腐熟羊粪有机肥配施无机肥对植烟土壤微生物群落结构和多样性的影响

为了探究腐熟羊粪有机肥与无机肥配施对洛阳烟区植烟土壤微生物群落结构和土壤肥力特性的影响，采用盆栽控制试验和高通量测序技术，研究了不施肥（CK）、100%无机氮肥（T0）、50%羊粪有机肥氮+50%无机氮肥（T50）和100%羊粪有机肥氮（T100）4个处理下植烟土壤细菌和真菌群落结构及多样性的差异，并结合土壤理化性质分析了土壤肥力指标与土壤微生物多样性的关系。结果表明：植烟土壤细菌优势菌门为变形菌门（Proteobacteria）、酸杆菌门（Acidobacteria）和放线菌门（Actinobacteria），土壤真菌优势菌门为子囊菌门（Ascomycota）和担子菌门（Basidiomycota）；T50处理的变形菌门、酸杆菌门和子囊菌门相对丰度最高，但放线菌门和担子菌门相对丰度最低；NMDS分析和相似性分析发现，T50处理的土壤微生物群落结构与其他处理差异显著；Alpha多样性分析也表明，T50处理土壤细菌和真菌群落的丰富度和多样性最高。不同处理土壤理化性质和土壤关键酶活性差异显著，以T50处理土壤养分含量和土壤碳氮代谢酶的活性最高；Pearson相关性分析显示，土壤碱解氮、有效磷、速效钾含量及蛋白酶和脲酶活性与植烟土壤微生物多样的关系最为密切。总之，采用腐熟羊粪有机肥和无机肥配施通过影响植烟土壤细菌和真菌群落结构和多样性，促进了土壤养分的释放和转化，有利于土壤养分的供应和土壤质量的提升，研究结果为洛阳烟区应用羊粪有机肥改良土壤提供了理论依据。     

榆林沙区典型林地不同植被类型对土壤微生物群落结构的影响

  目的  掌握榆林沙区典型林地土壤微生物特征,明确地上植被对土壤微生物群落结构的影响。  方法  采集榆林沙区四种林分类型土壤,分析其土壤微生物群落结构。  结果  测序共产生有效操作分类单元（OTU）15,509个,各林分间OTU及各类多样性指数没有显著性差异。优势菌种及其丰度土层间变化较大,但林分间优势细菌种一致,丰度排名前五的分别是变形菌门（Proteobacteria）、放线菌门（Actinobacteria）、酸杆菌门（Acidobacteria）、厚壁菌门（Firmicutes）和绿弯菌门（Chloroflexi）;樟子松、油松林下土壤中,酸杆菌门所占比例最大、分别达到22.32%和29.02%,而在沙柳和小叶锦鸡儿林下土壤中,变形菌门又成为优势菌,比例占到27.64%和28.51%。变形菌门和放线菌门在灌木林土壤中所占比例要高于乔木。  结论  虽然各优势种丰度在林分间略有差别,但差异不显著（P < 0.05）。说明在一定区域内微生物群落结构复杂程度是受土壤本底的影响,不同季节或者土壤温度、湿度的变化对微生物群落结构的影响在一段时间后消除,群落结构归于稳定。     

Response of microbial activity and microbial community composition in soils to long-term arsenic and cadmium exposure

Arsenic (As) and cadmium (Cd) in soils can affect soil microbial function and community composition and, therefore, may have effects on soil ecosystem functioning. The aim of our study was to assess the effects of long-term As and Cd contamination on soil microbial community composition and soil enzyme activities. We analyzed soils that have been contaminated 25 years ago and at present still show enhanced levels of either As, 18 and 39 mg kg⁻¹, or Cd, 34 and 134 mg kg⁻¹. Soil without heavy metal addition served as control. Polymerase chain reaction (PCR) followed by denaturing gradient gel electrophoresis (DGGE) showed that bacterial community composition in As and Cd contaminated soils differed from that in the control soil. The same was true for the microbial community composition assessed by analysis of respiratory quinones. Soil fungi and Proteobacteria appeared to be tolerant towards As and Cd, while other groups of bacteria were reduced. The decline in alkaline phosphatase, arylsulphatase, protease and urease activities in the As- and Cd-contaminated soils was correlated with a decrease of respiratory quinones occuring in Actinobacteria and Firmicutes. Xylanase activity was unaffected or elevated in the contaminated soils which was correlated with a higher abundance of fungal quinones, and quinones found in Proteobacteria.     

不同类型粪肥还田对土壤酶活性及微生物群落的影响

畜禽粪便作为有机肥还田可以维持和提高土壤有机质、改良土壤,有利于农业可持续发展。不同类型粪肥还田后对土壤生物学性状的影响不同,为探究这一问题,在内蒙古乌兰察布市设置田间试验,包括化肥(F)、羊粪(GM)、猪粪(PM)、牛粪(CM)4个处理,研究其对土壤养分、酶活性及微生物群落的影响。结果表明,施用粪肥较化肥具有增加土壤有机质、全氮、有效磷、铵态氮等养分含量的趋势。不同粪肥较化肥处理的土壤脲酶、蔗糖酶、碱性磷酸酶和过氧化氢酶活性最高增幅分别为32.4%、150.4%、26.8%和30.1%。牛粪处理的土壤微生物生物量碳氮显著提高,分别较化肥增加33.2%和33.4%。不同处理在细菌门水平上的优势种群较一致,放线菌门(Actinobacteria)、变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、绿弯菌门(Chloroflexi)、厚壁菌门(Firmicutes)是优势种群。本试验条件下,牛粪处理更能提高土壤微生物生物量碳氮,短期内施用不同粪肥对于提高土壤微生物群落多样性差异不显著,土壤pH、有效磷、铵态氮是影响土壤微生物群落结构的主要环境因子。     

Bacterial community structure in fumigated soil

Soil microbial biomass has been determined since the mid 1970's by the chloroform fumigation incubation technique as proposed by Jenkinson and Powlson (1976). The microbial biomass C can be determined by subtracting the CO₂ emitted from an unfumigated soil (mineralization of soil organic matter) from that emitted from a chloroform fumigated inoculated soil (mineralization of soil organic matter and killed soil microorganisms) and dividing the difference by a proportionality factor (k_C = 0.45). The question remained which microorganisms recolonized a fumigated soil. An arable soil was fumigated for one day with ethanol-free chloroform or left unfumigated and incubated aerobically after removal of the chloroform for 10 days. The bacterial population structures were determined in the fumigated and unfumigated soil after 0, 1, 5 and 10 days by means of 454 pyrosequencing of the 16S rRNA gene. Fumigating the arable soil reduced significantly the relative abundance of phylotypes belonging to different groups, but increased the relative abundance of only four genera belonging to two phyla (Actinobacteria and Firmicutes) and two orders (Actinomycetales and Bacillales). The relative abundance of phylotypes belonging to the Micromonospora (Micromonosporaceae) increased significantly from 0.2% in the unfumigated soil to 6.7% in the fumigated soil and that of Bacillus (Bacillaceae) from 3.6% to 40.8%, Cohnella (Paenibacillaceae) from undetectable amounts to 0.6% and Paenibacillus (Paenibacillaceae) from 0.3% to 4.2%. The relative percentage of phylotypes belonging to the Acidobacteria, Bacteroidetes, Chloroflexi, Gemmatimonadetes and Proteobacteria (α- β-, δ- and γ-Proteobacteria) were significantly lower in the fumigated than in the unfumigated soil and in most of them the relative abundance of different bacterial orders (i.e. Gp3, Gp4, Gp6, Sphingobacteriales, Gemmatimonadales, Rhodospirillales, Burkholderiales, Xanthomonadales) was reduced strongly (P < 0.001). It was found that the relative abundance of a wide range of bacteria was reduced shortly after fumigating an arable soil, but only a limited group of bacteria increased in a fumigated arable soil indicating a capacity to metabolize the killed soil microorganisms or recolonize a fumigated soil.     

基于高通量测序研究草莓根际微生物群落结构和多样性

研究草莓根际土壤微生物群落组成和结构,对健康草莓土壤生态系统的构建和保持具有重要意义。以不同地区草莓根际土壤为研究样本,利用MiSeq平台Illumina第二代高通量测序技术并结合相关生物信息学分析土壤细菌16S rRNA基因V4+V5区域和真菌ITS1+ITS2区域的丰富度和多样性指数以及群落结构。结果表明:从15个草莓根际土壤样本中获得4554个细菌分类操作单元OTU和1298个真菌OTU,草莓根际土壤的优势细菌门为变形菌门、厚壁菌门、放线菌门、酸杆菌门和绿弯菌门,主要的优势细菌属有16种;优势真菌门为子囊菌门、接合菌门和担子菌门,主要的优势真菌属有8种。冗余分析(RDA)显示,全氮和pH对土壤微生物群落结构的影响最大,共解释了61%的群落变化,各因子的贡献率大小依次为土壤全氮pH有效磷全钾全磷有机质速效钾碱解氮;相关性分析也表明,土壤理化指标均与不同优势菌门存在密切的相关关系。本研究结果加深了对草莓根际微生物群落结构和多样性的认识,为深入研究草莓根际微生物多样性及功能与环境因子之间的关系提供了借鉴。     

Combined effect of biochar and soil moisture on soil chemical properties and microbial community composition in microplastic-contaminated agricultural soil

Accumulation of microplastics (MPs) in agricultural environments has caused growing concern in recent years because of its detrimental impacts on soil quality, crop productivity and ecosystem function. This study was conducted to assess the impact of biochar on soil chemical and microbial properties in a MP-contaminated soil under two moisture regimes. Soil was contaminated with 1% (w/w) of low-density polyethylene MPs. Four types of standard biochar, that is, oil seed rape (OSR) biochar produced at 550°C (OSR 550) and 700°C (OSR 700) and soft wood pellet (SWP) biochar produced at 550°C (SWP 550) and 700°C (SWP 700), were applied at a rate of 5% (w/w). The control was maintained without MP addition. The samples were incubated in soil with two moisture regimes, that is, at 30% and 70% of the water holding capacity, and the soil chemical and microbiological properties were assessed after 100 days of incubation. OSR biochar application significantly increased soil pH (8.53–8.81) and electrical conductivity (0.51–0.58 dS/m) in both moisture regimes. The effect of biochar application on soil enzyme activity and microbial community composition did not show a clear trend. However, SWP 700 biochar improved soil enzyme activity compared with that of the control and improved bacterial diversity and evenness compared with those of other biochars, which was attributed to the high surface area available for microbial colonization. Low soil moisture content significantly reduced enzyme activity and bacterial richness even with biochar amendment, except for SWP 550 biochar. This study implies the suitability of biochar for improvement of soil quality in MP contaminated soil under both moisture regimes. However, further long-term studies are needed to get a clear understanding on the impact of different types of biochar on MP-contaminated soil.     

基于高通量测序的两种植被恢复类型根际土壤细菌多样性研究

为研究库布齐沙地生态恢复过程中不同植被恢复类型土壤微生物细菌群落结构、多样性的变化特征,以流动沙地为对照,运用高通量测序技术,对自然恢复的油蒿群落、人工种植的中间锦鸡儿群落根际和非根际土壤细菌多样性进行了研究,并分析了土壤理化性质对其分布的影响。结果表明:(1)与流沙对照相比,两种植被恢复类型对细菌多样性产生了正效应,细菌群落丰度、多样性和均匀度明显增加。其中,自然恢复的油蒿群落土壤细菌丰度高于人工种植的中间锦鸡儿群落;(2)变形菌门、酸杆菌门和放线菌门为研究区土壤中的优势细菌类群,其中变形菌门在各样地丰度比例最高,变形菌的4个亚群变化趋势一致,α-变形菌相对含量在油蒿和中间锦鸡儿群落根际土壤中明显增加,尤其是自然恢复的油蒿群落根际土壤中α-变形菌得到了很好的恢复;(3)土壤有机质、全氮、速效氮、速效钾含量和土壤含水量是影响土壤细菌群落丰度和多样性的主要土壤因子,典型相关分析表明土壤有机质、全氮、全钾、速效钾、速效氮含量对于研究区土壤细菌群落遗传多样性的变化起着重要作用。     

连作对再植枸杞根际细菌群落多样性和群落结构的影响研究

受枸杞道地产区土地资源等因素限制,连作障碍已成为影响枸杞产业发展的重要原因之一,导致严重的经济损失.研究连作条件下枸杞农田土壤生态系统微生物群落的演替规律对枸杞产业的可持续发展具有重要的理论意义.以宁夏银川市南梁农场连作多年的枸杞地为研究对象,利用Illumina MiSeq测序技术分析了连作对再植枸杞根际/非根际细菌群落的影响.结果表明,连作地显著抑制再植枸杞苗地径的增加,且其土壤pH较对照样地显著降低(p<0.05).测序结果证实,与对照样地相比,连作地再植枸杞根际土壤细菌物种数显著降低(p<0.05),细菌群落α多样性下降(p>0.05).主坐标分析表明,连作和对照样地间枸杞非根际细菌群落结构无明显差异,但连作显著改变再植枸杞根际细菌的群落结构.对细菌群落丰度的统计分析发现,连作地枸杞根际浮霉菌门、非根际假单胞菌门的相对丰度较对照样地显著降低(p<0.05).此外,冗余分析结果表明:枸杞园土壤pH和有效磷含量是影响枸杞非根际土壤细菌群落结构变化的主要因素,分别解释了41.8%和35.4%的群落结构变化(p<0.05),其他土壤因子无统计学意义,但土壤理化因子对再植枸杞根际细菌群落结构变化的影响均未达显著水平.这些结果证实连作能够显著抑制再植枸杞生长、影响再植枸杞根际细菌群落结构和多样性,干扰枸杞与土壤细菌群落间的互作关系.这些研究结果将为解析枸杞连作障碍机制提供理论基础.     

寡糖对土壤微生物多样性及群落结构的调节作用

土壤微生物种群类型,对土壤质量和作物生长具有重要影响,研究寡糖对土壤微生物种群的影响特征,有助于正确、高效及安全使用寡糖。本研究利用人工气候室进行土壤培养,土壤施加50mg·L−1的壳寡糖（CSOS）和纤维寡糖（COS）溶液,以清水（CK）为对照处理,培养6d后取样,利用高通量测序技术,分析土壤微生物群落结构组成及多样性分布特征。结果表明：壳寡糖（CSOS）和纤维寡糖（COS）处理均显著改变细菌、真菌的群落结构,提高细菌的物种观测数。变形菌门（Proteobacteria）、酸杆菌门（Acidobacteria）、放线菌门（Actinobacteria）、绿弯菌门（Chloroflexi）、芽单胞菌门（Gemmatimonadetes）和拟杆菌门（Bacteroidetes）为优势细菌门,子囊菌门（Ascomycota）、担子菌门（Basidiomycota）和被孢霉门（Mortierellomycota）为优势真菌门。通过组间群落组成比较分析可知,壳寡糖（CSOS）和纤维寡糖（COS）处理均不同程度降低酸杆菌门（Acidobacteria）的相对丰度,增加变形菌门（Proteobacteria）、放线菌门（Actinobacteria）、芽单胞菌门（Gemmatimonadetes）、壶菌门（Chytridiomycota）以及有益菌属溶杆菌属（Lysobacter）、硝化螺旋菌属（Nitrospira）、Haliangium、芽球菌属（Blastococcus）和链霉菌属（Streptomyces）的相对丰度,但与纤维寡糖（COS）相比,壳寡糖（CSOS）处理微生物群落组成的变化幅度更大。此外,壳寡糖（CSOS）和纤维寡糖（COS）处理在调节土壤微生物群落结构上存在一定差异。其中,壳寡糖（CSOS）处理有益菌属Talaromyces的相对丰度增加195%,纤维寡糖（COS）处理有益菌属假单胞菌属（Pseudomonas）的相对丰度增加215%。综上,壳寡糖和纤维寡糖处理均能优化土壤微生物群落的结构组成,其调控差异性有助于理解不同寡糖的调控机制,推动寡糖的应用与推广。     

Influence of grass species and soil type on rhizosphere microbial community structure in grassland soils

A number of studies have reported species specific selection of microbial communities in the rhizosphere by plants. It is hypothesised that plants influence microbial community structure in the rhizosphere through rhizodeposition. We examined to what extent the structure of bacterial and fungal communities in the rhizosphere of grasses is determined by the plant species and different soil types. Three grass species were planted in soil from one site, to identify plant-specific influences on rhizosphere microbial communities. To quantify the soil-specific effects on rhizosphere microbial community structure, we planted one grass species (Lolium perenne L.) into soils from three contrasting sites. Rhizosphere, non-rhizosphere (bulk) and control (non-planted) soil samples were collected at regular intervals, to examine the temporal changes in soil microbial communities. Rhizosphere soil samples were collected from both root bases and root tips, to investigate root associated spatial influences. Both fungal and bacterial communities were analysed by terminal restriction fragment length polymorphism (TRFLP). Both bacterial and fungal communities were influenced by the plant growth but there was no evidence for plant species selection of the soil microbial communities in the rhizosphere of the different grass species. For both fungal and bacterial communities, the major determinant of community structure in rhizospheres was soil type. This observation was confirmed by cloning and sequencing analysis of bacterial communities. In control soils, bacterial composition was dominated by Firmicutes and Actinobacteria but in the rhizosphere samples, the majority of bacteria belonged to Proteobacteria and Acidobacteria. Bacterial community compositions of rhizosphere soils from different plants were similar, indicating only a weak influence of plant species on rhizosphere microbial community structure.     

西北半干旱荒漠草原与耕地土壤可培养微生物多样性及分布特征比较

采用稀释涂布平板、分离培养和16S r DNA序列分析法对我国甘肃白银地区半干旱荒漠草原土壤可培养细菌、放线菌、真菌数量及群落分布特征进行了分析,比较了荒漠草原和耕地土壤微生物多样性。发现荒漠草原土壤可培养细菌、放线菌、真菌数量分别为1.23×106、0.19×106、0.18×106cfu·g-1,耕地三类微生物数量分别是3.03×106、0.53×106、0.05×106cfu·g-1。荒漠草原可培养细菌、放线菌数量明显低于耕地,而真菌数量高于耕地。从荒漠草原分离出14株细菌,分别属于γ-变形菌纲(γ-Proteobacteria)噬冷杆菌属(Psychrobacter),放线菌门(Actinobacteria)皮球菌属(Kytococcus),厚壁菌门(Firmicutes)芽孢杆菌属(Bacillus)、亮氨酸芽孢杆菌属(Lysinibacillus)、土壤芽孢杆菌属(Solibacillus)、气球菌属(Aerococcus),优势菌为芽孢杆菌属和噬冷杆菌属。耕地分离出可培养细菌19株,分别属于ɑ-变形菌纲(ɑ-Proteobacteri)根瘤菌属(Rhizobium)、中华根瘤菌属(Sinorhizobium),γ-变形菌纲(γ-Proteobacteria)假单胞菌属(Pseudomonas),厚壁菌门(Firmicutes)芽孢杆菌属(Bacillus),放线菌门(Actinobacteria)微杆菌属(Microbacterium)、节杆菌属(Arthrobacter)、微球菌属(Micrococcus)、考克氏菌属(Kocuria),以放线菌门细菌为主(占57.9%)。从荒漠草原分离放线菌共8株,分别属于链霉菌属(Atreptomyces)、小单孢菌属(Micromonspora)、间孢囊菌属(Intrasporangium),而耕地主要为链霉菌属(Atreptomyces)、小单孢菌属(Micromonspora)。荒漠草原真菌主要是交链孢霉属(Alternaria)、芽枝霉属(Cladosporium),耕地土壤真菌包括青霉属(Penicillium)、交链孢霉属(Alternaria)、曲霉属(Aspergillus)、毛霉属(Mucor)、链孢霉属(Coniothecium)。试验结果表明,荒漠草原与耕地土壤微生物都具有较丰富的多样性,但微生物群落结构存在一定差异,同一区域不同深度土壤中微生物数量和种类也存在差异,耕地土壤微生物多样性明显高于荒漠草原。     

Artificial neural network modeling of microbial community structures in the Atlantic Forest of Brazil

Microbial communities vary across the landscape in forest soils, but prediction of their biomass and composition is a difficult challenge due to the large numbers of variables that influence their community structures. Here we examine the use of artificial neural network (ANN) models for extraction of patterns among soil chemical variables and microbial community structures in forest soils from three regions of the Atlantic Forest of Brazil. At each location, variations in soil chemical properties and FAME profiles of microbial community structures were mapped at 20 × 20 m intervals within 10 ha parcels. Geostatistical analyses showed that spatial variability in soil physical and chemical variables could be mapped at scale distances of 20 m, but that FAME profiles representing the microbial communities were highly variable and had no spatial dependence at the same scale in most cases. RDA analysis showed that FAME signatures representing different microbial groups were positively associated with soil pH, OM, P and base cations concentrations, whereas microbial biomass was negatively associated with the same environmental factors. In contrast, ANN models revealed clear relationships between microbial community structures at each parcel location, and generated verifiable predictions of variations in FAME profiles in relation to soil pH, texture, and the relative abundances of base cations. The results suggest that ANN modeling provides a useful approach for describing the relationships between microbial community structures and soil properties in tropical forest soils that were not able to be captured using geostatistical and RDA analyses.     

三种城市景观竹土壤微生物群落结构特征

为探究城市景观竹林对土壤微生物群落的影响,选取上海辰山植物园种植的3种景观竹,利用Illumina MiSeq高通量测序结合土壤理化指标分析研究了不同景观竹林对土壤理化性质和微生物群落结构特征的影响。结果表明：不同竹林土壤全磷、有效磷、有机质、全氮、全钾含量和电导率存在显著差异(P<0.05);土壤细菌丰富度和多样性指数在毛竹林土壤中最高,在淡竹林土壤中最低;土壤细菌优势门为放线菌门(Actinobacteriota)、变形菌门(Proteobacteria)、酸杆菌门(Acidobacteriota)和绿弯菌门(Chloroflexi),真菌优势门为子囊菌门(Ascomycota),其中放线菌门和绿弯菌门在淡竹林土壤中的相对丰度最高,且显著高于其他竹林,而变形菌门、粘球菌门(Myxococcota)的相对丰度在毛竹林土壤中最高,土壤真菌优势门在不同竹林间的差异不显著。此外,土壤细菌群落结构和丰度受土壤p H、硝态氮、电导率、全磷、铵态氮、含水率、速效钾和微生物生物量碳影响显著,而真菌群落变化受全磷、电导率和全钾影响显著。     

Potentialities and soil impact analysis of rock phosphorus fertilization of perennial and annual legume crops

ABSTRACT

The objective of this study is to evaluate the impacts of the direct use of natural rock phosphates on two forage legumes, Vicia sativa, and Medicago sativa. Crops were grown in pots containing two P deficient soils and fertilized with Tunisian rock phosphate (TRP) or chemical triple superphosphate (TSP). TRP amendment significantly increased shoot dry yield (+53% to +162%), P-content, protein content, and chlorophyll content mainly at the rate of 150 kg P ha^?1. No significant effects were observed on mycorrhization and antioxidant enzyme activities. However, TSP negatively affected the frequency of root mycorrhization and stimulated antioxidant enzyme activities. TRP amendment induced an increase in fluorescein diacetate activity, alkaline phosphatase activity, and species richness. The stimulation involved mainly Actinobacteria, Proteobacteria, and Firmicutes with higher stimulation of Actinobacteria in the case of vetch. The present work indicated a better P-use efficiency of vetch. It suggested also that at least a part of the enhancement observed in plant-growth parameters following TRP fertilization is driven by changes in the structure of rhizosphere microbial communities. Nevertheless, the correlation between changes in microbial composition and phosphorus uses efficiency needs to be investigated further.     

Effects of atrazine on microbial activity in semiarid soil

The effect of an atrazine formulation on microbial biomass, microbial respiration, ATP content and dehydrogenase and urease activity in a semiarid soil and the influence of time on the response of soil microbial activity to the herbicide treatment were assessed. The atrazine formulation was added to soil as aqueous solutions of different concentrations of active ingredient to obtain a range of concentrations in the soil from 0.2 to 1000 mg kg⁻¹. Microcosms of soil with the different herbicide concentrations and untreated control soil were incubated for 6 h, 16 and 45 days. In general, an increase in the measured microbiological and biochemical parameters with atrazine concentration in soil was observed. The increase in microbial activity with atrazine pollution was noticeable after lengthy incubation.     

Soil microbial communities response to herbicide 2,4-dichlorophenoxyacetic acid butyl ester

2,4-Dichlorophenoxyacetic acid butyl ester (2,4-D butyl ester) is extensively applied for weed control in cultivation fields in China, but its effect on soil microbial community remains obscure. This study investigated the microbial response to 2,4-D butyl ester application at different concentrations (CK, 10, 100 and 1000 μg g^?1) in the soils with two fertility levels, using soil dilution plate method and phospholipid fatty acid (PLFA) analysis. Culturable microorganisms were affected by the herbicide in both soils, particularly at the higher concentration. After treating soil with 100 μg g^?1 herbicide, culturable bacteria and actinomycetes were significantly higher, compared to other treatments. Treatment of soil with 1000 μg g^?1 2,4-D butyl ester caused a decline in culturable microbial counts, with the exception of fungal numbers, which increased over the incubation time. PLFA profiles showed that fatty acids for Gram-negative (GN) bacteria, Gram-positive (GP) bacteria, total bacteria and total fungi, as well as total PLFAs, varied with herbicide concentration for both soil samples. As herbicide concentration increased, the GN/GP ratio decreased dramatically in the two soils. The higher stress level was in the treatments with high concentrations of herbicide (1000 μg g^?1) for both soils. Principal component analysis of PLFAs showed that the addition of 2,4-D butyl ester significantly shifted the microbial community structure in the two soils. These results showed that the herbicide 2,4-D butyl ester might have substantial effects on microbial population and microbial community structure in agricultural soils. In particular, the effects of 2,4-D butyl ester were greater in soil with low organic matter and fertility level than in soil with high organic matter and fertility level.     

Changes in enzyme activities and microbial biomass after “in situ” remediation of a heavy metal-contaminated soil

Microbial properties such as microbial biomass carbon (MBC), arylsulfatase, β-glucosidase and dehydrogenase activities, and microbial heterotrophic potential, together with several chemical properties such as pH, CaCl₂ soluble heavy metal concentrations, total organic carbon and hydrosoluble carbon were measured to evaluate changes in soil quality, after “in situ” remediation of a heavy metal-contaminated soil from the Aznalcóllar mine accident (Southern Spain, 1998). The experiment was carried out using containers, filled with soil from the affected area. Four organic amendments (a municipal waste compost, a biosolid compost, a leonardite and a litter) and an inorganic amendment (sugarbeet lime) were mixed with the top soil at the rate of 100 Mg ha⁻¹. Unamended soil was used as control. Agrostis stolonifera L. was sown in the containers. The soil was sampled twice: one month and six months after amendment application. In general, these amendments improved the soil chemical properties: soil pH, total organic carbon and hydrosoluble carbon increased in the amended soils, while soluble heavy metal concentrations diminished. At the same time, higher MBC, enzyme activities and maximum rate of glucose mineralization values were found in the organically amended soils. Plant cover was also important in restoring the soil chemical and microbial properties in all the soils, but mainly in those that were not amended organically. As a rule, remediation measures improved soil quality in the contaminated soils.     

Long-term effect of plastic film mulching and fertilization on bacterial communities in a brown soil revealed by high through-put sequencing

The objective of this study was to access the effect of long-term fertilization and film mulching application on soil properties and bacterial community structure. We used 16S rRNA gene to investigate soil bacterial community composition by high through-put sequencing. The results demonstrated that predominant groups in the bacterial community were: Proteobacteria, Bacteroidetes, Actinobacteria, Acidobacteria, Firmicutes, Planctomycetes, Gemmatimonadetes, Verrucomicrobia, Chloroflexi and Cyanobacteria. Long-term fertilization of combined manure and nitrogen application caused significant decrease in soil bacterial diversity and richness compared to non-fertilization control, though manure fertilization alone played a significant role in restoring bacterial diversity. Film mulching and manure fertilization significantly increased the relative abundances of soil bacterial groups mentioned above. Furthermore, film mulching played significant role in shaping the bacterial community structure regardless of fertilization. Redundancy analysis (RDA) showed that soil moisture content, pH, total N and soil organic C had significant (P < 0.05) influence on dominant bacterial groups. Altogether, plastic film mulching and manure application prevented loss of soil bacterial diversity and abundance during long-term fertilization. These findings showed the detrimental use of combined manure and nitrogen fertilization to soil microbes and the useful application of manure fertilization coupled with film mulching to soil biodiversity in long-term fertilization experiments.