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。     

一株氯嘧磺隆降解菌分离鉴定及降解条件优化

为解决氯嘧磺隆残留对土壤、水体污染及后茬敏感作物药害问题,为污染土壤微生物修复提供降解菌种资源,文中采用富集培养、逐级驯化等方法,从氯嘧磺隆污染土壤中分离到1株高效氯嘧磺隆降解菌T9DB-01,经形态特征、生理生化及16S rDNA序列分析,鉴定为假单胞菌Pseudomonas sp.。采用单因素实验探究温度、pH值、底物浓度、装液量和接种量对菌株T9DB-01降解氯嘧磺隆的影响,采用正交试验及验证,优化菌株T9DB-01对氯嘧磺隆降解条件。结果表明,在30℃,pH 8.0,底物浓度200 mg/L,装液量100 mL/250 mL,接种量4%的条件下,5 d后降解率达到93.7%。该降解菌株对氯嘧磺隆污染土壤原位生物修复具有一定的应用潜力。     

氯嘧磺隆与尿素对土壤微生物生物量碳、氮及无机氮的影响

通过室内培养试验,研究了不同浓度氯嘧磺隆(20、200、2000 μg·kg-1土)单一施用及与尿素(120 mg· kg-1土)配合施用情况下,土壤微生物生物量碳、氮和土壤铵态氮、硝态氮随时间的动态变化规律.结果表明:各浓度氯嘧磺隆单独处理在整个培养期(60 d)中对微生物生物量碳、氮均有抑制作用,且浓度越高,后期抑制作用越强;各浓度氯嘧磺隆处理在培养前期对硝态氮、铵态氮没有明显影响,中期(15 d)能显著提高土壤中铵态氮的含量,后期(30 d后)显著提高了土壤中硝态氮的含量.尿素单独施用及与氯嘧磺隆配施均能在短时间内增加微生物生物量碳、氮,但随后配施处理的促进作用减弱;尿素单独和配施均能持久增加土壤中铵态氮、硝态氮含量.     

森草净杀灭薇甘菊(Mikania micrantha)及其安全性

在深圳市内伶仃岛薇甘菊危害的不同群落生境中,设立9块样地81个小样方,用森草净(即70%嘧碘降水溶性粉剂)杀灭样地中的薇甘菊施量为0.0001～0.02g.m-2,结果表明:各浓度的森草净杀灭效果均较好,杀灭率随着用药量的增加而提高;在坡地和溪谷生境中,森草净用药量分别为0.05～0.1g.m-2、>0.2g.m-2能较彻底地杀灭薇甘菊。应用HPLC法检测样地土壤中嘧磺隆残留量,溪谷土壤中嘧磺隆半衰期C=C0.e-0.083T,T1/2=8.4,施药后37d消解95.9%,坡地高浓度级半衰期C=C0.e-0.046T,T1/2=15.1d,施药后37d消解85.0%,坡地低浓度级半衰期C=C0.e-0.090T,T1/2=7.7d,施药后15d消解74.2%。不同浓度森草净处理样地,施药后7、15、37d均可检测到嘧磺隆,并且含量越来越小,但施药后68d的土样,均未检测到嘧磺隆的存在。     

长白山温带森林不同演替阶段土壤化学性质及微生物群落结构的变化

土壤微生物是生态系统物质循环和能量流动的驱动者,其群落结构可以用来表征土壤生态过程及其对地上植被变化的响应机制。本研究采用时空替代法在长白山地区选取了阔叶红松林演替序列的5个不同阶段:杨桦幼龄林、杨桦中龄林、杨桦成熟林、阔叶红松成熟林和阔叶红松过熟林,采用磷脂脂肪酸法(PLFA)测定了土壤微生物群落组成和结构,分析了其随地上植被演替过程的变化,同时比较了不同演替阶段土壤化学性质差异。结果表明:随着演替的正向进行,土壤总有机碳、全碳、全氮、全磷含量显著提高,碳氮比逐渐下降。土壤微生物生物量、群落结构及组成发生明显变化:土壤微生物总PLFAs、细菌PLFAs、革兰氏阳性菌PLFAs、革兰氏阴性菌PLFAs含量显著增加;真菌PLFAs(18:2ω6c)先增加后减少,中期阶段的杨桦成熟林土壤真菌PLFA含量最高,同时细菌/真菌最小;革兰氏阳性菌/革兰氏阴性菌(G+/G-)随着演替的进行逐渐增大。土壤微生物生物量与土壤全碳、总有机碳、全氮、全磷含量呈显著正相关,与碳氮比呈显著负相关;冗余分析(RDA)结果显示,全碳、总有机碳、全氮和碳氮比是影响土壤微生物群落结构的主要因素。本研究表明,随着植被演替的正向进行,土壤质量逐渐提高;土壤微生物群落组成明显改变;土壤微生物群落结构与土壤理化性质显著相关。     

积雪覆盖变化对大兴安岭多年冻土区土壤微生物群落结构的影响

高纬度多年冻土区是全球变化的敏感区域,揭示不同雪被覆盖条件下土壤微生物群落结构的演变规律,对于预测寒区森林生态系统对全球变化的响应具有重要意义。以大兴安岭多年冻土区白桦次生林为研究对象,采用磷脂脂肪酸法(PLFA)对比分析自然积雪和遮雪处理土壤微生物群落结构的动态变化特征。结果表明:土壤总磷脂脂肪酸含量在植被生长季初期最高,积雪稳定期最低,其中含量较高的PLFA为18:2ω6,9c、a15:0、i16:0、17:1ω8c、18:1ω9c和16:1ω5c,不同时期各优势PLFA含量存在一定差异。遮雪显著降低积雪稳定期细菌PLFA含量、增加真菌PLFA含量(P0.05),但对其他时期土壤微生物群落结构和多样性均未产生显著影响。冗余分析(RDA)表明,土壤总磷脂脂肪酸、真菌、真菌/细菌和革兰氏阴性菌与土壤含水量、pH和铵态氮均呈显著正相关,细菌和革兰氏阳性菌受土壤总磷、总氮和硝态氮影响较大。     

氯嘧磺隆降解菌的分离鉴定及其降解特性

利用富集培养技术从长期施用氯嘧磺隆的土壤中分离得到1株能够降解氯嘧磺隆的细菌L-7。通过生理生化特性和16SrRNA序列分析,初步鉴定L-7为寡养单胞菌属(Stenotrophomonas sp.)。并分析了氯嘧磺隆的初始浓度、接种量、温度和pH值对L-7菌株降解氯嘧磺隆效果的影响,确定了最佳降解条件。结果表明,该菌在氯嘧磺隆浓度为100mg/L、接种量为5%、pH4.0、温度30°C条件下,接种5d后对氯嘧磺隆的降解效率达到80%以上。     

氯嘧磺隆降解菌株LW-3的分离及生物学特性研究

从长期受氯嘧磺隆污染的土壤中分离到一株氯嘧磺隆高效降解菌株,命名为LW-3,菌株LW-3可以氯嘧磺隆为唯一氮源生长,接种量为2%时,50 mg/L氯嘧磺隆经过7 d,降解率达70%～80%.生理生化实验和16S rDNA序列同源性分析,将菌株LW-3归属于假单胞茵属(Pseudomonas sp.);菌株的适宜降解条件为;温度30℃～35℃,pH 6.5～7.2,pH对菌株LW-3降解氯嘧磺隆有明显地影响.     

不同土壤细菌种群结构对氯嘧磺隆胁迫的响应及降解菌系的获得

应用PCR-DGGE、DNA测序等方法,在室内驯化条件下研究了8种来源于中国不同地区土壤样品细菌种群结构对氯嘧磺隆胁迫的响应。结果表明:在氯嘧磺隆100～500mg·L-1浓度梯度下,土壤细菌群落组成有明显的更迭现象,多样性发生明显变化,驯化至10周,绝大部分细菌种群消失,样品的细菌种群结构趋于简单并呈现趋同效应;DNA测序结果表明,在驯化第10周可培养Methylophilus sp.、Beta proteobacterium、uncultured bacterium成为优势菌属,所获细菌种群出现的16个优势种群中有10个与已知的具有有机污染物降解功能和有机污染环境修复功能细菌的相似性大于97%;其中5个与嗜甲基菌16S rDNA部分序列相似性达98%以上。获得了一组对氯嘧磺隆具有降解作用的细菌菌系,可在5d内将100mg·L-1氯嘧磺隆降解67%;其主要组成为嗜甲基菌属(Methylophilus sp.)、丛毛单胞菌属(Comamonas sp.)、鞘酯杆菌属(Sphingobacterium sp.)和嗜氢菌属(Hydrogenophi-lus sp.)。     

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。     

Characterization of the rhizosphere microbial community from different Arabidopsis thaliana genotypes using phospholipid fatty acids (PLFA) analysis

Total and culturable rhizosphere microbial communities structure from three different genotypes of Arabidopsis thaliana growing on three different substrates was studied with phospholipid fatty acid analysis (PLFA) and multivariate statistical analyses: correspondence analysis (CA) and distance based redundancy analyses (db-RDA). In addition, microbial biomass from different groups (total bacteria, Gram+, Gram? and fungi) was calculated from biomarkers PLFA peak area, both from total and culturable microbial community. db-RDA analysis showed significant differences between soils but not between plant genotypes for culturable microbial community structure. Total microbial community was significantly different between soils, and also between plant lines in each soil. Biomass of different bacterial groups showed significant higher values in soil two rhizosphere irrespective of the plant line. In addition, significant differences between plant lines were also found for microbial biomass of different bacterial groups both in total and culturable microbial community. Throughout the work we have demonstrated that PLFA analysis has been able to show a different behaviour of total microbial community with regard to the culturable fraction analyzed in this work under the influence of plant roots. Microbial biomass of different microbial groups calculated with PLFA biomarkers was a suitable tool to detect differences between soils irrespective of the plant line, and differences in the same soil between plant lines. According to this data, a previous study should be carried out before GMPs are used in field conditions to evaluate the potential alterations that may take place on rhizosphere microbial communities structure which may further affect soil productivity. In conclusion, based on data presented in this work, GMPs alter rhizosphere microbial communities structure and this effect is different depending on the soil. Furthermore, total microbial community is affected to a greater extent than the culturable fraction analyzed.     

Responses of soil nitrogen-fixing, ammonia-oxidizing, and denitrifying bacterial communities to long-term chlorimuron-ethyl stress in a continuously cropped soybean field in Northeast China

Chlorimuron-ethyl is a type of long-residual herbicide applied widely to soybean fields in China, but little information is available about the long-term impact of this herbicide on soil nitrogen-transforming microbial communities. Soil samples (0–20 cm) were collected from three treatments (no, 5-year and 10-year application of chlorimuron-ethyl) in a continuously cropped soybean field. Plate count (CFU), most probable number (MPN) count, and clone library analyses were conducted to investigate the abundance and composition of nitrogen-fixing, ammonia-oxidizing, and denitrifying bacterial communities, and a chlorate inhibition method was adopted to measure the soil nitrification potential. Long-term chlorimuron-ethyl application reduced the abundance of soil culturable nitrogen-fixing, ammonia-oxidizing, and denitrifying bacteria. Moreover, chlorimuron-ethyl decreased the diversity of nitrogen-fixing and ammonia-oxidizing bacteria but promoted that of denitrifying bacteria. Chlorimuron-ethyl restrained some uncultured nitrogen-fixing bacteria, ammonia-oxidizing bacteria Nitrosospira sp. cluster 3a and 3d, and some novel or putative denitrifying bacteria. The nitrogen-fixing bacteria were closely related to Bradyrhizobium sp., ammonia-oxidizing bacteria Nitrosospira sp. cluster 3b and 3c, and most denitrifying bacteria were resistant to chlorimuron-ethyl. There was a negative correlation between the nitrification potential and the residual amount of soil chlorimuron-ethyl (R²?=?0.88, n?=?3, P?<?0.05). Therefore, long-term application of chlorimuron-ethyl in the continuously cropped soybean field could seriously disturb soil N-transforming communities, and might impact soybean soil biological quality and soybean growth. Further studies should address rational amendment models of this herbicide to reduce the possible ecological risks of long-term application of this herbicide to soybean fields.     

基于PLFA指纹图谱表征浓香型酒糟醅微生物群落结构

以9株白酒酿造过程中常见的菌株为对象,研究了不同种属菌株的细胞膜特征组分磷酸脂肪酸(PLFA)的特征,以及检出量与菌株生物量之间的关系.结果表明:供试细菌、放线菌、霉菌和酵母菌的PLFA指纹图谱存在显著差异,各菌株的PLFA指纹图谱可作为区别种属的依据.不同供试菌株生物量在一定范围内与检出的总PLFA量或16:0含量呈线性关系.将不同生物量的革兰氏阳性菌G+、革兰氏阴性菌G-和真菌分别加入糟醅后,检出的PLFA相对含量与对照差异显著.基于PLFA的指纹图谱能够定量或半定量地表征糟醅微生物群落结构特征及动态变化.经对多家酿酒企业糟醅PLFA组成的检测及微生物群落结构的剖析,该方法具有普适性.     

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。     

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。     

Characterization of Microbial Consortia in Paddy Rice Soil by Phospholipid Analysis

Abstract Microbial biomass and community structure in paddy rice soil during the vegetation period of rice were estimated by analysis of their phospholipid fatty acids (PLFA), hydroxy fatty acids of lipopolysaccharides (LPS-HYFA), and phospholipid ether lipids (PLEL) directly extracted from the soil. A clear change in the composition of the community structure at different sampling periods was observed, indicated by the principal component analysis of the PLFA. A dramatic decline of ester-linked PLFA was observed in the soil samples taken at the second sampling time. In contrast to the ester-linked PLFA, the non-ester-linked PLFA composition did not change. The hydroxy fatty acids of lipopolysaccharides as well as ether lipids decreased consecutively during the observation period. Total microbial abundance was estimated to be (4.1–7.3) × 10⁹ cells g^-1 soil (dry weight). About 44% account for aerobic and 32% for facultative anaerobic bacteria, and 24% for archaea, on average. According to the profile and patterns of PLFA in the soil sample, it may be suggested that the paddy soil at the August sampling period contained more abundant facultative anaerobic bacteria (ca. 36%) and archaea (ca. 37%), but the total microbial biomass was significantly lower than in the remaining sampling periods. As the plant approached maturity, the microbial community structure in the soil changed to contain more abundant Gram-negative bacteria and methanotrophs. Received: 23 September 1999; Accepted: 28 February 2000; Online Publication: 12 May 2000     

Physiology and microbial community structure in soil at extreme water content

A sandy loam soil was brought to 6 water contents (13-100% WHC) to study the effects of extreme soil moistures on the physiological status of microbiota (represented by biomass characteristics, specific respiration, bacterial growth, and phospholipid fatty acid, PLFA, stress indicators) and microbial community structure (assessed using PLFA fingerprints). In dry soils, microbial biomass and activity declined as a consequence of water and/or nutrient deficiency (indicated by PLFA stress indicators). These microbial communities were dominated by G+ bacteria and actinomycetes. Oxygen deficits in water-saturated soils did not eliminate microbial activity but the enormous accumulation of poly-3-hydroxybutyrate by bacteria showed the unbalanced growth in excess carbon conditions. High soil water content favored G bacteria.     

The responses of soil organic carbon mineralization and microbial communities to fresh and aged biochar soil amendments

While biochar soil amendment has been widely proposed as a soil organic carbon (SOC) sequestration strategy to mitigate detrimental climate changes in global agriculture, the SOC sequestration was still not clearly understood for the different effects of fresh and aged biochar on SOC mineralization. In the present study of a two‐factorial experiment, topsoil samples from a rice paddy were laboratory‐incubated with and without fresh or aged biochar pyrolyzed of wheat residue and with and without crop residue‐derived dissolved organic matter (CRM) for monitoring soil organic matter decomposition under controlled conditions. The six treatments included soil with no biochar, with fresh biochar and with aged biochar treated with CRM, respectively. For fresh biochar treatment, the topsoil of a same rice paddy was amended with wheat biochar directly from a pyrolysis wheat straw, the soil with aged biochar was collected from the same soil 6 years following a single amendment of same biochar. Total CO₂ emission from the soil was monitored over a 64 day time span of laboratory incubation, while microbial biomass carbon and phospholipid fatty acid (PLFA) were determined at the end of incubation period. Without CRM, total organic carbon mineralization was significantly decreased by 38.8% with aged biochar but increased by 28.9% with fresh biochar, compared to no biochar. With CRM, however, the significantly highest net carbon mineralization occurred in the soil without biochar compared to the biochar‐amended soil. Compared to aged biochar, fresh biochar addition significantly increased the total PLFA concentration by 20.3%–33.8% and altered the microbial community structure by increasing 17:1ω8c (Gram‐negative bacteria) and i17:0 (Gram‐positive bacteria) mole percentages and by decreasing the ratio of fungi/bacteria. Furthermore, biochar amendment significantly lowered the metabolic quotient of SOC decomposition, thereby becoming greater with aged biochar than with fresh biochar. The finding here suggests that biochar amendment could improve carbon utilization efficiency by soil microbial community and SOC sequestration potential in paddy soil can be enhanced by the presence of biochar in soil over the long run.     

长期不同施肥方式对日光温室番茄土壤养分和微生物群落结构的影响

为了阐明长期不同施肥方式对日光温室番茄土壤养分和微生物群落结构的影响,以山东寿光12年番茄定位施肥试验土壤为对象,采用磷脂脂肪酸(PLFA)分析方法,研究了传统施氮(CN)、传统施氮+秸秆(CNS)、优化施氮(SN)、优化施氮+秸秆(SNS)、有机肥+秸秆(MNS)5种施肥方式对土壤主要理化性质和微生物群落结构的影响,...