Restoration of giant cardon cacti in barren desert soil amended with common compost and inoculated with Azospirillum brasilense

Barren desert soil that otherwise could not support perennial plant growth was amended with six levels of common agricultural compost. Seedlings of the giant cardon cactus, one of the primary plant species responsible for soil stabilization in the southern Sonoran Desert, were inoculated with the plant-growth-promoting bacterium Azospirillum brasilense Cd, planted, and grown for 18 months under nursery conditions typical for slow-growth cacti. Control plants were grown without compost amendment, without inoculation (negative control), or in fertile, rare “resource island” soil preferred by cardon seedlings (positive control). During the prolonged growth period, the decisive factor in seedling growth in barren soil was the addition of small amounts of common compost; 6 to 25% of the growth substrate volume gave the best growth response and, to a lesser extent, so did inoculation with A. brasilense Cd. Although the bacteria significantly affects plant growth when amended with “resource island” soil and added to barren soil, its effect on plant growth was far smaller than when compost alone was added. Compost added to barren soil significantly increased the dry weight parameters of the plant to almost similar levels obtained by the “resource island” soil; however, the compost amendment supports a more voluminous and greener plant with elevated pigment levels. This study shows that barren soil supplemented with compost can replace the rare “resource island” soil for cardon nurseries destined to abate soil erosion in the desert.     

设施种植模式对土壤细菌多样性及群落结构的影响

为了研究有机和常规设施种植模式及轮作对土壤细菌多样性和群落结构的影响,本研究采用Illumina平台Hiseq 2500高通量测序技术,于2016年6月(作物处于收获期)对北京市顺义区不同设施种植模式(分别为有机设施种植模式和常规设施种植模式下的叶菜连作、茄果连作和叶茄轮作)下土壤细菌进行16S r RNA测序。测序质控后共获得17 278个操作分类单元(operational taxonomic units,OTUs),共计318 851条有效序列。比较不同种植模式和轮作下土壤细菌多样性、细菌群落结构组成、相对丰度及土壤理化性质与细菌群落多样性关系的差异性。结果表明:土壤微生物群落结构在有机和常规设施种植模式下差异明显,有机设施种植土壤细菌多样性高于常规设施种植;有机设施种植下轮作与连作土壤细菌群落结构表现出明显差异,而常规设施种植下,两者没有明显差异;有机种植模式下,轮作土壤细菌群落多样性高于连作土壤;设施种植土壤细菌群落主要属于鞘氨醇单胞菌属(Sphingomonas,5.05%)和芽孢杆菌属(Bacillus,4.84%),相对丰度大于0.5%的共有14个属。有机设施种植土壤含有较多促进植物生长、有机质分解的细菌,常规设施种植土壤中降解化学杀虫剂、防治土壤病害、促进硝化过程的细菌较多。RDA分析结果显示土壤细菌群落主要受全磷、速效磷、有机质的影响。Tumebacillus、Candidatus Solibacter和Acidothermus都是分解有机质、利用碳源的细菌属,与土壤有机质含量呈正相关关系。由此可见,设施条件下,有机和常规种植土壤微生物群落结构的差异性主要源于肥料使用、有害生物防治措施和管理方式的不同。有机设施种植模式下,轮作更有利于发挥其改良土壤营养循环和防治土壤病虫害的作用。上述结果为在微生物水平上研究设施条件下不同种植模式的土壤生态质量差异提供了参考。     

毒死蜱对棉花根际土细菌群落多样性和结构的影响

通过室内盆栽试验模拟自然环境条件,采用高效液相色谱(HPLC)和末端限制性片段长度多态性(T-RFLP)技术,研究了土壤使用推荐剂量(5 mg·kg~(-1))及推荐剂量的2倍、3倍和4倍(10 mg·kg~(-1)、15 mg·kg~(-1)、20 mg·kg~(-1))毒死蜱对棉花根际土壤细菌群落多样性和结构的影响,以不施用毒死蜱的土壤为对照。结果表明,5 mg·kg~(-1)、10 mg·kg~(-1)、15 mg·kg~(-1)和20 mg·kg~(-1)毒死蜱在土壤中的半衰期分别为10.04 d、11.36 d、11.55 d和12.16 d,60 d时基本完全降解。毒死蜱处理60 d后,棉花生物量显著降低;毒死蜱浓度越高,棉花生物量越低。无毒死蜱条件下不同取样时间根际细菌多样性无显著差异,毒死蜱处理组前30 d细菌多样性均显著降低,60 d时毒死蜱处理组细菌多样性恢复到正常水平。研究发现毒死蜱浓度越高对细菌多样性抑制作用越显著,恢复越缓慢。主成分分析结果发现,第10 d、30 d和60 d毒死蜱处理组与对照组细菌群落结构差异显著,其中60 d时20 mg·kg~(-1)毒死蜱处理组差异最显著,即使土壤中毒死蜱完全降解,根际细菌群落结构仍不会恢复到正常水平。60 d时,被毒死蜱抑制的细菌有硝化刺菌属(Nitrospina sp.)和Cellulophaga sp.等,被激活的有芽孢杆菌属(Bacillus sp.)和链霉菌属(Streptomyces sp.)等。可见,毒死蜱的引入,重新构建了土壤细菌群落结构,显著影响棉花生长,对棉花根际土壤微生态环境冲击较大,应对其生态安全性予以重视。     

A single application of Cu to field soil has long-term effects on bacterial community structure, diversity, and soil processes

Long-term diversity-disturbance responses of soil bacterial communities to copper were determined from field-soils (Spalding; South Australia) exposed to Cu in doses ranging from 0 through to 4012 mg Cu kg⁻¹ soil. Nearly 6 years after application of Cu, the structure of the total bacterial community showed change over the Cu gradient (PCR-DGGE profiling). 16S rRNA clone libraries, generated from unexposed and exposed (1003 mg Cu added kg⁻¹ soil) treatments, had significantly different taxa composition. In particular, Acidobacteria were abundant in unexposed soil but were nearly absent from the Cu-exposed sample (P<0.05), which was dominated by Firmicute bacteria (P<0.05). Analysis of community profiles of Acidobacteria, Bacillus, Pseudomonas and Sphingomonas showed significant changes in structural composition with increasing soil Cu. The diversity (Simpsons index) of the Acidobacteria community was more sensitive to increasing concentrations of CaCl-extractable soil Cu (Cu_Ext) than other groups, with decline in diversity occurring at 0.13 Cu_Ext mg kg⁻¹ soil. In contrast, diversity in the Bacillus community increased until 10.4 Cu_Ext mg kg⁻¹ soil, showing that this group was 2 orders of magnitude more resistant to Cu than Acidobacteria. Sphingomonas was the most resistant to Cu; however, this group along with Pseudomonas represented only a small percentage of total soil bacteria. Changes in bacterial community structure, but not diversity, were concomitant with a decrease in catabolic function (BioLog). Reduction in function followed a dose-response pattern with Cu_Ext levels (R²=0.86). The EC₅₀ for functional loss was 0.21 Cu_Ext mg kg⁻¹ soil, which coincided with loss of Acidobacteria diversity. The microbial responses were confirmed as being due to Cu and not shifts in soil pH (from use of CuSO₄) as parallel Zn-based field plots (ZnSO₄) were dissimilar. Changes in the diversity of most bacterial groups with soil Cu followed a unimodal response - i.e. diversity initially increased with Cu addition until a critical value was reached, whereupon it sharply decreased. These responses are indicative of the intermediate-disturbance-hypothesis, a macroecological theory that has not been widely tested in environmental microbial ecosystems.     

Fluazifop-P-butyl (herbicide) affects richness and structure of soil bacterial communities

Fusilade (fluazifop-P-butyl) is an extensively used herbicide in legumes cropping systems; nevertheless, the extent of its ecological risk remains unknown. The aim of this work is to estimate the impact of this herbicide on some key enzyme activities, namely acid phosphatase, urease and FDA, and the effect on soil bacterial communities in the rhizosphere of pea (Pisum sativum). Acid phosphatase, urease and FDA showed a high decrease following the application of fluazifop-P-butyl mainly at ten-fold field rate. Grain yield was also severely affected at this concentration (−50%). T-RFLP analysis of 16S rDNA revealed a significant effect of fluazifop-P-butyl application on richness and structure of soil bacterial communities. Putative taxonomic assignation of induced TRFs revealed a marked pattern of acidogenic bacteria like as Staphylococcus, Escherichia, Desulfobacter, Clostridium, Paludibacter and other sulfate-reducing bacteria like as Desulfocella halophila and Desulfocurvus vexinensis. These bacteria may participate in the acidogenesis and the production of propionic acid from the fluazifop-P-butyl after hydrolytic cleavage. They are also known to play important roles in the global biogeochemical sulfur cycles. Unfortunately, some potential human pathogens were also stimulated. Moreover, many putative plant-growth-promoting bacteria belonging to the genera Streptomyces and Bacillus were found to be inhibited in the bulk soil and rhizosphere. The fluazifop-P-butyl effect on bacterial diversity was more pronounced in the rhizosphere compared to bulk soil or uncultivated soil which may suggest that a major component of this effect is mediated by the root system.     

Long-term fertilization regimes affect bacterial community structure and diversity of an agricultural soil in northern China

Background, Aims, and Scope  Knowledge about shifts of microbial community structure and diversity following different agricultural management practices could improve our understanding of soil processes and thus help us to develop sound management strategies. A long-term fertilization experiment was established in 1989 at Fengqiu (35°00′N, 114°24′E) in northern China. The soil (sandy loam) is classified as aquic inceptisols and has received continuous fertilization treatments since then. The fertilization treatments included control (CK, no fertilizer), chemical fertilizers nitrogen (N) and potassium (K) (NK), phosphorous (P) and K (PK), NP, NPK, organic manure (OM), and half chemical fertilizers NPK plus half organic manure (1/2NPKOM). The objective of this study was to examine if the microbial community structure and diversity were affected by the long-term fertilization regimes. Materials and Methods  Soil samples were collected from the long-term experimental plots with seven treatments and four replications in April 2006. Microbial DNAs were extracted from the soil samples and the 16S rRNA genes were PCR amplified. The PCR products were analyzed by DGGE, cloning and sequencing. The bacterial community structures and diversity were assessed using the DGGE profiles and the clone libraries constructed from the excised DGGE bands. Results  The bacterial community structure of the OM and PK treatments were significantly different from those of all other treatments. The bacterial community structures of the four Ncontaining treatments (NK, NP, NPK and 1/2NPKOM), as well as CK, were more similar to each other. The changes in bacterial community structures of the OM and PK treatments showed higher richness and diversity. Phylogenetic analyses indicated that Proteobacteria (30.5%) was the dominant taxonomic group of the soil, followed by Acidobacteria (15.3%), Gemmatimonadetes (12.7%), etc. Discussion  Irrespective of the two fertilization treatments of OM and PK, the cluster analysis showed that bacterial communities of the remaining five treatments of CK, NK, NP, NPK and 1/2NPKOM seemed to be more similar to each other, which indicated the relatively weak effects of the four N-containing treatments on soil bacterial communities. N fertilizer may be considered as a key factor to counteract the effects of other fertilizers on microbial communities. Conclusions  Our results show that long-term fertilization regimes can affect bacterial community structure and diversity of the agricultural soil. The OM and PK treatments showed a trend towards distinct community structures, higher richness and diversity when compared to the other treatments. Contrasting to the positive effects of OM and PK treatments on the bacterial communities, N fertilizer could be considered as a key factor in the soil to counteract the effects of other fertilizers on soil microbial communities. Recommendations and Perspectives  Because of the extremely high abundance and diversity of microorganisms in soil and the high heterogeneity of the soil, it is necessary to further examine the effects of fertilization regimes on microbial community and diversity in different type soils for comprehensively understanding their effects through the appropriate combination of molecular approaches. ESS-Submission Editor: Chengrong Chen, PhD (c.chen@griffith.edu.au)     

Microarray analysis of bacterial diversity and distribution in aggregates from a desert agricultural soil

Previous research has shown that soil structure can influence the distribution of bacteria in aggregates and, thereby, influence microbiological processes and diversity at small spatial scales. Here, we studied the microbial community structure of inner and outer fractions of microaggregates of a desert agricultural soil from the Imperial Valley of Southern California. To study the distribution of soil bacteria, 1,536 clones were identified using phylogenetic taxon probes to classify arrays of 16S rRNA genes. Among the predominant taxonomic groups were the α-Proteobacteria, Planctomycetes, and Acidobacteria. When compared across all phyla, the taxonomic compositions and distributions of bacterial taxa associated with the inner and outer fractions were nearly identical. Our results suggest that the ephemeral nature of soil aggregates in desert agricultural soils may reduce differences in the spatial distribution of bacterial populations as compared to that which occur in soils with more stable aggregates.     

不同深度土壤控水对稻田土壤微生物区系及细菌群落多样性的影响

为研究不同深度土壤控水对壤土稻田土壤水势、微生物区系和细菌群落多样性的影响,通过土培池栽试验,在水稻生育后期设置土壤深度0~5 cm（S05）、0~10 cm（S10）和0~15 cm（S15）控水处理,以保持水层为对照,分析了不同深度控水处理下5 cm、10 cm、15 cm深土壤水势与土壤微生物区系、细菌群落多样性的变化。结果表明：土壤5 cm、10 cm、15 cm深度的水势随着控水深度增加而降低,S05控水处理主要影响上层（5 cm）土壤水势,S10控水处理影响上、中层（10 cm）土壤水势,S15控水处理土壤水势随土层深度的增加而升高。花后8 d和32 d,S05控水处理上层土壤细菌数量显著高于S10、S15控水处理;花后16~24 d,S05控水处理中层、下层（15 cm）土壤细菌数量均显著高于S15控水处理;土壤水势与水稻生育后期中、下层土壤细菌数量呈极显著正相关关系。S05控水处理10 cm、15 cm土层的细菌丰富度Chao指数均显著高于S15控水处理及CK。3个控水处理中,5 cm土层细菌的多样性Shannon指数以S05控水处理最低。优势细菌菌群分析发现,优势群落主要为变形菌门、绿弯菌门、酸杆菌门、拟杆菌门,四者总相对丰度在80%以上;S15控水处理中层土壤变形菌门相对丰度低于S05和S10控水处理。3个控水处理土壤样品中优势纲（相对丰度大于2%）达15个,主要包括α-变形菌纲、β-变形菌纲、δ-变形菌纲、厌氧绳菌纲等,这4个纲的总相对丰度在47%以上,其中厌氧绳菌纲相对丰度最高;上层土壤中S05控水处理的β-变形菌纲相对丰度显著低于S10和S15控水处理。因此,不同深度土壤控水对壤土土壤水势、细菌数量存在影响,改变了细菌的多样性及丰富度,对土壤细菌优势菌种类无显著影响。     

Relative impact of soil, metal source and metal concentration on bacterial community structure and community tolerance

In a study to assess the sustainable use of sewage sludge application to land, the long-term effect of Zn and Cu contaminated sludge additions on the structure of the bacterial communities (using T-RFLP analysis) and their tolerance to additional metal exposure through pollution-induced community tolerance (PICT) assays was assessed. This used two soils that received metal-rich sludge cake (SC), liquid sludge (LS) or metal salts (MS) additions more than 10 years previously. Soil type had the predominant influence on bacterial community structure and PICT. The source of the metal contamination also had a large influence on community structure and PICT, greater than the effects due to metal concentrations. Nevertheless, in both Zn and Cu contaminated soils, PICT was observed and decreased in the order MS > LS > SC. Within a metal source and site, there was evidence of increased PICT with increasing Zn or Cu contamination, however few differences were significant as a result of high variability between sample replicates. These results highlight the importance of considering soil physico-chemical properties and the source of metal contamination as well as total metal concentrations when considering the long-term effects of metals on soil microbial communities. Further, the matrix that a metal is associated with prior to addition may play an important factor in determining levels of toxicity. This could have consequences for the interpretation and use of data from metal spiking experiments when considering metal limits for sludge application to land.     

Soil fertility is associated with fungal and bacterial richness,whereas pH is associated with community composition in polar soil microbial communities

Microbial activities in Arctic and Antarctic soils are of particular interest due to uncertainty surrounding the fate of the enormous polar soil organic matter (SOM) pools and the potential to lose unique and vulnerable micro-organisms from these ecosystems. We quantified richness, evenness and taxonomic composition of both fungi and bacteria in 223 Arctic and Antarctic soil samples across 8 locations to test the global applicability of hypotheses concerning edaphic drivers of soil microbial communities that have been primarily developed from studies of bacteria in temperate and tropical systems. We externally validated our model's conclusions with an independent dataset comprising 33 Arctic heath samples. We also explored if our system was responding to large scale climatic or biogeographical processes that we had not measured by evaluating model stability for one location, Mitchell Pennisula, that had been extensively sampled. Soil Fertility (defined as organic matter, nitrogen and chloride content) was the most important edaphic property associated with measures of α-diversity such as microbial richness and evenness (especially for fungi), whereas pH was primarily associated with measures of β-diversity such as phylogenetic structure and diversity (especially for bacteria). Surprisingly, phosphorus emerged as consistently the second most important driver of all facets of microbial community structure for both fungi and bacteria. Despite the clear importance of edaphic factors in controlling microbial communities, our analyses also indicated that fungal/bacterial interactions play a major, but causally unclear, role in structuring the soil microbial communities of which they are a part.     

苯醚甲环唑胁迫下农田土壤微生物群落和多样性分析

为研究三唑类杀菌剂苯醚甲环唑(dif)施用后土壤中微生物群落结构的响应变化,通过模拟生产中的施用剂量,监测35 d后土壤的理化性质并利用高通量测序技术分析土壤中细菌和真菌的群落组成变化,通过db-RDA分析建立环境因子与微生物群落响应关系。研究结果显示,随着dif施用浓度的增加,土壤中有机质(SOM)、总氮(TN)、碱解氮(AN)均有所提高,有效磷(AP)呈下降趋势,Alpha多样性指数中细菌Chao1、ACE和Shannon指数均低于未处理土壤,真菌Shannon指数高于未处理土壤;所有样本中细菌共检出31门、91纲、206目、315科、553属,真菌共检测出11门、34纲、79目、174科、370属;苯醚甲环唑施用浓度为50 mg/kg时,土壤细菌中变形菌门和拟杆菌门的相对丰度分别提高了14.2%和1.81%,酸杆菌门和绿弯菌门相对丰度分别降低了8.28%和3.44%,绝对优势菌属节杆菌属相对丰度下降7.85%,真菌中赤霉属和被孢霉属逐渐成为优势菌群;细菌的优势菌群中变形菌门与TN呈显著正相关,真菌优势菌门中子囊菌门与AN、TN和dif呈显著正相关,被孢霉门与SOM呈显著负相关;TN和AN对细菌的Shannon指数影响较大。以上说明,高浓度苯醚甲环唑的施用对土壤的理化性质以及微生物的丰富度和群落结构均产生了影响。     

土壤微生物群落结构与多样性对农田集约化管理程度降低的响应

Using a scheme of agricultural fields with progressively less intensive management （deintensification）, different management practices in six agroecosystems located near Goldsboro, NC, USA were tested in a large-scale experiment, including two cash-grain cropping systems employing either tillage （CT） or no-tillage （NT）, an organic farming system （OR）, an integrated cropping system with animals （IN）, a successional field （SU）, and a plantation woodlot （WO）. Microbial phospholipid fatty acid （PLFA） profiles and substrate utilization patterns （BIOLOG ECO plates） were measured to examine the effects of deintensification on the structure and diversity of soil microbial communities. Principle component analyses of PLFA and BIOLOG data showed that the microbial community structure diverged among the soils of the six systems.Lower microbial diversity was found in lowly managed ecosystem than that in intensive and moderately managed agroecosystems, and both fungal contribution to the total identified PLFAs and the ratio of microbial biomass C/N increased along with agricultural deintensification. Significantly higher ratios of C/N （P 〈 0.05） were found in the WO and SU systems, and for fungal/bacterial PLFAs in the WO system （P 〈 0.05）. There were also significant decreases （P 〈 0.05） along with agricultural deintensification for contributions of total bacterial and gram positive （G＋） bacterial PLFAs.Agricultural deintensification could facilitate the development of microbial communities that favor soil fungi over bacteria.     

Compaction of forest soils with heavy logging machinery affects soil bacterial community structure

Soil compaction is widespread but tends to be most prevalent where heavy machinery is used in landfill sites, agriculture and forestry. Three forest sites strongly disturbed by heavy logging machinery were chosen to test the physical effects of different levels of compaction on soil bacterial community structure and soil functions. Community analysis comprised microbial biomass C and T-RFLP genetic profiling. Machine passes, irrespective of the compaction level, considerably modified soil structural characteristics at two soil depths (5–10 cm; 15–20 cm). Total porosity decreased up to 17% in the severe compaction. Reflected in this overall decline were large decreases in macroporosity (>50 μm). Reduction in macroporosity was associated with higher water retention and restricted gas exchange in compacted soils. The strongest effect was observed in the severely compacted wheel tracks where air and water conductivities were reduced permanently to 10% or even lower of the original conductivities of undisturbed soils. Very slow drainage in combination with a dramatically reduced gas permeability led to unfavorable soil conditions in severely disturbed traffic lanes reflecting the changes in the total bacterial community structures at both soil depths. Additionally, microbial biomass C tended to be lower in compacted soil. Our results indicate that the type of severe treatments imposed at these forest sites may have strong adverse effects on long-term soil sustainability.     

马铃薯与玉米复合种植对土壤化感物质及土壤细菌群落结构的影响

为探究马铃薯与玉米复合种植对化感物质积累与细菌群落结构的影响,分析轮作、间作缓解连作障碍的机制,本研究以马铃薯连作、玉米连作、马铃薯||玉米间作、马铃薯-玉米轮作第8年的土壤为对象,利用GC-MS测定土壤中化感物质含量,并采用Illumina Miseq高通量测序技术对土壤细菌16Sr DNA V4-V5区域进行测序,分析土壤中细菌多样性和群落结构的变化,并对化感物质和优势菌属进行相关性分析。结果表明:玉米连作和马铃薯连作会导致化感物质的积累,玉米连作土壤积累了更多的油酸、亚油酸、花生酸、木焦油酸等脂肪酸,马铃薯连作土壤积累了更多的硬脂醇、二十烷醇等脂肪醇类物质。轮作降低了大部分化感物质的积累,间作降低的化感物质种类相对轮作较少。不同种植方式下土壤细菌群落结构发生了显著变化,相对于连作,间作和轮作Ace指数和Chao指数显著升高。在门水平上,轮作土壤放线菌丰度显著高于马铃薯连作土壤,间作土壤拟杆菌门丰度显著低于玉米连作土壤,两种连作土壤中酸杆菌门丰度都较轮作显著升高。在属水平上,一些有益细菌如节杆菌属、溶杆菌属等在复合种植土壤中相对丰度更高。通过相关性分析发现土微菌属、小梨形菌属与脂肪醇类物质呈显著正相关,黄杆菌属、溶杆菌属、微杆菌属等与脂肪酸类物质呈显著负相关。马铃薯与玉米复合种植降低了化感物质在土壤中的积累,从而抑制了土壤细菌丰度的降低,提高了有益菌属丰度,消减了连作障碍。     

菜-菌轮作模式下土壤微生物群落结构及多样性的变化

为明确菜-菌轮作模式对土壤微生物的影响,基于高通量测序技术,对4种轮作模式下的土壤微生物群落结构与多样性进行了研究,结果显示：不同轮作模式下土壤样品中真菌和细菌OUT总数分别是2298和15840条,相较于常规轮作模式A,菜-菌轮作模式B、C、D下真菌的OUT总数、ACE指数、Chao1指数、Shannon指数降低显著,Simpson指数增加明显,但细菌的各参数没有显著的差异;其土壤全氮、碱解氮、速效钾、有机质含量显著高于常规轮作模式A;真菌的优势群落是子囊菌门,相对丰度在66%以上,枝孢属、镰刀菌属两类致病菌群的丰度在菜-菌轮作模式B、C、D中降低明显;细菌的优势群落是变形菌门、放线菌门、绿弯菌门以及酸杆菌门,鞘氨醇单胞菌属、伯克霍尔德氏菌属丰度在菜-菌轮作模式B、C、D中显著提高;聚类分析表明菜-菌轮作模式C、D下微生物群落结构相似度最高,常规轮作模式A可划分为区别于菜-菌轮作模式B、C、D的单独类群。综上所述,菜-菌轮作可降低真菌群落丰度,改变土壤微生物群落的结构组成,同时提高土壤中有益菌群的丰度,降低有害菌群的丰度。     

不同施肥处理对农田土壤微生物区系和功能的影响

本研究采用DGGE和Biolog两种方法, 研究了五种不同施肥处理对土壤微生物区系组成和功能的影响。采用UPGMA对DGGE试验结果进行聚类分析得出,小麦季中不施肥对照（T1）和常规氮磷钾肥（T2）聚为一类,相似性为41%,常规氮磷钾肥+秸秆还田（T4）和常规氮磷钾肥+秸秆还田+秸秆腐熟剂（T5）聚为一类,相似性为52%,而70%常规氮磷钾肥+有机肥（T3）单独聚为一类;玉米季也得到类似的结果,只是T1和T2的相似性为68%,T4和T5相似性达78%,而T3依然归为一类,这说明不同施肥处理间土壤微生物区系存在相似性。经过切胶测序及BLAST比对,发现大多为不可培养细菌,可培养细菌中多为芽孢杆菌属和类芽孢杆菌属。Biolog试验采用主成份分析得出,T1和T2土壤微生物功能类似,T4和T5土壤微生物也具有相似的功能,而T3则单独分为一类。其中,T5和T4的土壤细菌群落对底物碳源的代谢活性最强,T3处理次之,T2和T1最低。通过这两种不同试验方法的分析可以看出,不同施肥处理对土壤微生物区系和功能的影响存在关联性。     

Impact of sulfadiazine and chlorotetracycline on soil bacterial community structure and respiratory activity

Veterinary medicines enter agricultural soils by the use of animal excrements as fertilizers. To study their impact on soil bacterial communities, microcosms containing orthic luvisol soil were spiked with the antimicrobial agents sulfadiazine (SDZ) and chlorotetracycline (CTC) at three different concentrations (1, 10, 50 mg kg⁻¹ soil) and incubated for 48 days at 20 °C. The impact on the microbial respiratory activity was measured continuously in a respirometer (Sapromat). Changes in bacterial community structure were visualized by means of PCR-denaturing gradient gel electrophoresis (DGGE) of 16S rDNA derived from soil samples after 1, 7, 11 and 48 days. Additionally, growth inhibitory effects of SDZ and CTC on bacteria previously isolated from the same soil were tested in agar diffusion tests. In microcosms with soil and antibiotics only, no effects could be observed, either on respiratory activity or on bacterial population structure. Therefore, further incubations were conducted in the presence of an additional assimilable carbon source (5 g glucose kg⁻¹ soil). In the presence of glucose, SDZ affected soil respiration as well as the bacterial community structure: Additional bands appeared and some bands already visible at the beginning of incubations increased in intensity. A clear relationship between SDZ concentrations and changes in DGGE patterns became visible. During 48 days of incubation, changes in DGGE patterns were minimal in microcosms with 50 mg SDZ kg⁻¹soil indicating an inhibition of strains, which were capable of growing on glucose in the presence of lower SDZ concentrations. Only a few soil bacterial isolates (5 out of 47 strains tested) were weakly inhibited by SDZ in agar diffusion disk tests. Contrastingly, CTC inhibited growth of 12 soil bacterial isolates significantly in disk tests, but no effects on soil respiration and bacterial community structure could be observed. In the presence of the soil matrix the growth inhibitory potential of CTC decreased due to adsorption or complexation. This was confirmed in growth inhibition experiments with soil suspensions and time-dependent sampling.     

玉米秸秆生物炭对褐土微生物功能多样性及 细菌群落的影响

生物炭施入土壤被认为是一种有效的固碳减排措施,可增加土壤有机碳及矿质养分含量,提高土壤的持水能力及保肥能力。为探明其施入土壤后对土壤微生物活性及多样性的影响,本文在盆栽试验条件下,采用Biolog与高通量测序相结合的方法,研究了CK(不施生物炭)和施用5 g·kg~(-1)、10 g·kg~(-1)、30 g·kg~(-1)、60 g·kg~(-1)玉米秸秆生物炭对土壤微生物碳源利用能力(AWCD)、功能多样性指数以及土壤细菌的丰度和多样性的影响。结果表明,随着生物炭施用量的增加,表征土壤微生物活性的AWCD值呈下降趋势,表现为:5 g·kg~(-1)处理≈CK10 g·kg~(-1)处理30 g·kg~(-1)处理60 g·kg~(-1)处理,其中CK和5 g·kg~(-1)处理间差异不显著(P0.05),而10 g·kg~(-1)、30 g·kg~(-1)和60 g·kg~(-1)处理在整个培养期间的AWCD值显著低于CK处理(P0.05);土壤微生物群落代谢功能多样性指数(H′)、碳源利用丰富度指数(S)均随生物炭施用量的增加而呈下降趋势,但均匀度指数(E)表现出相反趋势,5g·kg~(-1)、10 g·kg~(-1)、30 g·kg~(-1)、60 g·kg~(-1)各处理的H′较CK处理分别增加0.16%、-0.88%、-3.14%、-11.09%,S分别增加-2.82%、-11.27%、-18.31%、-47.89%,E分别增加1.14%、3.00%、3.73%和13.76%。主成分分析表明,与CK处理比较,5 g·kg~(-1)处理对土壤微生物群落碳源利用方式没有显著影响(P0.05),而10 g·kg~(-1)、30 g·kg~(-1)和60g·kg~(-1)处理对土壤微生物群落碳源利用方式影响显著(P0.05)。随着生物炭施用量的增加,土壤细菌OTU数目及丰富度指数(Chao1)呈增加趋势,5 g·kg~(-1)处理与CK处理差异不显著,而10 g·kg~(-1)、30 g·kg~(-1)、60 g·kg~(-1)处理的OTU数目较CK处理分别增加1.09%、5.26%、24.42%,Chao1分别增加5.73%、10.21%、37.68%。土壤中施用生物炭后土壤细菌变形菌门(Proteobacteria)的丰度在CK处理和5 g·kg~(-1)处理间差异不显著(P0.05),而10g·kg~(-1)、30 g·kg~(-1)、60 g·kg~(-1)处理较CK处理分别增加32.3%、21.1%、16.7%,拟杆菌门(Bacteroidetes)的丰度随着生物炭施用量的增加各处理较CK处理分别减少22.1%、55.3%、66.8%、50.5%。生物炭施入土壤后降低了土壤可培养微生物的活性,减少或改变了土壤微生物碳源利用的种类,使土壤原有微生物群落组分发生改变,生物炭也影响了土壤细菌各菌群在土壤中的丰度,使其分布的均匀性降低。为了不影响微生物群落结构和功能,石灰性褐土上生物炭一次还田量不能超过5 g·kg~(-1)(干土)。     

盐地碱蓬根际土壤细菌群落结构及其功能

盐地碱蓬作为生物改良盐碱地的理想材料,其根际土壤微生物对土壤改良发挥着重要作用。为了深入探索环渤海滨海盐碱地碱蓬根际土壤细菌群落结构组成及其功能,采用Illumina Misep高通量测序平台对环渤海地区滨海盐碱地盐地碱蓬根际土壤和裸地土壤进行测序。从16个样本中获得有效序列734 792条, 4 285个OTUs,归属于41门、100纲、282目、400科、892属、1 577种。盐地碱蓬根际土壤细菌群落由变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、绿弯曲门(Chloroflexi)、拟杆菌门(Bacteroidetes)、芽单胞菌门(Gemmatimonadetes)、酸杆菌门(Acidobacteria)、厚壁菌门(Firmicutes)、蓝藻细菌门(Cyanobacteria)、髌骨细菌门(Patescibacteria、浮霉菌门(Planctomycetes)组成。Alpha多样性计算结果表明,盐地碱蓬根际土壤细菌群落结构多样性高并与裸地土壤间差异显著;LEfSe(LDAEffectSize)分析发现,盐地碱蓬与裸地差异指示种明显不同。PCoA与相关性Heatmap表明,盐地碱蓬、速效氮、速效钾、速效磷、电导率是影响土壤细菌目类水平群落组成的主要因子。PICRUSt(Phylogenetic InvestigationofCommunitiesbyReconstructionofUnobserved States)分析表明微生物群落在新陈代谢等40个功能方面盐地碱蓬根际土壤比裸地土壤高。本研究表明盐地碱蓬覆盖能够降低土壤盐分,增加土壤养分,对土壤细菌群落多样性及其功能有积极作用。     

植烟土壤提取物质对烟株生长及根际土壤细菌多样性的影响

对盆栽烟草外源添加不同浓度植烟土壤提取物质(T1:40μg·mL-1;T2:120μg·mL-1;CK:蒸馏水对照),探讨植烟土壤提取物质对烟草生长及土壤细菌多样性的影响。结果表明,植烟土壤提取物处理使烟株生长受抑制,且随处理浓度的增加受抑制程度显著提高,具体表现为烟株变矮,叶面积变小,光合作用能力降低,且烟草的保护酶系统受到破坏,丙二醛含量随处理浓度加大而增加,T2处理的丙二醛含量是对照的3.44倍。对外源添加物质处理后烟草根际土壤微生物T-RFs分析发现,在对照检测到17个门24个纲,T1处理有14个门21个纲,T2有10个门17个纲;丰富度指数的变化也和门纲的变化一致,随着处理浓度的增加而显著降低。可见外源添加物质处理后,根际土壤细菌群落减少,多样性水平下降。对各处理的根际土壤微生物T-RFs变化与烟株生长变化进行相关性分析表明,在外源添加物质处理的土壤中存在较多的负相关T-RFs片段,且这些片段中较多为病原菌;而正相关的T-RFs片段主要存在于对照土壤中,其中有较多与土壤营养元素循环相关的微生物。本研究结果显示,在外源添加植烟土壤提取物质处理下,烟草的生长受抑制,烟草根际土壤的微生态受到破坏,且随浓度的提升而加重。因此,连作土壤中自毒物质的富集是造成烟草连作障碍的主要原因。关键词烟草连作障碍根际细菌自毒作用T-RFLP