高山林线土壤微生物群落结构对模拟增温的响应

研究土壤微生物群落结构对模拟增温的响应,对预测全球气候变化背景下土壤碳氮磷循环具有重要意义.采用开顶式生长室(OTC)模拟增温,研究了土壤有机质层和矿质土壤层真菌(F)、细菌(B)、革兰氏阳性菌(G⁺)和革兰氏阴性菌(G^-)PLFAs微生物量,以及真菌/细菌(F/B)和革兰氏阴性菌/革兰氏阳性菌(G^-/G⁺)比值对模拟增温的响应.结果表明: OTC模拟增温使空气温度增加0.87 ℃,土壤有机质层温度增加0.5 ℃,矿质土壤层温度增加0.23 ℃.土壤有机质层微生物群落组成比矿质土壤层对模拟增温的响应更敏感.细菌比真菌对模拟增温的响应更加敏感,模拟增温显著影响了土壤有机质层的F/B和G^-/G⁺比值,对矿质土壤层的所有PLFAs含量或比值均没有显著影响.微生物的PLFAs含量及真菌/细菌和G^-/G⁺比值总体呈现非生长季低于生长季前期和生长季后期.冗余分析表明,土壤中的碳含量(可溶性有机碳DOC 12.1%、凋落物可溶性碳DC 9.5%和全碳TC 3%)是微生物群落结构的决定性因素,可溶性组分(DOC和DC)对微生物群落结构的影响大于全量养分(全碳和总氮).     

采油微生物在微驱过程中的生长、运移及分布规律

[目的]深入了解现场微生物驱油机理、效果评价标准及影响因素.[方法]结合现场微生物驱油过程产出液的跟踪监测及室内物模实验对微生物在地层中的生长繁殖、运移及分布规律进行研究.[结果]结果表明,通过从水井注入的外源微生物在油藏中能够有效生长繁殖,而且注入的营养液也能够激活内源微生物,但由于地层渗透率及营养液浓度的影响,产出液菌浓要比注入菌浓低1-2个数量级;葡萄糖的快速降解以及地层对微生物的过滤及吸附作用使大量的微生物停留在近井地带,仅有部分微生物能够从生产井采出,而且其运移速度要比营养液慢.[结论]地层渗透率和产出液中营养物浓度是影响微生物数量及分布的两个关键因素,现场微生物驱油产出液中的菌浓一般很难达到106个/mL以上,该研究结果对微生物驱油技术的发展和应用具有重要意义.     

石油污染土壤中菲、蒽和正十六烷的微生物降解

在模拟自然条件下,考察了芳香烃(菲、蒽)和脂肪烃(正十六烷)单独存在以及共存于土壤中时,土著微生物对它们降解的影响。结果表明,土著微生物对它们的降解均符合一级反应动力学。菲、蒽或正十六烷单独存在于土壤中时,其微生物降解速率常数分别为0.0226、0.0283和0.0096 d^-1。菲和正十六烷共存时,正十六烷能够作为土著微生物降解菲的共代谢底物,促进菲的降解,使菲的半衰期比其单独存在于土壤中缩短44%;同时,正十六烷的加氧酶被菲诱导,使其活性提高而增强对正十六烷的降解作用,其微生物降解半衰期比其单独存在于土壤中缩短49%。菲和蒽共存促进了土著微生物对菲的降解,却抑制了对蒽的降解。     

盐碱地土壤微生物对不同改良方法的响应

实验室条件下分别模拟客土改良、排水改良、生物(苜蓿)改良等3种常用的盐碱地改良方法,以菜园土壤和盐碱土(未改良的原始土)作为对照,研究了土壤微生物生态特征对不同改良方法的响应。结果显示:①土壤微生物区系及生理生化指标均对不同改良方式有良好的响应,其中土壤脲酶活性随土壤盐碱胁迫加剧而提高;②3种改良方式中,客土法在短期内最能改善土壤微生物各指标,依次是排水法、生物法;③排水法对土壤微生物生态状况不利;④生物法对于改善土壤微生物生态状况速度相对较慢,但因其克服了客土法、排水法显见的缺点,相对有更好的应用前景。     

土壤微生物对不同种源乌桕生长的影响

【背景】土壤微生物对植物成功入侵具有重要影响,研究中国本地土壤微生物对美国入侵种乌桕的净生长效应有利于进一步理解乌桕成功入侵的机理。【方法】以本地和入侵2个种源多个种群的乌桕为试验材料,通过对土壤进行灭菌处理,研究土壤微生物对不同种源乌桕幼苗生长的影响。【结果】土壤微生物对入侵种乌桕具有正效应,与本地种相比,入侵种乌桕幼苗总生物量、相对生长速度、根冠比以及总叶面积、叶面积比、比叶面积等生物学指标在新鲜土壤中均表现出明显优势。种源和土壤处理间存在交互效应的指标显示土壤微生物去除对入侵种乌桕的抑制作用更为明显。这一结果与土壤自然天敌逃逸假说相背,入侵种乌桕在一定程度上具有较本地种更强的土壤微生物利用能力,在非根际土壤微生物的作用下入侵种乌桕具有较强的生长能力和明显的竞争优势。【结论与意义】本研究证实土壤微生物可能对乌桕成功入侵具有不可忽略的作用。由于该试验在乌桕起源地进行,结合在乌桕入侵地进行的研究有助于进一步分析土壤微生物对入侵的贡献作用;研究土壤微生物与入侵植物的互作关系有助于对入侵物种进行准确预测和有效控制,减少生物入侵对生态系统功能的破坏。     

岷江上游华山松林冬季土壤呼吸对模拟增温的短期响应

冬季的土壤呼吸是生态系统呼吸的重要组成部分, 对气候变化的响应可能更为敏感。该文采用红外辐射加热器模拟土壤增温, 研究了岷江上游华山松(Pinus armandii)人工林冬季的土壤呼吸、微生物生物量及无机氮库对模拟增温的响应。结果表明: 在冬季(2009年11月-翌年3月), 模拟增温往往能显著提高土壤呼吸速率, 平均增幅达31.4%; 同样模拟增温使土壤微生物生物量碳、氮分别增加23.2%和22.7%, 而对微生物生物量碳氮比没有影响, 温度升高显著促进了微生物的生长, 但没有改变微生物的群落结构; 增温样地土壤的NO₃ ^--N和NH₄ ⁺-N浓度较对照分别增加了38.5%和12.3%, 增温显著提高了土壤的可溶性无机氮含量。综上所述, 该区针叶林冬季土壤呼吸、微生物生长和养分矿化对未来气候变暖非常敏感。     

中亚热带几种针、阔叶树种凋落物混合分解对土壤微生物群落碳代谢多样性的影响

土壤微生物群落功能如何响应凋落物物种组成的改变,对于指导我国的人工林种植具有重要意义。通过小盆模拟试验,用BIOLOG微平板培养的方法研究了南方红壤丘陵区典型物种马尾松和湿地松的凋落物分别与白栎和青冈的凋落物混合,与单一针叶凋落物分解时相比,针阔混合凋落物分解过程中土壤微生物群落功能多样性的差异。结果表明：(1)针叶和阔叶混合分解提高了土壤微生物群落碳代谢的强度、丰富度以及多样性,针阔混合凋落物分解时土壤微生物群落的碳源代谢方式与单一针叶分解时差异显著;(2)凋落物分解前期土壤微生物群落碳代谢强度、丰富度和多样性与凋落物初始C/N呈显著负相关,分解后期土壤微生物群落碳代谢强度与凋落物初始木质素/N比呈显著负相关,碳代谢多样性与初始P含量呈显著正相关;(3)通过改变凋落物物种组成,进而改变凋落物元素组成(C/N、木质素/N和P),能显著影响土壤微生物群落的碳代谢强度和代谢多样性。在南方红壤丘陵区,针-阔叶混合的凋落物比单一针叶凋落物更有利于提高土壤微生物群落的功能。     

祁连山3种典型生态系统土壤微生物活性和生物量碳氮含量

 测定分析了祁连山高寒草甸、山地森林和干草原土壤中微生物活性、生物量碳氮含量。结果显示：就土壤微生物生物量碳含量,森林比干草原和高寒草甸中分别高60%和120%以上,干草原比高寒草甸中高40%以上(p<0.05)。就土壤微生物生物量氮含量,0～5 cm土层,森林比高寒草甸和干草原中分别高64%和111%以上,高寒草甸比干草原中高29%;5～15 cm土层,森林比干草原和高寒草甸中分别高7%和191%以上,干草原比高寒草甸中高171% 以上(p<0.05)。森林和干草原中土壤微生物生物量碳比例比高寒草甸中高32%以上,0～5和5～15 cm土层,森林和干草原中土壤微生物生物量氮比例比高寒草甸中高150%以上（p<0.05）。就土壤微生物活性,0～5和5～15 cm土层,森林和高寒草甸比干草原中高26%以上;15～35 cm土层,森林比干草原和高寒草甸中高28%以上 (p<0.05)。土壤微生物生物量碳氮含量与有机碳含量及微生物生物量氮含量和比例与微生物生物量碳含量和比例呈现正相关（r²>0.30,p<0.000 1）。土壤微生物生物量氮含量、微生物生物量碳氮含量比例、微生物活性与土壤pH值呈显著负相关,土壤微生物生物量碳氮含量及其比例、微生物活性与土壤湿度呈正相关。说明祁连山3种生态系统土壤中微生物生物量和活性受气候要素、植被、有机碳、pH值和湿度等因素 的共同影响。     

黄土丘陵区刺槐叶片-土壤-微生物碳氮磷化学计量学及其稳态性特征

明确植物和微生物在植被恢复过程中的内稳态特性,对反映生物随恢复环境变化的适应性和阐明生态系统养分循环规律有重要意义。以黄土丘陵区恢复5年、10年、20年、30年和45年的刺槐(Robinia pseudoacacia)人工林为研究对象,测定刺槐叶片、土壤和微生物生物量C、N、P含量及其化学计量学指标,重点揭示了叶片和微生物生物量养分在恢复过程中随土壤养分变化的稳态性特征。结果表明:(1)随着恢复年限的增加,土壤、叶片和微生物生物量C、N、P含量表现为增加趋势;(2)不同恢复年限叶片、土壤、微生物生物量C∶N分别为17.03—26.03、9.55—16.94、5.57—10.76、C∶P分别为465.04—634.48、19.89—65.81和39.64—110.53、N∶P分别为17.89—37.03、1.24—4.68和7.15—10.26,除叶片C∶N随恢复年限增加而降低外,其他指标均表现为随恢复年限增加而增加或先增加后降低;刺槐林生长后期可能面临P限制;(3)叶片和微生物生物量C、N、P及其计量比大部分指标与土壤指标的关系能够被内稳态模型很好地模拟(P0.01);其中叶片N∶P、微生物C、N对土壤养分变化较为敏感;其他指标比较稳定。研究表明植物和微生物在面对土壤养分变化时均会通过自我调节呈现内稳态性,说明刺槐在黄土丘陵区有较好的适应性;微生物对土壤养分的变化比植物更加敏感,其养分和计量比指标能较好地指示土壤恢复状况。     

长期模拟升温对崇明东滩湿地土壤微生物生物量的影响

以崇明东滩芦苇湿地为对象,采用开顶室生长箱(Open top chambers OTCs)原位模拟大气升温试验,研究了连续升温8a对崇明东滩湿地0—40cm土层土壤微生物生物量碳氮含量的影响。结果表明:连续升温显著提高了崇明东滩湿地土壤微生物生物量碳氮含量,从土壤表层到深层(0—10,10—20,20—30,30—40cm),微生物生物量碳分别增加了39.32%、70.79%、65.20%、74.09%,微生物生物量氮分别增加了66.46%、178.27%、47.24%、64.11%。但升温对土壤微生物生物量的影响因不同土层和不同季节并未表现出统一的规律,长期模拟升温显著提高4月0—20cm土层和7月0—40cm土层微生物生物量碳氮含量,对10月0—40cm土层微生物生物量碳含量没有影响,但是显著提高了10月0—40cm土层微生物生物量氮含量,同时,微生物生物量碳氮比在7月也显著提高。相关分析表明:无论在升温条件还是在对照条件下,土壤温度、含水量、总氮与土壤微生物生物量碳氮及微生物生物量碳氮比均无相关关系,升温条件下,有机碳与微生物生物量碳氮含量以及微生物生物量碳氮比呈显著正相关,但是在对照条件下有机碳与微生物生物量碳氮含量以及微生物生物量碳氮比呈显著负相关。因此,土壤有机碳是影响土壤微生物生物量碳氮含量对长期模拟升温响应的重要生态因子。     

Optimization of a two-stage recombinant fermentation process: the dilution rate effect

The effects of dilution rates on the performance of a two-stage fermentation system for a recombinant Escherichia coli culture were studied. Dilution rate determines the apparent or averaged specific growth rate of a heterogeneous population of cells in the recombinant culture. The specific growht rate affects the genetic parameters involved in product formation in the second stage, such as plasmid stability, plasmid content, and specific gene expression rate. Kinetic models and correlations were developed for these parameters based on experimental data. Simulations of plasmid stability in the first stage showed that for longer fermentation periods, plasmid stability is better at higher dilution rates. However, the plasmid content is lower at these dilution rates. The optimal apparent specific growth rate for maximum productivity in the second stage was determined using two methods: (1) direct search for a constant specific growth rate, and (2) dynamic optimization using the maximum principle for a time-dependent specific growth rate profile. The results of the calculations showed that the optimum constant apparent specific growth rate for maximum over-all productivity is 0.40 h(-1). This coincides with the optimal specific growht rate for maximum plasmid content in the expressed stage. A 3.5% increase in overall productivity can be obtained by using a linear time dependent apparent specific growth rate control, mu(2)(t) = 0.0007t, in the course of the fermentation time.     

Specific heat flow rate: an on-line monitor and potential control variable of specific metabolic rate in animal cell culture that combines microcalorimetry with dielectric spectroscopy

One of the requirements for enhanced productivity by the animal culture systems used in biotechnology is the direct assessment of the metabolic rate by on-line biosensors. Based on the fact that cell growth is associated with an enthalpy change, it is shown that the specific heat flow rate is stoichiometrically related to the net specific rates of substrates, products, and indeed to specific growth rate, and therefore a direct reflection of metabolic rate. Heat flow rate measured by conduction calorimetry has a technical advantage over estimates for many material flows which require assays at a minimum of two discrete times to give the rate. In order to make heat flow rate specific to the amount of the living cellular system, it would be advantageous to divide it by viable biomass. This requirement has been fulfilled by combining a continuous flow microcalorimeter ex situ with a dielectric spectroscope in situ, the latter measuring the viable cell mass volume fraction. The quality of the resulting biosensor for specific heat flow rate was illustrated using batch cultures of Chinese hamster ovary cells (CHO 320) producing recombinant human interferon-gamma (IFN-gamma) during growth in a stirred tank bioreactor under fully aerobic conditions. The measuring scatter of the probe was decreased significantly by applying the moving average technique to the two participant signals. It was demonstrated that the total metabolic rate of the cells, as indicated by the specific heat flow rate sensor, decreased with increasing time in batch culture, coincident with the decline in the two major substrates, glucose and glutamine, and the accumulation of the by-products, ammonia and lactate. Furthermore, the specific heat flow rate was an earlier indicator of substrate depletion than the flow rate alone. The calorimetric-respirometric ratio showed the intensive participation of anaerobic processes during growth and the related IFN-gamma production. Specific heat flow rate was monotonically related to specific cell growth rate and associated with specific IFN-gamma production. Specific heat flow rate is potentially a valid control variable for the growth of genetically engineered cell lines producing target proteins.     

Metabolite profiles and growth characteristics of Rhizobium meliloti cultivated at different specific growth rates

Rhizobium meliloti (ATCC 55340) was grown at different specific growth rates in a chemostat apparatus. Metabolic products, relating to the Embden-Meyerhof-Parnas (EMP) pathway and the tricarboxylic acid (TCA) cycle, were measured and quantified to probe the influence of specific growth rate on the distribution of important metabolites. The detection of propionate in the fermentation broth implies that the imbalance of reducing equivalents of FADH(2) and NADH + H(+) resulted in a partially reductive operation of the TCA cycle. Additionally, experimental results show that the specific growth rate plays an essential role in modulating the biomass concentration, the specific substrate uptake rate, the cell length, the specific exopolysaccharide (EPS) production rate, the distribution of EPS molecular weight, and the profiles of carbohydrate and organic acid. The specific EPS production rate (varying from 13.3 to 111 mg EPS/g-DW/h) follows a growth-associated pattern at the specific growth rate ranging from 0.06 to 0.20 h(-1) and switches into non-growth-associated mode when the specific growth rate is over 0.20 h(-1).     

Impact of flow velocity on the dynamic behaviour of biofilm bacteria

The impact of flow velocity (FV) on the growth dynamics of biofilms and bulk water heterotrophic plate count (HPC) bacteria in drinking water distribution systems was quantified and modeled by combining a logistic growth model with mass balance equations. The dynamic variations in the specific growth and release rates of biofilm bacteria were also quantified. The experimental results showed that the maximum biofilm biomass did not change when flow velocity was increased from 20 to 40 cm s(-1), but was significantly affected when flow velocity was further increased to 60 cm s(-1). Although the concentration of biofilm bacteria was substantially reduced by the higher shear stress, the concentration of bacteria in the bulk fluid was slightly increased. From this it is estimated that the specific growth rate and specific release rate of biofilm bacteria had doubled. The specific release (detachment) rate was dependent on the specific growth rate of the biofilm bacteria.     

Transient kinetics of hybridoma growth and monoclonal antibody production in serum-limited cultures

A quantitative study of the influence of initial serum concentration on hybridoma growth rate, maximum viable and total cell yield, and specific antibody production rate is presented. The specific growth rate varied in a Monod fashion with initial serum levels (2-10% FCS), giving K(m) = 1.6 v/v% and mu(max) = 0.92 d(-1). The maximum cell yields (total and viable) were linear with initial serum level, indicating stoichiometric as well as kinetic limitation by serum component(s). The specific antibody production rate for each individual run fitted well to a non-growth-associated model. However, the non-growth-associated parameter varied monotonically with initial serum concentration, suggesting the catalytic role of serum component(s) in antibody production. Also, glutamine was utilized inefficiently, with at least a third of it secreted back into the culture supernate in the form of glutamate. While very simple model equations describe the specific growth and product formation rate for an individual batch run, the larger picture indicates need for a more detailed unstructured or structured model.     

The effect of the dilution rate on CHO cell physiology and recombinant interferon-gamma production in glucose-limited chemostat culture

The physiology of a recombinant Chinese hamster ovary cell line in glucose-limited chemostat culture was studied over a range of dilution rates (D = 0.008 to 0.20 h(-1)). The specific growth rate (mu) deviated from D at low dilution rates due to an increased specific death rate. Extrapolation of these data suggested a minimum specific growth rate of 0.011 h(-1) (mu(max) = 0.025 h(-1)) The metabolism at each steady state was characterized by determining the metabolic quotients for glucose, lactate, ammonia, amino acids, and interferon-gamma (IFN-gamma). The specific rate of glucose uptake increased linearly with mu, and the saturation constant for glucose (K(s)) was calculated to be 59.6 muM. There was a linear increase in the rate of lactate production with a higher yield of lactate from glucose at high growth rates. The decline in the rate of production of lactate, alanine, and serine at low growth rate was consistent with the limitation of the glycolytic pathway by glucose. The specific rate of IFN-gamma production increased with mu in a manner indicative of a growth-related product. Despite changes in the IFN-gamma production rate and cell physiology, the pattern of IFN-gamma glycosylation was similar at all except the lowest growth rates where there was increased production of nonglycosylated IFN-gamma. (c) 1993 John Wiley & Sons, Inc.     

Continuous culture of Methanococcus jannaschii, an extremely thermophilic methanogen

Methanococcus jannaschii, an extremely thermophilic methanogen isolated from a deep-sea hydrothermal vent, was grown at 80 degrees C in continuous culture on a mineral salts medium gassed with H(2) and CO(2) at three different flow rates. The maximum specific growth rate was 0.56 h(-1), and the maximum specific methane productivity was 0.32 (mol g(-1) h(-1)). Uncoupling of growth and methane production was evidenced by an increase in teh non-growth-associated rate of methane formation, beta, with increasing gaseous input. The specific hydrogenase activity exhibited growth-assiciated behaviour at low growth rates, but showed no dependence on growth at higher growth rates. The growth dependence of hydrogenase activity is consistent with the pressure dependence of hydrogenase activity measured in previous experiments. In contrast, the specific protease activity was independent of the growth rate over the entire range of dilution rates studied. (c) 1994 John Wiley & Sons, Inc.     

Influence of specific growth rate on specific productivity and glycosylation of a recombinant avidin produced by a Pichia pastoris Mut+ strain

A recombinant avidin-producing Mut+ Pichia pastoris strain was used as a model organism to study the influence of the methanol feeding strategy on the specific product productivity (q(p)) and protein glycosylation. Fed-batch cultivations performed at various specific growth rates (micro) and residual methanol concentrations showed that the specific avidin productivity is growth-dependent. The specific productivity increases strongly with the specific growth rate for micro ranging from 0 to 0.02 h(-1), and increases only slightly with the specific growth rate above this limit. N-terminal glycosylation was also found to be influenced by the specific growth rate, since 9-mannose glycans were the most abundant form at low growth rates, whereas 10-mannose carbohydrate chains were favored at higher micro. These results show that culture parameters, such as the specific growth rate, may significantly affect the activity of glycoproteins produced in Pichia pastoris. In terms of process optimization, this suggests that a compromise on the specific growth rate may have to be found, in certain cases, to work with an acceptable productivity while avoiding the addition of many mannoses.     

On the interpretation of some planktonology equations

Summary The concept of the mean value of a function is used to interpret some population-dynamics equations. The well-known formula for the per capita growth rate r gives a precise mean value for any (not only exponentially growing) populations. This result is used to derive the birth and death rate equations of Paloheimo (1974) with minimal initial limitations.Abbreviations t time - N number of animals - E number of eggs - r specific (i.e. per capita) population growth rate - b specific birth rate - d specific death rate - D duration of embryonic development     

Over-expression of recombinant human interferon-gamma in high cell density fermentation of Escherichia coli

Human interferon-gamma (hIFN-gamma) was expressed in Escherichia coli BL21(DE3) under the control of the T7 promoter. Glucose was used as the sole source of carbon and energy with simple exponential feeding rate in fed-batch process. Cell density of recombinant E. coli was reached to 100 g dry wt l(-1) under both constant (0.12 h(-1)) and variable (0.12-0.52 h(-1)) specific growth rates. In the variable specific growth rate fed-batch process, plasmid stability and specific yield of rhIFN-gamma were greater than constant specific growth rate fed-batch process. The final specific yield and overall productivity of rhIFN-gamma were 0.35 +/- 0.02 g rhIFN-gamma g(-1) dry cell wt and 0.9 +/- 0.05 g rhIFN-gamma l(-1) h(-1) in the variable specific growth rate fed-batch process, respectively.