生物表面活性剂-微生物强化紫花苜蓿修复多环芳烃污染土壤

在温室盆栽条件下,通过单独或联合添加生物表面活性剂鼠李糖脂(RH)和接种多环芳烃专性降解菌(DB),研究了利用生物表面活性剂-微生物强化紫花苜蓿(Medicago sativa L.)修复多环芳烃(PAHs)长期污染土壤的效果.结果表明,添加鼠李糖脂和接种PAHs专性降解菌能促进紫花苜蓿的生长和土壤中PAHs的降解.培养90d后,RH、DB处理的PAHs的降解率分别为30.0%和49.6%,均高于对照处理(CK)(21.7%).RH+DB处理PAHs的降解率最高达53.9%,说明鼠李糖脂和PAHs专性降解菌协同作用显著.另外,随着PAHs苯环数的增加,其平均降解率逐渐降低,但是接种PAHs专性降解菌能够提高4环和5环PAHs的降解率.同时也发现土壤中脱氢酶活性和PAHs降解菌数量越高的处理,土壤PAHs的降解率也越高.所以添加鼠李糖脂和接种PAHs专性降解菌能够有效促进土壤多环芳烃降解.     

菇渣和鼠李糖脂联合强化苜蓿修复多环芳烃污染土壤

采用植物生物量、多环芳烃含量、土壤微生物数量、土壤酶活性和土壤微生物功能多样性等多个指标,通过菇渣、鼠李糖脂和植物的单独及联合作用的盆栽试验,评价了菇渣和鼠李糖脂联合强化苜蓿修复多环芳烃(PAHs)污染土壤的效果.结果表明,60d内,苜蓿单独修复(AL)的降解率仅为14.43%,菇渣、鼠李糖脂联合苜蓿修复(GZ+RH0.5+AL和GZ+RH1.0+AL)显著提高了PAHs降解率,达到了32.64%和36.95%,比AL处理提高了115.45%和156.06%.与AL相比,GZ+RH1.0+AL对植物生物量提高程度最大,地上和地下生物量分别达到了1.05g/盆和0.20g/盆.在修复过程,GZ+RH1.0+AL显著提高了土壤细菌和真菌的数量,分别达到了31.37×106CFU·g-1和5.86×106CFU·g-1,特别是多环芳烃降解菌数量达到了39.57×105MPN·g-1,分别是对照(CK)和植物单独处理(AL)的29倍和4倍.就土壤脱氢酶活性而言,菇渣和苜蓿联合作用(GZ+AL)处理的活性最高,GZ+RH0.5+AL和GZ+RH1.0+AL的活性次之,分别为90.57、67.56和21.02μg/(g·d).此外,与对照(CK)相比,菇渣、鼠李糖脂和苜蓿的联合作用(GZ+RH1.0+AL)显著提高了土壤微生物群落的功能多样性.因此,菇渣和鼠李糖脂联合强化苜蓿修复PAHs污染土壤达到了比较理想效果,大面积田间试验有待进一步验证该方法的可行性.     

生物源表面活性剂对焦化土壤中多环芳烃增溶效能研究

考察了单一生物源表面活性剂鼠李糖脂、黄腐酸、槐糖脂、烷基糖苷、无患子皂苷及鼠李糖脂与各非离子生物源表面活性剂复配体系对萘和菲的增溶效能,并将优选的淋洗剂体系应用于焦化场地土壤多环芳烃（PAHs）的去除.结果表明：相较其他生物源表面活性剂,鼠李糖脂和黄腐酸对萘的增溶效果更好;对于菲的增溶,槐糖脂的增溶效果更好,这与其较低的临界胶束浓度和多环芳烃的性质有关.生物源表面活性剂复配体系的增溶能力较单一表面活性剂明显增强,以3∶1复配时增溶效果最佳.60 mmol·L^-1的鼠李糖脂-黄腐酸复配体系（3∶1）对土壤中萘的去除率可达96.64%,分别是单一鼠李糖脂和黄腐酸的1.09倍和2.21倍;而对土壤中蒽、苯并（a）蒽和苯并（k）荧蒽的去除率远低于萘,分别为63.31%、42.51%和39.10%,这是由于PAHs与鼠李糖脂-黄腐酸复配体系的相互作用随苯环数的增加而减弱.本研究可为生物源表面活性剂及其复配体系高效修复PAHs污染土壤提供科学依据.     

多环芳烃类污染土壤强化生物修复技术研究及应用

针对多环芳烃类(PAHs)污染土壤,采用富集培养的方法分离筛选得到高效PAHs降解菌,并作为外源添加至高效复合微生物降解菌剂中运行过程中控制土壤含水率、微生物菌剂、营养物质的投加比例以及人工翻抛频率,开展强化生物修复试验。土壤中PAHs的降解主要出现在前3个月,之后进入生物降解拖尾阶段,土壤中各PAHs的浓度下降缓慢。其中,堆体土壤中低环PAHs、中环PAHs和高环PAHs的最高去除率分别为66.50%、71.78%和56.99%,16种PAHs的平均去除率为64.06%。分别从生物可利用性、微生物激活营养剂、提高供氧效率和微生物菌剂4个因素进行强化生物修复研究。运行结果显示:高密度投加PAHs高效降解菌,并提供适量的营养物质,同时添加生物表面活性剂进行强化,最终污染土壤中总PAHs降解率可以达到85.07%,且16种PAHs均得到显著降解。     

鼠李糖脂及其产生菌对原油生物降解影响研究

考察了外加鼠李糖脂生物表面活性剂和接种鼠李糖脂产生菌O-2-2对混合烃类降解菌降解原油的影响。结果表明,在降解体系中添加鼠李糖脂使原油20天的降解率由35.7%提高到57.6%。加入鼠李糖脂可同时提高烷烃和芳烃的降解率。在降解体系中接入菌株O-2-2能够快速利用石油烃中的烷烃类化合物并合成鼠李糖脂类生物表面活性剂,从而有效提高总石油烃的降解率。体系中菌株O-2-2的接入虽然使饱和烃的降解率大大提高,却降低了芳烃的降解率;这说明菌株O-2-2和其它烃类降解菌之间可能存在竞争生长关系。     

表面活性剂淋洗与投菌法联合修复2,4,6-三氯苯酚污染土壤

为了解决高浓度氯酚土壤污染问题,提出了一种表面活性剂淋洗与投菌法相结合的土壤修复方法。该方法采用表面活性剂淋洗污染土壤,使高浓度氯酚得到大部分去除后再加入微生物降解菌,从而实现低浓度氯酚的持续降解。通过2,4,6-三氯苯酚(2,4,6-TCP)的增溶实验、吸附实验以及表面活性剂的酶抑制实验,对蔗糖酯(SE)、鼠李糖脂(RL)、茶皂素(TS)、脂肪醇聚氧乙烯醚(AEO-9)4种表面活性剂进行了筛选。结果表明,SE增溶、解吸的综合效果较好,且具备良好的生物相容性,适用于淋洗有机污染土壤。淋洗-投菌联合实验表明,以0.5wt%蔗糖酯水溶液作为淋洗剂淋洗土壤并投菌降解,25 d内土壤中的2,4,6-三氯苯酚从190.4降至3.1 mg/kg,去除率达到98.4%。     

鼠李糖脂对铜绿假单胞菌降解颗粒有机质的影响

通过好氧降解实验研究了生物表面活性剂鼠李糖脂对1株铜绿假单胞菌(Pseudomonas Aeruginosa)降解颗粒有机质的影响,着重探讨了其作用方式,并与TritonX-100和SDS等2种化学表面活性剂作了对比.结果表明,浓度高于临界胶束浓度的鼠李糖脂在基质表面的等温吸附呈线性规律.鼠李糖脂在基质和微生物表面的吸附使菌体在基质表面的吸附性能减弱.鼠李糖脂和2种化学表面活性剂的物化作用使基质水分得以较长时间地保持,并加强了有机质在基质液相中的分散.在鼠李糖脂的作用下,有机质的降解从颗粒表面转移到液相,降解方式发生了改变.通过以上作用,鼠李糖脂促进了微生物的生长和有机质的降解.SDS和TritonX-100对有机质颗粒的降解也产生了一定的促进作用.     

鼠李糖脂在污染场地修复中的应用研究进展

微生物代谢产物鼠李糖脂(rhamnolipid)作为生物表面活性剂的代表之一,在土壤和地下水污染场地修复中有着良好的应用前景。通过文献调研等方法,总结了国内外关于鼠李糖脂应用于环境修复领域的研究现状以及修复机理,尤其是鼠李糖脂在促进疏水性有机物和重金属污染场地修复中的作用,提出目前研究可能存在的问题和未来的发展前景。     

鼠李糖脂对微生物降解正十六烷以及细胞表面性质的影响

使用大庆油田石油污染土壤中分离出的优势菌种（蜡状芽孢杆菌DQ01和芽孢杆菌DQ02）,在实验室可控条件下,研究了生物表面活性剂鼠李糖脂对微生物降解正十六烷以及降解菌生长、菌体表面疏水性和外表形态的影响.结果表明,鼠李糖脂可以提高正十六烷的降解率,48 h时蜡状芽孢杆菌DQ01和芽孢杆菌DQ02对正十六烷的降解率比未加鼠李糖脂的体系分别增加了8.1％和11.6％. 正十六烷培养基中的鼠李糖脂能引起菌体表面疏水性的明显增大,且低浓度鼠李糖脂的这种能力明显优于高浓度鼠李糖脂,蜡状芽孢杆菌DQ01和芽孢杆菌DQ02分别在加入0.4 mmol/L和0.2 mmol/L鼠李糖脂时疏水性最大. 尤其是芽孢杆菌DQ02在对数生长后期BATH（bacterial adherence to hydrocarbon）达到44％,远大于未加鼠李糖脂时的BATH. 但在葡萄糖培养基中鼠李糖脂并没有明显影响菌种的生物量和BATH. 另外,加入鼠李糖脂后, 2株菌的培养基接触角比未加鼠李糖脂的培养基接触角均减小了约一半. 未加入鼠李糖脂的菌体细胞表面比较光滑,且菌体之间独立生长,而加入鼠李糖脂后影响了降解菌的外表形态,菌体细胞表面略显粗糙且相互粘连,有助于细胞与疏水性有机物的接触.     

石油污染土壤的生物修复技术及微生物生态效应

利用投菌法和生物刺激法对陕北子长石油污染土壤进行微生物修复研究.通过利用红外分光光度法测定不同处理方法对石油烃的去除效果确定了修复陕北石油污染土壤的最佳方案.修复过程中利用最大可能计数法(MPN)、PCR-琼脂糖电泳法、PCR-DGGE法分别测定了石油烃降解菌数目、催化基因、土壤微生物多样性对土壤微生物生态效应进行研究.结果发现石油污染土壤不同生物处理修复效果为:生物刺激(加入N、P营养物质)生物强化(投加降解菌)其他.土壤中石油烃降解率与可降解石油烃的催化基因含量之间存在正相关关系,修复过程中土壤中的石油烃和烷烃降解菌数量显著多于多环芳烃降解菌数量,投加外源降解菌SZ-1可以显著提高土壤细菌群落的多样性.研究结果有助于深入理解生物修复石油土壤过程中的微生物生态效应变化.     

Effect of organic wastes on the plant-microbe remediation for removal of aged PAHs in soils

The effectiveness of in-situ bioremediation of polycyclic aromatic hydrocarbons (PAHs) may be inhibited by low nutrients and organic carbon. To evaluate the effect of organic wastes on the PAHs removal efficiency of a plant-microbe remediation system, contaminated agricultural soils were amended with different dosages of sewage sludge (SS) and cattle manure (CM) in the presence of alfalfa (Medicago sativa L.) and PAHs-degraders (Bacillus sp. and Flavobacterium sp.). The results indicated that the alfalfa mean biomasses varied from 0.56 to 2.23 g/pot in root dry weight and from 1.80 to 4.88 g/pot in shoot dry weight. Low dose amendments, with rates of SS at 0.1% and CM at 1%, had prominent effects on plant growth and soil PAHs degradation. After 60-day incubation, compared with about 5.6% in the control, 25.8% PAHs removal was observed for treatments in the presence of alfalfa and PAHs-degraders; furthermore, when amended with different dosages of SS and CM, the removed PAHs from soils increased by 35.5%-44.9% and 25.5%-42.3%, respectively. In particular, the degradation of high-molecular-weight PAHs was up to 42.4%. Dehydrogenase activities (DH) ranged between 0.41 and 1.83 μupg triphenylformazan/(g dry soil. hr) and the numbers of PAHs-degrading microbes (PDM) ranged from 1.14× 10⁶ to 16.6× 10⁶ most-probable-number/g dry soil. Further investigation of the underlying microbial mechanism revealed that both DH and PDM were stimulated by the addition of organic wastes and significantly correlated with the removal ratio of PAHs. In conclusion, the effect of organic waste application on soil PAHs removal to a great extent is dependent on the interactional effect of nutrients and dissolved organic matter in organic waste and soil microorganisms.     

Enhancing plant-microbe associated bioremediation of phenanthrene and pyrene contaminated soil by SDBS-Tween 80 mixed surfactants

The use of surfactants to enhance plant-microbe associated dissipation in soils contaminated with polycyclic aromatic hydrocarbons(PAHs) is a promising bioremediation technology. This comparative study was conducted on the effects of plant-microbe treatment on the removal of phenanthrene and pyrene from contaminated soil, in the presence of low concentration single anionic, nonionic and anionic-nonionic mixed surfactants. Sodium dodecyl benzene sulfonate(SDBS) and Tween 80 were chosen as representative anionic and nonionic surfactants, respectively. We found that mixed surfactants with concentrations less than 150 mg/kg were more effective in promoting plant-microbe associated bioremediation than the same amount of single surfactants. Only about(m/m) of mixed surfactants was needed to remove the same amount of phenanthrene and pyrene from either the planted or unplanted soils, when compared to Tween 80. Mixed surfactants( 150 mg/kg) better enhanced the degradation efficiency of phenanthrene and pyrene via microbe or plant-microbe routes in the soils. In the concentration range of 60–150 mg/kg, both ryegrass roots and shoots could accumulate 2–3 times the phenanthrene and pyrene with mixed surfactants than with Tween 80. These results may be explained by the lower sorption loss and reduced interfacial tension of mixed surfactants relative to Tween 80, which enhanced the bioavailability of PAHs in soil and the microbial degradation efficiency. The higher remediation efficiency of low dosage SDBS-Tween 80 mixed surfactants thus advanced the technology of surfactant-enhanced plant-microbe associated bioremediation.     

不同秸秆还田方式对旱地红壤细菌群落、有机碳矿化及玉米产量的影响

为探讨长期不同秸秆还田方式对旱地红壤细菌群落、有机碳矿化及玉米产量的影响,基于中国科学院鹰潭红壤生态实验站设置的不同秸秆还田方式长期定位试验(9 a),选取不施肥(CK)、单施化肥(N)、化肥+秸秆(NS)、化肥+秸秆猪粪配施(NSM)和化肥+生物质炭(NB)这5个处理,通过高通量测序技术研究旱地红壤细菌多样性和群落结构差异,揭示了细菌群落对土壤有机碳矿化和玉米产量的影响机制.结果表明:①不同秸秆还田处理下,红壤化学性质发生显著变化,其中NSM处理对旱地红壤肥力的综合提升效果最好,并显著提高了玉米产量.②相比于CK和N处理,秸秆还田处理均提高了细菌多样性.主成分分析发现,秸秆还田处理改变了土壤细菌群落结构.③秸秆还田处理提高了土壤有机碳矿化能力,NSM处理的土壤有机碳矿化速率和累积矿化量最高.④相关性分析表明土壤AN/AP比值显著改变了细菌群落结构,结构方程模型结果表明,土壤AN/AP比可能通过影响土壤细菌多样性和群落结构,间接提高了土壤有机碳矿化能力和玉米产量.本研究结果为协同提升旱地红壤生物多样性和耕地地力水平,保障健康红壤生态系统和粮食安全提供了科学依据.     

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.     

Soil fungistasis and its relations to soil microbial composition and diversity: a case study of a series of soils with different fungistasis

Fungistasis is one of the important approaches to control soil-borne plant pathogens.Some hypotheses about the mechanisms for soil fungistasis had been established,which mainly focused on the soil bacterial community composition,structure,diversity as well as function.In this study,the bacterial community composition and diversity of a series of soils treated by autoclaving,which coming from the same original soil sample and showing gradient fungistasis to the target soil-borne pathogen fungi Fusarium gr...     

岩性对喀斯特灌丛土壤固氮菌与丛枝菌根真菌群落结构及丰度的影响

运用末端限制性片段长度多态性(terminal restriction fragment length polymorphism,T-RFLP)和荧光定量PCR(real-time PCR)法,检测喀斯特灌丛生态系统中不同岩性条件下土壤固氮菌与丛枝菌根(arbuscular mycorrhizal,AM)真菌群落结构与丰度的变化,揭示岩性对灌丛生态系统土壤中固氮菌与AM真菌群落结构与丰度的影响.结果表明不同岩性条件下,土壤固氮菌与AM真菌丰度存在显著差异,其中,固氮菌与AM真菌丰度在石灰岩土壤中最大,白云岩土壤中最小,石灰岩-白云岩夹层土壤介于两者之间;同样,不同岩性土壤中固氮菌与AM真菌群落结构存在显著性差异.土壤Olsen-P、有机碳、黏粒含量与固氮菌丰度存在显著正相关关系,而土壤全氮、黏粒含量与AM真菌丰度存在显著正相关.RDA分析表明,植物均匀度影响固氮菌群落组成结构,而植物均匀度、香农多样性指数及丰富度指数影响AM真菌群落组成结构.以上的研究结果表明:岩性主要是通过影响植物与土壤养分来影响土壤固氮菌与AM真菌群落组成结构及丰度.     

DNA和cDNA水平对比研究施肥对稻田土壤细菌多样性的影响

依托长沙农业环境观测研究站不同施肥方式长期定位试验,直接从土壤中抽提总DNA和RNA,应用T-RFLP和RTPCR技术在DNA和c DNA水平对比研究施肥对亚热带稻田土壤细菌丰度和群落结构的影响.结果表明,施肥和秸秆还田显著改变了土壤细菌群落组成(DNA水平)和转录组成(c DNA水平)结构,且不同处理中DNA水平的T-RFLP图谱与cDNA水平相差很大.施肥和秸秆还田降低了土壤中存在的细菌多样性,对土壤中转录的细菌多样性没有影响.土壤16S rRNA基因丰度(DNA水平)平均是其转录丰度(c DNA水平)的337倍,与不施氮肥处理(N0)相比,平衡施肥(NPK)没有改变土壤细菌的数量,在平衡施肥的基础上进行秸秆还田增加了土壤细菌的数量,但高量与低量秸秆还田没有显著差异.RDA分析表明,稻田土壤中铵态氮含量是调控各处理土壤中存在及转录的细菌群落的关键因子.总体而言,不管在DNA还是c DNA水平,研究施肥对土壤细菌丰度的影响均是可行并可信的;但只有在c DNA水平开展研究,才能有效发现细菌群落对环境的适应性.