蚯蚓和丛枝菌根真菌对南瓜修复多环芳烃污染土壤的影响

在温室盆栽实验条件下,研究接种AM(arbuscular mycorrhiza)真菌、蚯蚓(Eisenia fetida)对南瓜(Cucurbita moschata)修复3环以上多环芳烃(PAHs)污染农田土壤的影响,试验设置单接AM真菌、单接蚯蚓、双接AM真菌和蚯蚓、不接种的对照共4个处理,播种10周后收获。结果表明,接种AM真菌和蚯蚓促进AM真菌侵染南瓜,增加南瓜生物量;显著提高南瓜修复土壤中Phe(菲)、An(t蒽)、Py(r芘)、BkF(苯并(k)荧蒽)、BaP(苯并(a)芘)、BPe(r苯并(g,h,i)苝)等PAHs污染物的效率,促进南瓜高效地吸收3~5环PAHs,尤其是AM真菌和蚯蚓共同接种条件下对南瓜修复土壤效果最优;AM真菌利于南瓜转移根系吸收的高浓度PAHs化合物至地上部,降低PAHs对根系的胁迫,增强南瓜在高浓度PAHs污染土壤中存活,有利于南瓜应用于高浓度PAHs污染土壤的高效修复;蚯蚓对南瓜地下部吸持3~5环高分子量的PAHs化合物有积极作用。因此,选用的AM真菌和蚯蚓在土壤中具有协同作用,促进南瓜高效修复PAHs污染土壤。     

混合菌降解土壤中多环芳烃的试验研究

PAHs生物降解程度受多种因素影响。通过筛选驯化PAHs降解菌,研究混合菌对土壤中菲、芘、苯并（a）蒽、苯并（b）荧蒽、苯并（k）荧蒽、茚并（1,2,3-cd）芘的生物降解性能,并考察污染时间对土壤中PAHs降解效果的影响。结果表明,筛选的混合菌具有很强的PAHs降解能力,缩短了PAHs生物降解的半衰期,且PAHs起始降解速率较快,之后趋于平缓。27d内土壤中的菲、芘、苯并（a）蒽、苯并（b）荧蒽、苯并（k）荧蒽、茚并（1,2,3-cd）芘的平均降解率分别为98.14%、89.97%、88.47%、63.55%、65.24%、60.49%,其中菲在5d之内的降解率高于93%。污染210d的土壤中各PAHs的起始降解速率高于污染50d的土壤,因此污染时间越长,PAHs生物降解的停滞期越短。     

南京有机污染风险区农田土壤多环芳烃污染状况研究

采集南京地区不同有机污染风险区农田表层土壤,用超快速液相色谱仪检测样品中15种EPA优控的多环芳烃(PAHs)含量。结果表明,被检农田土壤多环芳烃总量分布于306.0~1251.3μg kg~(-1)之间,均值682.0μg kg~(-1),四环以上高环多环芳烃占较大比例(80%)。根据欧洲土壤质量标准,所检土壤样本已达污染水平。不同风险污染区农田土壤PAHs的含量由高至低为:钢铁工业区、有机垃圾处理区、化工工业区及炼油工业区。钢铁工业区附近主要的污染物为荧蒽、芘、屈和苯并[a]蒽,分别占到污染物总量的16%、13%、10%和10%。采用荧蒽/(荧蒽+芘)与茚并[1,2,3-cd]芘/(茚并[1,2,3-cd]芘+苯并[g,h,i]苝)比值对各地污染物来源进行分析,结果发现调查区域的PAHs污染物以燃烧源为主,生物质燃料为主要污染物,部分地区同时有石油燃烧污染。     

东莞市土壤中多环芳烃的含量、代表物及其来源

2002年10~12月采集东莞市各地土壤样品64个,采用气相色谱-质谱仪对土壤中的16种优先控制的多环芳烃(PAHs)进行分析。结果显示,东莞市土壤中16种PAHs平均含量413μg/kg,含量较高的几种组分分别是菲、萤蒽、屈、苯并[b]萤蒽、芘。与PAHs总含量相关性较好的7种组分分别为屈、苯并[a]蒽、苯并[b]萤蒽、苯并[k]萤蒽、苯并[a]芘、茚并[1,2,3]芘、二苯并[a,h]蒽;所有相关系数R2均超过0.9000,可作为东莞市土壤中PAHs的代表物。北部平原水乡的PAHs污染明显高于南部丘陵山区、水乡上游保护区以及中部过渡带。化石燃料高温燃烧产物的大气沉降为东莞市土壤中PAHs的主要来源。东莞市农业土壤中PAHs含量显著低于天津市受污染严重的农业土壤。     

多环芳烃高效降解菌的筛选

以多环芳烃(PAHs)菲、蒽、芘、■和苯并(a)芘为供试物,对土著混合菌和引进菌同时进行筛选实验。结果表明,引进菌和土著混合菌经过驯化后对菲、蒽、芘、■和苯并(a)芘均具有一定的降解能力。其中,在pH=6时,混合菌U03在48h内对5种PAHs的降解率均相对较高,分别为:72.38%;64.46%;65.77%;66.49%和64.77%,并且其的降解速率在各菌剂中同样最快,通过SPSS数理统计分析软件对数据进行处理后得出,混合菌U03可在较短时间内达到较好的降解目的。室内模拟试验证明混合菌U03具有较强的降解PAHs的能力,混合菌的协同作用有利于污染土壤中PAHs的降解。     

活性炭对土壤中多环芳烃生物有效性的影响

多环芳烃(PAHs)是土壤中污染较为严重的一类持久性有机污染物，对农产品安全和人体健康构成严重威胁。本研究在某场地采集不同土层土壤，通过室内培养实验，研究添加5%活性炭对土壤中PAHs残留的影响，并利用羟丙基-β-环糊精提取方法评价土壤中PAHs生物有效性的动态变化，以期为治理石油泄露等突发性环境污染事件造成的土壤PAHs污染提供修复技术参考。结果表明，在8个月内，添加活性炭组土壤中菲、芘的残留量显著高于对照组，苯并[b]荧蒽的残留量显著低于对照组，苯并[ghi]芘残留量在两处理组之间无显著差异。在第1个月时，活性炭组土壤中菲、芘、苯并[b]荧蒽生物有效性较对照组分别降低了35.06%、37.73%、39.60%，苯并[ghi]芘生物有效性仅降低了1.37%。随着时间延长活性炭对土壤中PAHs生物有效性的阻控效果呈减弱趋势。因此，土壤添加5%活性炭能够有效降低PAHs生物有效性，减少其环境风险，并且在添加活性炭1个月内阻控效果最佳。     

小麦秸秆生物炭对石油烃污染土壤的修复作用

以小麦秸秆为原材料,在300℃下缺氧裂解3、6、8 h制备生物炭,比较了3种生物炭的产率、pH值、灰分以及C、H、N元素含量,表征了300℃、6 h生物炭的表面形态,并用其作为修复材料,对大港油田的石油污染土壤进行修复。结果表明,随裂解时间的延长,生物炭产率下降,pH值升高,灰分含量增加,H/C值下降,但产率、pH值、灰分和H/C值都是从3 h到6 h差异显著,6 h到8 h差异不显著。C元素含量先升高后下降。石油污染土壤经生物炭修复14 d和28d后,总石油烃降解率分别为45.48%和46.88%,均显著高于对照组。修复14 d后土壤中的萘、苊、苯并[a]蒽、屈、苯并[b]荧蒽、苯并[k]荧蒽、苯并[a]芘、茚并[1,2,3-cd]芘也都有不同程度的下降,其中苯并[a]芘含量下降幅度达98.18%,其他几种PAH的降解率也都高于对照组,28 d后这些PAH的含量又有上升趋势。这说明小麦秸秆裂解时间对生物炭的性质有影响;300℃、6h生物炭可以用来修复石油污染的土壤。     

甜菜与牧草间作对多环芳烃污染土壤的修复作用

通过盆栽试验方法,选择经济作物甜菜和牧草类黑麦草、苏丹草、香根草为供试植物,研究了甜菜与3种牧草分别间作及各自单作对多环芳烃（PAHs）菲、荧蒽、芘和苯并[a]芘污染土壤修复作用。结果显示：经6个月连续两茬种植试验后,所有种植植物的处理中土壤PAHs的去除率均高于无植物种植组,间作种植土壤PAHs的去除率高于单作,黑麦草、苏丹草、香根草与甜菜间作对土壤PAHs的去除率分别达到84.85%、79.96%、84.11%;在土壤污染条件下,间作模式更有利于甜菜生长;种植植物增强了土壤中多酚氧化酶和过氧化氢酶的活性,间作模式下二者活性高于单作4.37%~43.07%,过氧化氢酶较多酚氧化酶对PAHs土壤污染更敏感,可作为关键酶用于评价土壤PAHs污染状况。在不影响农业生产的前提下,修复植物牧草和经济作物甜菜间作种植模式显著提高了土壤PAHs的降解率。     

某氮肥厂场地土壤PAHs污染特征研究

采用现场采样及室内测试方法对广州某氮肥厂原料车间和油库区土壤中16种优控多环芳烃（PAHs）的含量进行调查研究,分析了EPAHs含量及其组成特征和垂直分布特征,并在此基础上进行了源解析。结果表明,分析样品中∑PAHs范围在10-7795μg·kg,原料车间土壤中的∑PAHs小于油库区土壤中的,菲、芘、荧蒽、并（b）荧蒽、苯并（a）芘为主要污染物;油库土壤0-40cm的样品中16种PAHs均有检出,∑PAHs和单体分布基本一致;原料车间土壤∑PAHs和单体浓度随着地面深度的增加而减少。通过对单组分比值（菲／蒽,荧蒽／芘）的分析可以看出油库区土壤中PAHs来源于石油和燃烧源,而原料车间污染源主要为燃烧源。     

江苏省典型生态示范区土壤中多环芳烃的含量与风险评价

采集了江苏省某典型生态示范区内58个监测单元的土壤样品,并对样品中的多环芳烃(PAHs)进行了定量分析。结果表明,生态示范区土壤中的PAHs平均含量较低,但不同土壤样品之间PAHs的变异系数较大,以二苯[a,h]并蒽、苯并[a]蒽、苯并[b]荧蒽、苯并[k]荧蒽为主要组成成分。从8个生态类型区分析,工矿企业搬迁区PAHs含量最高,达49.196μg/kg,其次是化工区周边,农业科技园土壤中的PAHs含量最低。工业区的PAHs多来源于石油输入,而以农业为主的生产基地、科技园等,PAHs主要来源于化石燃料的不完全燃烧。生态风险评价结果显示生态示范区内的PAHs处于较低毒性水平,尚未对生物造成不利的影响。     

高产漆酶平菇的筛选及其在降解黄曲霉毒素B1中的应用

通过选择培养基平板培养法和液体发酵培养法筛选得到2株高产漆酶的平菇菌株P1和P2,并对平菇菌株产漆酶的培养基进行筛选,得到产漆酶的最适培养基为最低盐MSM培养基。菌株P1不仅产漆酶能力最高,而且降解黄曲霉毒素的能力也最好。在MSM培养基中培养10d时,产漆酶量高达769.44U/L,在800μl的反应体系中,790μl粗酶液可以将1000ng黄曲霉毒素B₁降解到222.62ng,降解率为77.74%,并且平菇粗酶液降解黄曲霉毒素B₁的能力与其中漆酶的含量呈一定的正相关性。     

Laccases: toward disentangling their diversity and functions in relation to soil organic matter cycling

Degradation of the recalcitrant polyphenolic plant residue lignin is a bottleneck of element turnover in terrestrial ecosystems. Consequently, there is a great interest to understand underlying mechanisms and dynamics, considering the possible ecological roles of soils as sinks or sources of carbon dioxide. The present review provides a critical, holistic view of the ecological importance of the degradation of recalcitrant residues attributed to laccase-producing soil microbes and laccase activity under different environmental conditions. We synthesize and discuss the results of previous classical ecological, enzymatic, and molecular-ecological studies to point out discrepancies between gene detection, enzyme activity, and substrate degradability. We single out major hindrances to current research and outline a progression toward a better understanding of laccase activity by fungi in soil ecosystems.     

Fungal communities in bulk soil and stone compartments of different forest and soil types as revealed by a barcoding ITS rDNA and a functional laccase encoding gene marker

The soil fungal diversity and community partitioning between the bulk soil and stone compartments was investigated using PCR based approaches targeting the barcoding internal transcribed spacer (ITS) of rDNA and the laccase encoding functional gene as genetic markers. Soil samples were collected from the B-horizon of spruce and beech forests at the Hainich Biodiversity Exploratory, central Germany. The targeted markers were amplified from the respective DNA extracts using general fungal primers and basidiomycete laccase gene specific primers, cloned and sequenced. Differences in the fungal community composition between the two forest types and the soil compartments were indicated by both markers. When the effects of ecological factors were considered, the two markers produced different patterns of results. The ITS rDNA marker revealed communities principally influenced by forest type, while those detected with the functional marker were mainly affected by soil pH. The fungal communities detected by the functional marker in particular, differed significantly between soils and stones, indicating that laccase-producing fungi are specifically adapted to degrade organic matter in soils rather than weathering of stones. The study underlines the fact that coherent and complementary results may be obtained with both genetic markers used.     

Lyophyllum sp. strain Karsten alleviates pH pressure in acid soil and enhances the survival of Variovorax paradoxus HB44 and other bacteria in the mycosphere

In previous work, Variovorax paradoxus strain HB44, next to Burkholderia terrae BS001 and Dyella japonica BS003, were found to be selected in the mycosphere of the tricholomataceous fungi Laccaria proxima and Lyophyllum sp. strain Karsten in an acid soil denoted G. V. paradoxus HB44 showed poor survival in G bulk soil, irrespective of prior soil sterilization, and this poor survival also occurred for B. terrae BS001 and D. japonica BS003. In contrast, the survival rate of strain HB44 in two other soils, with pH values > 5.5, was significantly raised. Also, significantly enhanced strain HB44 survival in G soil was found if the pH was raised to 5.5 or 6.5, and it was even shown to grow (in the presence of the exogenous carbon source glycerol) at such pH values in the sterile G soil. This behaviour was similar to that of the V. paradoxus type strain. Strikingly, Lyophyllum sp. strain Karsten, when colonizing the sterilized G soil, significantly raised the soil pH from about 4.6 to ≥5.0. The pH raise was dependent on time, hyphal development, as well as on initial soil pH, but was consistent throughout. The modulated soil pH conditions were shown to be permissive for the survival and growth of strain HB44, and this was extended to strains BS001 and BS003. These findings corroborate the hypothesis that L. sp. strain Karsten provides a suitable habitat for acid-sensitive strains like HB44, BS001 and BS003 in its mycosphere in acid soil, which is strongly defined by the establishment of a growth-permissive pH.     

Effect of seed inoculation with the rhizopseudomonad strain 7NSK2 on the root microbiota of maize (Zea mays) and barley (Hordeum vulgare)

Summary The addition of sugars or amino acids to the soil gave rise to the development of different groups of microorganisms. The increase in the number of different groups of microorganisms in the soil had an influence on the microbiota in the rhizoplane and endorhizosphere of maize and barley grown in that soil. Furthermore, growth of maize and barley decreased with increasing microbial activity and density in soil. This effect could be counteracted effectively by the rhizopseudomonad strain 7NSK2. The beneficial effect of the strain 7NSK2 correlated inversely with the microbial activity, as measured by soil respiration, in the bulk-pretreated soil.The effect of seed inoculation with the rhizopseudomonad strain 7NSK2 on the root microbiota of maize and barley was evaluated. The strain 7NSK2 was capable of colonizing the rhizoplane and endorhizosphere of the maize cultivar Beaupré and barley cultivar Than very effectively and of considerably altering their composition. The number of total bacteria, fungi, pseudomonads and coliform bacteria in the rhizoplane and endorhizosphere of both plants was strongly reduced by inoculating the seeds with the strain 7NSK2.     

硅酸盐细菌NBT菌株解钾效能及对钾的吸持作用

在摇瓶和土壤耗竭条件下研究了硅酸盐细菌NBT菌株的解钾作用以及对作物生长的促进作用。结果表明 ,在摇瓶条件下 ,培养 120h ,NBT菌株可以从钾长石中释放K 159.1mg/L ,比接灭活菌对照 (48.8mg/L)增加 226.02% ;耗竭条件下NBT菌株在未灭菌土壤中的解钾作用与在灭菌土壤中的解钾作用相当。在未灭菌土壤中 ,NBT菌株释放的矿物钾占植株吸钾量的 14.4 %～ 4 3.1% ;不接菌或接灭活菌处理土壤中矿物钾的释放量为零或极少。NBT菌株的解钾效能与土壤中速效性钾及有机质含量密切相关。土柱试验表明 ,供试土壤接种硅酸盐细菌后的土壤滤液中流失的钾比接种灭活硅酸盐细菌后的土壤滤液中流失钾减少 29.6 %～56.5% ;硅酸盐细菌NBT菌株荚膜多糖吸附钾的量占加入钾的量的 31.8%～ 69.4 %。NBT菌株的吸钾作用与NBT菌株本身及荚膜多糖的多少密切相关。     

湖南邵阳地区高液限红黏土干缩裂隙演化过程的量化分析

红黏土对环境湿度变化非常敏感,在干燥环境中极易开裂,纵横交错的裂缝网络损害了土壤结构的完整性,很容易诱发红黏土边坡失稳和崩塌,导致农田水利设施的破坏,甚至加剧整个生态环境的水旱灾害。为探究红黏土裂隙的演化规律,该研究采用自制试验装置和三维应变测量系统开展了自然湿热条件下的红黏土泥浆样干燥试验,通过采集土体水分和土表位移、应变和裂隙的变化,定量分析脱湿过程中土体表面裂隙形态和应变场的演变特征,并进一步探讨水分变化对裂隙形态和应变场的影响。结果表明：1）土样表面干缩裂隙的演化一共经历了6个阶段,后阶段裂隙分割前阶段裂隙围成的区域,且不同阶段裂隙的交叉角接近90o;2）裂隙产生初始,裂隙尖端处拉应变约为0.5%,土表面大部分区域处于受拉状态;随着裂隙进一步发展,裂隙周边土体逐渐由拉应变状态向压应变状态转变;当所有裂隙发育完成,裂隙周边土体处于压应变状态;3）裂隙演化阶段与界限含水率有关,当泥浆土样的含水率接近液限时（67.7%）,土体表面裂隙开始发育,裂隙迅速张开和延伸;当土的含水率达到塑限时（28.3%）,裂隙发展的速率逐渐变缓;当含水率小于缩限时（18.8%）,裂隙变化已经很小,裂隙发育接近完成;4）在裂隙演化过程中,早期裂隙的发展持续时间和裂隙宽度均超过后期裂隙;土表不同位置的位移和应变均不相同,土块中心竖向收缩大于边缘竖向收缩,而土块中心位移及应变均小于土块边缘,研究可为红黏土开裂引发的工程地质灾害的预防及治理提供参考。     

Promotion of plant growth and in situ degradation of phenol by an engineered Pseudomonas fluorescens strain in different contaminated environments

We show that Pseudomonas fluorescens strain P13, a plant growth-promoting bacterium, enhanced the growth of corn in uncontaminated soil but not in contaminated soil, perhaps because of its inability to reduce phytotoxicity. Another bacterial strain, Pseudomonas aeruginosa strain SZH16, showed in situ phenol-degrading activity and contained a plasmid loaded with a gene encoding for catechol 2, 3-dioxygenase, an important enzyme in the degradation pathway of aromatic compounds. We implanted this biodegradation ability into strain P13, using horizontal gene transfer techniques using strain SZH16 as the donor and P13 as the recipient, to generate a phenol-degrading transconjugant which obtained the effective plasmid from strain SZH16. Introduction of the transconjugant P13 strain into an artificially phenol-spiked soil promoted the growth of corn and in situ phenol degradation, and the increase in plant biomass correlated with the decrease in soil phenol content. Furthermore, the transconjugant P13 strain was also found to stimulate corn growth and reduce phenol concentration in water containing phenol and in historically contaminated field soils, indicating that the transconjugant strain could promote plant growth in both contaminated and uncontaminated environments. The transconjugant P13 strain was more efficient than either strain P13 or SZH16, and shows how plant growth-promoting bacteria which show no, or only limited, ability to degrade organic pollutants may be modified. This technique is attractive for many environmental remediation and agronomic applications.     

Quantification of the biocontrol agent Pseudomonas fluorescens Pf153 in soil using a quantitative competitive PCR assay unaffected by variability in cell lysis- and DNA-extraction efficiency

Although often neglected, variability in cell lysis efficiency and DNA extraction yield represents the major hurdles of any polymerase chain reaction (PCR)-based quantification protocol in soil and other natural environments. In this study we developed a technique that minimizes the effects of these constraints, providing at the same time a reliable internal control to distinguish between PCR-inhibition and negative results. We used Pseudomonas fluorescens Pf153, a root-colonizing bacterium that shows biocontrol activity against tobacco and cucumber black root rot, as the target organism for PCR quantification. Prior to DNA extraction, the genetically engineered, cognate reference strain P. fluorescens CHA0/c2 was inoculated in a reference soil. CHA0/c2 in the reference soil and Pf153 in the soil sample were lysed in parallel and afterward the lysates were mixed in known proportions. CHA0/c2 carries the plasmid pME6031-cmp2 that contains an allelic variant (competitor) of the Pf153 specific sequence Pf153_2. In a quantitative competitive PCR (QC-PCR) assay the competitor allows the quantification of the target strain down to 0.66 Pf153 CFU/mg soil. Processing the reference strain in the same way as Pf153 enables the exact quantification of the target strain in biocontrol assays performed in natural soil, overcoming differences in DNA extraction efficiency and PCR amplification from different soil environments. This technique is easily adaptable to other Pseudomonas strains simply by replacing the competitor used here with one derived from a SCAR-marker which is specific for the strain of choice.     

Survival of myxobacter strain 8 in natural soil in the presence and absence of host cells

Myxobacter strain 8 is one component of a sequence of three predatory bacteria that develop in soil when Micrococcus luteus host cells are added to the soil. The survival of strain 8 in the presence and absence of added host cells in natural soil not allowed to dry out was examined. Strain 8 vegetative cells died relatively rapidly in unamended soil. Death was faster and occurred to a greater extent in acidic than in neutral pH soil. However, in both cases death was accompanied by formation of sonication-resistant myxospores so that they comprised the ultimate population. These myxospores survived for prolonged periods in both acidic and neutral pH soils.Vegetative cells added in high numbers to soil did not multiply under any of the conditions tested. They did multiply, however, when they were added in low numbers to soil (including acidic soil) receiving sequential (additive) amendments of heart infusion broth or living M. luteus cells. This multiplication produced strain 8 cell numbers approximating those in the above experiments receiving high strain 8 cell number inoculations. Possibly, this represents a maximum vegetative cell number for soil.Germination of the myxospores in soil, followed by growth, seemed to require an approximately neutral pH and the presence of a proper host organism. Germination occurred with M. luteus as host, but not with Escherichia coli. A delayed germination occurred when sequential amendments of heart infusion broth, instead of M. luteus host cells, were made, but this could reflect a growth response by some indigenous components of the soil microflora that then served as host cells for germination.