had an antagonistic effect on the inhibition of root elongation of the three crops. However, acetochlor and Pb had significantly （P〈0.05） synergic effects on the inhibition of root elongation when concentration of added Pb^2＋ was up to 1000 mg/kg. At the low concentration of added Pb, joint toxicity of acetochlor and Pb was more dependent on the concentration of Pb. Among the three crops, wheat was the most sensitive to the toxicity of Pb and Chinese cabbage was the most sensitive to the toxicity of acetochlor.     

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.     

添加猪粪对不同施肥历史土壤细菌群落的影响

为探讨粪肥携带的微生物对土壤微生物的影响，以长期施用粪肥和化肥的土壤为研究对象，耦合传统微生物学培养方法和现代分子微生物学方法，分析了猪粪和灭菌猪粪对2种土壤的细菌数量、群落结构和热代谢活性的影响.结果表明，以9.0t/hm²的用量施入时，猪粪和灭菌猪粪对两种土壤中的可培养细菌数量以及细菌16S-rRNA拷贝数均无明显影响；土壤中的土著微生物对猪粪携带的外源细菌在土壤中定殖具有明显的抑制作用，将2种土壤灭菌后添加猪粪，可培养细菌平均数量较未灭菌土壤分别增加45.96和96.21倍.粪肥输入的化学物质未导致土壤细菌群落结构和相对丰度发生明显变化；而粪肥输入的外源细菌Marinilabiaceae、Porphyromonadaceae、Bacteroidale和Pseudomonadaceae则可以在长期施用化肥的土壤中定殖并对相应的细菌丰度造成影响，但是在长期施用粪肥的土壤中，这些细菌在最开始就被抑制.添加粪肥提高了长期施用化肥土壤的微生物热代谢活性，但对长期施用粪肥的土壤无明显影响.长期施用化肥的土壤受粪源细菌的影响较大，而长期施用粪肥的土壤对粪肥的敏感度降低，对粪肥携带的外源细菌的抑制作用增强.     

Impact of acetochlor on ammonia-oxidizing bacteria in microcosm soils

Acetochlor is an increasingly used herbicide on corn in North China. Currently, the effect of acetochlor on soil ammonia-oxidizing bacteria (AOB) communities is not well documented. Here, we studied the diversity and community composition of AOB in soil amended with three concentrations of acetochlor (50, 150, 250 mg/kg) and the control (0 mg acetochlor/kg soil) in a microcosm experiment by PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) and the phylogenetic analysis of excised ...     

Degradation of chlorpyrifos alone and in combination with chlorothalonil and their effects on soil microbial populations

In practice, pesticides are usually applied simultaneously or one after another for crop protection, and this type of pesticide application often leads to a combined contamination of pesticide residues in the soil environment. A laboratory study was conducted to investigate the influence of chlorothalonil on chlorpyrifos degradation and its effects on soil bacterial, fungal, and actinomycete populations. Under the experimental conditions here, the half-lives of chlorpyrifos alone, and in combination with chlorothalonil, at the recommended and double dosages, were measured to be 3.24, 2.77, and 2.63 d, respectively. Chlorpyrifos degradation was not significantly altered by its combination with chlorothalonil. However, the inhibitory effect of chlorpyrifos on soil microorganisms was increased by its combination with chlorothalonil, and the increase was related to the levels of chlorothalonil added. Compared to those in the controls, the populations of bacteria, fungi, and actinomycetes were significantly reduced by 44.1%, 61.1%, and 72.8%, respectively, on the first day after treatment (DAT) by chlorpyrifos alone. With the addition of chlorothalonil, the inhibition was increased to 55.2%, 79.3%, and 85.8% at the recommended dosage, and 86.0%, 94.1%, and 90.8% at the double dosage, at one DAT, respectively. The results suggested that combined effects should be taken into account to assess the actual impacts of pesticide applications.