Methane production and growth of microorganisms under different moisture conditions in soils with added chitin and without it

The limits of soil moisture providing the possibility of methane production and growth of microorganisms in soils with added chitin and without it were determined. Samples of gray forest, soddy-podzolic, gley taiga, chestnut, and chernozemic soils were studied. It was found that methane emission increases significantly under a high soil moisture content in the presence of chitin. The increase of the soil moisture up to the maximum water-holding capacity enhanced the emission of methane by two-six times. The dynamics of the methane emission from the soils in the course of microbial successions initiated by the addition of chitin or by the soil moistening to different levels were studied by the gas-chromatographic method. The population density and biomass of fungal, bacterial, and actinomycetic complexes under different moister levels were studied by the method of luminescent microscopy. It was determined that many microorganisms participate in the transformation of chitin in the soil under conditions of oxygen deficiency (upon the increased moisture content). Prokaryotes dominated by actinomycetes were the group that increased its biomass most actively (the biomass doubling took place).     

Microbial destruction of chitin in soils under different moisture conditions

The most favorable moisture conditions for the microbial destruction of chitin in soils are close to the total water capacity. The water content has the most pronounced effect on chitin destruction in soils in comparison with other studied substrates. It was found using gas-chromatographic and luminescent-microscopic methods that the maximum specific activity of the respiration of the chitinolytic community was at a rather low redox potential with the soil moisture close to the total water capacity. The range of moisture values under which the most intense microbial transformation of chitin occurred was wider in clayey and clay loamy soils as compared with sandy ones. The increase was observed due to the contribution of mycelial bacteria and actinomycetes in the chitinolytic complex as the soil moisture increased.     

Resistance of microbial populations in DDT-contaminated and uncontaminated soils

One DDT-contaminated soil and two uncontaminated soils were used to enumerate DDT-resistant microbes (bacteria, actinomycetes and fungi) by using soil dilution agar plates in media either with 150 μg DDT ml⁻¹ or without DDT at different temperatures (25, 37 and 55°C). Microbial populations in this study were significantly (p<0.001) affected by DDT in the growth medium. However, the numbers of microbes in long-term contaminated and uncontaminated soils were similar, presumably indicating that DDT-resistant microbes had developed over a long time exposure. The tolerance of isolated soil microbes to DDT varied in the order fungi>actinomycetes>bacteria. Bacteria from contaminated soil were more resistant to DDT than bacteria from uncontaminated soils. Microbes isolated at different temperatures also demonstrated varying degrees of DDT resistance. For example, bacteria and actinomycetes isolated at all incubation temperatures were sensitive to DDT. Conversely fungi isolated at all temperatures were unaffected by DDT.     

Specificity of soil hydrolytically microbial complex under different moisture conditions

It has been established that soil moisture has a significant impact on the activity of chitinolytic microbial processes, rather than pectinolytic processes. The degradation of polysaccharides with an increase in soil moisture in microbial complex markedly increases the role of prokaryotic microorganisms, especially actinomycetes. For the first time, using the FISH method, the amount of detected phylogenetic composition of a metabolically active hydrolytic complex of humus horizons of grey forest and gley and weakly podzolic soil and humus has been estimated depending on the humidity. At optimum moisture, phylogenetic groups Actinobacteria and Firmicutes dominated in the chitinolytic process. An increase in the proportion of proteobacteria is observed with an increase in humidity. The role of gamma- and alphaproteobacteria and actino-bacteria is heightened with the drying of soil in the hydrolytic complex. A quantitative estimate of the rate of degradation of polysaccharides (pectin and chitin) in different types of soils at different levels of moisture is given. The dependence of the phylogenetic composition of an active microbial hydrolytic complex of humus horizons of grey forest and gley, weakly podzolic soils and humus on humidity is revealed.     

The influence of aeration and temperature on the structure of bacterial complexes in high-moor peat soil

The number and taxonomic structure of the heterotrophic block of aerobic and facultative anaerobic bacteria were studied in monoliths from a high-moor peat (stored at room temperature and in a refrigerator) and in the peat horizons mixed in laboratory vessels. The monitoring lasted for a year. In the T0 horizon, spirilla predominated at room and low temperatures; in the T1 and T2 horizons, bacilli were the dominants. The continuous mixing of the peat layers increased the oxygen concentration and the peat decomposition; hence, the shares of actinomycetes and bacilli (bacteria of the hydrolytic complex) increased. In the peat studied, the bacilli were in the active state; i.e., vegetative cells predominated, whose amount ranged from 65 to 90%. The representatives of the main species of bacilli (the facultative anaerobic forms prevailed) hydrolyzed starch, pectin, and carboxymethylcellulose. Thus, precisely sporiferous bacteria can actively participate in the decomposition of plant polysaccharides in high-moor peat soils that are characterized by low temperatures and an oxygen deficit. The development of actinomycetes is inhibited by low temperatures; they can develop only under elevated temperature and better aeration.     

不同种类微生物及其碳酸酐酶对土壤-灰岩系统钙镁锌元素迁移作用的土柱模拟实验研究

通过土柱模拟实验研究了筛选自西南岩溶区的典型细菌、真菌、放线菌对石灰岩的溶蚀动态及作用效果,并研究了这3种类群微生物及其碳酸酐酶(CA)对土壤-灰岩系统Ca、Mg、Zn元素迁移的影响。结果表明:①实验所用的典型细菌、真菌、放线菌均对石灰岩有较强的溶蚀作用,但是不同类群的微生物对石灰岩的溶蚀效果不同,溶蚀后石灰岩的表面形貌变化不同,其中,真菌的溶蚀效果最显著,放线菌和细菌次之;②典型细菌、真菌、放线菌对不同元素迁移的影响不同,对于Zn2 、Ca2 的迁移影响而言,真菌>放线菌>细菌>无菌对照组;对于Mg2 的迁移影响而言,细菌>真菌>放线菌>无菌对照组;③一定范围内温度增加对Ca、Mg、Zn3种元素的迁移有不同程度的促进作用;④土柱淋出液中CA平均活性与Ca、Mg、Zn3种元素的总淋失量之间存在一定的正相关关系,说明CA对这3种元素的迁移具有一定的影响,且因不同元素而异。本研究结果对于评价不同类群微生物在生物岩溶及岩溶系统元素迁移中的作用和地位提供了一定的科学依据。     

The origin and nature of organic nitrogen in soil as assessed by acidic and alkaline hydrolysis

The origin and nature of much of the organic nitrogen in soil is unknown or speculative. Alkaline hydrolysis was used to fractionate soils into alkali‐soluble and insoluble fractions. Alkali‐insoluble residues from a wide variety of soils contained largely fixed ammonium or chitin or both. Acid hydrolysis of alkali‐insoluble residues from soils, microbes, insects and well‐rotted plant litter confirmed the presence of hexosamine‐N. The hexosamine‐N derived from estimates of soil biomass accounted for less than 10% of the chemically determined hexosamine‐N values. It is suggested that most of the hexosamines are held in complexes formed between chitin complexed with other materials, e.g. pigments, that occur predominantly in the cell walls of microbes and insects. These complexes show resistance to enzymes during the life of soil organisms that persists after their death.     

DDT Resistance and Transformation by Different Microbial Strains Isolated from DDT-Contaminated Soils and Compost Materials

Bioremediation is a potential option to treat 1, 1, 1-trichloro-2, 2-bis (4-chlorophenyl) ethane (DDT) contaminated sites. In areas where suitable microbes are not present, the use of DDT resistant microbial inoculants may be necessary. It is vital that such inoculants do not produce recalcitrant breakdown products e.g. 1, 1-dichloro-2, 2-bis (4-chlorophenyl) ethylene (DDE). Therefore, this work aimed to screen DDT-contaminated soil and compost materials for the presence of DDT-resistant microbes for use as potential inoculants. Four compost amended soils, contaminated with different concentrations of DDT, were used to isolate DDT-resistant microbes in media containing 150 mg l^?1 DDT at three temperatures (25, 37 and 55°C). In all soils, bacteria were more sensitive to DDT than actinomycetes and fungi. Bacteria isolated at 55°C from any source were the most DDT sensitive. However DDT-resistant bacterial strains showed more promise in degrading DDT than isolated fungal strains, as 1, 1-dichloro 2, 2-bis (4-chlorophenyl) ethane (DDD) was a major bacterial transformation product, while fungi tended to produce more DDE. Further studies on selected bacterial isolates found that the most promising bacterial strain (Bacillus sp. BHD-4) could remove 51% of DDT from liquid culture after 7 days growth. Of the amount transformed, 6% was found as DDD and 3% as DDE suggesting that further transformation of DDT and its metabolites occurred.     

不同土壤管理方式对梨园土壤微生物及养分含量的影响

通过田间试验研究不同土壤管理方式对梨园表层（0 ~ 20 cm）和亚表层土壤（20 ~ 40 cm）细菌、真菌、放线菌数量和土壤养分含量的影响。结果表明：梨园土壤微生物数量具有垂直分布的特征,土壤微生物数量随土层的加深而递减。梨园表层和亚表层土壤中微生物数量均细菌最多,放线菌次之,真菌最少。与覆草1年相比较,覆草3年的梨园土壤除表层土壤细菌含量无显著差异外,表层和亚表层土壤中真菌和放线菌数量均显著增加。与自然生草、覆膜、常规灌溉处理相比较,覆草与覆膜结合能最大限度地增加土壤微生物数量。连续覆草3年显著提高了梨园土壤有机质、碱解氮、速效磷、速效钾含量,而覆膜处理使土壤速效养分含量下降,覆膜结合覆草处理能显著提高土壤有机质、速效磷、速效钾含量。     

Amino sugars and muramic acid—biomarkers for soil microbial community structure analysis

Characterizing functional and phylogenetic microbial community structure in soil is important for understanding the fate of microbially-derived compounds during the decomposition and turn-over of soil organic matter. This study was conducted to test whether amino sugars and muramic acid are suitable biomarkers to trace bacterial, fungal, and actinomycetal residues in soil. For this aim, we investigated the pattern, amounts, and dynamics of three amino sugars (glucosamine, mannosamine and galactosamine) and muramic acid in the total microbial biomass and selectively cultivated bacteria, fungi, and actinomycetes of five different soils amended with and without glucose. Our results revealed that total amino sugar and muramic acid concentrations in microbial biomass, extracted from soil after chloroform fumigation varied between 1 and 27 mg kg⁻¹ soil. In all soils investigated, glucose addition resulted in a 50-360% increase of these values. In reference to soil microbial biomass-C, the total amino sugar- and muramic acid-C concentrations ranged from 1-71 g C kg⁻¹ biomass-C. After an initial lag phase, the cultivated microbes revealed similar amino sugar concentrations of about 35, 27 and 17 g glucosamine-C kg⁻¹ TOC in bacteria, fungi, and actinomycetes, respectively. Mannosamine and galactosamine concentrations were lower than those for glucosamine. Mannosamine was not found in actinomycete cultures. The highest muramic acid concentrations were found in bacteria, but small amounts were also found in actinomycete cultures. The concentrations of the three amino sugars studied and muramic acid differed significantly between bacteria and the other phylogenetic microbial groups under investigation (fungi and actinomycetes). Comparison between the amino sugar and muramic acid concentrations in soil microbial biomass, extracted after chloroform fumigation, and total concentrations in the soil showed that living microbial biomass contributed negligible amounts to total amino sugar contents in the soil, being at least two orders of magnitude greater in the soils than in the soil inherent microbial biomass. Thus, amino sugars are significantly stabilized in soil.     

Psychrotolerant actinomycetes of plants and organic horizons in tundra and taiga soils

It has been revealed that in organic horizons and plants of the tundra and taiga ecosystems under low temperatures, actinomycetal complexes form. The population density of psychrotolerant actinomycetes in organic horizons and plants reaches tens and hundreds of thousands CFU/g of substrate or soil, and decreases in the sequence litters > plants > soils > undecomposed plant remains > moss growths. The mycelium length of psychrotolerant actinomycetes reaches 220 m/g of substrate. Application of the FISH method has demonstrated that metabolically active psychrotolerant bacteria of the phylum Actinobacteria constitute 30% of all metabolically active psychrotolerant representatives of the Bacterià domain of the prokaryotic microbial community of soils and plants. Psychrotolerant actinomycetes in tundra and taiga ecosystems possess antimicrobial properties.     

Cucumber Productivity and Soil Degradation in Recropping System in Greenhouse

Soil degradation resulting from recropping of greenhouse-cultivated cucumber (Cucumis statirus L.) has become a serious problem for production. The objective was to determine the soil degradation processes and make suggestions for prevention. Experiments were conducted in a solar greenhouse 4.0 m long and 0.7 m wide during 2001–2005 in northern China. In 2001, every fourth plot was filled with continuously cropped typical sandy soils from the winter–spring cucumber season for 0, 1, 4, or 8 years, respectively. Soil particle composition, nutrient content, and species of soil microbes were analyzed. A grafted cucumber with squash root was transplanted to the continuously cropped soil. Results indicated that cucumber fruit productivity increased with increases of recropping year up to the fourth year and then decreased thereafter with continuous cropping. Soil particle percentages did not change significantly as recropping years increased. Soil nutrients were not the main factor for decreased productivity. As the number of recropping years increased, the soil available nitrogen (N), available phosphorus (P), available potassium (K), and organic content increased. The soil bacteria and fungi increased after 3 or 5 years of recropping, and soil actinomycetes decreased significantly after 5 years of recropping. The ratio of bacteria–fungi–actinomycetes changed, populations of fungi and bacteria increased, and the ratio of actinomycetes decreased, which led to an increase in the occurrence of pathogenic and soil-borne microbes and a decrease in fruit productivity, which were assumed to be the main factors of soil degradation. To stop recropping from influencing populations of fungi, bacteria, and actinomycetes, recropping should not be utilized after 4 years of recropping in order to continue with vegetable productivity in greenhouses.     

土壤质地对秸秆分解的影响及其微生物机制

本研究以河南省封丘县的砂质、壤质、黏质潮土为对象,基于网袋秸秆埋设法,比较分析了经半年埋设后小麦和玉米秸秆分解速率变化,及基于磷脂脂肪酸(PLFA)的微生物组成差异。结果表明:秸秆分解率在不同质地潮土间无显著性差异;残留秸秆中碳含量与原始秸秆相当,表明秸秆分解过程中碳按比例减少;但氮和磷含量显著增加,尤其在质地黏重的土壤处理中更为明显。经半年埋设,小麦和玉米秸秆中钾的释放比例分别高达99%和97%。PLFA结果显示,两种秸秆中的细菌、真菌、放线菌在砂、壤、黏土处理间无显著差异,表明影响秸秆分解的微生物生物量在不同土壤质地处理间相当,这可能是导致秸秆在不同质地潮土中分解率无显著差异的原因之一;但PCA分析表明,参与小麦和玉米秸秆分解的微生物种类有所不同,其差别主要与革兰氏阳性菌14:0、i14:0、3OH-16:0、i17:0,革兰氏阴性菌cy17:0以及放线菌10Me18:0有关。     

干旱胁迫下枸溶性钾肥配施对烤烟土壤理化性质、微生物数量及根系生长的影响

为探明枸溶性钾肥与硫酸钾肥配施对土壤改良和烤烟生长的影响,通过盆栽烟株进行干旱处理,研究枸溶性钾肥不同配施比例在干旱胁迫条件下对土壤理化性质(土壤容重、土壤孔隙度、土壤钾含量),土壤微生物(真菌、细菌、放线菌)活性以及根系生长(根系生物量、根系构型参数、根系活力)的影响。结果表明,施加枸溶性钾肥可使土壤容重降低、孔隙度增大、速效钾和缓效钾含量增加、微生物数量提高,促进烤烟根系生长。在正常水分调控下,以75%枸溶性钾肥+25%的硫酸钾肥的处理最佳;在干旱胁迫条件下,以50%枸溶性钾肥+50%硫酸钾的处理最佳。在烤烟实际生产中,建议在保水能力差的土壤或降雨量少的烟区配施枸溶性钾肥。本研究对改善植烟土壤和提高烟叶品质具有指导意义。     

Response of the chitinolytic microbial community to chitin amendments of dune soils

 The dynamics of culturable chitin-degrading microorganisms were studied during a 16-week incubation of chitin-amended coastal dune soils that differed in acidity. Soil samples were incubated at normal (5% w/w) and high (15% w/w) moisture levels. More than half of the added chitin was decomposed within 4 weeks of incubation in most soils. This rapid degradation was most likely due to fast-growing chitinolytic fungi (mainly Mortierella spp. and Fusarium spp.) at both moisture levels, as dense hyphal networks of these fungi were observed during the first 4 weeks of incubation. Chitin N mineralization was inhibited by cycloheximide, and fast-growing fungal isolates were capable of rapid chitin decomposition in sterile sand, further suggesting that these fungi play an important role in initial chitin degradation. The strong increase in fast-growing fungi in chitin-amended dune soils was only detected by direct observation. Plate counts and microscopic quantification of stained hyphae failed to reveal such an increase. During the first part of the incubation, numbers of unicellular chitinolytic bacteria also increased, but their contribution to chitin degradation was indicated to be of minor importance. During prolonged incubation, colony forming units (CFU) of chitinolytic streptomycetes and/or slow-growing fungi increased strongly in several soils, especially at the 5% moisture level. Hence, the general trend observed was a succession from fast-growing fungi and unicellular bacteria to actinomycetes and slow-growing fungi. Yet, the composition of chitinolytic CFU over time differed strongly between chitin-amended dune soils, and also between the two moisture levels. These differences could not be attributed to pH, organic matter or initial microbial composition. The possible consequence of such unpredictable variation in microbial community composition for the use of chitin-amendments as a biocontrol measure is discussed. Received: 10 March 1998     

植烟土壤中微生物特性及氨化、亚硝化菌分离鉴定与活性研究

针对烤烟生产中施用有机肥后植烟土壤有机态氮素分解速率不易调控的问题,并为配制有机氮分解微生物菌剂提供优良菌株,本研究以化肥、牛粪、玉米秸秆、油枯处理下的烤烟根区土壤为材料,分离筛选出高活性氨化、亚硝化土著菌,并分别测定其对有机氮降解及氨氮分解的效果。结果表明:在烤烟生长旺长期,施用有机肥处理的土壤可培养细菌、真菌、放线菌、氨化菌数量均高于单施化肥处理;在烤烟成熟期,施用有机肥处理,烤烟根区土壤氨化菌、亚硝化菌、真菌、放线菌数量均高于单施化肥。通过各类菌株的土样来源分析,氨化作用强度较高的优势菌株均来源于有机肥处理的土样,植烟土壤施用有机肥较单施化肥能够获得较高活性的氨化细菌。各类菌活性研究显示,短小芽孢杆菌(Bacillus pumilus)、嗜热脂肪地芽孢杆菌(Geobacillus stearothermophilus)、巨大芽孢菌(B.megaterium)培养液有机氮含量降幅最大,较初始有机氮含量分别降低84.74%、92.74%、79.52%;寡养单胞菌(Stenotrophomonas sp.)兼具有亚硝化作用及硝化作用,培养7 d后,培养液中硝态氮含量增加了0.617 mg·L~(-1);嗜麦芽寡单胞菌(S.maltophilia)亚硝化作用最强,培养7 d后,培养液中亚硝态氮含量为0.518 mg·L~(-1)。氨化菌培养48 h后有机氮分解速率降低,具有亚硝化及硝化作用的菌株培养到第7 d活性仍处于较高水平。因此,试验分离出的不同功能细菌在配制解氮复合微生物菌剂时需在不同时间加入菌株进行发酵,以获得高活性微生物菌剂。根据分解有机氮、解氨试验及相关文献中各菌株作用功能分析,筛选出纳西杆菌(Naxibacter sp.)、寡养单胞菌、短小芽孢杆菌、嗜热脂肪地芽孢杆菌、同温层芽孢杆菌(B.stratosphericus)、纤维菌(Cellulosimicrobium cellulans)、高地芽孢杆菌(B.altitudinis)和巨大芽孢菌8株高效氮素功能菌株用于有机氮分解微生物菌剂的配制。     

A simple method for detection of lipolytic microorganisms in soils

Media selective for the isolation of bacteria, actinomycetes and fungi were amended with 0.1% sunflower oil emulsified with 0.01% Tween 80. Lipase-producing microorganisms produced clear zones on these media. When lipase-producing bacteria were cultured on a polycarbonate membrane laid on the selective medium for bacteria, clear zones were produced on the medium when the membrane along with bacteria was removed. The agar disc cut from the clear zone also produced a clear zone when placed on the fresh medium, indicating that clear zone formation is the result of the activity of extracellular lipases. The largest population of lipase-producing microorganisms in an agricultural soil was actinomycetes followed by bacteria and fungi. Ranging from 12 to 75% of bacteria, actinomycetes and fungi isolates from soils collected from three different locations were capable of producing lipases. In general, relatively small percentages of soil bacteria were lipase producers, and lipase producers were more common among soil actinomycetes and fungi. These three groups of microorganisms appear to be all important in decomposition of oils in organic matters in soils.     

不同施肥制度对稻田土壤微生物的影响研究

湖南省进行了18年土壤肥力与施肥效益监测的长期定位试验,就3个定位点不同施肥制度对稻田土壤微生物的影响进行了研究。结果表明,长期施肥土壤的有机质含量与细菌总数及放线菌总数之间无直接相关,土壤有机质含量的高低并不能作为评价土壤微生物多少的指标;真菌数量可反映土壤的肥力水平,土壤真菌数量可用来评价不同施肥制度的效果。施用30%有机肥和习惯施肥处理的氮素养分和硝化细菌数量较多,硝化细菌数量与有机质含量也无相关;长期施用大量有机肥(60%)的土壤反硝化细菌数量高于其它施肥处理,这可能与长期使用大量有机肥后土壤氧化还原电位降低有关。硫化细菌和反硫化细菌数量与土壤有机质含量无直接相关性,施肥可影响土壤硫素的供应。施用单质氮肥多的土壤,氨化细菌数量较多,而有机肥施用量高的土壤,氨化细菌数量较少。无肥区的土壤微生物活度最低,60%有机肥处理的土壤微生物活度最高。土壤有机质含量高,土壤微生物活度也高,两者呈正相关。保持农业土壤的健康要从多方面来进行联合调控,而不是单纯的采用某一农业技术来进行。     

温度水分对秸秆降解微生物群落功能多样性影响

秸秆腐解微生物群落结构受环境因子(温度和水分)影响显著。本试验利用BIOLOG技术,以秸秆腐解微生物碳源利用的平均颜色变化率(averagewell colordevelopment,AWCD)为指标,研究了不同温度(15℃,25℃和35℃和水分(40%,70%和90%田间持水量)条件下小麦和玉米秸秆腐解过程中微生物碳源代谢多样性的差异试验结果表明不同处理的AWCD随培育温度升高而降低,即15℃25℃35℃。随着培育时间的增加,其降低的趋势更加明显。同样地,小麦玉米秸秆腐解微生物的物种丰富度指数Shannon-Wiener(H)和优势度指数Simpson(D)也呈现出随温度升高而降低的趋势。微生物主要利用糖类和脂类物质。主成分分析结果表明在不同腐解时间,腐解微生物代谢多样性在不同温度下差异显著,而在不同水分下差异不显著。     

铅锌矿区重金属污染对微生物数量及放线菌群落结构的影响

采集四川省汉源县富泉乡万顺铅锌矿区5个不同重金属浓度的土壤样品,进行了微生物数量及放线菌多样性的研究。经分离、纯化得到43株不同的放线菌,然后对其进行BOXAIR-PCR和16SrDNAPCR-RFLP分析。结果表明,铅锌矿区重金属复合污染对土壤微生物数量有较大的影响,随着铅锌矿区重金属污染程度的加剧,土壤微生物的总数下降。相关性分析表明,重金属含量与细菌数量呈极显著负相关（P〈0.01）,与放线菌数量、真菌数量呈显著负相关（P〈0.05）。供试菌株的16SrDNA用HaeⅢ、HinfⅠ和TaqⅠ酶切后具有32种遗传图谱类型。BOXAIR-PCR的聚类结果表明在86%的水平上,所有菌株分为10个遗传类型,结果基本与16SrDNAPCR-RFLP聚类差异不大。来源于高重金属的含量样品的菌株基本聚在一起,可能是重金属含量影响了放线菌的分布。同时,16SrDNA序列聚类分析结合系统发育树分析表明链霉菌属是汉源铅锌矿区主要的放线菌属并且具有遗传多样性。