放牧与围栏内蒙古针茅草原土壤微生物生物量碳、氮变化及微生物群落结构PLFA分析

以内蒙古贝加尔针茅草原、大针茅草原和克氏针茅草原为研究对象,采用氯仿熏蒸法和磷脂脂肪酸（PLFA）分析方法研究了放牧与围栏条件下内蒙古针茅草原土壤微生物生物量和群落结构特征的变化情况。研究表明放牧与围栏草地土壤微生物生物量和群落结构差异显著。氯仿熏蒸法分析结果表明内蒙古针茅草原土壤微生物生物量碳的含量介于166.6-703.5mg·kg^-1之间,微生物生物量氮含量介于30.34-92.15mg·kg^-1之间,其中贝加尔针茅草原土壤微生物生物量碳、氮最高,大针茅草原次之,克氏针茅草原则最低。放牧条件下,贝加尔针茅草原、大针茅草原土壤微生物生物量碳、氮显著低于围栏草地,克氏针茅草原则无显著变化。PLFA分析结果显示,内蒙古针茅草原土壤微生物群落PLFAs种类、含量丰富,共检测出28种PLFA生物标记磷脂脂肪酸,并且以直链饱和脂肪酸和支链饱和脂肪酸为主,相对含量占总量的2/3左右,其中贝加尔针茅草原土壤微生物含量最丰富,其围栏样地土壤的PLFA含量达到27.3nmol·g-1,大针茅草原和克氏针茅草原依次降低。围栏条件下,各类型草原土壤细菌脂肪酸与总PLFA含量均显著高于放牧草地,真菌脂肪酸含量则因草原类型不同各有差异;放牧导致各类型草原革兰氏阳性细菌PLFAs/革兰氏阴性细菌PLFAs（GPPLFAs/GNPLFAs）比值显著降低,而除了克氏针茅草原,细菌PLFAs/真菌PLFAs比值则显著升高。PLFAs主成分分析表明,放牧和围栏处理对内蒙古针茅草原土壤微生物群落结构产生影响,且围栏处理的影响程度大于放牧处理。经相关分析表明,氯仿熏蒸法和PLFA分析方法之间有很好的一致性,且土壤微生物PLFAs与土壤有机质、全磷、硝态氮显著相关。     

氧调控下人工湿地微生物群落结构变化研究

运用磷脂脂肪酸(PLFAs)技术研究了氧调控下复合垂直流人工湿地(IVCW)微生物群落结构及活性变化,结果表明,不曝气IVCW基质表层以好氧微生物为优势种群,但绝大部分微生物都集中在0～20cm基质层,系统的净化空间受到限制;氧调控下微生物群落向基质纵深发展,表征好氧微生物的单不饱和脂肪酸的含量显著增加,曝气系统下行池表层各类群微生物的生物量为不曝气系统的2～6倍,表征微生物活性的PLFAs总不饱和度水平也明显升高;革兰氏阴性细菌成为曝气IVCW基质微生物群落的优势种群,群落具有更高的活性和专一性,提高了污染物的去除效果。因此,进一步研究微生物的这种适应机制可以为优化湿地系统提供一定的理论基础。     

寒温带兴安落叶松林不同林型土壤微生物群落特征

为了研究不同林型兴安落叶松林土壤微生物生物量和群落结构特征,探讨土壤微生物生物量、群落结构的分布规律以及影响因子,选择了4个典型兴安落叶松林,即藓类兴安落叶松林、杜香兴安落叶松林、草类兴安落叶松林和杜鹃兴安落叶松林,采用磷脂脂肪酸生物标记法分析4个林型的土壤微生物量和群落组成。结果表明:4个林型中共检测出10种类型51种不完全分布的磷脂脂肪酸(PLFAs)生物标记,微生物量由多到少的排序为细菌、真菌、放线菌。4个林型中微生物总生物量、细菌生物量是藓类兴安落叶松林最高,草类兴安落叶松林最低;真菌生物量为杜香兴安落叶松林最高,草类兴安落叶松林最低;放线菌生物量为杜鹃兴安落叶松林最高,草类兴安落叶松林最低。冗余分析显示,土壤含水率对细菌影响最大,全钾、速效钾质量分数对真菌影响最大,土壤有机碳和碱解氮质量分数对放线菌影响最大。从而表明,寒温带兴安落叶松林不同林型的土壤微生物生物量和群落存在差异,土壤微生物群落与土壤性质之间具有相关性。     

Negligible contribution from roots to soil-borne phospholipid fatty acid fungal biomarkers 18:2ω6,9 and 18:1ω9

The phospholipid fatty acid biomarkers 18:1ω9, 18:2ω6,9 and 18:3ω3,6,9 are commonly used as fungal biomarkers in soils. They have, however, also been found to occur in plant tissues, such as roots. Thus, the use of these PLFAs as fungal biomarkers in sieved soil, which may still contain small remains of roots, has been questioned. We used data from a recent beech tree girdling experiment to calculate the contribution of roots to these biomarkers and were able to demonstrate that not more than 0.61% of 18:1ω9 and 18:2ω6,9 in sieved soil samples originated from roots (but 4% of 18:3ω3,6,9). Additionally, the abundance of the biomarker 18:2ω6,9 in the soil was found to be highly correlated to ectomycorrhizal root colonization, which further corroborates its fungal origin. PLFA biomarkers were substantially reduced in vital roots from girdled trees compared to roots of control trees (by up to 76%), indicating that the major part of PLFAs measured in roots may actually originate from ectomycorrhizal fungi growing inside the roots. We calculated, that even a near to 50% reduction in fine root biomass - as observed in the girdling treatment - accounted for only 0.8% of the measured decrease of 18:2ω6,9. Our results demonstrate that both 18:1ω9 and 18:2ω6,9 are suitable biomarkers for detecting fungal dynamics in soils and that especially 18:2ω6,9 is a reliable biomarker to study mycorrhizal dynamics in beech forests.     

Microorganisms as driving factors for the community structure of testate amoebae along an altitudinal transect in tropical mountain rain forests

We investigated microbial biomass, fungal biomass and microbial community structure at three altitudes (1000, 2000 and 3000 m) and in two soil layers [L/F layer (Layer I) and underlying H/Ah layer (Layer II)] of tropical mountain rain forests in southern Ecuador. Basal respiration, microbial biomass and concentration of ergosterol generally declined from Layer I to Layer II and peaked at 2000 m. Compared to temperate forest ecosystems microbial biomass and ergosterol concentrations were generally low. Patterns in phospholipid fatty acids indicated that the composition of microbial communities markedly changed from Layer I to Layer II. These differences between layers decreased with increasing altitude. The concentration of the arbuscular mycorrhizal fungal marker PLFA 16:1ω5c decreased with altitude in Layer I but increased in Layer II. The fungal-to-bacterial ratio increased with altitude and was higher in Layer I than in Layer II. Presumably, low microbial biomass in soils of tropical forest ecosystems is due to high temperature associated with high respiration but also low litter quality, with the latter declining with altitude. These conclusions are supported by the fact that at higher altitude the microbial community changed from a bacterial-dominated to a fungal-dominated system. CCA showed that microbial biomass correlated closely with density of a number of putatively bacterial feeding testate amoeba species including Corythion dubium Taranek, 1871, Euglypha cristata Leidy, 1879, Trigonopyxis arcula Penard, 1912, Tracheleuglypha dentata Deflandre, 1928 and Trinema lineare Penard, 1890. Ergosterol concentrations, but not the PLFA 18:2ω6c, strongly correlated with the putatively fungal feeding species Phryganella acropodia (Hertwig, Lesser, 1874) Hopkinson, 1909. Generally, parallel to microbial biomass and ergosterol concentrations the density of testate amoebae peaked at 2000 m. However, compared to microbial parameters changes in testate amoebae communities between two layers were less pronounced. The data suggest that density and community structure of testate amoebae are driven by the availability of food resources (bacteria and fungi) which at high altitude decrease with increasing moisture and decreasing pH.     

Microbial response to rhizodeposition depending on water regimes in paddy soils

Rhizodeposit-carbon (rhizo-C) serves as a primary energy and C source for microorganisms in the rhizosphere. Despite important progress in understanding the fate of rhizo-C in upland soils, little is known about microbial community dynamics associated with rhizo-C in flooded soils, especially depending on water regimes in rice systems. In this study, rice grown under non-flooded, continuously flooded and alternating water regimes was pulse labeled with ¹³CO₂ and the incorporation of rhizo-C into specific microbial groups was determined by ¹³C in phospholipid fatty acids (PLFAs) at day 2 and 14 after the labeling.A decreased C released from roots under continuously flooded condition was accompanied with lower total ¹³C incorporation into microorganisms compared to the non-flooded and alternating water regimes treatments. Continuous flooding caused a relative increase of ¹³C incorporation in Gram positive bacteria (i14:0, i15:0, a15:0, i16:0, i17:0, a17:0). In contrast, Gram negative bacteria (16:1ω7c, 18:1ω7c, cy17:0, cy 19:0) and fungi (18:2ω6, 9c, 18:1ω9c) showed greater rhizo-C incorporation coupled with a higher turnover under non-flooded and alternating water regimes treatments. These observations suggest that microbial groups processing rhizo-C differed among rice systems with varying water regimes. In contrast to non-flooded and alternating water regimes, there was little to no temporal ¹³C change in most microbial groups under continuous flooding condition between day 2 and 14 after the labeling, which may demonstrate slower microbial processing turnover. In summary, our findings indicate that belowground C input by rhizodeposition and its biological cycling was significantly influenced by water regimes in rice systems.     

Influence of an indigenous European alder (Alnus glutinosa (L.) Gaertn) rhizobacterium (Bacillus pumilus) on the growth of alder and its rhizosphere microbial community structure in two soils

Europeanalder seedlings were inoculated with a suspension of the putative plant growthpromoting rhizobacterium (PGPR) Bacillus pumilus (CECT5105), or left non-inoculated (controls) in two different soils, and grownundercontrolled conditions. Soil A showed a thick texture, slightly acidic with ahigh mineral nitrogen content, while soil B showed a thin texture, basic andwith a lower nitrogen content. At each sampling time, over an 8-week period,shoot and root systems of the plants were measured, nodules counted, and shootand root length and surface were determined. In addition, changes in themicrobial rhizosphere structure were evaluated by the phospholipid fatty acid(PLFA) profile extracted directly from the rhizosphere soil. The increasesdetected in shoot surface were significant only in soil A, while the rootsystemwas affected in both soils. In soil A, inoculation with B.pumilus caused a perturbation that subsequently disappeared, whilethe rhizosphere community structure was seriously altered in soil B. Allbiometric parameters were enhanced to a greater extent in soil A, in which theinoculum did not alter the existing rhizosphere communities and nutrientavailability was better.     

On silica-based solid phase extraction techniques for isolating microbial membrane phospholipids: Ensuring quantitative recovery of phosphatidylcholine-derived fatty acids

A silica-based solid phase extraction (SPE) protocol commonly used to isolate phospholipids from total lipid extracts failed to quantitatively recover phosphatidylcholines (PC) from three commercial SPE columns because a polar eluent volume of 5 mL methanol per 0.5 g silica was shown to be insufficient. Phosphatidylcholines, which are an important component of some fungal and bacterial cell membranes, were completely recovered when a larger ratio of 20:1 v/w methanol (mL) to silica (g) was used. Separation of phospholipids from a soil sample showed that a methanol:silica ratio of 20:1 recovered substantially greater amounts of phospholipids and resulted in a different phospholipid fatty acid (PLFA) structural profile than a 10:1 ratio. This study also confirmed that methanol preconditioning of the manufactured SPE columns studied is necessary for quantitative recovery of phospholipids.     

不同种植模式对土壤微生物区系及烟叶产量与质量的影响

为探明不同烤烟(nicotiana tobacumL.)种植模式对土壤微生物区系及烟叶产量和质量的影响,采用磷脂脂肪酸(phospholipid fatty acids,PLFAs)方法研究了连作烟、轮作烟、麦套烟和麦茬烟等4种烤烟种植模式下耕层(0～25cm)土壤微生物的群落动态,并探讨了烟叶产质量的变化。研究结果表明,在绿肥翻压期,土壤微生物总PLFAs的含量以麦茬烟处理最低;在烟草团棵期,轮作烟处理最高且与其他处理差异显著;而在现蕾期和成熟期,各处理之间无显著差异。各微生物类群的PLFAs含量及其在不同处理间的差异均随生育期而异,其中真菌PLFAs含量在各处理间的差异不显著。真菌和细菌的PLFAs比值及革兰氏阳性细菌与阴性细菌的比值显示,整个土壤微生物及细菌部分的群落结构均受烤烟种植模式和生育期的影响。其中,土壤微生物群落结构的变化以麦茬烟处理最明显。各微生物类群在成熟期的主成分分析结果显示,烤烟种植模式之间存在明显分异,与连作烟相比,麦茬烟的土壤微生物群落结构与之类似,而轮作烟和套作烟则变化较大。此外,不同种植模式显著影响烤后烟叶的经济性状及化学成分。与连作烟相比,种植模式改变后有利于烟叶等级结构、均价及产值的提高,并显著增加烟叶中的钾含量,但不利于烟叶中糖分的积累,各植烟模式中以麦套烟和麦茬烟的综合效益较好。该研究结果有助于了解烤烟种植对土壤生物环境的影响,并为中国北方烟区烤烟种植制度的优化提供依据。     

Methane uptake in soils from Pinus radiata plantations, a reverting shrubland and adjacent pastures: Effects of land-use change, and soil texture, water and mineral nitrogen

Afforestation and reforestation of pastures are key land-use changes in New Zealand that help sequester carbon (C) to offset its carbon dioxide (CO₂) emissions under the Kyoto Protocol. However, relatively little attention has been given so far to associated changes in trace gas fluxes. Here, we measure methane (CH₄) fluxes and CO₂ production, as well as microbial C, nitrogen (N) and mineral-N, in intact, gradually dried (ca. 2 months at 20 °C) cores of a volcanic soil and a heavier textured, non-volcanic soil collected within plantations of Pinus radiata D. Don (pine) and adjacent permanent pastures. CH₄ fluxes and CO₂ production were also measured in cores of another volcanic soil under reverting shrubland (mainly Kunzea var. ericoides (A. Rich) J. Thompson) and an adjacent pasture. CH₄ uptake in the pine and shrubland cores of the volcanic soils at field capacity averaged about 35 and 14 μg CH₄-C m⁻² h⁻¹, respectively, and was significantly higher than in the pasture cores (about 21 and 6 μg CH₄-C m⁻² h⁻¹, respectively). In the non-volcanic soil, however, CH₄-C uptake was similar in most cores of the pine and pasture soils, averaging about 7-9 μg m⁻² h⁻¹, except in very wet samples. In contrast, rates of CO₂ production and microbial C and N concentrations were significantly lower under pine than under pasture. In the air-dry cores, microbial C and N had declined in the volcanic soil, but not in the non-volcanic soil; ammonium-N, and especially nitrate-N, had increased significantly in all samples. CH₄ uptake was, with few exceptions, not significantly influenced by initial concentrations of ammonium-N or nitrate-N, nor by their changes on air-drying. A combination of phospholipid fatty acid (PLFA) and stable isotope probing (SIP) analyses of only the pine and pasture soils showed that different methanotrophic communities were probably active in soils under the different vegetations. The C18 PLFAs (type II methanotrophs) predominated under pine and C16 PLFAs (type I methanotrophs) predominated under pasture. Overall, vegetation, soil texture, and water-filled pore space influenced CH₄-C uptake more than did soil mineral-N concentrations.