两种代表性蚯蚓对设施菜地土壤微生物群落结构及理化性质的影响

不同生态型蚯蚓的取食偏好和生境有所差异,因此蚯蚓的生态型差异可能关乎其对土壤性质的不同影响;有关不同生态型蚯蚓对土壤性质尤其是微生物学性质影响的研究有助于了解蚯蚓生态功能的作用机制。在野外调控试验的第4年采集土壤,研究了牛粪混施和表施处理下内层种威廉腔环蚓(Metaphire guillelmi)和表层种赤子爱胜蚓(Eisenia foetida)对设施菜地土壤微生物群落结构和主要理化性质的影响。结果表明,土壤微生物群落结构同时受到蚯蚓种类和牛粪施用方式的影响。牛粪表施时,两种蚯蚓均显著降低了菌根真菌、真菌生物量和原生动物生物量(P0.05);牛粪混施时,不同蚯蚓的影响有所差异,威廉腔环蚓明显增加了菌根真菌、真菌生物量和放线菌生物量,而赤子爱胜蚓的作用不明显。此外,两种蚯蚓均提高了土壤孔隙度、团聚体稳定性和土壤p H、矿质氮以及微生物生物量碳氮水平,但提高幅度取决于蚯蚓种类和牛粪施用方式。冗余分析表明蚯蚓影响下土壤微生物群落结构的变化与团聚体稳定性、p H、速效磷、矿质氮呈正相关,而与土壤容重呈负相关。     

蚯蚓在我国南方土壤修复中的应用

蚯蚓作为生物量最大的土壤动物, 对土壤生态系统和环境质量影响深远。本研究介绍了华南地区主要应用的皮质远盲蚓(Amynthas corticis)、毛利远盲蚓(A. morrisi)、壮伟远盲蚓(A. robustus)、参状远盲蚓(A. aspergillum)、南美岸蚓(Pontoscolex corethrurus)和赤子爱胜蚓(Eisenia fetida)的生态特征, 阐述了它们与土壤pH值、酶活性、金属富集和有效性改变、孔道和微团聚体形成之间的紧密关系: (1)蚯蚓生存的土壤酸碱性范围较广(pH为3.8-7.9), 其存活率与土壤类型、有机质含量和成分、土壤污染程度和蚯蚓种类相关; (2)肠道内、蚓粪和蚓触圈的酶活性分别表征了蚯蚓取食喜好、土壤养分循环及微生物种群特征; (3)蚯蚓能够富集不同种类的金属并改变其有效性, 这些变化具有蚓种间、金属种类间和土壤类型之间的差异; (4)蚯蚓活动及其生产的蚓粪能改变土体结构、产生孔道、影响土壤团聚体数量、大小和分布。蚯蚓的上述作用使其在解决中国南方红壤酸化、土壤金属污染、茶园土壤养分不平衡、高速公路建设临时用地土壤损毁等方面具有广阔的应用前景。目前, 由于华南远盲蚓的生理特征差异研究较少, 远盲蚓繁育技术的缺乏一定程度上限制了这些蚯蚓在中型和大型尺度下应用技术的研究和推广。有必要进一步挖掘蚯蚓在土壤修复中的潜力, 进行蚯蚓主导的相关技术研发, 深入探讨其影响机制。     

砷矿区农田土壤微生物群落碳源代谢多样性

采用Biolog方法研究了砷(As)矿区农田土壤微生物碳源利用多样性及其与土壤化学性质的关系.结果表明： 3种土壤的N、P、K、有机质(OM)、Cu和Zn全量依次为中As > 高As > 低As土壤.中As和高As土壤微生物平均吸光度以及多样性指数(H′、D、U)显著高于低As土壤.主成分分析与生理碳代谢指纹图谱分析表明,中As和高As土壤微生物对糖类、氨基酸类等碳源的利用程度显著高于低As土壤.典范对应分析显示,影响土壤微生物群落碳源代谢的主要有全P、OM、全Pb、全Zn、全N和pH,全As并不是最主要的影响因子.可见,养分是影响长期污染土壤微生物群落结构和功能多样性的主要因素.     

4种抗生素对蚯蚓-污泥系统细菌群落结构的影响

蚯蚓可摄食污泥中的有机物,其肠道微生物群落在其分解过程中起着主要的作用。利用赤子爱胜蚓(Eisenia foetida)和人工湿地基质构建蚯蚓-污泥系统,添加氯霉素、四环素、链霉素和青霉素4种抗生素,研究不同抗生素对污泥和蚓粪的细菌群落结构的影响。采用高通量测序技术比较分析污泥和蚓粪的细菌多样性及群落结构变化。结果表明,外加抗生素能够导致污泥的Chao1和ACE指数降低,同时降低拟杆菌门和变形菌门的相对丰度,加入氯霉素和青霉素会增加厚壁菌门的相对丰度,降低酸杆菌门、放线菌门和绿弯菌门的相对丰度,加入四环素和链霉素则与之相反。蚓粪样品中,添加氯霉素和链霉素导致Chao1和ACE指数降低,而添加四环素和青霉素则导致Chao1和ACE指数升高,外加抗生素可降低拟杆菌门的相对丰度,增加放线菌门、变形菌门和疣微菌门的相对丰度。主成分分析(PCA)和聚类分析表明,氯霉素和青霉素对污泥细菌群落影响作用相似,四环素与链霉素效果类似;氯霉素对蚓粪群落结构的影响小于其他抗生素。研究结果显示,抗生素可影响污泥和蚓粪的细菌多样性及群落结构,不同抗生素对污泥和蚓粪的影响程度存在差异。     

高砷含水层参与腐殖酸-铁矿物转化的关键微生物群落及其对砷迁移转化的影响

【目的】探究不同腐殖酸浓度下参与含砷水铁矿转化的微生物类群组成和丰度变化及对砷释放的影响,预测原位高砷含水层中功能微生物群参与有机质—含砷铁矿物转化过程对砷转化释放的作用。【方法】对河套平原高砷地下水和同深度高砷沉积物中的铁还原功能群落进行富集培养,构建室内厌氧微宇宙体系,将富集菌群分别加入到实验室条件下合成的不同浓度腐殖酸(0、1.5、7、14 mg C/L)-含砷水铁矿体系中,通过体系中砷、铁形态及浓度的变化分析,结合高通量测序技术、X-射线衍射(X-ray diffractometer, XRD),探究不同条件下砷的释放固定和群落的演替。【结果】高砷地下水组(G组)和沉积物组(S组)富集得到的铁还原功能群落具有明显差异,G组中以Aeromonadaceae为特殊优势菌群,而S组中以Shewanellaceae为特殊优势菌群。微宇宙实验结果显示,S组的铁还原量相对较高且速率较快;G组与S组中液相砷形态存在明显差异,整个培养期内G组均以As(Ⅴ)为主,而S组中前期以As(Ⅴ)为主,当反应到达20 d时液相As(Ⅲ)高达3.4μmol/L,推测此时具有砷还原功能的群落占优势地位。当反应...     

砷矿区农田土壤微生物群落碳源代谢多样性

采用Biolog方法研究了砷(As)矿区农田土壤微生物碳源利用多样性及其与土壤化学性质的关系.结果表明:3种土壤的N、P、K、有机质(OM)、Cu和Zn全量依次为中As＞高As＞低As土壤.中As和高As土壤微生物平均吸光度以及多样性指数(H'、D、U)显著高于低As土壤.主成分分析与生理碳代谢指纹图谱分析表明,中As和高As土壤微生物对糖类、氨基酸类等碳源的利用程度显著高于低As土壤.典范对应分析显示,影响土壤微生物群落碳源代谢的主要有全P、OM、全Pb、全Zn、全N和pH,全As并不是最主要的影响因子.可见,养分是影响长期污染土壤微生物群落结构和功能多样性的主要因素.     

外源砷胁迫对两种土壤中细菌和古菌群落的影响

采矿和冶炼等活动会导致土壤中砷的累积,给农产品质量安全和土壤微生物带来不利影响。本文研究了外源砷进入黄壤(YS)和紫色砂页岩发育土壤(RS)后有效砷含量随时间的变化,并采用MiSeq高通量测序研究土壤中细菌和古菌在未加砷和外源砷胁迫1、30、360 d后的群落变化,探讨砷胁迫下土壤细菌和古菌的适应机制。结果表明: 外源砷进入土壤后,土壤有效砷含量随时间推移逐渐降低,并显著影响土壤细菌和古菌的群落组成。土壤细菌的优势菌群丰度变化显著;而古菌中仅丰度较低的菌群发生显著改变,优势菌群丰度变化较小,推测古菌群落具有高耐砷性和稳定性。与砷胁迫时间相比,土壤砷的有效性对细菌和古菌群落结构的影响更大。研究结果可为砷污染农田的安全利用及微生物修复等提供参考。     

耐砷促生菌对超富集植物蜈蚣草砷吸收及根际微生物群落的调控作用

【目的】分析添加外源促植物生长微生物(plant growth-promoting bacteria,PGPB)对植物生长、砷富集和根际微生物的影响,为植物-微生物联合修复砷污染土壤提供参考。【方法】通过添加外源PGPB,研究蜈蚣草生物量和砷富集量与外源微生物促植物生长特性的关系,采用高通量测序技术分析蜈蚣草根际微生物群落在外源PGPB干预下的变化规律。【结果】2株根际菌(假单胞菌PG12、芽孢杆菌R19)和1株内生菌(恶臭假单胞菌S6)具备典型的促植物生长特性,对蜈蚣草的促生作用顺序为：PG12>S6>R19,与对照组相比,生物量分别提高了234%(P<0.01)、136%(P<0.01)和67%;添加外源PGPB后,蜈蚣草砷含量从对照的18.50 mg提高到了31.25-46.95 mg,增幅高达153%(PG12)和139%(S6),对应的蜈蚣草砷浓度从2616.34mg/kg降至1348.04-2 156.23 mg/kg,呈现出典型的砷“稀释效应”;α多样性指数Sobs、Chao和Ace显示,仅R19处理显著提高了根际微生物的群落多样性,而β多样性指数...     

中国林蛙蝌蚪肠道及皮肤微生物多样性分析

肠道及皮肤微生物群落对宿主的健康有着至关重要的影响。本研究使用16S rRNA基因测序技术研究中国林蛙(Rana chensinensis)Gosner38期蝌蚪肠道和皮肤共生微生物群落组成之间的差异。在门水平上,林蛙蝌蚪肠道中的优势门为变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidetes),皮肤中的优势门为拟杆菌门、变形菌门和疣微菌门(Verrucomicrobia)。此外,显著性检验结果显示,肠道中厚壁菌门和放线菌门(Actinobacteria)的丰度显著偏高,皮肤中拟杆菌门的丰度显著偏高。肠道微生物群落多样性高于皮肤,而肠道和皮肤的物种丰富度之间无显著差异。在对KEGG通路的丰度进行比较时,结果显示肠道"环境信息处理"功能的丰度显著高于皮肤,而皮肤中"遗传信息处理"及"新陈代谢"功能显著偏高。本研究提示林蛙蝌蚪肠道及皮肤微生物群落组成之间存在显著差异。     

新疆石油污染土壤苯并(a)芘降解微生物多样性研究

为研究新疆石油开采区污染土壤中苯并(a)芘降解微生物的群落结构特点,采集克拉玛依石油开采区受污染程度不同的土壤样品, 以苯并(a)芘为唯一碳源、氮源的无机盐培养基五代富集培养, 采用PCR-DGGE 技术对第五代富集培养物的微生物群落结构开展研究, 根据DGGE 指纹图谱分析它们的遗传多样性。研究显示: 三个不同污染样品微生物多样性指数(H) 丰度(S)和均匀度(EH)均有所不同, 重度污染土样降解苯并(a)芘的微生物类群最为丰富, 其次是中度污染土样, 最少的是轻度污染土样; 并且三个样品微生物类群种类差异较大, 菌落相似性低。对 DGGE 的优势条带序列分析, 同源性最高的微生物分别属于变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)、放线菌门(Actinobacteria)和厚壁菌门(Firmicutes)。研究表明新疆石油开采区污染程度不同土壤中降解苯并(a)芘的微生物群落结构不同, 提示污染严重的环境里可能蕴藏着高效降解苯并(a)芘的微生物资源。     

A comparative study of gut microbiota profiles of earthworms fed in three different substrates

The gut microbiome of earthworms has a complex interdependence with the host. When the soil minerals pass through earthworm’s gut, they may affect the gut microbiota. To gain insight into the response of gut microbiota to the passed minerals, we fed earthworm (Eisenia fetida) on nutrient-poor soil and ore powder, and used high throughput sequencing to characterize the earthworm intestinal microbial community to find evidence for a core bacterial community of the E. fetida. The results showed that earthworms’ gut maintained a core microbiome that appeared in all samples. These core microbiota may play a significant role in a species’ environmental interactions. The composition of intestinal microbiomes varied with substrates. The earthworm guts from two nutrient-poor substrates had similar microbial communities and they were different from nutrient-rich substrate. Proteobacteria and Bacteroidetes were more abundant in the gut of earthworms kept on a nutrient-poor substrate such as ore powder or mineral soil than in the gut of earthworms kept in organic-rich compost soil; some of these microorganisms may help earthworms survive in nutrient-poor substrates.     

Earthworm invasions of ecosystems devoid of earthworms: effects on soil microbes

Recent studies document North American earthworm invasions and their profound effects on the structure of the soil profile, which is the habitat for soil microorganisms (mainly fungi and bacteria). Dramatic alterations made to these layers during earthworm invasion significantly change microbial community structure and therefore microbial activities such as C transformations. Understanding the impacts of earthworm invasion on the microbes themselves will give insight into earthworm effects on microbial activities. Bacterial and actinomycete communities in earthworm guts and casts have not been studied in environments recently invaded by earthworms. Earthworm invasion tended to decrease fungal species density and fungal species diversity and richness. The presence of earthworms decreased zygomycete species abundance probably due to disruption of fungal hyphae. Physical disruption of hyphae may also explain decreased mycorrhizal colonization rates, decreased mycorrhizal abundance and altered mycorrhizal morphology in the presence of earthworms. Mixing of organic layers into mineral soil during earthworm invasion tended to decrease microbial biomass in forest floor materials while increasing it in mineral soil. In newly invaded forest soils, microbial respiration and the metabolic quotient tended to decline. In forests where either the microbial community has had time to adapt to earthworm activities, or where the destruction of the forest floor is complete, as in invasions by the Asian Amynthas hawayanus, the presence of earthworms tends to increase the metabolic quotient indicating a shift to a smaller, more active microbial community.     

Viability of gut microbes as a complementary earthworm biomarker of metal exposure

Earthworms are standard species used in soil ecotoxicology to evaluate the adverse effects of soil contaminants. This study proposes the assessment of the viability of earthworm gut microbes as an indicator in a site-specific test of soil toxicity. Using slow centrifugation, the microbial community was extracted from the guts of earthworms that had been exposed to copper (Cu)- or nickel (Ni)-contaminated soil. Microbial cell viability was assessed using calcein acetoxymethyl ester staining and flow cytometric analysis. We confirmed a metal concentration-dependent decrease in the cell viability of the gut microbial community. The general endpoints, including survival, abnormalities, coelomocyte activity, and metal bioaccumulation, showed a metal concentration-dependent response, and were strongly associated with gut microbial viability in the Ni-exposure group. In contrast, the general endpoints in the Cu-exposure group were significantly different from those in the former group, because the soil penetration rate of the earthworms was very low on the Cu-contaminated soil. Our results indicated that the gut microbial community viability assay holds potential for assessing the toxicity of soil to field worms by simply and rapidly monitoring the viability of the earthworm gut microbial community.     

The effect of anthropogenic arsenic contamination on the earthworm microbiome

Earthworms are globally distributed and perform essential roles for soil health and microbial structure. We have investigated the effect of an anthropogenic contamination gradient on the bacterial community of the keystone ecological species Lumbricus rubellus through utilizing 16S rRNA pyrosequencing for the first time to establish the microbiome of the host and surrounding soil. The earthworm‐associated microbiome differs from the surrounding environment which appears to be a result of both filtering and stimulation likely linked to the altered environment associated with the gut micro‐habitat (neutral pH, anoxia and increased carbon substrates). We identified a core earthworm community comprising Proteobacteria (～50%) and Actinobacteria (～30%), with lower abundances of Bacteroidetes (～6%) and Acidobacteria (～3%). In addition to the known earthworm symbiont (Verminephrobacter sp.), we identified a potential host‐associated Gammaproteobacteria species (Serratia sp.) that was absent from soil yet observed in most earthworms. Although a distinct bacterial community defines these earthworms, clear family‐ and species‐level modification were observed along an arsenic and iron contamination gradient. Several taxa observed in uncontaminated control microbiomes are suppressed by metal/metalloid field exposure, including eradication of the hereto ubiquitously associated Verminephrobacter symbiont, which raises implications to its functional role in the earthworm microbiome.     

蚯蚓在植物修复芘污染土壤中的作用

采用盆栽试验法,研究了蚯蚓(Pheretima hupeiensis)在植物修复芘污染土壤中的作用。结果显示,试验浓度(20.24-321.42 mg/kg) 范围内,蚯蚓活动促进了芘污染土壤中修复植物黑麦草(Lolium multiforum)黑麦草的生长,其根冠比明显增大。添加蚯蚓72 d后,种植黑麦草的土壤中芘的去除率高达60.01%-86.26%,其平均去除率(74.66%)比无蚯蚓活动的土壤-植物系统(64.55%)提高10.11%,比无植物对照组(18.24%)提高56.42%。各种生物、非生物修复因子中,植物-微生物交互作用对芘去除的平均贡献率(51.75%)最为突出,比无蚯蚓活动时(44.94%)提高6.81%。说明蚯蚓活动可强化土壤-植物系统对土壤芘污染的修复作用。     

Changes in diversity, protein content, and amino acid composition of earthworms from a paddy soil under different long-term fertilizations in the Tai Lake Region, China

Influence of the agricultural management practices on soil quality and the ecosystem functioning has been an increasing concern in soil science and ecology with sustainable agriculture. This study deals with the changes of soil earthworm communityfrom a paddy soil under different long-term fertilizations. The soil earthworms were collected and counted from different fertilizer treated plots in the field after the rape harvest in May 2004, and their taxonomic groups were determined under a binocular stereoscope at the laboratory. The body of the earthworm (Metaphire californica) was crushed by a cell crusher to collect protein, and the protein molecules with different sizes were analyzed by electrophoresis. Furthermore, the Metaphire californica collected was hydrolyzed and the aliquots were subject to an amino acid auto-analyzer. The results showed that totally seven species of earthworms were recognized in the paddy field with the number varying with different fertilization treatments. The structure of earthworm communities was dramatically affected by the fertilization practice. Under chemical fertilization only, both the number of earthworm species and the quantity of individuals were significantly smaller than those under other treatments, or even than those under no fertilization. Furthermore, there was an obvious decrease in the total amino acid and the contents of most individual amino acids of Metaphire californica under chemical fertilization only, compared with those under the combined fertilization of chemical and organic fertilizers. Although chemical fertilizers in combination with rice straw return increased earthworm amino acid content, long-term pig manure application tended to increase earthworm protein content. As a molecular footprint, long-term chemical fertilization caused a reduction in the content of protein with MW less than 25 kd, but a significant increase in that of protein with molecule size around 33 kd. Our study demonstrated that different fertilizations affected not only earthworm population but also diversity and richness in the paddy soil after 16 years of treatment, and that long-term chemical fertilization may impact the soil animal community and, thus, influence the paddy ecosystem functioning for yield stability. This study implicated that not only the community structure but also the amino acid metabolism for life functioning of earthworms in cropland soils may pose significant responses to the agricultural management practices.     

Specific features of nitrogen transformation in the gut and coprolites of earthworms

It has been found that nitrogenase activity in the guts and coprolites of three earthworm species (Lumbricus terrestris, Aporrectodea rosea, and Aporrectodea caliginosa) is one to three orders of magnitude higher than that in the control soil. In A. caliginosa earthworms, the actual nitrogenase activity remains high upon keeping them in soils of different types. However, it reaches a peak in the gut lumen and decreases to a minimum on its wall, which is evidence for a major role of transitional microflora in intestinal nitrogen fixation. Nitrogen fixation activity in freshly excreted coprolites decreases to one-half or one-third of that in the gut and then increases again, reaching a peak on days 3–5 of exposure in the soil. According to the results of multisubstrate testing, the functional diversity of nitrogen-fixing soil microorganisms increases in the course of passage through the earthworm gut. Thus, the microbial community in coprolites retains its functional potential and, within a few days, shows the second peak of activity in the soil. Due to a short-term increase in the rate of nitrogen fixation, coprolites contain a pool of bound amino acids, which become involved in the formation of new humus substances.     

Comparative Assessment of the Aerobic and Anaerobic Microfloras of Earthworm Guts and Forest Soils

Aerobic and anaerobic microbial potentials of guts from earthworms (Lumbricus rubellus Hoffmeister and Octolasium lacteum (Oerl.)) collected from a beech forest were evaluated. On the basis of enumeration studies, microbes capable of growth under both aerobic and anaerobic conditions were more numerous in the earthworm intestine than in the beech forest soil from which the worms were obtained. The intestine of worms displayed nearly equivalent aerobic and anaerobic microbial growth potentials; in comparison, soils displayed greater aerobic than anaerobic microbial growth potentials. Hence, the ratio of microbes capable of growth under obligately anaerobic conditions to those capable of growth under aerobic conditions was higher with the worm intestine than with the soil. Process level studies corroborated these population differentials: (i) under anaerobic conditions, worm gut homogenates consumed glucose, cellobiose, or ferulate more readily than did soil homogenates; and (ii) under aerobic conditions, worm gut homogenates consumed cellobiose or oxygen more readily than did soil homogenates. Collectively, these results reinforce the general concept that the earthworm gut is not microbiologically equivalent to soil and also suggest that the earthworm gut might constitute a microhabitat enriched in microbes capable of anaerobic growth and activity.     

Eisenia fetida (Oligochaeta, Lumbricidae) Activates Fungal Growth, Triggering Cellulose Decomposition During Vermicomposting

Cellulose is the most abundant polymer in nature and constitutes a large pool of carbon for microorganisms, the main agents responsible for soil organic matter decomposition. Cellulolysis occurs as the result of the combined action of fungi and bacteria with different requirements. Earthworms influence decomposition indirectly by affecting microbial population structure and dynamics and also directly because the guts of some species possess cellulolytic activity. Here we assess whether the earthworm Eisenia fetida (Savigny 1826) digests cellulose directly (i.e., with its associated gut microbiota) and also whether the effects of E. fetida on microbial biomass and activity lead to a change in the equilibrium between fungi and bacteria. By enhancing fungal communities, E. fetida would presumably trigger more efficient cellulose decomposition. To evaluate the role of E. fetida in cellulose decomposition, we carried out an experiment in which pig slurry, a microbial-rich substrate, was treated in small-scale vermireactors with and without earthworms. The presence of earthworms in vermireactors significantly increased the rate of cellulose decomposition (0.43 and 0.26% cellulose loss day⁻¹, with and without earthworms, respectively). However, the direct contribution of E. fetida to degradation of cellulose was not significant, although its presence increased microbial biomass (C_mic) and enzyme activity (cellulase and β-glucosidase). Surprisingly, as fungi may be part of the diet of earthworms, the activity of E. fetida triggered fungal growth during vermicomposting. We suggest that this activation is a key step leading to more intense and efficient cellulolysis during vermicomposting of organic wastes.