Tree species influences soil microbial community diversity but not biomass in a karst forest in southwestern China

中国西南喀斯特森林树种对土壤微生物群落多样性和生物量的影响 陆地生态系统中植物种对土壤微生物群落结构的影响不一，而喀斯特生态系统中植物种对土壤微生物群落结构影响的研究尚未见报道。本研究利用磷酸脂肪酸(PLFA)法，分析了黔中高原型喀斯特常绿落叶阔叶混交林5个优势树种—窄叶石栎(Lithocarpus confinis Huang)、圆果化香(Platycarya longipes Wu)、滇鼠刺(Itea yunnanensis Franch.)、安顺润楠(Machilus cavaleriei H. Lév.)、云贵鹅耳枥(Carpinus pubescens Burkill)—与土壤理化性质对土壤微生物群落组成和生物量的影响。在测试的212个土壤样品 中共检测出132种PLFA，每个样品土壤微生物平均PLFA数量和生物量分别为65.97和11.22 µg g^–1。土壤表层(0–10 cm)的土壤微生物PLFA数量与下层(10–20 cm)土壤接近，但前者土壤微生物生物量显著高于后者(P < 0.05)。树种影响土壤微生物PLFA数量，但对土壤微生物生物量没有影响。云贵鹅耳枥附近的土壤微生物PLFA数量显著高于其他树种(P < 0.05)，而其他树种土壤微生物PLFA数量接近。土壤微生物 生物量与表层土壤的理化性质无显著相关，但与下层土壤的有机碳、全氮和全磷含量呈显著正相关 (P < 0.05)。总之，黔中高原型喀斯特森林真菌/细菌生物量比率低，微生物总生物量低，但微生物群落多样性高。树种对土壤微生物群落多样性产生影响。     

施磷对干旱胁迫下箭竹根际土壤养分及微生物群落的影响

以箭竹及其根际土壤作为研究对象,采用两因素随机区组实验,设置2种水分处理（正常浇水和干旱胁迫）和2种施磷量处理（施磷和不施磷）,探究施磷对干旱胁迫下箭竹根际土壤养分及微生物群落结构和多样性的影响。结果表明：（1）干旱胁迫显著降低了箭竹根际土壤中微生物量碳、可溶性有机氮和有效磷的含量,虽对箭竹根际土壤微生物群落的多样性无显著影响,但显著降低了箭竹根际土壤中总PLFA（phospholipid fatty acid contents）的含量和真菌、细菌、革兰氏阳性菌与革兰氏阴性菌的PLFA含量以及革兰氏阳性菌/革兰氏阴性菌的PLFA比值,显著改变了箭竹根际土壤微生物群落结构,结果显著降低了箭竹的生物量。（2）施磷显著增加了受旱箭竹根际土壤中微生物量碳和有效磷的含量,虽大体上对受旱箭竹根际土壤微生物群落的多样性无显著影响,但显著增加了受旱箭竹根际土壤中总PLFA和真菌PLFA的含量,并在一定程度上增加了细菌、革兰氏阳性菌、革兰氏阴性菌和放线菌的PLFA含量以及革兰氏阳性菌/革兰氏阴性菌和真菌/细菌的PLFA比值,也在一定程度上改善了受旱箭竹根际土壤微生物群落结构,从而改善受旱箭竹的生长。（3）主成分分析表明,干旱对箭竹根际土壤微生物群落结构的影响显著,而施磷的影响不明显。（4）相关分析发现,箭竹根际土壤微生物群落结构与箭竹根际土壤微生物量碳、可溶性有机氮及箭竹生物量呈显著正相关。综上,干旱降低了箭竹根际土壤养分含量和微生物生物量,改变了箭竹根际土壤微生物群落结构,抑制了箭竹的生长;施磷能增加受旱箭竹根际土壤养分含量和微生物生物量,改善受旱箭竹根际土壤微生物群落结构,进而改善受旱箭竹的生长。     

南亚热带红椎和格木人工幼龄林土壤微生物群落结构特征

采用氯仿熏蒸浸提法和磷脂脂肪酸法(Phospholipids fatty acid,PLFA)研究了我国南亚热带地区非固氮树种红椎(Castanopsis hystrix)和固氮树种格木(Erythrophleum fordii)人工幼龄林土壤微生物生物量与微生物群落结构特征。结果表明,在旱季和雨季,红椎幼龄林土壤微生物总PLFAs量,细菌PLFAs量、放线菌PLFAs量及丛枝菌根真菌PLFAs量均大于格木幼龄林。红椎幼龄林土壤PLFA Shannon多样性指数(H_(PLFA))在旱季和雨季均大于格木幼龄林。主成分分析表明,土壤微生物群落结构组成受到林分类型和季节的双重影响。冗余分析表明,土壤硝态氮(NO_3~--N)含量、土壤含水量、p H及土壤微生物生物量氮(MBN)与特征磷脂脂肪酸之间呈显著相关关系。以上结果表明固氮树种格木与非固氮树种红椎人工幼龄林对土壤微生物生物量和群落结构的影响存在显著差异。     

长白山土壤微生物群落结构及酶活性随海拔的分布特征与影响因子

长白山是由火山喷发形成的山地生态系统,是研究生态系统重建和演替过程的天然实验室。以长白山西坡垂直带中的亚高山针叶林带、亚高山岳桦林带、高山草甸带、高山苔原带土壤为研究对象,采用磷脂脂肪酸法(PLFA)和微孔板法分别测定土壤微生物群落结构及酶活性,探讨(1)长白山西坡微生物群落结构及酶活性沿海拔的分布特征;(2)垂直带植被因子、土壤因子对微生物群落结构及酶活性的影响。结果表明:土壤有机碳、全氮、全磷含量均随海拔升高先增加再减少,有机碳和全氮最高值出现在岳桦林带;随海拔升高,土壤微生物总PLFA和各类群微生物PLFA呈现出先增加后减少的变化特征,表现为亚高山岳桦林带亚高山针叶林带高山草甸带高山苔原带;在对土壤微生物总PLFA的贡献率上,表现为细菌真菌放线菌,G~-G~+;微生物PLFA相关性方面,细菌、放线菌、G~+、G~-之间的关联性较大,真菌与这四者之间的关联性较小;土壤全氮含量与各微生物类群均表现为显著正相关,而C/N则与各微生物类群均表现为负相关,二者是调控土壤微生物沿海拔变化的主要因子;土壤水解酶βG和AP活性随海拔升高而逐渐增加,其中AP活性对高山苔原带生态系统表现出很好的响应;土壤含水量、C/N和土壤温度是调控土壤酶活性垂直变化的主要因子;高山苔原带草甸化过程对土壤含水量、全磷含量、水解酶AP活性产生重要影响,而对土壤微生物PLFA含量和其他酶活性影响不大。长白山垂直带土壤微生物群落结构和酶活性除了受到土壤环境因子和水热条件的影响,还与植被群落组成及凋落物性质具有紧密联系。     

磷脂脂肪酸方法在土壤微生物分析中的应用*

磷脂脂肪酸(PLYA)是活体微生物细胞膜的重要组分,不同类群的微生物能通过不同的生化途径合成不同的PLFA,部分PLFA可以作为分析微生物量和微生物群落结构等变化的生物标记。在土壤微生物分析中,越来越多地采用了PLFA方法。主要对PLFA方法在土壤微生物分析中的应用做一综述。     

闽江河口红树林土壤微生物群落对互花米草入侵的响应

采用磷脂脂肪酸标记法(PLFA)研究外来入侵植物互花米草对闽江河口湿地红树林土壤微生物群落结构的影响,并探讨其主要影响因素。结果表明:从3种不同植被群落土壤(红树林群落MC、红树林-互花米草混生群落MS、互花米草群落SC)共检测到22种PLFA生物标记,MS土壤微生物PLFA生物标记总量明显高于其他植被群落,3种植被群落土壤理化性质和酶活性的变化趋势为:MCMSSC,表明互花米草入侵后土壤微生物量增加,而理化性质和酶活性均有明显下降,红树林湿地土壤质量发生了明显退化。3种植被群落土壤中含量最高的PLFA生物标记是16:0,16:1w7c,9Me15:0w,18:1w12c。土壤中特征微生物相对生物量存在明显差异,细菌分布量最大,其次是真菌和放线菌,原生动物分布量最小。群落多样性指数呈相似规律,MS土壤微生物类群多样性指数均小于MC,表明互花米草入侵后土壤微生物群落多样性指数均有下降。通过主成分分析,基本能区分出3种不同植被群落微生物群落的特征。土壤理化性质、酶活性间存在相关性,有机碳、全氮、蔗糖酶、过氧化氢酶与革兰氏阴性菌、放线菌呈显著或极显著正相关。研究结果表明互花米草入侵在一定程度上具有影响红树林群落土壤营养代谢循环的潜力,特别是关于碳、氮、磷等的循环及酶活性,改变部分有利于自身生长的土壤环境相关的微生物类群含量,竞争有利环境,迅速扩张实现入侵。     

氯嘧磺隆对土壤微生物群落结构的影响

采用磷脂脂肪酸(PLFA)法研究了黑龙江省苇河地区不同氯嘧磺隆施药历史下土壤微生物群落结构的差异, 并测定了土壤中氯嘧磺隆的残留量.结果表明：不同施药历史下氯嘧磺隆在土壤中的残留量均很低;随着施用氯嘧磺隆年限的增加,土壤微生物的PLFA总量减少, 革兰氏阴性菌/革兰氏阳性菌（GN/GP）和真菌/细菌比降低,土壤微生物的压力指数增加.主成分分析表明,氯嘧磺隆显著改变了大豆田土壤微生物群落结构.     

采煤塌陷地不同施肥处理对土壤微生物群落结构的影响

采用磷脂脂肪酸(PLFA)分析方法,对山西长治襄垣五阳煤矿采煤沉陷区复垦土壤的微生物群落结构进行了研究。结果表明:施肥处理能够不同程度的增加土壤微生物的PLFA总量、细菌PLFA量和真菌PLFA量,其中化肥+有机肥处理下的土壤微生物PLFA总量,细菌PLFA量和真菌PLFA量增加作用比较明显,差异显著于对照处理;对不同施肥处理的复垦土壤微生物群落PLFA进行主成分分析也可得出化肥+有机肥处理下的土壤微生物的群落结构变化比较大,从因子载荷图上进一步分析可知,化肥+有机肥处理下的土壤中代表真菌的不饱和脂肪酸C18∶2ω6t、C18∶3ω6、18∶1ω9t、18∶3ω3的含量较高,这些脂肪酸将化肥+有机肥处理与单施化肥和对照处理区分开来,产生了明显的优势种群。土壤PLFA总量与土壤有机质、碱解氮、速效磷和速效钾含量都有很好的相关性,相关系数分别为0.76,0.85,0.67和0.67。     

喀斯特地区三种人工林土壤微生物群落结构特征

为揭示不同人工植被修复模式对喀斯特土壤微生物群落的影响,采用氯仿熏蒸提取法和磷脂脂肪酸(phospholipid fatty acid, PLFA)法研究人工构建的降香黄檀(Dalbergia odorifera)纯林(PDOP)、顶果木(Acrocarpus fraxinifolius)纯林(PAFP)、顶果木×降香黄檀混交林(MADP)对土壤微生物生物量及土壤微生物群落结构的影响。结果表明:(1)PDOP的土壤微生物生物量碳(MBC)和微生物生物量氮(MBN)含量显著高于PAFP和MADP,PAFP显著高于MADP。(2)三种人工林土壤真菌、丛枝菌根真菌和总PLFA含量无显著差异,但PDOP土壤细菌、放线菌、丛枝菌根真菌和总PLFA含量均高于PAFP和MADP,PAFP高于MADP。PDOP的土壤细菌、革兰氏阳性菌、革兰氏阴性菌、放线菌的PLFA含量显著高于MADP。MADP的真菌细菌比显著高于PDOP,但与PAFP无显著差异。(3)冗余分析表明,土壤阳离子交换量、pH和C:N是影响土壤微生物群落组成的最主要影响因子。从三种人工林的土壤微生物生物量及微生物群落结构来看,在喀斯特地区MADP并未显示出酸性土地区混交林提高土壤微生物生物量、改善土壤微生物群落结构的优势,但混交林的真菌细菌比最高,更有利于提高土壤生态系统的稳定性。     

羊草基因型数目与氮添加对土壤微生物群落的交互影响

物种多样性(或同一物种遗传多样性)减少和氮富集都是影响陆地生态系统进程的主要因素,它们之间的交互作用是否对土壤微生物群落产生显著影响已成为研究者关心的主要科学问题。研究羊草基因型数目(1、2、4三种基因型数目组合)和氮添加(无氮添加、低氮添加和高氮添加3种水平)对土壤微生物群落的总磷脂脂肪酸(PLFA,Phospholipid Fatty Acid)含量、细菌PLFA生物标记含量、真菌PLFA生物标记含量、真菌/细菌比、以及基于每个PLFA生物标记相对含量百分比所得微生物群落的Shannon-Wiener多样性指数和Simpson优势度指数的影响。结果表明:氮添加对细菌PLFA生物标记含量,以及土壤微生物PLFA生物标记的Shannon-Wiener多样性指数和Simpson优势度指数具有显著影响(P0.05);基因型数目对所测变量无显著影响(P0.05),但基因型数目和氮添加的交互作用对细菌PLFA生物标记含量和真菌/细菌比具有显著影响(P0.05)。研究结果为全球变化背景下氮沉降及重要物种种群数量减少对土壤微生物群落的影响提供了科学数据,为合理解释群落动态变化提供了数据支持。     

滩涂围垦和土地利用对土壤微生物群落的影响

土壤微生物在生态系统营养物质循环过程,特别是碳、氮循环过程中扮演着重要的角色。上海市崇明岛位于长江入海口,因其土壤发育时间较短、土地利用历史背景清晰、土壤本底均一,不同土壤围垦年代的土壤,代表了土壤发育年代的不同时期。以空间变化代替时间变化,对崇明岛稻田和旱地6个不同围垦年代土壤的磷酸脂肪酸(PLFA)指纹图谱研究表明,湿地滩涂围垦16a后土壤微生物总PLFA、细菌PLFA、革兰氏阳性菌(G+)PLFA和革兰氏阴性菌(G-)PLFA含量显著降低。随着围垦时间的逐步增加,PLFA含量逐步上升。经过长时间的农业种植,G+PLFA在围垦120a和300a稻田和旱地土壤中没有显著性差异;而总PLFA、细菌和G-PLFA在围垦75、120a和300a的土壤中含量趋于稳定且没有显著性差异。围垦16a和40a稻田土壤中总PLFA和G+PLFA显著高于旱地土壤;围垦40a稻田土壤中细菌和G-PLFA显著高于旱地土壤。不同围垦年代土壤总PLFA、细菌PLFA与土壤总氮、粘土含量成显著的正相关关系。河口湿地围垦后微生物数量的变化与土壤营养含量存在强烈相关关系,提示土壤围垦及演替过程中微生物与土壤肥力之间的紧密关系,对探讨土壤演替过程中微生物群落的变化具有重要意义。     

罕山土壤微生物群落组成对植被类型的响应

选取分布在中国东北部地区的阔叶林-针叶林-亚高山草甸这一明显的植被垂直带谱来研究植被类型对土壤微生物群落组成的影响。选取5种植被类型-山杨(Populus davidiana)(1250—1300 m),山杨(P.davidiana)与白桦(Betula platyphylla)的混交林(1370—1550 m),白桦(B.platyphylla)(1550—1720 m),落叶松(Larix principis-rupprechtii)(1840—1890 m),亚高山草甸(1900—1951 m),采用磷脂脂肪酸(Phopholipid Fatty Acids,PLFAs)分析方法测定不同植被类型下的土壤微生物群落组成。分别采用主成分分析(Principal Components Analysis,PCA)以及冗余分析(Redundancy Analysis,RDA)来解释单种特征PLFAs的分异以及土壤理化指标与微生物PLFAs指标间的相关性。结果表明不同植被类型下土壤有机碳(SOC)对土壤微生物PLFAs总量,各类群(真菌(f)、细菌(b)、革兰氏阳性菌(G+)、革兰氏阴性菌(G-))生物量以及群落结构影响显著;土壤微生物PLFAs总量及各类群的生物量随土层加深总体上表现降低趋势,G+/G-和f/b分别随土层加深总体上表现升高趋势。不同植被类型下,阔叶混交林土壤PLFAs总量及各类群生物量总体上最高;针叶林比阔叶林下的f/b和G+/G-高;亚高山草甸下低的p H值对有机碳的可利用性有一定的抑制作用,导致f/b和G+/G-的值相对较高。总之,不同植被类型下SOC对土壤微生物群落组成的影响最为显著,而较低的p H对有机碳的可利用性有一定的抑制作用;真菌对植被类型的变化比细菌更敏感,而细菌更易受可利用性养分和p H变异的影响,这对预测不同林型下的土壤微生物群落组成有重要的启示作用。     

Standardizing methylation method during phospholipid fatty acid analysis to profile soil microbial communities

Phospholipid fatty acid (PLFA) as biomarkers, is widely used to profile microbial communities in environmental samples. However, PLFA extraction and derivatization protocols are not standardized and have widely varied among published studies. Specifically investigators have used either HCl/MeOH or KOH/MeOH or both for the methylation step of PLFA analysis, without justification or research to support either one. It seems likely that each method could have very different outcomes and conclusions for PLFA based studies. Therefore, the objective of this study was to determine the effect of catalyst type for methylation on detecting PLFAs and implications for interpreting microbial profiling in soil. Fatty acid samples extracted from soils obtained from a wetland, an intermittently flooded site, and an adjacent upland site were subjected to HCl/MeOH or KOH/MeOH catalyzed methylation procedures during PLFA analyses. The methylation method using HCl/MeOH resulted in significantly higher concentrations of most PLFAs than the KOH/MeOH method. Another important outcome was that fatty acids with a methyl group (18:1ω,7c 11Me, TBSA 10Me 18:0, 10Me 18:0, 17:0 10Me and 16:0 10Me being an actinomycetes biomarker) could not be detected by HCl/MeOH catalyzed methylation but were found in appreciable concentrations with KOH/MeOH method. From our results, because the HCl/MeOH method did not detect the fatty acids containing methyl groups that could strongly influence the microbial community profile, we recommend that the KOH/MeOH catalyzed transesterification method should become the standard procedure for PLFA profiling of soil microbial communities.     

Dynamics of a microbial community associated with manure hot spots as revealed by phospholipid fatty acid analyses.

Microbial community dynamics associated with manure hot spots were studied by using a model system consisting of a gel-stabilized mixture of soil and manure, placed between layers of soil, during a 3-week incubation period. The microbial biomass, measured as the total amount of phospholipid fatty acids (PLFA), had doubled within a 2-mm distance from the soil-manure interface after 3 days. Principal-component analyses demonstrated that this increase was accompanied by reproducible changes in the composition of PLFA, indicating changes in the microbial community structure. The effect of the manure was strongest in the 2-mm-thick soil layer closest to the interface, in which the PLFA composition was statistically significantly different (P < 0.05) from that of the unaffected soil layers throughout the incubation period. An effect was also observed in the soil layer 2 to 4 mm from the interface. The changes in microbial biomass and community structure were mainly attributed to the diffusion of dissolved organic carbon from the manure. During the initial period of microbial growth, PLFA, which were already more abundant in the manure than in the soil, increased in the manure core and in the 2-mm soil layer closest to the interface. After day 3, the PLFA composition of these layers gradually became more similar to that of the soil. The dynamics of individual PLFA suggested that both taxonomic and physiological changes occurred during growth. Examples of the latter were decreases in the ratios of 16:1 omega 7t to 16:1 omega 7c and of cyclopropyl fatty acids to their respective precursors, indicating a more active bacterial community. An inverse relationship between bacterial PLFA and the eucaryotic 20:4 PLFA (arachidonic acid) suggested that grazing was important.     

Responses of soil microbial communities to prescribed burning in two paired vegetation sites in southern China

Prescribed burning is a common site preparation practice for forest plantation in southern China. However, the effects of prescribed burning on soil microbial communities are poorly understood. This study examined changes in microbial community structure, measured by phospholipid fatty acids (PLFAs), after a single prescribed burning in two paired vegetation sites in southern China. The results showed that the total amount of PLFA (totPLFA) was similar under two vegetation types in the wet season but differed among vegetation type in the dry season, and was affected significantly by burning treatment only in the wet season. Bacterial PLFA (bactPLFA) and fungal PLFA (fungPLFA) in burned plots all decreased compared to the unburned plots in both seasons (P = 0.059). Fungi appeared more sensitive to prescribed burning than bacteria. Both G⁺ bacterial PLFA and G⁻ bacterial PLFA were decreased by the burning treatment in both dry and wet seasons. Principal component analysis of PLFAs showed that the burning treatment induced a shift in soil microbial community structure. The variation in soil microbial community structure was correlated significantly to soil organic carbon, total nitrogen, available phosphorus and exchangeable potassium. Our results suggest that prescribed burning results in short-term changes in soil microbial communities but the long-term effects of prescribed burning on soil microbial community remain unknown and merit further investigation.     

Phospholipid Fatty Acid Composition and Heavy Metal Tolerance of Soil Microbial Communities along Two Heavy Metal-Polluted Gradients in Coniferous Forests

The effects of long-term heavy metal deposition on microbial community structure and the level of bacterial community tolerance were studied along two different gradients in Scandinavian coniferous forest soils. One was near the Harjavalta smelter in Finland, and one was at Ronnskar in Sweden. Phospholipid fatty acid (PLFA) analysis revealed a gradual change in soil microbial communities along both pollution gradients, and most of the individual PLFAs changed similarly to metal pollution at both sites. The relative quantities of the PLFAs br18:0, br17:0, i16:0, and i16:1 increased with increasing heavy metal concentration, while those of 20:4 and 18:2(omega)6, which is a predominant PLFA in many fungi, decreased. The fungal part of the microbial biomass was found to be more sensitive to heavy metals. This resulted in a decreased fungal/bacterial biomass ratio along the pollution gradient towards the smelters. The thymidine incorporation technique was used to study the heavy metal tolerance of the bacteria. The bacterial community at the Harjavalta smelter, exposed mainly to Cu deposition, exhibited an increased tolerance to Cu but not to Cd, Ni, and Zn. At the Ronnskar smelter the deposition consisting of a mixture of metals increased the bacterial community tolerance to all tested metals. Both the PLFA pattern and the bacterial community tolerance were affected at lower soil metal concentrations than were bacterial counts and bacterial activities. At Harjavalta the increased Cu tolerance of the bacteria and the change in the PLFA pattern of the microbial community were found at the same soil Cu concentrations. This indicated that the altered PLFA pattern was at least partly due to an altered, more metal-tolerant bacterial community. At Ronnskar, where the PLFA data varied more, a correlation between bacterial community tolerance and an altered PLFA pattern was found up to 10 to 15 km from the smelter. Farther away changes in the PLFA pattern could not be explained by an increased community tolerance to metals.     

Comparison of Whole-Cell Fatty Acid (MIDI) or Phospholipid Fatty Acid (PLFA) Extractants as Biomarkers to Profile Soil Microbial Communities

The whole-cell lipid extraction to profile microbial communities on soils using fatty acid (FA) biomarkers is commonly done with the two extractants associated with the phospholipid fatty acid (PLFA) or Microbial IDentification Inc. (MIDI) methods. These extractants have very different chemistry and lipid separation procedures, but often shown a similar ability to discriminate soils from various management and vegetation systems. However, the mechanism and the chemistry of the exact suite of FAs extracted by these two methods are poorly understood. Therefore, the objective was to qualitatively and quantitatively compare the MIDI and PLFA microbial profiling methods for detecting microbial community shifts due to soil type or management. Twenty-nine soil samples were collected from a wide range of soil types across Oregon and extracted FAs by each method were analyzed by gas chromatography (GC) and GC-mass spectrometry. Unlike PLFA profiles, which were highly related to microbial FAs, the overall MIDI-FA profiles were highly related to the plant-derived FAs. Plant-associated compounds were quantitatively related to particulate organic matter (POM) and qualitatively related to the standing vegetation at sampling. These FAs were negatively correlated to respiration rate normalized to POM (Resp_POM), which increased in systems under more intensive management. A strong negative correlation was found between MIDI-FA to PLFA ratios and total organic carbon (TOC). When the reagents used in MIDI procedure were tested for the limited recovery of MIDI-FAs from soil with high organic matter, the recovery of MIDI-FA microbial signatures sharply decreased with increasing ratios of soil to extractant. Hence, the MIDI method should be used with great caution for interpreting changes in FA profiles due to shifts in microbial communities.