高羊茅草对盐碱地原油污染土壤微生物的影响

采用常规土壤理化分析和Biolog-Eco微孔板鉴定系统,研究了松嫩平原盐碱土地区高羊茅草的生长对原油污染土壤pH、总盐含量和微生物群落的影响.结果表明: 石油污染导致表征土壤微生物代谢活性的平均颜色变化率(AWCD)、物种多样性指数和碳源利用数增加, 微生物碳源利用模式发生改变.高羊茅草对石油污染土壤有较好的修复作用,降低了土壤pH和石油烃(TPH)含量,提高了土壤含水率.高羊茅草根际土壤的AWCD和碳源利用丰富度指数明显高于裸地土壤,为其根际微生物的生长发育提供了适宜的环境.     

Assessment of Pollution-Induced Microbial Community Tolerance to Heavy Metals in Soil Using Ammonia-Oxidizing Bacteria and Biolog Assay

This study attempted to investigate if the tolerance of soil bacterial communities in general, and autotrophic ammonia-oxidizing bacteria (AOB) in particular, evolved as a result of prolonged exposure to metals, and could be used as an indigenous bioindicator for soil metal pollution. A soil contaminated with copper, chromium, and arsenic (CCA) was mixed with an uncontaminated garden soil (GS3) to make five test soils with different metal concentrations. A modified potential ammonium oxidation assay was used to determine the metal tolerance of the AOB community. Tolerance to Cr, Cu, and As was tested at the beginning and after up to 13 months of incubation. Compared with the reference GS3 soil, the five CCA soils showed significantly higher tolerance to Cr no matter which form of Cr (Cr³⁺, CrO₄ ^2?, or Cr₂O₇ ^2?) was tested, and the Cr tolerance correlated with the total soil Cr concentration. However, the tolerance to Cu²⁺, As³⁺, and As⁵⁺ did not differ significantly between the GS3 soil and the five CCA soils. Community level physiological profiles using Biolog microtiter plates were also used to examine the chromate tolerance of the bacterial communities extracted after six months of exposure. Our results showed that the bacterial community tolerance was altered and increased as the soil Cr concentration was increased, indicating that the culturable microbial community and the AOB community responded in a similar manner.     

Functional Diversity of Culturable Bacterial Communities in the Rhizosphere in Relation to Fine-root and Soil Parameters in Alder Stands on Forest, Abandoned Agricultural, and Oil-shale Mining Areas

Grey alder (Alnus incana) and black alder (Alnus glutinosa) stands on forest land, abandoned agricultural, and reclaimed oil-shale mining areas were investigated with the aim of analysing the functional diversity and activity of microbial communities in the soil–root interface and in the bulk soil in relation to fine-root parameters, alder species, and soil type. Biolog Ecoplates were used to determine community-level physiological profiles (CLPP) of culturable bacteria in soil–root interface and bulk soil samples. CLPP were summarized as AWCD (average well color development, OD 48 h⁻¹) and by Shannon diversity index, which varied between 4.3 and 4.6 for soil–root interface. The soil–root interface/bulk soil ratio of AWCD was estimated. Substrate-induced respiration (SIR) and basal respiration (BAS) of bulk soil samples were measured and metabolic quotient (Q = BAS/SIR) was calculated. SIR and Q varied from 0.24 to 2.89 mg C g⁻¹ and from 0.12 to 0.51, respectively. Short-root morphological studies were carried out by WinRHIZO^TM Pro 2003b; mean specific root area (SRA) varied for grey alder and black alder from 69 to 103 and from 54 to 155 m² kg⁻¹, respectively. The greatest differences between AWCD values of culturable bacterial communities in soil–root interface and bulk soil were found for the young alder stands on oil-shale mining spoil and on abandoned agricultural land. Soil–root interface/bulk soil AWCD ratio, ratio for Shannon diversity indices, and SRA were positively correlated. Foliar assimilation efficiency (FOE) was negatively correlated with soil–root interface/bulk soil AWCD ratio. The impact of soil and alder species on short-root morphology was significant; short-root tip volume and mass were greater for black alder than grey alder. For the investigated microbiological characteristics, no alder-species-related differences were revealed.     

外来物种刺槐对土壤微生物功能多样性的影响

以延河流域不同植被区内人工刺槐(Robinia pseudoacacia)群落和乡土植物群落的土壤微生物为研究对象,利用Biolog微平板技术对土壤微生物功能多样性进行测定,分析人工引种刺槐在不同环境梯度(3个植被区)下对土壤微生物功能多样性的影响。结果表明:刺槐对不同环境梯度下的土壤微生物的影响明显不同。从草原区到森林区,刺槐林之间土壤微生物群落的平均颜色变化率(AWCD)和土壤微生物功能多样性指数均没有显著变化;但与乡土植物群落比较,草原区、森林草原区和森林区土壤AWCD分别表现为刺槐乡土植物、刺槐乡土植物、刺槐乡土植物;在草原区和森林草原区刺槐林土壤微生物群落的群落丰富度指数(S)、Shannon-Wiener指数(H)、Simpson指数(D)、Mc Intosh指数(U)均大于乡土植物,森林区刺槐林群落丰富度指数(S)、Shannon指数(H)、Simpson指数(D)均小于乡土植物;刺槐林和乡土植物群落下土壤微生物碳源利用存在差异,主要体现在对糖类、氨基酸类的利用上。PCA分析显示主成分1贡献较大的碳源有24种,在主成分分离中起主要贡献作用的是糖类、氨基酸类和羧酸类。土壤碳氮含量能影响土壤微生物功能多样性指数,土壤含水量和温湿度能够影响碳源的利用类型。刺槐对土壤微生物功能多样性的影响存在区域差异,在评价刺槐对土壤生态过程与功能的影响时必须要考虑这种空间差异性。     

Evaluation of Biolog system for identification of strains of Paenibacillus azotofixans

Biolog system was evaluated for the identification of strains of Paenibacillus azotofixans as no data concerning this species were in the list of Bacillus currently identified using Biolog data base. The P. azotofixans type strain P3L5 was first tested with the results recorded manually or using the automatic plate reader. In both cases, P3L5 utilized 22 carbon sources and when the results obtained were compared to data of Biolog software (Release 3.50), P3L5 was identified as B. azotoformans with a similarity coefficient of 0.913 (data recorded manually) and of 0.791 (data recorded automatically). Metabolic profiles of P3L5 were also compared after readings of 4 and 24 h using the computer-driven automatic plate reader. No significant difference was observed in both cases and P3L5 was identified again as B. azotoformans with indices of similarity considered only for excellent identification. Besides P3L5, other 15 P. azotofixans strains were tested with the Biolog system and all were identified as B. azotoformans with similarity coefficients varying from 0.511 to 0.927. Phenotypic and genetic characteristics of B. azotoformans were compared to those described for P. azotofixans to explain the misidentification of the latter species. We could conclude that these two species are quite different and that data of Biolog software are from P. azotofixans and not from B. azotoformans.     

深松对乌拉尔甘草根际土壤养分以及微生物群落功能多样性的影响

通过大田试验和室内分析相结合,研究了深松对乌拉尔甘草根际土壤养分和微生物群落功能多样性的影响,以期为乌拉尔甘草人工种植地土壤耕作措施优化和土壤环境改良提供依据。结果表明,与未深松(常规耕作)处理相比,深松处理对乌拉尔甘草根际土壤0—20 cm耕层土壤养分含量无显著性影响,可显著提高乌拉尔甘草根际土壤20—40 cm耕层有机质、全氮、全磷和全钾的含量,分别提高了60.8%、65.3%、48.9%和86.8%;明显增加了0—20 cm和20—40 cm耕层细菌、真菌和放线菌的数量(P0.05),3种类型的微生物数量均呈现出上层大于下层,深松大于未深松的变化趋势。在156 h的微生物温育期内,深松处理下不同土层的平均颜色变化率(AWCD)均显著高于未深松处理,并显著提高了AWCD的利用率(72 h,P0.05),较未深松分别提高了35.5%和130.8%。与未深松处理相比,深松处理显著提高了土壤微生物的多样性指数(H、S、D)。主成分分析(PCA)表明,深松优化了乌拉尔甘草根际土壤微生物的群落组成;聚合物、羧酸类化合物、氨基酸和碳水化合物是深松处理下根际土壤微生物利用的主要碳源。总而言之,深松处理显著提高乌拉尔甘草根际土壤养分含量、微生物数量和微生物多样性指数,改变了微生物群落功能多样性,造成这种差异的主要原因可能是深松改变了土壤耕层结构,改善了微生物的生存环境。因此,深松对乌拉尔甘草人工种植地土壤质量的改良有积极作用。     

猫儿山不同海拔植被带土壤微生物群落功能多样性

为研究中亚热带森林土壤微生物群落功能多样性特征及其随海拔梯度的变化,应用Biolog微平板技术,对猫儿山不同海拔植被带(常绿阔叶林(EBF)、落叶阔叶混交林(DBF)、针阔混交林(CBF))土壤微生物群落功能多样性差异进行了比较。结果表明,不同海拔植被带土壤微生物群落功能多样性差异显著。土壤平均颜色变化率(AWCD)随培养时间延长而逐渐增加,随着海拔升高,土壤AWCD值逐渐降低,大小顺序为EBFDBFCBF。土壤微生物群落Shannon指数和丰富度指数的总体趋势为EBF最高,DBF次之,CBF最低。不同海拔植被带土壤微生物群落均匀度指数之间差异不显著。不同海拔植被带土壤微生物对不同碳源的利用能力存在差异,其中EBF利用率最高,CBF利用率最低,氨基酸类、胺类和酯类碳源为各海拔植被带土壤微生物利用的主要碳源。主成分分析结果表明,主成分1和主成分2分别能解释变量方差的40.42%和15.97%,在主成分分离中起主要贡献作用的是酯类、胺类和氨基酸类碳源。土壤理化性质与土壤微生物群落功能多样性之间的相关性分析结果表明,微生物群落多样性的Shannon指数与全钾(TK)呈极显著正相关(P0.01),与含水量呈极显著负相关(P0.01),与总有机碳(TOC)、全氮(TN)、速效氮(AN)、有效P(AP)之间的相关性显著(P0.05)或极显著(P0.01),且为负相关。土壤TK含量和含水量可能是造成不同海拔土壤微生物群落功能多样性差异的主要原因。     

生物炭对菜园土壤微生物功能多样性的影响

研究生物炭的施用及其与不同肥料混施对菜园土壤中微生物群落功能多样性的影响,为农业废弃物的合理利用和菜园土优化培肥提供科学依据和理论指导。以清远市连州县代表性菜园土(属肥熟旱耕人为土)为研究对象,通过盆栽试验,利用BIOLOG方法对10个施肥处理(对照CK(0%生物碳+无肥)、T1(0%生物碳+0.1%商品有机肥)、T2(0.1%生物碳+无肥)、T3(0.25%生物碳+无肥)、T4(0.5%生物碳+无肥)、T5(1%生物碳+无肥)、T6(100(N)+30(P_2O_5)+75(K_2O)mg/kg干土)、T7(0.1%生物碳+0.1%商品有机肥)、T8(0.1%生物碳+100(N)+0(P_2O_5)+75(K_2O)mg/kg干土)、T9(0.1%生物碳+100(N)+30(P_2O_5)+75(K_2O)mg/kg干土)、T10(0.1%生物碳+0.1%商品有机肥+100(N)+0(P_2O_5)+75(K_2O)mg/kg干土))的土壤微生物群落功能多样性进行分析。结果表明:(1)T1和T3处理比其它处理显著提高土壤微生物对碳源的利用率(P0.05),但生物炭施用量增加会降低平均颜色变化率(AWCD值);(2)T1处理可以显著提高土壤微生物的群落物种均匀度(Mclntosh指数),而T3处理显著提高土壤微生物的物种丰富度和均匀度(Shannon和Mclntosh指数);(3)T1和T3处理对聚合物类、碳水化合物类、羧酸类、氨基酸类和酚类碳源利用率最高;(4)添加化肥处理中磷肥的施用可以提高土壤微生物活性,增加土壤微生物碳源利用能力,而氮肥和钾肥的添加显著降低了土壤微生物的碳源利用能力;(5)主成分分析表明,T1、T2和T3处理的微生物碳代谢功能群结构相似;单施有机肥或适量生物炭对土壤微生物群落结构的影响较混合施用更为显著;化学磷肥的添加及在施用化肥的基础上配施适量生物炭改变了土壤微生物对碳源种类的利用。     

Screening and identification of extracellular lipase-producing thermophilic bacteria from a Malaysian hot spring

Seven lipase-producing thermophilic bacteria (ST 1, ST 4, ST 6, ST 7, ST 8, ST 9 and ST 10) were isolated from the Setapak hot spring in Malaysia. The crude extracellular lipases recovered by ultrafiltration of cell-free culture supernatant were reacted in an olive oil mixture and their lipolytic activities were compared. Identification of the bacteria was carried out using the Biolog system and biochemical tests. Strain ST 7 that exhibited the highest lipolytic activity of 4.58 U/ml was identified as belonging to the Bacillus genus. Strain ST 6 with an activity of 3.51 U/ml, was identified as Ralstonia paucula. The lipolytic activities of strains ST 1, ST 4, ST 8, ST 9 and ST 10 were 2.39, 1.84, 2.38, 1.80 and 2.62 U/ml respectively. Strains ST 1, ST 4, and ST 10 were identified as Ralstonia paucula while strains ST 8 and ST 9 were Bacillus spp. Strains ST 7 and ST 9 were tentatively identified as Bacillus thermoglucosidasius, Bacillus stearothermophilus or Bacillus coagulans, whereas strain ST 8 was tentatively identified as Bacillus subtilis.     

车八岭山地常绿阔叶林冰灾后土壤微生物群落功能多样性

研究了我国南方冰灾后常绿阔叶林林冠开度及土壤养分的空间异质性对土壤微生物功能多样性的影响.在受冰灾影响的粤北车八岭山地常绿阔叶林2 hm2固定样地中按照冠层受损程度选取16个20 m×20 m的样方,用半球面影像技术获取林冠开度,并取0～20 cm的表层土壤混合样品分析土壤的理化性质,同时应用Biolog技术分析微生物功能多样性.按林冠开度梯度对各样方土壤微生物群落利用单一碳源的分析发现,林冠开度大的样方土壤微生物的活性、丰富度、多样性和均匀度都较低,反之则较高.聚类分析的结果与林冠开度的梯度有高度的一致性.主成分分析表明各样方土壤微生物功能多样性具有显著差异(第一轴p<0.005;第二轴p<0.001),其结果与聚类结果基本吻合.冗余分析揭示了土壤全磷、全钾、全氮、速效氮、有机碳、容重、总孔隙度和林冠开度的综合作用对土壤微生物功能多样性有显著影响(p<0.005),其中林冠开度与土壤微生物群落功能多样性的关系最密切.土壤微生物功能多样性受土壤养分的影响,具体表现为:与土壤有机碳呈明显的正相关;与全氮正相关;与速效氮、全磷负相关.研究说明冰灾所造成林冠开度和土壤养分的空间异质性会影响到土壤微生物功能多样性,而土壤微生物功能多样性可用于对生境恢复的指示和评价.