Salt-tolerant phenol-degrading microorganisms isolated from Amazonian soil samples

Two phenol-degrading microorganisms were isolated from Amazonian rain forest soil samples after enrichment in the presence of phenol and a high salt concentration. The yeast Candida tropicalis and the bacterium Alcaligenes faecoalis were identified using several techniques, including staining, morphological observation and biochemical tests, fatty acid profiles and 16S/18S rRNA sequencing. Both isolates, A. faecalis and C. tropicalis, were used in phenol degradation assays, with Rhodococcus erythropolis as a reference phenol-degrading bacterium, and compared to microbial populations from wastewater samples collected from phenol-contaminated environments. C. tropicalis tolerated higher concentrations of phenol and salt (16 mM and 15%, respectively) than A. faecalis (12 mM and 5.6%). The yeast also tolerated a wider pH range (3-9) during phenol degradation than A. faecalis (pH 7-9). Phenol degradation was repressed in C. tropicalis by acetate and glucose, but not by lactate. Glucose and acetate had little effect, while lactate stimulated phenol degradation in A. faecalis. To our knowledge, these soils had never been contaminated with man-made phenolic compounds and this is the first report of phenol-degrading microorganisms from Amazonian forest soil samples. The results support the idea that natural uncontaminated environments contain sufficient genetic diversity to make them valid choices for the isolation of microorganisms useful in bioremediation.     

Isolation and partial purification of fungal ligninolytic enzymes from the forest soil fungi isolated from Bhadra Wildlife Sanctuary

Screening was done for the isolation of effective lignin degraders from the forest soil samples, by providing lignin as a carbon source through the enrichment method, which leads to the isolation of 8 effective fungal isolates among 14 isolates. Submerged fermentation was done for the production of ligninolytic enzymes with the effective microorganisms by providing Guiaicol as a carbon source. The assay of laccase, lignin peroxidise activity and specific activity was done after the incubation intervals of 2, 4, 6, 7, 8, 10 and 12 days at 27±2℃ under shake culture condition. Partially purified protein content was estimated by using Lowry＇s method. Pleurotus sp. and Phanerochaetae chrysosporium are more effective at the 2nd and 7th days of incubation for the production of laccase and lignin peroxidases among the effective isolates.     

亚热带人工林下植被根际土壤酶化学计量特征

为探讨林下植被根际土壤酶化学计量特征及其对林分类型和季节的响应, 该研究以江西省泰和县千烟洲试验站典型人工杉木(Cunninghamia lanceolata)、马尾松(Pinus massoniana)和湿地松(Pinus elliottii)林林下优势灌草檵木(Loropetalum chinense)、杨桐(Adinandra millettii)、格药柃(Eurya muricata)、狗脊蕨(Woodwardia japonica)和暗鳞鳞毛蕨(Dryopteris atrata)为对象, 在植被生长初期(4月)和旺盛期(7月)测定优势灌草根际土壤与碳(C)循环相关的β-1,4-葡萄糖苷酶(BG)、与氮(N)循环相关的β-1,4-N-乙酰葡糖氨糖苷酶(NAG)和亮氨酸氨基肽酶(LAP)、与磷(P)循环相关的酸性磷酸酶(AP)活性、酶化学计量比及土壤理化性质。结果发现: (1)根际土壤与C和N循环相关的酶活性以及BG:AP (酶C:P)在不同林下植被之间存在显著差异, 而与P循环相关的酶活性差异不显著。林分类型和取样季节显著影响BG:(NAG+LAP)(酶C:N), 且林下植被类型、林分类型和取样季节交互影响酶C:P。主成分分析表明, 根际土壤酶的活性及计量比在不同林下植被(檵木不同于格药柃, 且二者显著区别于其他物种)、林分类型(杉木林区别于马尾松、湿地松林)和取样季节之间均存在显著差异。土壤硝态氮(NO₃ ^--N)、铵态氮(NH₄ ⁺-N)、可溶性有机碳(DOC)含量和碳氮比(C:N)是影响林下植被根际土壤酶的活性及化学计量比的主要因素。(2)标准主轴回归分析表明, 林下植被根际土壤lg(BG)、lg(NAG+LAP)和lg(AP)之间存在显著线性关系, lgBG:lg(NAG+LAP):lgAP (酶C:N:P)约为1:1:1.3, 酶C:P及(NAG+LAP):AP (酶N:P)分别为0.14和0.15。AP远大于BG和NAG+LAP的活性, 导致lg(BG)和lg(NAG+LAP)与lg(AP)的回归斜率极显著偏离1。说明林下植被根际土壤酶的活性及计量比受植被种类、林分类型及取样季节影响, 且基质有效性在其中发挥重要作用。相较于C循环和N循环, 微生物会分配更多资源用于P循环相关酶的生产, 暗示亚热带人工林林下植被根际土壤微生物生长和活性更易受P限制。     

短期放牧对草甸草原土壤微生物与土壤酶活性的影响

【目的】为呼伦贝尔草甸草原生态系统的保护、恢复及重建提供微生物学基础数据。了解草原土壤微生物和酶活性对放牧强度的响应。【方法】分别采集六个不同放牧强度的土壤样品,测定土壤微生物数量、土壤微生物量和土壤酶活性,分析短时期不同放牧强度土壤微生物数量、土壤微生物量和土壤酶活性的变化特征及其相互关系。【结果】不同放牧强度下,菌群数量分布为细菌>放线菌>真菌;土壤微生物数量、微生物量均表现为放牧区高于对照区;在土壤表层(0 10 cm),土壤过氧化氢酶、转化酶和蛋白酶活性表现出随放牧强度的增加先上升后略降的趋势,且放牧区均高于对照区,与土壤表层比较,在较深层(10 cm 20 cm),土壤细菌、真菌的数量和微生物量碳、氮下降幅度随放牧强度的增大而增大。土壤微生物数量、微生物量及土壤酶活性的垂直分布为0 10 cm>10 cm 20 cm。相关分析结果表明:放牧干扰条件下,土壤微生物数量与微生物量之间均存在显著或极显著的相关性。土壤酶活性与微生物数量、微生物量密切相关,过氧化氢酶、转化酶与细菌、放线菌极显著相关(P<0.01)、与微生物量碳显著相关(P<0.05);蛋白酶与真菌及微生物量碳、氮极显著相关(P<0.01),与细菌显著相关(P<0.05)。【结论】适度放牧可使土壤微生物数量、微生物量和土壤酶活性增加。土壤微生物数量、微生物量与土壤酶活性之间具有密切关系。     

罗浮栲林土壤微生物碳利用效率对短期氮添加的响应

作为调节土壤碳矿化过程的重要参数,微生物碳利用效率(CUE)对理解陆地生态系统中的碳循环至关重要。本研究在戴云山罗浮栲林设置对照(0 kg N·hm^-2·a^-1)、低氮(40 kg N·hm^-2·a^-1)和高氮(80 kg N·hm^-2·a^-1) 3个氮添加水平以模拟氮沉降,测定了表层(0～10 cm)土壤基本理化性质、有机碳组分、微生物生物量和酶活性;并利用¹⁸O标记水方法测定土壤微生物CUE,以更好地理解氮沉降加剧对微生物CUE的影响及其影响因素。结果表明: 短期氮添加显著降低了土壤微生物的呼吸速率、碳和氮获取酶活性,但显著增加了土壤微生物CUE。β-N-乙酰氨基酸葡糖苷酶(NAG)/微生物生物量碳(MBC)、微生物呼吸速率、β-葡萄糖苷酶(BG)/MBC、纤维素水解酶(CBH)/MBC和土壤有机碳含量是影响CUE的主要因素,且CUE与NAG/MBC、微生物呼吸速率、BG/MBC和CBH/MBC呈显著负相关,与土壤有机碳呈显著正相关。综上,短期氮添加导致土壤微生物获取碳和氮的成本降低,减少微生物呼吸,从而提高了土壤微生物CUE,这将有助于提高罗浮栲林土壤碳固存潜力。     

Factors determining rock phosphate solubilization by microorganisms isolated from soil

Forty two soil isolates (31 bacteria and 11 fungi) were studied for their ability to solubilize rock phosphate and calcium phosphate in culture medium. Eight bacteria and 8 fungi possessed solubilizing ability. Pseudomonas cepacia and Penicillium purpurogenum showed the highest activity. There was a correlation between final pH value and titratable acidity (r=–0.29 to –0.87) and between titratable acidity and soluble phosphate (r=0.22 to 0.99). Correlation values were functions of insoluble phosphate and of the group of microorganisms considered. A high correlation was observed between final pH and soluble phosphate only for the rock phosphates inoculated with the highest concentration of solubilizing bacteria (r=–0.73 to –0.98).     

Temperature sensitivity of soil enzyme kinetics under N‐fertilization in two temperate forests

Soil microbes produce extracellular enzymes that degrade carbon (C)‐containing polymers in soil organic matter. Because extracellular enzyme activities may be sensitive to both increased nitrogen (N) and temperature change, we measured the effect of long‐term N addition and short‐term temperature variation on enzyme kinetics in soils from hardwood forests at Bear Brook, Maine, and Fernow Forest, West Virginia. We determined the V_max and K_m parameters for five hydrolytic enzymes: α‐glucosidase, β‐glucosidase, β‐xylosidase, cellobiohydrolase, and N‐acetyl‐glucosaminidase. Temperature sensitivities of V_max and K_m were assessed within soil samples subjected to a range of temperatures. We hypothesized that (1) N additions would cause microbial C limitation, leading to higher enzyme V_max values and lower K_m values; and (2) both V_max and K_m would increase at higher temperatures. Finally, we tested whether or not temperature sensitivity of enzyme kinetics is mediated by N addition. Nitrogen addition significantly or marginally significantly increased V_max values for all enzymes, particularly at Fernow. Nitrogen fertilization led to significantly lower K_m values for all enzymes at Bear Brook, but variable K_m responses at Fernow Forest. Both V_max and K_m were temperature sensitive, with Q₁₀ values ranging from 1.64–2.27 for enzyme V_max and 1.04–1.93 for enzyme K_m. No enzyme showed a significant interaction between N and temperature sensitivity for V_max, and only β‐xylosidase showed a significant interaction between N and temperature sensitivity for K_m. Our study is the first to experimentally demonstrate a positive relationship between K_m and temperature for soil enzymes. Higher temperature sensitivities for V_max relative to K_m imply that substrate degradation will increase with temperature. In addition, the V_max and K_m responses to N indicate greater substrate degradation under N addition. Our results suggest that increasing temperatures and N availability in forests of the northeastern US will lead to increased hydrolytic enzyme activity, despite the positive temperature sensitivity of K_m.     

秦岭太白山不同林带土壤微生物呼吸速率及其影响因素

为探究森林土壤微生物呼吸对温度的敏感性及其影响因素,在太白山选取典型的4个不同海拔的林带（锐齿栎林、辽东栎林、红桦林、牛皮桦林）的0-10 cm表层土壤为对象,分别在15、25、35℃下进行控温培养实验并测量其土壤呼吸速率、微生物量和胞外酶活性等指标。结果表明：1）在1-20 d与20-72 d时的微生物呼吸速率分别呈现波动下降趋势与缓慢下降趋势,相比于其初始速率平均下降了68%与90%;表明高温在短期内促进土壤呼吸;2）太白山地区土壤温度敏感系数（Q₁₀）随温度的升高而降低;3）在培养过程中,出现15℃和25℃下微生物量先增多后减少,35℃下微生物量一直减少的现象,并且胞外酶是影响土壤微生物呼吸的重要因素,其中BG（β-葡萄糖苷酶）是胞外酶中最重要的影响因子;4）培养72 d以后,BG已无法为微生物生长繁殖提供充足的碳,在25℃和35℃下,由BX（β-木糖苷酶）提供的碳已成为微生物生长繁殖的重要碳源之一。在15℃和25℃下,N是培养前期限制土壤呼吸的因素,C是后期限制因素;在35℃下,N一直是限制土壤呼吸的因素。在15℃和35℃下,土壤呼吸不存在P限制;在25℃的培养前期,P是限制土壤呼吸的因子,而在培养后期不存在P限制。本研究结果阐明抑制土壤碳排放的关键在于抑制土壤微生物呼吸,揭示了在胞外酶驱动下的土壤碳循环特征,为准确预测全球未来气候变化的趋势提供理论基础。     

New perspectives on forest soil carbon and nitrogen cycling processes: Roles of arbuscular mycorrhizal versus ectomycorrhizal tree species

Nearly all tree species develop symbiotic relationships with either arbuscular mycorrhizal (AM) or ectomycorrhizal (EM) fungi to acquire nutrients from soils, and hence influence soil carbon (C) and nitrogen (N) cycles in terrestrial ecosystems. It is crucial to understand the differences in soil C and N cycles between AM and EM forests and the underlying mechanisms. In this review, we first compared the differences in the soil C and N cycles between AM and EM forests, and synthesized the underlying mechanisms from perspectives of the inputs, stabilization, and outputs of soil C and N in forest ecosystems. We also compared the responses of soil C and N cycles between AM and EM forests to global changes. In this field, one major research priority is comparing the structure and function (including the soil C and N cycles) between AM and EM forest ecosystems to provide theoretical basis and solid data for improving forest productivity and ecosystem services. The second research focus is deepening the understanding of the effects of interactions between aboveground litter and belowground mycorrhiza and free-living microbes on soil C and N cycles to reveal the potential underlying mechanisms in forests with different mycorrhizal symbioses. Third, the research methodology and new techniques need refining and applying to explicitly focus on scaling up the fine-scale measurements to better expound and predict the C and N cycles in forest ecosystems. Finally, more studies on the stability of soil organic matter among different mycorrhizal forests are needed to precisely assess responses of the structure and function of forest ecosystems to global changes.     

Responses of soil extracellular enzyme activities to carbon input alteration and warming in a subtropical evergreen broad-leaved forest

土壤胞外酶来源于土壤微生物、植物和动物, 是土壤生物地球化学过程的积极参与者, 在森林生态系统的物质循环和能量流动过程中扮演着重要角色。为探明土壤胞外酶活性对碳输入变化及增温的响应, 该研究基于长期增温、去除地表凋落物(以下简称去凋)和切根处理的云南哀牢山亚热带常绿阔叶林控制实验平台, 研究了不同处理(对照、去凋、切根、切根并增温)下表层矿质土壤(0-5和5-10 cm)与碳氮磷获取相关的胞外酶活性, 包括多酚氧化酶(POX)、过氧化物酶(PER)、β-葡萄糖苷酶(BG)、β-1,4-N-乙酰氨基葡萄糖苷酶(NAG)和酸性磷酸酶(AP)。结合铵态氮(NH₄⁺-N)含量、硝态氮(NO₃^--N)含量、溶解有机碳(DOC)含量、溶解总氮(DN)含量、土壤含水量(SWC)等相关指标, 探讨凋落物碳输入、根系碳输入和温度变化对土壤胞外酶活性及其生态化学计量特征的影响。研究结果表明: 在对照样方, 除POX外其余酶活性均为0-5 cm层显著高于5-10 cm层。与对照相比, 长期的凋落物去除显著降低了0-5 cm层土壤AP和BG活性, 对NAG、PER和POX活性则无显著影响; 长期切根处理显著降低了0-5 cm层土壤BG活性, 但提高了两个土层PER活性; 长期切根并增温处理显著降低了0-5 cm层AP和BG活性, 对其余酶活性无显著影响。冗余分析结果显示SWC和NH₄⁺-N含量是驱动土壤酶活性变化的重要因子。本研究为该生态系统土壤碳氮磷生物地球化学关键过程对全球变化的响应提供了土壤酶学的依据。     

Oligotrophic bacteria from rendzina forest soil

Oligotrophic bacteria were shown to exist abundantly in all layers of a rendzina forest soil throughout the year. Two-hundred-three oligotrophic bacteria were isolated from forest soil (Aoba, Sendai) at different layers (L, F, H and A layers) throughout the year, and their morphological and physiological characteristics were examined. A high proportion (95%) of the isolated oligotrophs were Gram-negative, non-spore forming bacteria. Based on the cell shape, the isolates were divided into four groups: regular rods, curved/spiral bacteria, irregular rods, and buddin and/or prosthecate bacteria. Each group of bacteria is discussed in relation to the physiological characteristics. Notably oligotrophic bacteria of different cell types showed a marked zonal distribution in respect to profile depth.     

土壤增温对亚热带杉木幼树不同深度土壤微生物胞外酶活性的影响

土壤微生物胞外酶可有效反映气候变暖对土壤微生物功能和土壤有机质分解的影响.目前关于气候变暖对土壤微生物胞外酶活性(EEAs)影响的相关研究主要关注有机碳含量较丰富的表层土壤(0～20 cm),而对深层土壤(>20 cm)EEAs的研究仍较缺乏.因此,本研究关注土壤增温对亚热带不同深度(0～10 cm、10～20 cm、20～40 cm和40～60 cm)EEAs的影响及主要调控因素,其中微生物胞外酶包括参与碳循环的β-葡萄糖苷酶(BG)、纤维二糖水解酶(CBH)、酚氧化酶(PHO)和过氧化物氧化酶(PEO).结果表明: 土壤增温提高了0～10 cm和10～20 cm土壤所有胞外酶的活性(18%～69%).在20 cm以下的深层土壤中,土壤增温仅显著提高了20～40 cm的PHO(10%),而对其余胞外酶的活性无显著影响或有一定的抑制作用(13%～31%).冗余分析(RDA)结果表明: 在微生物可利用有机碳较丰富的表层土壤中,铵态氮(NH₄⁺-N)和土壤含水率(M)是调控EEAs的主要因素,增温增强了微生物与植物之间的养分竞争,因而提高EEAs以获取微生物所需的养分NH₄⁺-N;而在微生物底物有效性较低的深层土壤中,EEAs主要受可溶性有机质(可溶性有机碳和可溶性有机氮)和微生物生物量(MBC)的影响,增温提高深层土壤可溶性有机质的含量,为微生物提供更多的底物,减少微生物对EEAs的需求,进而降低EEAs.本研究发现,不同深度EEAs对土壤增温具有不同响应,且土壤增温条件下表层和深层土壤的EEAs具有明显不同的调控因素.因此,加强不同深度土壤微生物的研究对于准确评估生态系统碳循环对全球变暖的响应具有重要意义.     

亚热带常绿阔叶林土壤胞外酶活性对碳输入变化及增温的响应

土壤胞外酶来源于土壤微生物、植物和动物, 是土壤生物地球化学过程的积极参与者, 在森林生态系统的物质循环和能量流动过程中扮演着重要角色。为探明土壤胞外酶活性对碳输入变化及增温的响应, 该研究基于长期增温、去除地表凋落物(以下简称去凋)和切根处理的云南哀牢山亚热带常绿阔叶林控制实验平台, 研究了不同处理(对照、去凋、切根、切根并增温)下表层矿质土壤(0-5和5-10 cm)与碳氮磷获取相关的胞外酶活性, 包括多酚氧化酶(POX)、过氧化物酶(PER)、β-葡萄糖苷酶(BG)、β-1,4-N-乙酰氨基葡萄糖苷酶(NAG)和酸性磷酸酶(AP)。结合铵态氮(NH₄⁺-N)含量、硝态氮(NO₃^--N)含量、溶解有机碳(DOC)含量、溶解总氮(DN)含量、土壤含水量(SWC)等相关指标, 探讨凋落物碳输入、根系碳输入和温度变化对土壤胞外酶活性及其生态化学计量特征的影响。研究结果表明: 在对照样方, 除POX外其余酶活性均为0-5 cm层显著高于5-10 cm层。与对照相比, 长期的凋落物去除显著降低了0-5 cm层土壤AP和BG活性, 对NAG、PER和POX活性则无显著影响; 长期切根处理显著降低了0-5 cm层土壤BG活性, 但提高了两个土层PER活性; 长期切根并增温处理显著降低了0-5 cm层AP和BG活性, 对其余酶活性无显著影响。冗余分析结果显示SWC和NH₄⁺-N含量是驱动土壤酶活性变化的重要因子。本研究为该生态系统土壤碳氮磷生物地球化学关键过程对全球变化的响应提供了土壤酶学的依据。     

模拟氮沉降增加条件下土壤团聚体对酶活性的影响

氮沉降增加改变了森林土壤生态系统物质输入,影响土壤生物及酶活性,而土壤团聚体内相对稳定的微域生境可能减弱或延缓土壤生物和酶对氮沉降增加的响应强度。以广东省东莞大岭山森林公园荷木人工林为研究对象,用模拟N沉降方法,分析了2011年12月到2012年11月一年内氮沉降增加条件下表层混合土壤和土壤团聚体内脲酶、蔗糖酶和酸性磷酸酶活性的变化及影响因素,旨在理解氮沉降增加条件下土壤团聚体对酶活性的影响。结果表明:氮沉降增加对表层混合土壤中脲酶和蔗糖酶的抑制作用不显著,而酸性磷酸酶受氮沉降显著影响,表现为低氮(50 kg N hm-2a-1)促进,高氮(300 kg N hm-2a-1)抑制的规律。表层土壤团聚体内脲酶活性随氮沉降增加而降低,N300处理显著低于对照;蔗糖酶和酸性磷酸酶活性随氮沉降增加先降低后增加,N100处理最低,分别比其他处理降低了6.46%—25.53%和42.33%—68.25%。试验区内各粒径土壤团聚体内酶活性高于混合土壤,且随团聚体粒径增加酶活性均为先增加后降低。不同粒径土壤团聚体的3种酶活性均以2—5 mm最高,但脲酶、酸性磷酸酶在各团聚体粒径间差异不显著,蔗糖酶活性2—5 mm显著高于5—8 mm。土壤酶相对活性指数和相对活性综合指数结果显示,超过85%的团聚体粒径内的相对酶活性指数大于1,而土壤酶相对活性综合指数均大于1。以上结果表明,氮沉降增加条件下土壤团聚体对其团聚体内的土壤酶活性有隔离保护作用,但其隔离保护效果与酶的种类和土壤团聚体粒径有关。     

土壤酶计量揭示了武夷山黄山松林土壤微生物沿海拔梯度的碳磷限制变化

了解土壤胞外酶活性和酶计量的变化对评估山地生态系统土壤养分有效性和微生物的营养限制状况具有重要意义.然而,亚热带山地森林土壤微生物的营养限制状况对海拔梯度变化的响应及其驱动因素尚不清楚.本研究以武夷山不同海拔(1200～2000 m)黄山松林为对象,测定了土壤基本性质、微生物生物量以及与碳(C)、氮(N)、磷(P)循环...     

不同林龄杉木林土壤细菌群落结构与土壤酶活性变化研究

探究不同林龄杉木人工纯林土壤中的微生物的群落演变与结构特征与酶活性变化,为杉木人工林可持续经营管理提供依据。以福建省南平市的五片不同林龄杉木林表层土壤作为研究对象,通过16SrDNA测定细菌的群落组成,分析与土壤质量密切相关的四种土壤酶活性变化,揭示细菌群落与土壤酶活性的变化机理。结果表明,微生物的多样性指数与OTU都随着林龄的增加而增加,且幼龄林、中龄林、近熟林、成熟林土壤微生物结构差异较大;不同林龄杉木人工林中包含了29个细菌门,其中酸杆菌门与变形菌门为优势菌群,根据各种群相对丰度变化以及冗余分析,放线菌门、浮霉菌门与疣微菌门等均随林龄增长出现较大变化,且与土壤可溶性有机质以及速效养分有显著相关性（P<0.05）,说明这几种细菌群落对土壤养分变化较敏感;土壤养分变化会影响土壤酶活性,蔗糖酶与全碳呈显著正相关（P<0.05）,与速效钾呈显著负相关（P<0.05）,与放线菌门呈极显著负相关（P<0.01）。脲酶与速效氮呈显著负相关（P<0.05）,脲酶与变形菌门、绿弯菌门、放线菌门、硝化螺旋菌门以及拟杆菌门均存在较强相关性。综上,不同的土壤细菌种群与酶活性对各养分变化的响应程度不一,细菌群落结构与酶活性能反映不同林龄杉木林土壤的质量变化,适量延长杉木人工林种植年限有益于土壤质量恢复。本研究结果对指导杉木人工林优质经营有重要意义。     

天津东郊盐碱土微生物分布及土壤酶活性

天津东郊盐碱土微生物分布及土壤酶活性靳素英,崔明学,蔺继尚（中国科学院沈阳应用生态研究所,沈阳１１００１５）Ｄｉｓｔｒｉｂｕｔｉｏｎｏｆｍｉｃｒｏｏｒｇａｎｉｓｍｓｉｎｓａｌｉｎｅ－ａｌｋａｌｉｓｏｉｌａｔｃａｓｔｅｒｎｓｕｂｕｒｂｓｏｆＴｉａｎｊｉ...