绵羊瘤胃微生物胞外蛋白酶性质的研究

本试验测定了pH、温度和EDTA对纯化的绵羊的瘤胃微生物胞外蛋白酶活性的影响,并用一已知氨基酸排列顺序的肽段(由15个氨基酸组成)作为底物,进一步测定了蛋白酶的酶切位点。结果表明,(1)纯化蛋白酶的最适pH在6.0～6.5之间,最适温度为40℃左右。(2)不同浓度的EDTA(浓度分别为1,10,25,50,75和100mM)对蛋白酶活性没有影响,因而该酶属于丝氨酸蛋白酶类。(3)纯化蛋白酶是一种内肽酶,其酶切位点在组氨酸—酪氨酸(?)、天冬氨酸—丙氨酸(?)、亮氨酸—赖氨酸(?)和(或)缬氨酸—赖氨酸(?)相连的肽键。(4)胰蛋白酶和纯化的蛋白酶虽同属丝氨酸蛋白酶类,但前者对底物具有高度的专一性,后者对底物的专一性不强,这是瘤胃内饲料蛋白质强烈降解的主要原因之     

秸秆还田种类对稻田N2O排放及硝化反硝化微生物的影响

以太湖流域典型单季稻田的原状土柱为研究对象,通过设置温室土柱试验,同步监测3种秸秆(水稻秸秆RS、小麦秸秆WS、玉米秸秆MS)施用下水稻各生长期N₂O排放、水稻产量和土壤理化因子,同时定量化分析多个N₂O排放相关菌群及功能基因的丰度,以阐明N₂O排放对不同种类秸秆施用引发的微生物响应机制,筛选控制单季稻田N₂O减排增效最佳的秸秆种类。结果表明：与对照相比,RS、WS和MS处理下水稻生长期N₂O排放量分别增加162.32%、107.11%和9.48%,其中RS处理显著高于MS处理。水稻生育期内,土壤氨氧化菌(AOA、AOB)和反硝化菌群落(nir S、nos Z)丰度均呈现先上升后下降的变化趋势。与对照相比,拔节期RS处理显著增加AOA、AOB、nirS和nosZ拷贝数,MS和WS处理对上述功能基因丰度均无显著影响。各生育期土壤NH₄⁺-N含量整体高于NO₃^–-N含量,二者均在水稻分蘖期达到峰...     

冻融交替对土壤氮素转化及相关微生物学特性的影响

土壤冻融是发生在中、高纬度及高海拔地区的一种常见的自然现象。冻融作用通过影响土壤物理性质及生物学性状进而对土壤氮素转化过程产生重要影响,但目前关于冻融作用对土壤氮素转化过程影响的研究结果还不尽一致,对于冻融作用下土壤微生物学特性的研究相对较少。本文着重论述了冻融作用对土壤氮素转化过程(有效氮素含量变化、氮素净矿化速率、氮素损失途径等)的影响,并对冻融作用下土壤微生物生理和代谢特性进行了归纳和总结,简要指出目前研究过程中存在的问题,并对未来研究方向提出展望。     

Kinetics of soil microbial uptake of free amino acids

 Amino acids and proteins typically form the biggest input of organic-N into most soils and provide a readily available source of C and N for soil microorganisms. Amino acids can also be taken up directly by plant roots, providing an alternative source of available soil N. However, the degree to which plants can compete against the soil microbial population for amino acids in soil solution remains poorly understood. The aim of this study was to measure the rate of microbial uptake of three contrastingly charged ¹⁴C-labelled amino acids (glutamate^1–, glycine⁰, lysine^0.9+) over a wide concentration range (0.1–5 mM) and in two contrastingly managed soils varying in their degree of erosion, organic-C content and microbial biomass. Amino acid uptake was concentration dependent and conformed to a single Michaelis-Menten equation. The mean maximum amino acid uptake rate (V _max) for the non-eroded (control) soil (high organic-C, high biomass) was 0.13±0.02 mmol kg^–1 h^–1, while half maximal uptake occurred at a concentration (K _m) of 2.63±0.07 mM. Typically, V _max was fourfold lower and K _m twofold lower in the eroded soil (low available organic-C, low biomass) compared to the non-eroded (control) soil. Amino acid substrate concentration had little effect on the proportion of amino acid utilized in catabolic versus anabolic metabolism and was similar for both. While the results obtained here represent the summation of kinetics for a mixed soil population, they indicate that amino acid uptake is saturated at concentrations within the millimolar range. Because the affinity constants also were similar to those described for plant roots, we hypothesized that competition for amino acids between plants and microbes will be strong in soil but highly dependent upon the spatial distribution of roots and microbes in soil. Received: 2 March 2000     

Application of 16S rDNA Sequence Analysis Technique in Microbial Detection

16srDNA既具有保守性区域。又具有高变性区域,是生物的种属鉴定和系统进化关系的重要分子基础。以16srDNA为目的物的现代分子生物学技术能精确地揭示微生物种类和遗传的多样性,从而16srDNA序列分析成了微生物分类鉴定主要依据。该方法克服了传统微生物培养方法的限制,操作方便、检测快速准确且灵敏度高,已被广泛应用到菌种鉴定、群落对比分析、群落中系统发育及种群多样性的评估等领域,是一种客观和可信度较高的分类方法。     

嗜金微生物研究概况

在广泛文献检索基础上,对嗜金微生物的种属、富集黄金的机理、工业应用价值及潜力进行了概述,为进一步对其开发利用提供科学资料。     

污染土壤的微生物修复原理与技术进展

微生物修复是土壤环境生物修复技术的重要组成部分,是最具发展和应用前景的生物学环保新技术.本文以土壤中重金属及典型有机污染物为对象,从土壤微生物与污染物质的相互作用入手,较为系统地综合评述国内外污染土壤的微生物修复原理与技术,并结合当前土壤污染的新特点,提出了需进一步认识和解决微生物修复过程中出现的新现象和新问题,以不断丰富和拓展土壤环境微生物修复范畴与内涵.     

Effects of antibiotics on soil microorganisms: time and nutrients influence pollution-induced community tolerance

A method for the detection of the effects of antibiotics on soil microbial communities was optimised in the present study. We investigated the influence of measurement time and nutrient status on the pollution-induced community tolerance (PICT), using the sulfonamide sulfachloropyridazine (SCP) as model compound. The tolerance development in soil microcosms that were exposed to SCP under different conditions was compared with the background tolerance in SCP-unexposed microcosms. The tolerance of bacterial extracts from the soil microcosms was determined in Biolog^® multiwell plates as the SCP sensitivity of a range of physiological processes. The background tolerance was not affected by soil nutrient amendments, but an influence of the inoculum density in the microtiter plates was observed. Still, the variance of the background tolerance was low, which is in favour of the usage of community tolerance measurements for a selective detection of a toxicant impact. In line with the assumptions of the PICT concept, soil amendment with SCP led to an increase in community tolerance. This tolerance development was enhanced upon additional soil amendment with fresh pig slurry, and less by alfalfa meal addition. Tolerance increases were observed after a soil exposure to SCP of only 7 days, possibly because nutrient input facilitates the fast adaptation of the soil microbial community. However, a further increase in exposure time led to variable changes in the observed tolerance. Prolonged tolerance detection in the microtiter plates (11 days) enabled a clearer differentiation between different soil treatments, as it better resolved the EC₅₀ values of processes with a high tolerance to bacteriostatic antimicrobial compounds. For the detection of antibiotic effects on soil microbial communities, it is therefore recommended to use nutrient amendments (possibly fresh pig slurry), to standardize the soil exposure time, and to extend the period of Biolog plate measurement beyond 7 days.     

Inoculation of root microorganisms for sustainable wheat-rice and wheat-black gram rotations in India

The scarcity of non-renewable resources such as soils and fertilizers and the consequences of climate change can dramatically influence the food security of future generations. Mutualistic root microorganisms such as plant growth-promoting rhizobacteria (PGPR) and arbuscular mycorrhizal fungi (AMF) can improve plant fitness. We tested the growth response of wheat (Triticum aestivum [L.]), rice (Oriza sativa [L.]) and black gram (Vigna mungo [L.], Hepper) to an inoculation of AMF and PGPR alone or in combination over two years at seven locations in a region extending from the Himalayan foothills to the Indo-Gangetic plain. The AMF applied consisted of a consortium of different strains, the PGPR of two fluorescent Pseudomonas strains (Pseudomonas jessenii, R62; Pseudomonas synxantha, R81), derived from wheat rhizosphere from one test region. We found that dual inoculation of wheat with PGPR and AMF increased grain yield by 41% as compared to un-inoculated controls. Yield responses to the inoculants were highest at locations with previously low yields. AMF or PGPR alone augmented wheat grain yield by 29% and 31%, respectively. The bio-inoculants were effective both at Zero and at farmers’ practice fertilization level (70 kg N ha⁻¹, 11 kg P ha⁻¹ in mineral form to wheat crop). Also raw protein (nitrogen × 5.7) and mineral nutrient concentration of wheat grains (phosphorus, potassium, copper, iron, zinc, manganese) were higher after inoculation (+6% to +53%). Phosphorus use efficiency of wheat grains [kg P grain kg⁻¹ P fertilizer] was increased by 95%. AMF and PGPR application also improved soil quality as indicated by increased soil enzyme activities of alkaline and acid phosphatase, urease and dehydrogenase. Effects on rice and black gram yields were far less pronounced over two cropping seasons, suggesting that AMF and PGPR isolated from the target crop were more efficient. We conclude that mutualistic root microorganisms have a high potential for contributing to food security and for improving nutrition status in southern countries, while safeguarding natural resources such as P stocks.     

微生物菌群之间配比协调关系的研究

对乳酸杆菌、乳酸球菌、枯草芽胞杆菌、酵母菌、大肠杆菌等菌种进行单独培养和不同比例的混合培养,测定了发酵过程中pH值、活菌数量及光密度的变化情况。结果表明,与含有25%混合菌液的发酵麸皮相比,含50%混合菌液的发酵麸皮,在发酵24 h后的需氧或厌氧活菌总数都有升高趋势(P>0.05),发酵1周后其pH值的下降趋势明显(P<0.05),说明乳酸菌数越多使pH下降越大。利用需氧(LB)和厌氧(MRS)培养液分别交叉培养需氧菌和厌氧菌,均能在相应的培养基上生长。充分说明厌氧菌和需氧菌混合培养在理论上是可行的。但是,如果用滤菌膜过滤后的培养过乳酸菌的MRS培养基来培养需氧菌时,这些菌种均不能在滤液中生长。