Cluster formation in liverwort-associated methylobacteria and its implications

Pink-pigmented methylotropic bacteria of the genus Methylobacterium inhabit the surfaces of plant organs. In bryophytes, these methylobacteria enhance cell growth, but the nature of this plant–microbe interaction is largely unknown. In this study, methylobacteria were isolated from the upper surface of the free-living thalli of the liverwort Marchantia polymorpha L. Identification of one strain by 16S ribosomal RNA (rRNA) gene-targeted polymerase chain reaction (PCR) and other data show that these microbes represent an undescribed species of the genus Methylobacterium (Methylobacterium sp.). The growth-promoting activity of these wild-type methylobacteria was tested and compared with that of the type strain Methylobacterium mesophilicum. Both types of methylobacteria stimulated surface expansion of isolated gemmae from Marchantia polymorpha by about 350%. When suspended in water, the liverwort-associated bacteria (Methylobacterium sp.) formed dense clusters of up to 600 cells. In liquid cultures of Methylobacterium mesophilicum, single cells were observed, but no clustering occurred. We suggest that the liverwort-associated methylobacteria are co-evolved symbionts of the plants: Cluster formation may be a behavior that enhances the survival of the epiphytic microbes during periods of drought of these desiccation-tolerant lower plants.     

Growth in liverworts of the Marchantiales is promoted by epiphytic methylobacteria

Liverworts, the most basal lineage of extant land plants, have been used as model systems in the reconstruction of adaptations to life on land. In this study, we used gemmae (specialized propagules) that were isolated from mature gemma cups of two distantly related species of liverworts, Marchantia polymorpha L. and Lunularia cruciata L. (order Marchantiales). We show that methylobacteria (genus Methylobacterium), microbes that inhabit the surfaces of land plants where they secrete phytohormones (cytokinines), promote the growth of isolated gemmae cultivated on agar plates. As a control, two species of higher plants, maize (Zea mays L.) and sunflower (Helianthus annuus L.) were raised aseptically from sterile seeds (i.e., caryopses, achenes). Inoculation of these propagules with methylobacteria was without effect on growth in the above-ground phytosphere (expansion of stems and leaves). We conclude that normal development in Marchantia and Lunularia is dependent on (and possibly regulated by) epiphytic methylobacteria, whereas representative higher plants grow at optimal rates in the absence of these prokaryotic epiphytes.     

Moss-associated methylobacteria as phytosymbionts: an experimental study

Methylotrophic bacteria inhabit the surface of plant organs, but the interaction between these microbes and their host cells is largely unknown. Protonemata (gametophytes) of the moss Funaria hygrometrica were cultivated in vitro under axenic conditions and the growth of the protonemal filaments recorded. In the presence of methylobacteria (different strains of Methylobacterium), average cell length and the number of cells per filament were both enhanced. We tested the hypothesis that auxin (indole-3-acetic acid, IAA), secreted by the epiphytic bacteria and taken up by the plant cells, may in part be responsible for this promotion of protonema development. The antiauxin parachlorophenoxyisobutyric acid (PCIB) was used as a tool to analyze the role of IAA and methylobacteria in the regulation of cell growth. In the presence of PCIB, cell elongation and protonema differentiation were both inhibited. This effect was compensated for by the addition of different Methylobacterium strains to the culture medium. Biosynthesis and secretion of IAA by methylobacteria maintained in liquid culture was documented via a colorimetric assay and thin layer chromatography. Our results support the hypothesis that the development of Funaria protonemata is promoted by beneficial phytohormone-producing methylobacteria, which can be classified as phytosymbionts.     

铜胁迫对小麦根系微域微生物群落的影响

小麦是我国主要的粮食作物,土壤重金属污染会严重威胁作物生长和粮食安全.大量研究表明根际微生物在调控作物发育和抗逆性方面具有非常重要的作用.因此,本研究通过高通量测序技术研究铜胁迫条件下小麦根际微生物群落的变化,从微生物学角度揭示铜污染胁迫对小麦根际的影响作用.采用盆栽培养试验对铜污染处理中小麦非根际、根际以及根内微生物进行测序后,分别比较小麦不同根系分区环境中的微生物群落结构和多样性.结果发现,所有处理中小麦的根内微生物多样性均显著（P<0.001）低于根际和非根际微生物多样性,表明根表作为微生物进入根内环境的门户,对根内微生物的定殖起着过滤和筛选的作用.铜污染胁迫降低了根际土环境中的微生物多样性,且差异显著（P<0.05）;而在非根际和根内环境中,尽管铜污染胁迫降低了对应根系分区中的微生物多样性,但差异不显著（P>0.05）.变形菌门（Proteobacteria）和放线菌门（Actinobacteria）是小麦根际和非根际环境中的共有优势菌群,通过比较发现铜污染胁迫对这两种优势菌门的影响作用较小.另外,芽孢杆菌属（Bacillus）、假黄色单胞菌属（Pseudoxanthomonas）和鞘氨醇单胞菌属（Sphingomonas）等菌属具有较强的抗逆性,能够在铜污染胁迫下存活且能够为植物提供营养物质.     

Effect of temperature switchover on the degradation of antibiotic chloramphenicol by biocathode bioelectrochemical system

Exposure to chloramphenicol(CAP),a chlorinated nitroaromatic antibiotic,can induce CAP-resistant bacteria/genes in diverse environments. A biocathode bioelectrochemical system(BES) was applied to reduce CAP under switched operational temperatures.When switching from 25 to 10°C,the CAP reduction rate(kCAP) and the maximum amount of the dechlorinated reduced amine product(AMCl,with no antibacterial activity) by the biocathode communities were both markedly decreased. The acetate and ethanol yield from cathodophilic microbial glucose fermentation(with release of electrons) was also reduced. Formation of the product AMCl was enhanced by the biocathode dechloridation reaction compared with that produced from pure electrochemical or microbial dechloridation processes. The electrochemical and morphological analyses of cathode biofilms demonstrated that some cathodophilic microbes could adapt to low temperature and play a key role in CAP degradation. The resilient biocathode BES has a potential for the treatment of CAP-containing wastewater in temperature fluctuating environments.     

典型氧化还原环境中微塑料表面的微生物群落组成特征与构建机制

微塑料广泛分布在微生物驱动的生物地球化学循环中,其表面会富集独特特征的微生物群落,构成微塑料圈（plastisphere）.自然环境中广泛存在的多种氧化还原环境不仅会影响微塑料圈中微生物群落的组成,还会影响微塑料的最终归宿.为探究典型氧化还原环境中微塑料表面的微生物群落组成特征与构建机制,将3种微塑料PHA （聚羟基脂肪酸酯）、PLA （聚乳酸）和PVC （聚氯乙烯）放置于好氧、硝酸盐还原、铁氧化物还原、硫酸盐还原和产甲烷这5种典型氧化还原环境中,利用污泥作为接种物,进行微宇宙模拟培养实验.结果表明,在分类学和系统发育学上,微塑料因子分别影响了微塑料表面18.94%和46.67%的微生物群落,氧化还原环境因子分别影响了微塑料表面31.04%和90.00%的微生物群落.与污泥相比较,3种微塑料表面富集的微生物群落丰富度和均匀度均降低,其中降低最明显的是更易降解的PHA微塑料,而难降解的PLA和PVC微塑料表面的微生物群落变化特征相似.PHA微塑料表面富集的微生物中,Anaerocolumna（26.44%）为其优势菌种,较少富集与氧化还原反应相关的特征菌群;PLA和PVC中,Clostridium_sensu_stricto_7 （15.49%和11.87%）为其优势菌种,且显著富集与氧化还原反应相关的特征菌群,表明与氧化还原反应相关的特征菌群更易于富集在难降解的微塑料表面,进而可能对生物地球化学循环速率造成影响.     

矿物油污染土壤中芳烃组分的生物降解与微生物生长动态

以石油污染土壤中分离的细菌和真菌为供试微生物,研究不同微生物组合对矿物油芳烃组分降解及降解率与微生物生长间的关系.将不同组合的微生物接种到加有柴油浓度为1 000mg/L的液体培养基中,25～30℃经摇床连续培养100d.于0～60h内连续取样,进行微生物生长动态检测;于5～100d定期取样,研究微生物生长状况与矿物油降解率动态变化.同时,以草甸棕壤(0～20cm)制备的土壤悬浮液为土壤微生物对照,以灭菌培养基为非生物降解对照.结果表明,试验前期(约20d)外源菌的降解效果优于土壤微生物,试验中、后期,土壤微生物降解优势增大且保持持续.试验结束时,土壤微生物处理的液体培养基中芳烃降解率最高达79.24%,显著高于其它3组处理.     

夏秋季节北京市居家室内微生物多样性的变化特征

室内空气中过高浓度的生物粒子有害人体健康.空气中的微生物通常与灰尘结合在一起,然而目前对家庭室内灰尘微生物多样性及其随季节变化特征的报道较少.本研究在北京市选择1户家庭,在夏季和秋季定期采集灰尘样品,通过高通量测序研究细菌和真菌群落组成及多样性,并分析细菌-真菌的网络互作特征.结果显示,室内灰尘细菌和真菌群落Shannon指数及Chao1指数夏季均显著高于秋季（p<0.05）.此外,室内灰尘细菌群落结构夏季与秋季无显著差异（p>0.05）,主要类群为假单胞菌属（Pseudomonas）、考克氏菌属（Kocuria）和芽孢杆菌属（Bacillus）;真菌群落结构夏季与秋季则明显不同（p<0.05）,夏季优势类群为曲霉属（Aspergillus）、链格孢属（Alternaria）和裂褶菌属（Schizophyllum）,而秋季曲霉属占绝对主导地位.室内灰尘微生物网络互作具有明显的季节特征,夏季微生物互作主要为细菌-细菌,以及细菌-真菌的正相互作用;而秋季微生物互作比夏季更紧密,以细菌-细菌的正相互作用为主.这些结果可为构建健康的居家环境提供参考依据.     

大庆聚驱后油藏内源微生物群落结构解析与分布特征研究

利用变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)技术和主成分分析法(principal component analysis,PCA)解析了大庆油田聚驱后油藏的细菌和古菌群落结构组成及分布特征.结果表明,注水井中的细菌主要以好氧的假单胞菌属和不动杆菌属为主,注水井近井地带以兼性厌氧的肠杆菌属为主,各采油井中的细菌则包括陶厄氏菌属、梭菌纲、假单胞菌属、油杆菌属及大量的未培养细菌;各注水井及注水井近井地带检测到的古菌主要是乙酸型产甲烷的甲烷鬃菌,各采油井中古菌则以甲烷微菌属、甲烷螺菌属及甲烷杆菌属等为主.总体上,该聚驱区块从注水井到采油井,细菌优势菌群依次呈好氧细菌-兼性厌氧细菌-严格厌氧细菌分布;古菌的分布受环境因素及微生物代谢产物影响,注水井和采油井中的优势菌群差异显著.     

梯度负荷下果蔬垃圾厌氧消化性能及微生物群落结构的研究

为优化果蔬垃圾厌氧消化工艺,提高厌氧消化性能,本文通过逐级提高CSTR反应器进料负荷,研究不同负荷下的厌氧消化性能及相应的微生物群落结构变化规律.结果表明,随着进料负荷的增高,容积产气率、甲烷产气量、氨氮、碱度、TCOD、SCOD均逐渐增高,在最高负荷(负荷以VS计)2.50g·L-·1d-1时分别达到最大值:1.22L·L-1d-1,5.10L·d-1,1563.86mg·L-1,7572.23mg·L-1,13283.26mg·L-1,2075.03mg·L-1,甲烷含量及VFA分别稳定在52.46%～54.59%和(879.30±18.69)mg·L-1;同时利用PCR-DGGE技术系统分析了厌氧消化中细菌与古细菌的群落结构,测序结果表明,整个过程中拟杆菌(Bacteroidetes)、甲烷鬃菌(Methanosaeta)及甲烷螺菌(Methanospirillum)为优势微生物,随着负荷的提高,甲烷鬃毛菌(Methanosaeta)活性逐渐降低;聚类分析及主成分分析表明,低负荷条件下(1.50g·L-·1d-1、1.75g·L-·1d-1),微生物种类(细菌、古细菌)差别不明显,且基本处于同一阶段.     

再生水灌溉对土壤化学性质及可培养微生物的影响

通过室内土柱模拟实验,探讨再生水灌溉对土壤化学性质和可培养微生物的影响,从而为再生水回用评价提供数据支持.结果表明,在土壤化学性质方面,再生水灌溉可显著提高土壤有机质(OM)和全氮(TN)含量,而对土壤总磷(TP)、速效磷(AP)和pH值无显著影响.在微生物数量方面,再生水灌溉可显著提高表层0~20 cm细菌和放线菌数量,而对20~40 cm和40~60 cm土层三大微生物类群数量影响较小.在微生物种类方面,不动杆菌属(Acinetobacter)是再生水灌区的优势菌属,芽孢杆菌属(Bacillus)是自来水灌区的优势菌属;自来水灌区特有种属4个,再生水灌区特有种属6个;再生水灌溉对表层0~20cm土壤微生物群落Shannon多样性无影响,使土壤微生物群落Pielou均匀度降低,可提高土壤微生物群落Margalef丰富度.通过SPSS 17.0对土壤微生物数量和土壤化学性质进行相关性分析表明,土壤微生物数量与OM、TN、TP和AP含量呈正相关,与土壤pH值、含水量(SWC)呈负相关;通过CANOCO 4.5对土壤微生物种类与土壤化学性质进行DCA去趋势分析和RDA冗余分析表明,AP含量与微生物群落相关性最强(P=0.002),TP和TN对链球菌属(Streptococcus)、气球菌属(Aerococcus)和奈瑟氏球菌属(Neisseria)的影响较大,OM和AP对气单胞菌属(Aeromonas)、动性球菌属(Planococcus)和盐杆菌属(Halobacterium)影响较大.     

Release of Pb adsorbed on graphene oxide surfaces under conditions of Shewanella putrefaciens metabolism

In this study, Pb(II) was used as a target heavy metal pollutant, and the metabolism of Shewanella putrefaciens (S. putrefaciens) was applied to achieve reducing conditions to study the effect of microbial reduction on lead that was preadsorbed on graphene oxide (GO) surfaces. The results showed that GO was transformed to its reduced form (r-GO) by bacteria, and this process induced the release of Pb(II) adsorbed on the GO surfaces. After 72 hr of exposure in an S. putrefaciens system, 5.76% of the total adsorbed Pb(II) was stably dispersed in solution in the form of a Pb(II)-extracellular polymer substance (EPS) complex, while another portion of Pb(II) released from GO-Pb(II) was observed as lead phosphate hydroxide (Pb₁₀(PO₄)₆(OH)₂) precipitates or adsorbed species on the surface of the cell. Additionally, increasing pH induced the stripping of oxidative debris (OD) and elevated the content of dispersible Pb(II) in aqueous solution under the conditions of S. putrefaciens metabolism. These research results provide valuable information regarding the migration of heavy metals adsorbed on GO under reducing conditions due to microbial metabolism.     

水环境中化能自养微生物种群及固碳功能研究

化能自养微生物可通过还原性物质的氧化获取化学能以固定无机碳,在缺乏光照的海洋深处、水体沉积物等植物无法生存的环境中固碳,对吸收大气、海洋、湿地中的CO₂具有重要作用。本文基于化能自养微生物的研究现状,概述了其主要固碳途径及参与化能自养固碳的主要微生物类群,着重阐述了化能自养微生物在不同水环境中的固碳功能。同时,本文对水环境中化能自养微生物研究的趋势进行了展望,固碳量的精确计算与化能自养固定的有机碳的存在形式与去向应得到更多研究与探讨,以期增进对碳循环的相关认识。     

青岛近海生物气溶胶中可培养微生物浓度及群落多样性的季节变化

为了研究近海生物气溶胶中可培养微生物浓度和群落多样性,于2009年7月~2010年6月在青岛两个采样点连续采集生物气溶胶样品,分析了其中陆源细菌、海源细菌、陆源真菌和海源真菌的浓度,并计算了Shannon-Weiner指数、Simpson’s指数和Pielou指数.结果表明,陆源细菌和海源细菌月均浓度分别为12~436 CFU·m-3和25~561 CFU·m-3,陆源真菌和海源真菌月均浓度分别为0~817 CFU·m-3和11~1 346 CFU·m-3之间.陆源细菌、海源细菌、陆源真菌和海源真菌浓度在冬季月份较低,2月达到最低值,在春夏月份较高.海源微生物对总可培养类微生物的贡献高于陆源,平均占63%.可培养微生物物种数在17~102之间,与微生物浓度具有一定的相关性,但并未呈现出明显的季节变化.3种指数表明,生物气溶胶中陆源细菌、海源细菌、陆源真菌和海源真菌的群落结构在2月最简单,1月、11月和5月群落多样性较高,群落多样性与浓度的季节变化特征并不一致,而且不同类别的微生物群落存在季节和空间差异.     

牵牛花对石油污染盐碱土壤微生物群落与石油烃降解的影响

利用磷脂脂肪酸（PLFAs）生物标记法分析野生型牵牛花（Pharbitis nil （Linn.） Choisy）根际土壤微生物群落结构,探讨牵牛花生长对石油烃污染土壤微生物群落与石油烃降解的影响.结果表明,供试土壤微生物群落中,先后出现了24种PLFAs,包括标记细菌的饱和脂肪酸（SAT）、革兰氏阳性菌（G⁺）的末端支链饱和脂肪酸（TBSAT）、革兰氏阴性菌（G^-）的单不饱和脂肪酸（MONO）和环丙脂肪酸（CYCLO）、真菌的多不饱和脂肪酸（PUFA）和放线菌的中间型支链饱和脂肪酸（MBSAT）等六大类型.PLFAs的主成分分析（PCA）表明,牵牛花根际与对照组（未种牵牛花,CK）土壤微生物群落具有明显的差异,微生物多样性在春季增加83%、夏季增加140%、秋季增加50%;微生物的生物量在春季增加97.6%,夏季增加116.3%,秋季增加60.3%.牵牛花根际与对照组相比土壤中石油烃降解率明显提高,在春、夏、秋季分别提高了7.5%、34.2%和19.7%;并且,在牵牛花生长的不同季节石油烃的降解率有明显的差异,春季为22.3%,夏季为51.8%,秋季为38.0%.相关性分析表明,石油烃降解与土壤微生物总生物量具有中等程度的相关性（|r|=0.75）;与G⁺细菌、甲烷氧化菌的生物量具有高度相关性,相关系数|r|>0.8;与G^-的生物量具有中度相关性,相关系数为|r|=0.74;与真菌的生物量具有低度相关性,相关系数为|r|=0.36,与放线菌没有相关性,相关系数为|r|<0.30.     

Assessing microbial and chemical exposure risks of Giardia in indoor swimming pool water disinfected by chlorine

Swimming pools adopt chlorination to ensure microbial safety. Giardia has attracted attention in swimming pool water because of its occurrence, pathogenicity, and chlorine resistance. To control Giardia concentrations in pool water and reduce the microbial risk, higher chlorine doses are required during disinfection. Unfortunately, this process produces carcinogenic disinfection byproducts that increase the risk of chemical exposure. Therefore, quantitatively evaluating the comparative microbial...     

北京市居家空气微生物污染特征

在北京市选取31户有1岁至10岁儿童的家庭进行空气微生物取样,系统研究了室内家庭空气微生物污染特征.结果表明,北京市居家环境空气微生物总浓度变化范围为269～13066 CFU·m-3,均值为2658 CFU· m-3,空气细菌浓度变化范围为47 ～ 12341 CFU·m-3,均值为1821 CFU·m-3,空气真菌浓度变化范围为62～3498 CFU·m-3,均值为837 CFU·m-3.空气细菌和真菌浓度百分比分别为61.0％和39.0％,细菌浓度明显高于真菌浓度.居家环境优势细菌属依次为微球菌属(Micrococcus)、芽孢杆菌属(Bacillus)、葡萄球菌属(Staphylococcus)和库克菌属(Kocuria),4属细菌百分比约占63.1％ ～70.9％,优势真菌属为青霉属(Penicillium)、枝孢属(Cladosporium)、曲霉属(Aspergillus)、链格孢属(Alternaria)和茎点霉属(Phoma),分别约占总数的36.0％、17.8％、9.3％、5.3％和3.6％.文中最后针对北京市居家环境空气微生物污染的现状及其来源,从宠物饲养、空调清理、室内外优良环境的保持及垃圾处理、室内花卉种植等方面提出了治理建议.     

Two selenium tolerant Lysinibacillus sp. strains are capable of reducing selenite to elemental Se efficiently under aerobic conditions

Microbes play important roles in the transport and transformation of selenium (Se) in the environment, thereby influencing plant resistance to Se and Se accumulation in plant. The objectives are to characterize the bacteria with high Se tolerance and reduction capacity and explore the significance of microbial origins on their Se tolerance, reduction rate and efficiency. Two bacterial strains were isolated from a naturally occurred Se-rich soil at tea orchard in southern Anhui Province, China. The reduction kinetics of selenite was investigated and the reducing product was characterized using scanning electron microscopy and transmission electron microscopy-energy dispersive spectroscopy. The bacteria were identified as Lysinibacillus xylanilyticus and Lysinibacillus macrolides, respectively, using morphological, physiological and molecular methods. The results showed that the minimal inhibitory concentrations (MICs) of selenite for L. xylanilyticus and L. macrolides were 120 and 220 mmol/L, respectively, while MICs of selenate for L. xylanilyticus and L. macrolides were 800 and 700 mmol/L, respectively. Both strains aerobically reduced selenite with an initial concentration of 1.0 mmol/L to elemental Se nanoparticles (SeNPs) completely within 36 hr. Biogenic SeNPs were observed both inside and outside the cells suggesting either an intra- or extracellular reduction process. Our study implied that the microbes from Se-rich environments were more tolerant to Se and generally quicker and more efficient than those from Se-free habitats in the reduction of Se oxyanions. The bacterial strains with high Se reduction capacity and the biological synthesized SeNPs would have potential applications in agriculture, food, environment and medicine.     

Arsenic release from microbial reduction of scorodite in the presence of electron shuttle in flooded soil

Scorodite (FeAsO₄·H₂O) is a common arsenic-bearing (As-bearing) iron mineral in nearsurface environments that could immobilize or store As in a bound state.In flooded soils,microbe induced Fe(Ⅲ) or As(Ⅴ) reduction can increase the mobility and bioavailability of As.Additionally,humic substances can act as electron shuttles to promote this process.The dynamics of As release and diversity of putative As(Ⅴ)-reducing bacteria during scorodite reduction have yet to be investigat...     

中亚热带马尾松林凋落物分解过程中的微生物与酶活性动态

采用凋落物原位分解法,研究了中亚热带马尾松林中马尾松、槲栎凋落叶单独及混合(自然质量比8∶2)分解过程中的微生物数量与酶活性动态.结果表明:①3类凋落物的年分解常数K的大小为:混合凋落物(0.94)>槲栎凋落物(0.86)>马尾松凋落物(0.67);②3类凋落物真菌数量和微生物数量均在夏季(135 d时)达到最大值,而此时细菌和放线菌数量最低;③3类凋落物纤维素酶活性、酸性磷酸酶活性均与凋落物干重剩余率呈显著正相关(P<0.05),而马尾松与混合型凋落物中的多酚氧化酶活性同凋落物干重剩余率呈极显著负相关(P<0.01);④微生物数量和多酚氧化酶活性均总体表现为槲栎凋落物>混合凋落物>马尾松凋落物,酸性磷酸酶活性多表现为槲栎凋落物最低,与分解常数K排列有一定的差异,说明凋落物分解是微生物和多种酶共同作用的结果.整体研究表明,凋落物质量和季节气候的差异显著影响微生物群落及其调控的生态过程,与纯马尾松凋落叶相比,针阔混合使微生物数量和多酚氧化酶活性显著提高,这可能是导致分解加快的重要原因.