基于pufM基因的乌梁素海富营养化湖区好氧不产氧光合细菌系统发育多样性分析

好氧不产氧光合细菌(AAPB)的多样性在海洋中已经广泛研究,但在富营养化湖泊中却研究甚少。通过构建和分析AAPB光合中心合成中的关键基因pufM克隆文库,以期揭示乌梁素海富营养化湖区AAPB分布及其系统发育多样性,探讨其在富营养化湖泊中的功能和作用。对乌梁素海红圪卜湖区水体文库中的52个克隆子进行分析,产生了28个OTU,文库覆盖度达到71.4%,反映出文库有较好的代表性。同源性和系统发育分析结果表明,乌梁素海红圪卜湖区AAPB有较高的多样性,与我们之前所发现的同一湖区总细菌多样性较低形成鲜明对比。所获得的序列分属7个亚群,即γ-Proteobacteria(44.2%,含Group-1,-2和-3共3个亚群)、β-Proteobacteria(21.2%)、Rhodobacter-like(7.69%)及2个未知亚群unknown Group-1(21.2%)和Group-2(5.77%)。其中γ-Proteobacteria占到总克隆的44.2%,在低盐的乌梁素海环境中出现高比例的γ-Proteobacteria之前并未见类似报道,并且乌梁素海中存在一些可能是富营养化湖泊特有的AAPB。这表明AAPB可能在富营养化湖泊生态系统的维持和稳定中有重要作用。     

玛珥湖好氧不产氧光合细菌pufM基因DNA和mRNA的定量及多样性分析

【目的】湖光岩玛珥湖是一类特殊的火山口湖,它完全封闭,地质年代久远,尚未受人类活动的剧烈影响,孕育着丰富而特殊的微生物种群。好氧不产氧光合细菌(AAPB)是以其在有氧情况下能行使光合功能而定义的一类专性异养细菌,其生理生态特征独特,进化年代久远,在水生生态系统的上层水体中广泛分布。目前,AAPB在玛珥湖水体中是否有分布仍是未知。【方法】构建和比较夏季湖水1 m、5 m、12 m三个水层的总DNA和总RNA的AAPB光合中心合成的关键基因pufM的6个克隆文库,并结合定量PCR技术,分析了不同水层AAPB的分布、系统发育多样性及其在总细菌中的比重。【结果】6个文库覆盖率和稀释曲线显示样本初步揭示了各水层优势AAPB类群的多样性。BLAST核苷酸同源性介于80%93%;多样性指数表明,湖光岩表层和底层多样性相当,中间层最低,总RNA的多样性高于总DNA。系统发育分析结果表明,OTU21 24所含的序列(占总序列的49.43%)与β-变形细菌的进化距离最接近,是湖光岩玛珥湖的优势AAPB菌群。定量PCR结果显示1 m水层中AAPB在总细菌中的比重最高,可达38.06%;而5 m水层中AAPB所占的比重最低,仅为0.85%;12 m为9.54%。【结论】湖光岩玛珥湖孕育着丰富而多样的AAPB类群。     

不产氧光合细菌光合色素的光氧调控机制

[目的]为不产氧光合细菌光合色素研究提供可行的较系统规范的研究方法和数据,揭示固氮红细菌(Rhodobacter azotoformans 134K20)光合色素光氧适应性机制.[方法]采用光谱法和色谱法对光和氧调控下的类胡萝卜素和细菌叶绿素合成代谢进行了研究.[结果]134K20菌株光照好氧时细胞得率最高.光照厌氧时主要合成3黄、1红、1紫、2绿、2蓝9种色素,黄色素大量表达.有氧时红色素大量表达,且启动2种新的红色素和1种新的紫色素表达,而黄色和蓝绿色素则受氧抑制.黑暗好氧主要合成2黄、3红、2紫、1绿、1蓝9种色素,但不同于光照厌氧.光照好氧时黄色素减少到1种,紫色素含量增加,其余同黑暗好氧.[结论]固氮红细菌(Rhodobacter azotoformans 134K20)是通过PpsR调节途径来调节光合基因表达的.黄色和红色素属于类胡萝卜素.黄色素1属于球形烯系列,其余两种黄色素是新的类胡萝卜素组分.红色素为新的球形烯酮组分,3种红色素极性、峰形和峰位差别显著,正己烷能显示其精细结构.紫色为极性较大的细菌脱镁叶绿素,绿色和蓝色为4种极性不同的细菌叶绿素a中间产物.乙醚甲醇法适合类胡萝卜素的提取,丙酮甲醇冰冻研磨法能快速有效完全提取光合色素.溶剂效应可有效鉴别细菌叶绿素a中间产物.     

不产氧光合细菌的分类学进展

不产氧光合细菌(APB)一直是研究生命起源与进化、光合作用和固氮机理的良好模式生物。近年来,APB资源研究发展迅速,不断发现新的物种、模式种及特殊功能物种,不断提出新的分类单元,APB分类系统发生了较大变化,同时引起了一定混乱。本文对不产氧光合细菌定义、分类系统和分类指征最新进展进行了系统述评,对分类系统中存在的问题和发展趋势也进行了评述。     

不产氧光合细菌色素蛋白复合体研究进展

摘要: 色素蛋白复合体是光合生物进行光合作用维持生命活动最重要结构基础。目前不产氧光合细菌色素蛋白复合体仍是最具前沿研究领域。本文概述了不产氧光合细菌各种属色素蛋白复合体研究现状,着重对光反应中心色素蛋白复合体（reaction center,RC）和捕光色素蛋白复合体（light-harvesting complex,LH）,尤其是新型捕光色素蛋白复合体LH3和LH4的组成、精细结构、蛋白同源性和功能进行了述评,并就研究中存在的问题和发展趋势进行了讨论。     

黄河滩地和稻田土中铁还原菌、不产氧光合细菌分布机制

【目的】探讨沿黄流域土壤中铁还原菌(ferric reducing bacteria, Fe RB)、不产氧光合细菌(anoxygenic phototrophic bacteria, An PB)的分布机制。【方法】以沿黄流域(原阳段)为研究对象,采集黄河滩地和稻田土样,利用16Sr RNA基因高通量测序和实时荧光定量分析技术,结合统计学分析,揭示Fe RB、An PB菌群结构、丰度和主要环境影响因子。【结果】二者中的优势Fe RB在科(属)水平为Hydrogenophilaceae(Thiobacillus)、 Bacillaceae(Bacillus)、 Clostridiaceae、Rhodobactereace(Rhodobacter)、 Geobacteraceae(Geobacter),优势An PB为Rhodobactereace(Rhodobacter)、 Chloroflexaceae(Chloronema)、 Acetobacteraceae(Roseomonas)。An PB中Rhodobacteraceae与Fe RB中Bacillaceae、 Clostri...     

锡林河流域潜在不产氧光合细菌陆向分异特征及影响因素

【目的】为探明锡林河流域潜在不产氧光合细菌（anoxygenic photosynthetic bacteria,AnPB）的陆向分异特征及影响因素。【方法】本研究沿着陆向梯度依次采集水生湍流带、缓流带、滞流带、水偏湿生样带、湿偏旱生样带、旱生样带土壤样品。基于文献建立AnPB在科水平的数据库,运用16S rRNA基因高通量测序筛选科水平潜在AnPB类群及其组成丰度的陆向分异,运用皮尔逊相关性及冗余分析等研究土壤理化因子对潜在AnPB陆向分异的影响。【结果】紫色硫细菌（外硫红螺菌科）和紫色非硫细菌（红杆菌科、红环菌科、醋酸杆菌科、丛毛单胞菌科、全噬菌科）主要分布在水生及水偏湿生生境,其相对丰度与湿度呈显著（P<0.05）或极显著（P<0.01）正相关关系;紫色非硫细菌（红螺菌科、慢生根瘤菌科、生丝微菌科、红菌科）、芽单胞菌科、酸杆菌科、绿色非硫细菌（蔷薇菌科）等主要分布在湿偏旱生和旱生环境中,其相对丰度与盐度和全氮含量呈显著（P<0.05）或极显著（P<0.01）正相关关系;多元回归树分析显示,盐度、湿度、全氮对潜在AnPB陆向分异的总解释度分别为62.39%、...     

一株好氧反硝化细菌的分离鉴定及反硝化特性研究

通过对采集水样进行富集培养,利用溴百里酚蓝( BTB)选择性培养基初筛和活性测定复筛得到一株好氧反硝化细菌N22’,发现该菌在好氧条件下能有效去除培养液中的NO3--N.硝酸盐氮初始浓度为125 mg/L,培养40h后硝酸盐氮去除率达86.39％;扫描电镜照片显示,菌株N22’为短杆菌,无鞭毛,大小约为(0.75-1.25)μm×(0.5-0.75) μm范围内,菌落表面呈乳白色.通过形态、生理生化特征及16S rRNA基因序列分析,初步判断菌株N22’为不动杆菌属Acinetobacter sp..反硝化性能测试结果表明,该菌反硝化作用的最适温度为25-30℃,pH值7.0.     

栎黄掌舟蛾幼虫肠道好氧菌群分析及产纤维素酶菌的筛选

栎黄掌舟蛾Phalera assimilis是柞树主要叶部害虫之一。本研究以栎黄掌舟蛾幼虫为材料,从幼虫肠道中分离出好氧细菌23株,通过对16S r DNA扩增产物ARDRA分析后,代表性菌株测序结果表明,23株肠道好氧细菌分别属于葡萄球菌属Staphylococcus sp.、短小杆菌属Curtobacterium sp.、赖氨酸芽孢杆菌属Lysinibacillus sp.、肠球菌属Enterococcus sp.和阿特拉津降解菌属Arthrobacter sp.5个属的细菌,其中,以葡萄球菌属及短小杆菌属细菌种类最多。通过筛选培养基从23株菌中筛选出产纤维素酶菌6株。本研究可为深入了解栎黄掌舟蛾肠道菌群结构、寻找产纤维素酶菌及新的微生物资源等奠定了理论基础。     

产碱菌Alcaligenes sp. qz-1 对铬污染土壤中玉米生长和铬累积的影响研究

通过盆栽试验研究了铬(Cr₂O₇^2-)污染土壤中接种铬耐受细菌Alcaligenes sp. qz-1 对玉米株高、茎粗、叶绿素含量、地上部分和地下部分干重以及铬含量的影响。结果表明, 在重金属铬(Cr₂O₇^2-) 胁迫(50 mg·kg^–1, 100 mg·kg^–1)下,玉米生长显著受到抑制(P<0.05)。与对照相比, 株高分别减少9.92%和18.97%, 茎粗分别降低了13.36%和14.24%, 叶绿素含量分别降低了10.46%和13.25%。接种Alcaligenes sp. qz-1, 可以显著缓解重金属胁迫对玉米生长的不利影响(P<0.05)。铬(Cr₂O₇^2-) 胁迫(50 mg·kg^–1, 100 mg·kg^–1)下, 接种Alcaligenes sp. qz-1 的植株与未接种的玉米植株相比, 株高分别增加了6.36%和10.91%, 茎粗分别增加了10.17%和8.01%, 叶绿素含量分别提高了9.16%和2.64%。同时, 接种Alcaligenes sp. qz-1, 可显著提高玉米(尤其是根系部分)对土壤铬的吸收量, 吸收总量较未接种的增加13.9%-36.9%。综上所述, 接种Alcaligenes sp. qz-1 改善了铬(Cr₂O₇^2-)胁迫下玉米生长特性, 增加了玉米根部对铬(Cr₂O₇^2-)的吸收, 降低了铬(Cr₂O₇^2-)对玉米的毒害, 为该菌株进一步应用农田重金属污染进行治理提供一定理论依据和实践指导。     

Abundant presence of the gamma-like Proteobacterial pufM gene in oxic seawater

Known anoxygenic photosynthetic bacteria (APB) affiliated to Gammaproteobacteria usually use anaerobic metabolism and are restricted to oxygen-free habitats. Here, we report abundant (average of 34.5%) presence of diverse APB related to gamma-like Proteobacteria in oxic oceanic surface water as indicated by the pufM gene, that encodes the M subunit of the light reaction centre complex. Thus, our sequences were most likely derived from aerobic anoxygenic phototrophs (AAnP). Two genetically distinct genotypes were revealed: one was from the oligotrophic North Pacific Ocean Gyre and the other, was from the trophic East China Sea and Bering Sea. The discovery of abundant presence of novel gamma-like Proteobacterial pufM gene in the oxic seawater extends the functional ecotypes of AAnP.     

赤杆菌科微生物分类研究进展

赤杆菌科微生物隶属于细菌域(Bacteria)、变形菌门(Proteobacteria)、α-变形菌纲(Alphaproteobacteria)、鞘脂单胞菌目(Sphingomonadales)。依据16S rRNA基因序列的系统发育分析结果,2005年赤杆菌科被提议建立,成立之初由3个属5个种组成;在过去10年中,该科的物种数量增加迅速;截止2015年10月,赤杆菌科合格发表6个属44个种。赤杆菌科微生物在土壤、热泉、海水和沉积物等环境中广泛分布,其在环境修复、食品工程及生物医药等领域具有重要的应用前景。此外,部分菌株具有细菌叶绿素a,营好氧异养生长兼具光合作用功能,是好养不产氧光合异养菌的一个重要类群,近年来其在全球海洋碳循环和能量代谢过程中的重要作用日益凸显,得到了国内外学者的关注。本文综述了赤杆菌科微生物的分类特征、生态分布和应用基础等方向的研究进展,并对其分类研究趋势做了分析。     

Anoxygenic phototrophic bacterial community of Lake Shira (Khakassia)

The anoxygenic phototrophic bacterial community of the brackish meromictic Lake Shira (Khakassia) was investigated in August 2001, July 2002, and February–March 2003. In all the periods of investigation, the prevailing microorganisms were purple sulfur bacteria similar to Lamprocystis purpurea in morphology and pigment composition. Their highest number (3 × 10⁵ cells/ml) was recorded in July 2002 at the depth of 15 m. According to 16S rRNA gene analysis, the strain of purple sulfur bacteria isolated in 2001 and designated ShAm01 exhibited 98.6% similarity to the type strain of Thiocapsa roseopersicina and 97.1–94.4% similarity to the type strains of Tca. pendens, Tca. litoralis, and Tca. rosea. The minor microorganisms of the anoxygenic phototrophic bacterial community within the period of investigation were nonsulfur purple bacteria phylogenetically close to Rhodovulum strictum (98.3% similarity, strain ShRb01), Ahrensia kielensis (of 93.9% similarity, strain ShRb02), Rhodomicrobium vannieli (of 99.7% similarity, strain ShRmc01), and green sulfur bacteria, phylogenetically close to Chlorobium limicola (of 98.7% similarity, strain ShCl03).     

Salinity and DOC Influence the Distribution of Free-living and Particle-attached Aerobic Anoxygenic Phototrophic Bacteria in the Qinghai–Tibetan Lakes

Aerobic anoxygenic phototrophic bacteria (AAPB) are widespread and play an important role in carbon cycling in the lakes of the Qinghai–Tibetan Plateau. However, little is known about how free-living and particle-attached AAPB distribute with salinity in the Qinghai–Tibetan lakes. In the present study, the abundance and diversity of free-living and particle-attached AAPB were investigated in seven Qinghai–Tibetan lakes with salinity ranging from freshwater to almost saturation (1.2 g L^?1 241.1 g L^?1). An integrated approach was employed including pufL-M gene (encoding the photosynthetic reaction center of AAPB) based quantitative polymerase chain reaction (qPCR) and PCR-cloning phylogenetic analysis. The qPCR data showed that the ratio between particle-attached and free-living AAPB was positively correlated (R² = 0.73, p < 0.01) with increasing salinity. The phylogenetic analysis of pufL-M gene showed that the composition of AAPB population varied with salinity in the studied Qinghai–Tibetan lakes. These results suggested that salinity may be the important factor shaping the AAPB distribution in the studied Qinghai–Tibetan lakes, and AAPB may be adapted to the harsh conditions in the Qinghai–Tibetan lakes by attaching to particles.     

10种常见细菌基因检测膜条的研制

以16S rRNA基因为检测靶基因,设计10种常见细菌的DNA探针,将探针固定于硝酸纤维素膜条;PCR扩增细菌的16S rRNA基因片段并标记生物素后与膜条杂交;采用碱性磷酸酶标记的链亲和素检测生物素标记,以NBT/BCIP显色。该膜条不仅能单独检测10种细菌中的任何一种,也能同时检测5种细菌。该方法具备高通量、低成本、快速、准确等特点,具有良好的临床应用前景。     

Seasonal changes in the structure of the anoxygenic photosynthetic bacterial community in Lake Shunet,Khakassia

Seasonal studies of the anoxygenic phototrophic bacterial community of the water column of the saline eutrophic meromictic Lake Shunet (Khakassia) were performed in 2002 (June) and 2003 (February–March and August). From the redox zone down, the lake water was of dark green color. Green sulfur bacteria predominated in every season. The maximum number of green sulfur bacteria was 10⁷ cells/ml in summer and 10⁶ cells/ml in winter. A multi-syringe stratification sampler was applied for the study of the fine vertical distribution of phototrophs in August 2003; the sampling was performed every 5 cm. A 5-cm-thick pink-colored water layer inhabited by purple sulfur bacteria was shown to be located above the layer of green bacteria. The species composition and ratio of purple bacterial species depended on the sampling depth and on the season. In summer, the number of purple sulfur bacteria in the layer of pink water was 1.6 × 10⁸ cells/ml. Their number in winter was 3 × 10⁵ cells/ml. In the upper oxygen-containing layer of the chemocline the cells of purple nonsulfur bacteria were detected in summer. The maximum number of nonsulfur purple bacteria, 5 × 10² cells/ml, was recorded in August 2003. According to the results of the phylogenetic analysis of pure cultures of the isolated phototrophic bacteria, which were based on 16S rDNA sequencing, green sulfur bacteria were close to Prosthecochloris vibrioformis, purple sulfur bacteria, to Thiocapsa and Halochromatium species, and purple nonsulfur bacteria, to Rhodovulum euryhalinum and Pinkicyclus mahoneyensis.