塔克拉玛干沙尘暴源区空气微生物群落的代谢特征

采用BIOLOG生态微平板(BIOLOG EcoPlateTM)技术,对采集自塔克拉玛干沙漠南北缘5个地区的空气样品进行研究,目的是了解空气微生物群落碳代谢功能的特点与差异.结果表明,5个地区空气样品的平均吸光度(AWCD)在培养10 d没有达到饱和且差异显著,AWCD值最高的为莎车县,和田县最低,分别为0.24和0.1;碳源利用水平表明这5个地区空气微生物对聚合物类碳源利用率最高,其次是碳水化合物、氨基酸类、羧酸类碳源,而酚类和胺类最低;主成分分析(PCA)发现有20种碳源与PC1显著相关,12种与PC2显著相关,并且这5个地区也被分为两类,和田和皮山聚为一类,乌恰、莎车和轮台聚为一类;典范对应分析(CCA)和相关分析表明一些非生物因素对空气微生物群落碳代谢强度和单一碳源的利用能力具有不同程度的影响,如风速、海拔、湿度等;进一步的研究也表明非生物因素对空气微生物群落利用的能力影响显著,其中β-甲基D-葡萄糖苷、D-半乳糖醛酸和腐胺的光密度值与纬度显著正相关(P<0.05),2-羟苯甲酸和α-D-乳糖与风速显著正相关(P<0.05),D-葡萄胺酸与气压显著正相关(P<0.05)且与海拔为显著负相关(P<0.05).综上可知,塔克拉玛干沙尘暴源区周边地区空气微生物群落碳代谢能力较低,呈现区域性特征,非生物因素显著影响空气微生物群落碳代谢强度及微生物群落对31种单一碳源的利用.     

溶解氧变化对底泥酶活性及微生物多样性的影响

采用室内静态实验考察了溶解氧变化对底泥酶活性影响,同时采用BIOLOG ECO微平板构造了不同溶解氧水平下底泥微生物多样性指数。结果表明:高溶解氧条件下,底泥多酚氧化酶及过氧化物酶活性显著高于缺氧条件及厌氧条件(P<0.01),脱氢酶活性高于背景值;厌氧条件下脱氢酶、脲酶及碱性磷酸酶活性显著高于高溶解氧组、缺氧组及背景值(P<0.05);缺氧条件下硝酸盐还原酶活性显著高于高溶解氧组及厌氧组(P<0.05);蛋白酶活性受溶解氧水平变化的影响较小。BIOLOG ECO微平板法分析表明,溶解氧变化对底泥微生物多样性有不同程度的负面影响,微生物多样性的降低程度从大到小依次为高溶解氧组>厌氧组>缺氧组。主成分分析结果表明:缺氧条件及厌氧条件下,底泥微生物对大分子有机物的利用出现不同程度的降低,相对地,高溶解氧条件下微生物对大分子有机物的利用程度呈现先下降后上升的趋势;高溶解氧组及缺氧组底泥微生物对碳水化合物利用程度升高,厌氧组对碳水化合物利用程度降低。总之,高溶解氧条件更有利于形成底泥有机物完整的代谢循环,提高大分子有机物的利用程度。     

雄黄矿区不同砷污染土壤中微生物群落结构及碳源利用特征

土壤砷污染是我国乃至世界范围内比较严重的环境问题.有关砷污染的生态毒理效应有很多研究,但对砷污染土壤中微生物群落的关注相对较少.本文以我国湖南省石门县雄黄矿地区长期砷污染土壤为例,采用PLFA及BIOLOG微平板技术考察了7个砷(As)污染程度不同的样点以及一个对照样点土壤微生物群落结构及碳源利用特征.结果发现,雄黄矿区存在多种重金属复合污染,除As含量较高以外,镉(Cd)和镍(Ni)的含量也超过了国家土壤环境质量三级标准.雄黄矿区土壤微生物的群落结构受到土壤有机碳(SOC)、有效磷(AP)、p H以及镁(Mg)、Cd、铁(Fe)、铜(Cu)含量的显著影响.各样点土壤微生物群落均以细菌为主,占微生物总量的71.54%~80.66%,真菌次之,放线菌最少.雄黄矿区土壤中的有效砷对微生物造成了较严重的胁迫.严重砷污染降低了微生物对于碳源利用的多样性以及均匀度.各样点微生物对于碳源的利用也表现出明显的差异.31种碳源中微生物利用较多的只有7类,分别属于糖类、胺类、羧酸类与多聚物,并且以糖类为主.     

硫酸铵施用量和温度对红壤稻田土硝化作用及微生物特性的影响

以红壤稻田土为供试土壤,设置不同硫酸铵(简称硫铵)施用量,分别在15、25、35℃条件下培养,研究短期内土壤硝化作用、微生物生物量和微生物功能多样性的变化。结果表明,在相同硫铵施用量下,不同温度处理土壤NH4+含量差异不显著;温度变化对于对照和常规硫铵用量(折纯N 120 mg·kg-1)处理土壤NO3-含量有显著影响,但对中、高量(折纯N 600和1 200 mg·kg-1)处理无显著影响。对照和常规硫铵用量处理土壤硝化速率均随温度升高而显著增加;中、高量硫铵处理土壤硝化速率普遍较低,且不同温度之间差异不显著;相同温度条件下,硝化速率随硫铵施用量的升高而降低。中、髙量硫铵处理对土壤微生物生物量碳有显著影响,且土壤微生物生物量碳随硫铵施用量的增加而显著降低;相同硫铵施用量下,不同温度处理土壤微生物生物量碳由高到低大致为25、15和35℃。BIOLOG分析显示,中、高量硫铵处理平均吸光值和多样性指数均较低,各处理中以25℃时对照处理的平均吸光值和Shannon、Simpson、McIntosh指数最大,其次为25℃时常规硫铵用量处理。过量施用硫铵有可能造成土壤生物结构和功能衰变。     

恩诺沙星对土壤微生物群落代谢功能多样性的影响（Effects of Enrofloxacin on the Functional Diversity of Soil Microbial Communities）

为了评价恩诺沙星(Enrofloxacin)对土壤微生物群落的影响,借助BIOLOG检测法比较了不同浓度恩诺沙星影响下的土壤微生物的群落特征.结果表明,加药组土壤微生物的丰富度指数和多样性指数显著低于空白对照组,且药物浓度越高丰富度和多样性越小.用药后第3d、14d,恩诺沙星含量0.1～100μg·g-1使土壤微生物群落代谢功能多样性显著降低(p<0.05);第35d,恩诺沙星含量10～100μg·g-1使土壤微生物群落代谢功能多样性显著降低,随着药物作用的时间延长,药物含量0.01～1μg·g-1组土壤微生物群落代谢功能多样性与空白对照组之间的差异变小.     

铅锌尾矿砂污染下的岩溶土壤微生物群落碳源代谢特征

以广西阳朔思的村铅锌矿尾砂污染的水稻田、玉米地、柑橘园以及对照(旱地)土壤为研究对象,采用碳素利用法和18S rRNA变性梯度凝胶电泳技术探究岩溶区重金属污染下土壤微生物群落碳源代谢特征以及土壤真菌与土壤有机碳的交互关系.结果表明,广西阳朔思的村铅锌矿尾砂污染区土壤重金属总体含量(Pb、Zn、Cu、Cd)依次为:水稻田玉米地柑橘园对照;同时地累积综合指数显示该区重金属污染以Pb、Cd为主,其中Cd带来的环境风险最高.受Pb、Zn、Cu、Cd等重金属复合污染物影响,土壤微生物生物量碳、微生物熵和碳源代谢速率表现出随总体重金属质量分数增加而降低的变化规律,但土壤总有机碳质量分数却表现为:水稻田对照柑橘园或玉米地.DGGE结果进一步显示对照组土壤存在担子菌门多孔菌目Pycnoporus sp.ZW02.30,柑橘园和玉米地土壤存在镰刀菌属Fusarium solani和Fusarium oxysporum以及柑橘园土壤存在青霉属Penicillium decumbens.然而,上述参与淀粉、纤维素、半纤维素、木质素等糖类降解过程的真菌没有在水稻田土壤检出.加之,水稻田土壤微生物功能多样性指数和微生物熵最低而土壤有机碳质量分数最高,说明铅锌尾矿砂通过影响不同样地土壤微生物糖类代谢过程,进而调节土壤碳在微生物特别是真菌作用下的矿化速率.     

Substrate-induced changes in microbial community-level physiological profiles and their application to discriminate soil microbial communities

The addition of simple substrates could affect the microbial respiration in soils. This substrate-induced respiration is widely used to estimate the soil microbial biomass, but little attention has been paid to its influence on the changes of community-level physiological profiles. In this study, the process of microbial communities responding to the added substrate using sole-carbon-source utilization （BIOLOG） was investigated. BIOLOG is biased toward fast-growing bacteria; this advantage was taken to detect the prompt response of the active microbial communities to the added substrate. Four soil samples from agricultural fields adjacent to heavy metal mines were amended with L-arginine, citric acid, or D-glucose. Substrate amendments could, generally, not only increase the metabolic activity of the microbial communities, but also change the metabolic diverse patterns compared with no-substrate control. By tracking the process, it was found that the variance between substrate-induced treatment and control fluctuated greatly during the incubation course, and the influences of these three substrates were different. In addition, the application of these induced changes to discriminate soil microbial communities was tested. The distance among all samples was greatly increased, which further showed the functional variance among microbial communities in soils. This can be very useful in the discrimination of microbial communities even with high similarity.     

Abundance and diversity of the phyllosphere bacterial communities of Mediterranean perennial plants that differ in leaf chemistry

Summary.  We studied the epiphytic bacterial communities of the summer leaves of eight perennial species naturally occurring in a Mediterranean ecosystem. The species differ in essential-oil content (from rich in essential oil to non-producers) and composition, and also in life form (from herbaceous species to tall shrubs). We compared the epiphytic bacterial communities on the basis of (i) their abundance, (ii) their metabolic profile (derived by use of the BIOLOG Ecoplate system) and (iii) richness and diversity of substrates that they use, as a measure of functional diversity. Among all species, the aromatic Melissa officinalis was the most abundantly colonized. The bacterial communities on the leaves of the aromatic Myrtus communis, Calamintha nepeta and Melissa officinalis, and also of Cistus incanus catabolized all 31 substrates offered; those on the evergreen-sclerophyllous species, Arbutus unedo and Quercus coccifera, catabolized only 14 and 17 substrates, respectively. Carbohydrates were consistently used abundantly by all communities, whereas carboxylic acids were most variably used. On average, the group of aromatic plants scored higher regarding bacterial abundance, and richness and diversity of substrates used by the bacterial communities on their leaves; the lowest values for both substrate-use indices were recorded in A. unedo. Bacterial abundance or richness or diversity of substrates used did not vary with leaf oil content. Abundance was positively correlated with both substrate-use indices. Results support claims that the antimicrobial effects of essential oils are not exerted so much under natural conditions as reports based on biassays with pathogens usually show. Although essential oils play a part in the microbial colonization of the phyllosphere, it is not likely that inhibition of phyllosphere bacteria is essential oils’ primary role, at least in the Mediterranean environment.     

Structural and metabolic responses of microbial community to sewage-borne chlorpyrifos in constructed wetlands

Long-term use of chlorpyrifos poses a potential threat to the environment that cannot be ignored, yet little is known about the succession of substrate microbial communities in constructed wetlands(CWs) under chlorpyrifos stress. Six pilot-scale CW systems receiving artificial wastewater containing 1 mg/L chlorpyrifos were established to investigate the effects of chlorpyrifos and wetland vegetation on the microbial metabolism pattern of carbon sources and community structure, using BIOLOG and denaturing gradient gel electrophoresis(DGGE) approaches. Based on our samples, BIOLOG showed that Shannon diversity(HV) and richness(S) values distinctly increased after 30 days when chlorpyrifos was added. At the same time, differences between the vegetated and the non-vegetated systems disappeared. DGGE profiles indicated that H Vand S had no significant differences among four different treatments. The effect of chlorpyrifos on the microbial community was mainly reflected at the physiological level. Principal component analysis(PCA) of both BIOLOG and DGGE showed that added chlorpyrifos made a difference on test results.Meanwhile, there was no difference between the vegetation and no-vegetation treatments after addition of chlorpyrifos at the physiological level. Moreover, the vegetation had no significant effect on the microbial community at the genetic level. Comparisons were made between bacteria in this experiment and other known chlorpyrifos-degrading bacteria. The potential chlorpyrifos-degrading ability of bacteria in situ may be considerable.     

Bacterial diversity in soils around a lead and zinc mine

Five samples of soil collected from a lead and zinc mine were used to assess the effect of combined contamination of heavy metals on soil bacterial communities using a polyphasic approach including characterization of isolates by culture method, community level catabolic profiling in BIOLOG GN microplates, and genetic community fingerprinting by denaturing gradient gel electrophoresis of 16S rDNA fragments amplified by PCR from community DNA （PCR-DGGE）. The structure of the bacterial community was affected to a certain extent by heavy metals. The PCR-DGGE analysis of 16S rRNA genes showed that there were significant differences in the structure of the microbial community among the soil samples, which were related to the contamination levels. The number of bacteria and the number of denaturing gradient gel electrophoresis （DGGE） bands in the soils increased with increasing distance from the lead and zinc mine tailing, whereas the concentration of lead （Pb） and cadmium （Cd） was decreased. Heavily polluted soils could be characterized by a community that differs from those of lightly polluted soils in richness and structure of dominating bacterial populations. The clustering analysis of the DGGE profiles showed that the bacteria in all the five samples of soil belonged to three clusters. The data from the BIOLOG analysis also showed the same result. This study showed that heavy metal contamination decreased both the biomass and diversity of the bacterial community in soil.