磷脂脂肪酸(PLFA)法检测蒙古沙冬青根围土壤微生物群落结构

为了评价和管理干旱和半干旱地区的植物生长,需要了解荒漠土壤资源空间分布和土壤微生物功能.本试验在乌海、磴口和阿拉善采取蒙古沙冬青(Ammopiptanthus mongolicus)根围土壤样品,采用磷脂脂肪酸(PLFA)法结合Sherlock微生物鉴定系统,研究了蒙古沙冬青根围土壤微生物群落空间分布特征.结果表明,蒙古沙冬青根围土壤微生物PLFA有较高的多样性,3样地土壤共检测到41、31和48种磷脂脂肪酸,土壤优势PLFA为16:0、16:0 10-methy1、18:1ω9c和16:1ω7c,均以16:0(表征细菌)含量最大,16:0 10-methy1、18:1ω9c和16:1ω7c在各样地含量有所差异.蒙古沙冬青根围土壤微生物群落结构有明显空间异质性:土壤微生物以革兰氏阳性细菌(G+)为主,AM真菌、革兰氏阳性菌、革兰氏阴性菌(G-)和真菌均表现为阿拉善乌海磴口,而放线菌PLFA总含量表现为乌海阿拉善磴口.在土壤真菌生物量中,AM真菌所占比重最大,尤其在磴口和阿拉善AM真菌占到真菌生物量91%和92%,说明AM真菌是荒漠土壤微生物系统中重要组成成分.RDA分析表明,AM真菌、革兰氏阳性菌、革兰氏阴性菌、真菌和放线菌与土壤磷酸酶、总球囊霉素、氨氮和p H正相关.G+/G-与脲酶、有机碳和易提取球囊霉显著负相关,而真/细菌与易提取球囊霉素,脲酶和有机碳显著正相关.研究说明土壤磷酸酶、总球囊霉素、氨氮和p H是影响土壤微生物PLFA变化的重要因子.同时,土壤微生物群落变化规律可用于检测土地荒漠化和土壤退化状况.     

缙云山柑橘林土壤微生物磷脂脂肪酸(PLFAs)及酶活性的季节变化特征

土壤微生物和土壤酶作为土壤生态环境最重要的组成成分,对环境变化敏感.本文以缙云山柑橘林为研究对象,采用磷脂脂肪酸法并结合主成分分析方法,分析季节更替对柑橘林土壤表层(0~20 cm)的土壤微生物数量、群落结构特征及酶活性等的影响.结果表明:1季节更替对土壤微生物有显著影响(P0.05),16:0、i17:0、16:1 2OH、18:0、cy19:0ω8c、i17:1ω9c或16:0 10-methyl为4个季节共有PLFAs,含量之和分别占PLFAs的49.57%、41.63%、35.41%和38.05%.各微生物种类中,细菌PLFA比例最高,其次为真菌PLFA,放线菌PLFA比例最低,且均具有显著的季节变化特征,柑橘林土壤PLFAs总量变幅为6.868~24.085 nmol·g-1,大小顺序为春季秋季冬季夏季,细菌PLFAs、G-、G+及放线菌PLFAs也呈现一致的变化规律,但真菌PLFAs则表现为秋季最高,其次是冬季和夏季,春季最低.季节更替对微生物群落多样性指数亦产生显著影响,丰富度指数(R)随季节变化依次为春冬秋夏,多样性指数(H')随季节变化表现出冬秋春夏,均匀度指数(J)表现为夏秋冬春,优势度指数(D)则随季节表现为直线升高的变化趋势.2土壤脲酶随季节变化表现为夏季春季秋季冬季;土壤蔗糖酶、土壤过氧化氢酶和酸性磷酸酶活性随季节变化均表现为秋季最高,其次是春季、夏季,冬季最低.3主成分分析结果表明细菌PLFAs、G+、G-、放线菌PLFAs和总PLFAs对土壤肥力贡献最大,其次是蔗糖酶、过氧化氢酶、酸性磷酸酶和真菌PLFAs,贡献最小为脲酶.     

多环芳烃污染对桑园土壤微生物结构及种群多样性的影响

采用磷脂脂肪酸(PLFAs)分析法和变性梯度凝胶电泳(DGGE)考察了受多环芳烃(PAHs)污染的桑园3个区域的土壤微生物群落结构及种群多样性的变化.PLFAs分析结果表明,区域2中微生物PLFA总量最高,主要为细菌和真菌;聚类分析揭示,土壤中微生物的PLFAs主要分为3大类群;冗余分析表明,土壤PAHs污染程度对土壤微生物群落结构有一定的影响.DGGE指纹图谱分析结果显示,在PAHs污染较高区域,其电泳条带较多,且3个区域中Shannon指数和Simpson优势度差异达显著水平,区域2种群优势度较高;主成分分析表明,不同区域微生物的种群结构存在显著性差异.     

土壤微生物群落对玉米根茬和茎叶残体碳的利用特征

以沈阳农业大学连续29a棕壤长期定位施肥试验为基础,以¹³C标记的玉米植株为试验试材,采用田间原位培养方法和磷脂脂肪酸-稳定同位素示踪联用（PLFA-SIP）技术,研究土壤不同粒级团聚体中微生物群落对残体碳的同化状况,及土壤有机碳的固定这一关键生物地球化学过程.结果表明：植物残体添加显著增加了全土及各粒级团聚体中各微生物群落PLFAs含量,其中以真菌PLFAs含量增幅最高,细菌中以革兰氏阴性菌含量增幅最高;但不同残体类型并未对全土中各群落PLFAs含量产生显著影响.茎叶碳与根茬碳的矿化率无显著差异;根茬碳对总PLFAs碳库的贡献是茎叶碳的3.9倍,说明根茬碳更有利于通过微生物合成的方式贡献于土壤有机碳库.残体碳占不同微生物群落PLFAs碳库的比例以真菌最高,表明真菌对植物残体碳具有最强的同化能力;而残体碳对PLFAs的贡献在革兰氏阳性菌和革兰氏阴性菌之间却差异不大.残体碳含量、PLFAs含量和残体碳占微生物PLFAs碳库的比例均在较小粒级的团聚体中（0.25~1mm和<0.25mm）更高,而细菌/真菌比在较大粒级团聚体（>2mm和1~2mm）中更高,说明较小粒级团聚体已经成为微生物对残体进行同化固定的主要位点.植物残体在土壤中的腐解过程与残体类型、土壤团聚体组成和微生物群落密切相关.     

土壤微生物群落对玉米根茬和茎叶残体碳的利用特征

以沈阳农业大学连续29a棕壤长期定位施肥试验为基础,以¹³C标记的玉米植株为试验试材,采用田间原位培养方法和磷脂脂肪酸-稳定同位素示踪联用（PLFA-SIP）技术,研究土壤不同粒级团聚体中微生物群落对残体碳的同化状况,及土壤有机碳的固定这一关键生物地球化学过程.结果表明：植物残体添加显著增加了全土及各粒级团聚体中各微生物群落PLFAs含量,其中以真菌PLFAs含量增幅最高,细菌中以革兰氏阴性菌含量增幅最高;但不同残体类型并未对全土中各群落PLFAs含量产生显著影响.茎叶碳与根茬碳的矿化率无显著差异;根茬碳对总PLFAs碳库的贡献是茎叶碳的3.9倍,说明根茬碳更有利于通过微生物合成的方式贡献于土壤有机碳库.残体碳占不同微生物群落PLFAs碳库的比例以真菌最高,表明真菌对植物残体碳具有最强的同化能力;而残体碳对PLFAs的贡献在革兰氏阳性菌和革兰氏阴性菌之间却差异不大.残体碳含量、PLFAs含量和残体碳占微生物PLFAs碳库的比例均在较小粒级的团聚体中（0.25~1mm和<0.25mm）更高,而细菌/真菌比在较大粒级团聚体（>2mm和1~2mm）中更高,说明较小粒级团聚体已经成为微生物对残体进行同化固定的主要位点.植物残体在土壤中的腐解过程与残体类型、土壤团聚体组成和微生物群落密切相关.     

紫茉莉对石油污染盐碱土壤微生物群落与石油烃降解的影响

以石油污染盐碱土壤为研究对象,利用磷脂脂肪酸(PLFAs)活性微生物标记法,分析紫茉莉(Mirabilis jalapa Linn.)根际土壤微生物群落结构的动态变化,探讨紫茉莉生长对根际土壤微生物与石油烃(TPH)降解的影响.结果表明,供试土壤中先后出现了22种微生物PLFAs,包括标识细菌的饱和脂肪酸(SAT)、标识革兰氏阳性菌(G+)的末端支链型饱和脂肪酸(TBSAT)、标识革兰氏阴性菌(G-)的单不饱和脂肪酸(MONO)和环丙脂肪酸(CYCLO)、标识真菌的多不饱和脂肪酸(PUFA)和标识放线菌的中间型支链型饱和脂肪酸(MBSAT)等六大类型.与未种紫茉莉土壤(CK)相比,根际土壤微生物PLFAs种类变异率在春、夏、秋季分别为71.4%、69.2%和33.3%;TPH降解率在春、夏、秋季分别提高了47.6%、28.3%、18.9%.相关性分析表明,石油烃的降解在CK土壤中与77.8%的PLFAs具有正相关关系(r0),55.6%的种类具有高度正相关关系(r≥0.8),其中,与SAT和MONO类群的相对含量正相关,相关系数分别为0.92、0.60;根际土壤中仅与42.1%的PLFAs正相关,21.1%的种类高度正相关,与TBSAT、MONO和CYCLO类群的相对含量正相关,相关系数分别为0.56、0.50、0.07.说明紫茉莉生长对根际土壤微生物群落结构及TPH降解速率均具有较大影响,且随生长季节的不同而有很大差异.该结果将为开展石油污染盐碱土壤的植物修复技术研究提供理论依据.     

不同添加量凋落物及生物质炭对土壤微生物群落结构的影响

以亚热带杉木人工林土壤为研究对象,采用培养试验,研究了杉木凋落物及其生物质炭(Biochar,BC)在不同添加量条件下(0、1%、2%、3%、4%、5%土壤质量分数)对土壤微生物群落结构的影响.结果表明:凋落物处理的土壤微生物总磷脂脂肪酸(PLFA)、细菌、真菌含量均随添加量的增加而增加,而丛枝菌根真菌(AMF)和放线菌(ACT)含量在不同添加量处理间则无显著差异.对于BC处理,在3%添加量下土壤微生物总PLFA、革兰氏阳性细菌(GP)、革兰氏阴性细菌(GN)、真菌、AMF和ACT含量均达到最大,表明BC对土壤微生物活性的促进作用与凋落物截然不同.此外,凋落物添加对真菌的影响较大,但对ACT的影响则显著低于BC处理.主成分分析表明,凋落物和BC添加均显著改变了土壤微生物群落结构,且前者的影响更为明显,而添加量对其影响则较小.冗余分析发现,土壤全氮与C/N比值分别为影响凋落物与BC处理中微生物群落结构变化的主要因子.     

铅-苄嘧磺隆对土壤微生物活性与群落结构的影响

选用苄嘧磺隆除草剂和重金属铅组成复合污染体系,以土壤微生物生物量、微生物代谢熵和磷脂脂肪酸为测试指标,在25℃暗培养条件下研究其对土壤微生物活性和微生物群落结构的影响.结果表明,苄嘧磺隆-铅对土壤微生物活性均有抑制作用,交互作用达显著水平.在低浓度时表现为简单加和作用,高浓度时表现为拮抗作用,浓度是影响交互作用类型的重要因子之一.磷脂脂肪酸图谱分析表明,丛枝菌根真菌的特征磷脂脂肪酸16:1ω5c的相对含量随污染物浓度升高而减少,而真菌特征磷脂脂肪酸18:2ω6,9c和放线菌类特征磷脂脂肪酸10Me17:0和10Me18:0的相对含量均随着污染物浓度升高而增加,2种污染物对微生物群落结构的影响存在较大的差异.     

多环芳烃污染土壤真菌群落组成及驱动因子

研究多环芳烃(PAHs)污染土壤中真菌群落结构及其驱动因子,可为PAHs污染土壤微生物修复技术研发提供技术支持。该文通过对华北某焦化厂5个点位(每个点位取6个深度)土壤样品中环境因子(PAHs含量、理化性质)和真菌群落结构进行了定量分析,研究了PAHs污染土壤中真菌群落结构与环境因子之间的关系。结果表明:(1)焦化厂土壤样品中微生物的主要真菌类群(属)为葡萄球菌属(Subulicystidium)、柄孢壳菌(Podospora)、土赤壳属(Ilyonectria)、介球菌属(Plectosphaerella),其和占真菌总量的20.22%～48.95%;(2)PAHs污染程度对真菌群落α多样性指数无显著影响(p0.05),但对真菌群落(属)有显著影响(p0.05);(3)冗余分析结果表明总有机碳和TPAHs是该区域真菌群落结构变化的主要驱动因子(p0.05)。     

生物质炭对邻苯二甲酸二丁酯污染土壤微生物群落结构多样性的影响

在邻苯二甲酸二丁酯(DBP)污染的不同类型土壤(有机质含量低的新垦红壤、有机质含量高的熟化红壤)中添加不同种类(稻草炭、毛竹炭)以及不同用量(0%、0.5%和2%)的生物质炭,温室种植上海青并在56 d后采集土样,采用磷脂脂肪酸法(PLFA)考察了土壤类型、生物质炭种类以及用量对土壤微生物群落结构多样性的影响.结果表明:对细菌、真菌及微生物总PLFA这三者的含量而言,熟化红壤显著(p0.05)高于新垦红壤,熟化红壤中添加2%稻草炭使其显著(p0.05)增加,新垦红壤中添加毛竹炭使其显著(p0.05)降低.新垦红壤中添加2%稻草炭对革兰氏阴性菌/革兰氏阳性菌比值的增加效果最显著(p0.05),添加2%毛竹炭对土壤微生物群落Shannon指数的降低效果最显著(p0.05).添加2%稻草炭对DBP污染土壤中微生物压力指数降低效果最显著(p0.05).生物质炭对熟化红壤中真菌/细菌、革兰氏阴性菌/革兰氏阳性菌及微生物群落Shannon指数均无显著影响.PCA分析表明,土壤有机质含量以及生物质炭的种类和用量均会对土壤微生物群落结构产生一定影响,且生物质炭的影响与土壤有机质含量密切相关.     

Influence of soil type and genotype on Cd bioavailability and uptake by rice and implications for food safety

Cadmium (Cd) entering the human body via the food chain is of increasing concern. This study investigates the effects of soil type and genotype on variations in the Cd concentrations of different organs of nine rice plants grown on two types of soils with two Cd levels. Cd concentrations in nine rice cultivars varied significantly with genotype and soil type (P < 0.01). The Cd concentration was higher in red paddy soil (RP) than in yellow clayey paddy soil (YP). The average Cd concentrations of different organs in three rice types were indica > hybrid > japonica for the Cd treatments and controls. The polished grain concentration in YP and RP soils had a range of 0.055-0.23 mg/kg and 0.13-0.36 mg/kg in the Cd treatment, respectively. Two rice cultivars in YP soil and five rice cultivars in RP soil exceeded the concentration limits in the Chinese Food Hygiene Standard (0.2 mg/kg). The Cd concentrations in roots, stems, and leaves were all significantly and positively correlated to that in polished grain in a single test. The Cd concentrations in polished grain were positively and significantly (P < 0.01) correlated with the calculated transfer factors of stem to grain and leaf to grain Cd transfer. The results indicated that the variations of Cd concentration in grain were related to Cd uptake and the remobilization of Cd from stem and leaf to grain. Also, the cultivars with a strong tendency for Cd-accumulation should be avoided in paddy soil with low soil pH and low organic matter content to reduce the risks to human health from high Cd levels in rice.     

Effect of copper on phospholipid fatty acid composition of microbial communities in two red soils

The phospholipid fatty acid （PLFA） composition was analyzed in two red soils experimentally contaminated with copper at different concentrations. The total amounts ofphospholipid fatty acids （PLFAs） in both red soils were significantly correlated with soil microbial biomass C and N, which decreased consistently with increasing levels of copper. The relative quantities of the PLFAs 17：0 （10 Me）, i16：0, il 5：0 and 16：1w5c, decreased with increasing heavy metal concentration, while those of cyl7：0, which is an indicator of gram-negative bacteria, increased. The Shannon index calculated from the PLFA data indicated that Cu addition in the red soils decreased the population diversity of soil microbial communities. Multivariate analysis of PLFA data demonstrated that high levels of Cu application had a significant impact on microbial community structure and there is a threshold metal concentration for PLFA composition. Comparatively higher toxic effect on microbial biomass and community structure were found in the red sandy soil than those in the red clayey soil. The differential effect of Cu addition on microbial communities in the two soils may be due to differences in soil texture and cation exchange capacity.     

重金属铜、锌、镉、铅复合污染对土壤环境微生物群落的影响

以铜锌冶炼厂附近的水稻土为例 ,研究了重金属复合污染对土壤微生物群落的影响 .结果表明 ,有效铜、锌、镉、铅与微生物生物量碳、微生物生物量氮、微生物商、微生物生物量氮 全氮均呈显著负相关 .重金属污染均能降低细菌、真菌和放线菌的数量 .用BIOLOG生态盘研究了重金属污染对微生物群落结构的影响 ,发现重金属污染明显影响了微生物群落结构 ,反映在典型变量 1(CV1)与重金属元素含量呈极显著正相关 ,因此认为典型变量 1是反映重金属污染程度的有效指标 .经逐步回归分析发现 ,有效铜是影响典型变量 1最主要的因素 .     

Phospholipid fatty acid patterns of microbial communities in paddy soil under different fertilizer treatments

The microbial communities under irrigated rice cropping with different fertilizer treatments, including control （CK）, PK, NK, NP, NPK fertilization, were investigated using phospholipid fatty acid （PLFA） profile method. The results of this study revealed that the fertilizer practice had an impact on the community structure of specific microbial groups. The principal components analysis （PCA） showed that proportion of the actinomycete PLFAs （10Me 18：0 and 10Me 16：0） were the lowest in the PK treatment and the highest in the NPK treatment, which means that soil nitrogen status affected the diversity of actinomycetes, whereas nitrogen cycling was related to the actinomycets. Under CK treatment, the ratio of Gram-positive to Gram-negative bacteria was lower compared with that in fertilizer addition treatments, indicating that fertilizer application stimulated Gram-positive bacterial population in paddy soil. The fatty acid 18：2to6,9, which is considered to be predominantly of fungal origin, was at low level in all the treatments. The ratio of cyl9：0 to 18：1 to7, which has been proposed as an indicator of stress conditions, decreased in PK treatment. Changes of soil microbial community under different fertilizer treatments of paddy soil were detected in this study; however, the causes that lead to changes in the microbial community still needs further study.     

Characterization of contamination, source and degradation of petroleum between upland and paddy fields based on geochemical characteristics and phospholipid fatty acids

To evaluate contamination caused by petroleum, surface soil samples were collected from both upland and paddy fields along the irrigation canals in the Hunpu wastewater irrigation region in northeast China. N-alkanes, terpanes, steranes, and phospholipid fatty acids (PLFA) in the surface soil samples were analyzed. The aliphatic hydrocarbon concentration was highest in the samples obtained from the upland field near an operational oil well; it was lowest at I-3P where wastewater irrigation promoted the downward movement of hydrocarbons. The Hunpu region was found contaminated by heavy petroleum from oxic lacustrine fresh water or marine deltaic source rocks. Geochemical parameters also indicated significantly heavier contamination and degradation in the upland fields compared with the paddy fields. Principal component analysis based on PLFA showed various microbial communities between petroleum contaminated upland and paddy fields. Gram-negative bacteria indicated by 15:0, 3OH 12:0, and 16:1(9) were significantly higher in the paddy fields, whereas Gram-positive bacteria indicated by i16:0 and 18:1(9)c were significantly higher in the upland fields (p < 0.05). These PLFAs were related to petroleum contamination. Poly-unsaturated PLFA (18:2ωup6, 9; indicative of hydrocarbon-degrading bacteria and fungi) was also significantly elevated in the upland fields. This paper recommends more sensitive indicators of contamination and degradation of petroleum in soil. The results also provide guidelines on soil pollution control and remediation in the Hunpu region and other similar regions.     

缙云山马尾松林和柑橘林土壤微生物PLFA沿海拔梯度的变化

土壤微生物群落是土壤生态系统的重要组成部分,对环境变化敏感.本文运用磷脂脂肪酸法(PLFA)研究缙云山马尾松林和柑橘林土壤微生物群落结构沿海拔梯度的变化特征.结果表明,从6个海拔土壤中共检测到48种PLFA,其中i16:0、10Me17:0、10Me18:0 TBSA在6个海拔中含量均最高,且柑橘林土壤PLFA种类和含量明显高于马尾松林.随着海拔升高马尾松林土壤微生物种类和含量逐渐增加,柑橘林则逐渐降低,各海拔间细菌、真菌、放线菌、革兰氏阴性菌(G~-)及革兰氏阳性菌(G~+)含量差异显著.土壤微生物群落多样性研究结果表明,马尾松林低海拔丰富度指数(R),多样性指数(H')、均匀度指数(J)均显著高于高海拔,而柑橘林R在低海拔最高,H'、J则在高海拔最高.不同海拔土壤细菌、放线菌、G~-及G~+与土壤酶和环境因子之间存在相关性,细菌、放线菌、G~-及G~+与脲酶(Ure)、转化酶(Ive)、过氧化氢酶(CAT)、林分类型均呈极显著正相关,真菌与Ure、Ive、CAT呈显著正相关;而细菌、真菌、放线菌、G~-及G~+与海拔呈极显著或显著负相关.Ure、Ive、CAT、林分及海拔是影响土壤微生物PLFA变化的重要因子.     

稻田与旱地土壤中真菌和细菌对秸秆碳的利用特征

微生物将植物残体矿化为CO₂和同化为微生物细胞组成部分是新鲜有机物料转化为土壤有机质的关键环节.以亚热带两种典型农业利用（稻田和旱地）土壤为对象,采用40 d室内模拟培养试验结合磷脂脂肪酸-稳定同位素示踪联用(¹³C-PLFA-SIP)技术,研究¹³C标记秸秆的矿化特征以及参与秸秆降解的细菌和真菌类群变化规律.结果表明,培养前期（0.25～1 d）,秸秆碳在稻田土壤中的矿化速率高于旱地土壤,中期（2～20 d）以稻田土壤低于旱地土壤（P<0.05）,后期（21～40 d）两者矿化速率相当.培养结束时,秸秆碳在稻田土壤中的累积矿化率（11%）约为旱地（20%）的一半.尽管稻田土壤中总微生物量（PLFA总量）比旱地高,但两种土壤中秸秆碳被微生物同化为细胞组分的量(¹³C-PLFA)相当,且稻田中秸秆碳的富集比例(PLFA中¹³C占总碳量的百分比)低于旱地,说明稻田土壤中参与秸秆碳降解的活性微生物占比少.整个培养期内,稻田土壤中秸秆碳被微生物利用的优势类群为细菌(占^1...     

Field released transgenic papaya effect on soil microbial communities and enzyme activities

Soil properties, microbial communities and enzyme activities were studied in soil amended with replicase （RP）-transgenic or non-transgenic papaya under field conditions. Compared with non-transgenic papaya, significant differences （P〈0.05） were observed in total nitrogen in soils grown with transgenic papaya. There were also significant differences （P〈0.05） in the total number of colony forming units （CFUs） of bacteria, actinomycetes and fungi between soils amended with RP-transgenic plants and non-transgenic plants. Compared with non-transgenic papaya, the total CFUs of bacteria, actinomycetes and fungi in soil with transgenic papaya increased by 0.43-1.1, 0.21-0.80 and 0.46-0.73 times respectively. Significantly higher （P〈0.05） CFUs of bacteria, actinomycetes and fungi resistant to kanamycin （Km） were obtained in soils with RP-transgenic papaya than those with non-transgenic papaya in all concentrations of Km. Higher resistance quotients for Km＇ （kanamycin resistant） bacteria, actinomycetes and fungi were found in soil planted with RP-transgenic papaya, and the resistance quotients for Km＇ bacteria, actinomycetes and fungi in soils with transgenic papaya increased 1.6-4.46, 0.63-2.5 and 0.75-2.30 times. RP-transgenic papaya and non-transgenic papaya produced significantly different enzyme activities in arylsulfatase （5.4-5.9x）, polyphenol oxidase （0.7-1.4x）, invertase （0.5-0.79x）, cellulase （0.23-0.35x） and phosphodiesterase （0.16-0.2x）. The former three soil enzymes appeared to be more sensitive to the transgenic papaya than the others, and could be useful parameters in assessing the effects of transgenic papaya. Transgenic papaya could alter soil chemical properties, enzyme activities and microbial communities.