东天山地区庙儿沟雪坑中微生物多样性、群落结构与环境关系研究

通过荧光计数、恢复培养和变形梯度凝胶电泳(DGGE),分析了东天山地区庙儿沟不同深度积雪中的可培养细菌数量、多样性及其群落结构。结果表明,东天山地区冰雪微生物数量和多样性指数与气候环境替代指标钙离子、镁离子、氯离子等具有相关性。该地区雪坑中细菌隶属于4个不同系统发育群:proteobacteria(α-,β-,γ-),CFB,HGC和LGC,其中CFB类和proteobacteria类为主要类群。与青藏高原、南北极冰雪中微生物的比较分析发现Paracoccu和Aquasalina属是该地区的特殊微生物类群。直接培养和DGGE分析发现不同深度雪坑中微生物数量和群落结构都有明显的变化。结果表明,由于东天山地区的特殊地理位置,该地区冰雪微生物具有其特殊性。     

Bacterial diversity in the snow over Tibetan Plateau Glaciers

Bacterial diversity and cell abundance in the snow of the four glaciers (Guoqu, Zadang, East Rongbuk and Palong No. 4) located in different climatic zones of the Tibetan Plateau were investigated through culture-independent molecular analysis of 16S rRNA gene clone library and flow cytometry approaches. Cell abundance ranged from 0.68 × 10³ to 720 × 10³ cells mL⁻¹, with higher values in the northern glaciers than in the southern ones. Bacterial diversity was unexpectedly high in the snow habitats of the world’s highest plateau, with 15 common genera distributed widely among the glaciers. The bacterial diversity in the snow at different glaciers was related to the surrounding environments. The Guoqu Glacier, to the north near the desert zone and with the lowest temperature, preserved more bacteria closely related to a cold environment and soil than the other glaciers. However, in the Palong No. 4 Glacier located in the south warm region around vegetation, most bacteria were phylogenetically related to plant-associated bacteria.     

北极海洋沉积物中锰细菌的分离与系统发育

对中国第二次北极科学考察采集的北极海洋沉积物中的锰细菌进行了筛选、分离和系统发育分析。根据其在筛选平板上菌落的形态学特征,分别从站位P11和S11采集的沉积物中分离到了21株和19株锰细菌。系统发育分析表明,两个站位的锰细菌群落组成有着明显的差别。站位P11分离的可培养锰细菌主要由细菌域（Bacteria）中变形杆菌门的γ-变形杆菌纲（γ-Proteobacteria）和放线菌纲（Actinobacteria）组成,二者分别占86％和14％;γ-变形杆菌纲主要包括嗜冷杆菌属（Psychrobacter）、希瓦氏菌属（Shewanella）、假交替单胞菌属（Pseudoaheromonas）、不动杆菌属（Acinetobacter）、海杆菌属（Marinobacter）,其中以嗜冷杆菌属为主,其比例可达67％。从站位S11分离到的可培养锰细菌主要包括细菌域中变形杆菌门的α-变形杆菌纲（α-Proteobacteria）和γ-变形杆菌纲以及拟杆菌门（Bacteroides）中的黄杆菌纲（Flavobaeteria）;γ-变形杆菌纲主要包括希瓦氏菌属、海单胞菌属（Marinomonas）和交替单胞菌属（Aheromonas）,α-变形杆菌纲主要为鞘氨醇单胞菌属（Sphingomonas）。实验菌株均对Mn^2＋有着较强的抗性,其中以菌株Marinomonas sp．S11-S-4耐受性最高。     

Phylogenetic and Physiological Diversity of Bacteria Isolated from Puruogangri Ice Core

The microbial abundance, the percentage of viable bacteria, and the diversity of bacterial isolates from different regions of a 83.45-m ice core from the Puruogangri glacier on the Tibetan Plateau (China) have been investigated. Small subunit 16S rRNA sequences and phylogenetic relationships have been studied for 108 bacterial isolates recovered under aerobic growth conditions from different regions of the ice core. The genomic fingerprints based on ERIC (enterobacterial repetitive intergenic consensus)-polymerase chain reaction and physiological heterogeneity of the closely evolutionary related bacterial strains isolated from different ice core depths were analyzed as well. The results showed that the total microbial cell, percentages of live cells, and the bacterial CFU ranged from 10⁴ to 10⁵ cell ml⁻¹ (Mean, 9.47 × 10⁴; SD, 5.7 × 10⁴, n = 20), 25–81%, and 0–760 cfu ml⁻¹, respectively. The majority of the isolates had 16S rRNA sequences similar to previously determined sequences, ranging from 92 to 99% identical to database sequences. Based on their 16S rRNA sequences, 42.6% of the isolates were high-G + C-content (HGC) gram-positive bacteria, 35.2% were low-G + C (LGC) gram-positive bacteria, 16.6% were Proteobacteria, and 5.6% were CFB group. There were clear differences in the depth distribution of the bacterial isolates. The isolates tested exhibited unique phenotypic properties and high genetic heterogeneity, which showed no clear correlation with depths of bacterial isolation. This layered distribution and high heterogeneity of bacterial isolates presumably reflect the diverse bacterial sources and the differences in bacteria inhabiting the glacier’s surface under different past climate conditions.     

不同降压过程对深海海水中可培养细菌群落组成的影响

【目的】控制不同的压力变化过程,比较对深海水样中可培养细菌组成的影响,探讨马里亚纳海沟深海水样中可培养细菌在不同降压处理过程下的丰度变化和群落组成。【方法】利用保压技术采集无污染、深度6001 m的深海水样后,模拟缓慢降压和快速降压过程,通过2216E培养基及2216E加氧化三甲胺(TMAO)富集培养基,对分离得到的可培养菌株进行16S rRNA基因测序分析和丰度检测。【结果】通过缓慢降压和快速降压处理后,深海海水样品中可培养细菌的丰度和群落组成差异较大。其中,在缓慢降压处理的样品中,平均丰度约为190 CFU/mL,且种群组成单一,以Bacillus属为主(占总菌落数的96%);而快速降压处理的样品中,平均丰度约为437 CFU/mL,主要分布在4个属中:Bacillus (占总菌落数的27.8%)、Achromobacter (24.4%)、Microbacterium (34.4%)和Pseudomonas (13.7%)。值得一提的是,添加TMAO后,2种降压过程处理的样品中,可培养细菌的平均丰度均没有明显提升,但样品中的可培养细菌种类明显提升,部分种属的丰度也发生了明显的变化。此外,一些种属仅在特定的压力和底物存在的条件下出现。【结论】不同的降压方式能够影响深海海水中可培养细菌的丰度和群落组成,添加TMAO的富集实验表明可以增加分离到的细菌的种类,为下一步的深入研究提供良好的研究基础。     

Isolation and phylogenetic analysis of cultivable manganese bacteria in sediments from the Arctic Ocean

Isolation, molecular identification and phylogenetic analysis were carried out to investigate the biodiversity of manganese bacteria in sediments which were collected from the Arctic Ocean during the 2nd Chinese Arctic Scientific Expedition. 21 and 19 species of cultivable strains were isolated from sediments at Stations P11 and S11, respectively, according to their distinct morphological character on the screening plate of manganese medium. Molecular identification and phylogenetic analysis showed that the cultivable manganese bacteria from Station P11 were basically composed of γ-Proteobacteria (γ subgroup of the Proteobacteria branch of the domain Bacteria) and Actinobacteria, which accounted for 86% and 14%, respectively. The isolates of γ-Proteobacteria mainly included Psychrobacter, Shewanella, Acinetobacter and Marinobacter, of which Psychrobacter was the major genus, which accounted for 67% of the γ-Proteobacteria. The cultivable manganese bacteria from Station S11 included α-Proteobacteria, γ-Proteobacteria and Flavobacteria of Bacteroides. The γ-Proteobacteria mainly included Shewanella, Marinomonas and Alteromonas. The majority of α-Proteobacteria was Sphingomonas. The phylogenetic analysis indicated that bacteria from sediments at Stations P11 and S11 had different cultivable manganese microbial communities. All tested strains had higher resistance to Mn2+, of which Marinomonas sp. S11-S-4 had the highest resistant ability.     

Differential enumeration and in situ localization of microorganisms in the hindgut of the lower termite Mastotermes darwiniensis by hybridization with rRNA-targeted probes

We examined the abundance and spatial distribution of major phylogenetic groups of the domain Bacteria in hindguts of the Australian lower termite Mastotermes darwiniensis by using in situ hybridization with group-specific, fluorescently labeled, rRNA-targeted oligonucleotide probes. Between 32.0 ± 7.2% and 52.3 ± 8.2% of the DAPI-stained cells in different hindgut fractions were detected with probe EUB338, specific for members of the domain Bacteria. About 85% of the prokaryotic cells were associated with the flagellates of the thin-walled anterior region (P3a) and the thick wall of the posterior region (P3b/P4) of the hindgut, as shown by DAPI staining. At most, half of the EUB338-detected cells hybridized with one of the other probes that targeted a smaller assemblage within the bacterial domain. In most fractions, cells were found in varying numbers with probe ALF1b, which targeted members of the α-Proteobacteria, whereas substantial amounts of sulfate-reducing bacteria, gram-positive bacteria with a high DNA G+C content and members of the Cytophaga-Flavobacterium cluster of the Cytophaga-Flavobacterium-Bacteroides (CFB) phylum could be detected only in the wall fraction of P3b/P4. This clearly indicates that the hindgut microhabitats differ in the composition of their microbial community. In situ hybridization of cryosections through the hindgut showed only low numbers of bacteria attached to the P3a wall. In contrast, the wall of P3b was densely colonized by rod- and coccus-shaped bacteria, which could be assigned to the Cytophaga-Flavobacterium cluster of the CFB phylum and to the group of gram-positive bacteria with a high DNA G+C content, respectively. Oxygen concentration profiles determined with microelectrodes revealed steep oxygen gradients both in P3a and P3b. Oxygen was consumed within 100 μm below the gut surface, and anoxic conditions prevailed in the central portions of both gut regions, indicating that oxygen consumption in the hindgut does not depend on the presence of a biofilm on the hindgut wall. Received: 17 May 1999 / Accepted: 16 September 1999     

崇测冰帽不同的冰川样品可培养细菌群落结构差异北大核心CSCD

【目的】研究细菌群落组成在西昆仑崇测冰帽冰川雪样、冰碛物和土样中的差异。【方法】通过传统的纯培养和菌株16S r RNA基因序列鉴定,分析菌株在门水平和属水平的群落结构。【结果】冰川细菌由Actinobacteria、Firmicutes、Proteobacteria和Bacteroidetes 4个门组成。雪样以Proteobacteria为优势,而土样和冰碛物则以Actinobacteria为优势。在属的水平上,冰川土样中的优势属仅有Arthrobacter,雪样中的优势属主要有Methylobacterium、Modestobacter、Hymenobacter、Brevundimonas、Bacillus这5种。雪环境的细菌群落结构与冰碛物和土样的差异性较大,而冰碛物和土样之间的差异性不大。Skermanella可能为崇测冰帽所特有的细菌。【结论】初步说明了在冰川退缩的气候环境下,冰川雪样细菌多样性的脆弱性,以及冰川雪环境细菌资源保护的重要性。     

Bacterial diversity of the Inner Mongolian Baer Soda Lake as revealed by 16S rRNA gene sequence analyses

Bacterial diversity associated with Baer Soda Lake in Inner Mongolia of China was investigated using a culture-independent method. Bacterial 16S rRNA gene libraries were generated using bacterial oligonucleotide primers, and 16S rRNA gene sequences of 58 clones were analyzed phylogenetically. The library was dominated by 16S rDNAs of Gram-negative bacteria (24% -Proteobacteria, 31% -Proteobacteria, 33% -Proteobacteria, and 2% -Proteobacteria), with a lower percentage of clones corresponding to Gram-positive bacteria. Forty cloned sequences were similar to that of known bacterial isolates (>97% sequence similarity), represented by the species of the genera Brevundimonas, Comamonas, Alcaligenes, Stenotrophomonas, and Klebsiella. Eighteen cloned sequences showed less affiliation with known taxa (<97% sequence similarity) and may represent novel taxa.Communicated by K. Horikoshi     

Microbial diversity in the snow,a moraine lake and a stream in Himalayan glacier

The microbial diversity and abundance in surface snow at different altitudes (5300 and 5504 m above sea level), a moraine lake and a glacial stream in the Yala Glacier on the southern slope of the Himalayas were investigated through a 16S rRNA gene clone library and flow cytometry approaches. Cell abundance in different habitats changed from 1.1 × 10⁴ to 25 × 10⁴ cells mL⁻¹, with the highest abundance in the moraine lake and the lowest abundance in the snow at 5504 m. Microbial communities in the snow were significantly different from those in the moraine lake and stream, although they were similar within snow and within the aquatic habitats. The two snow libraries were both dominated by Cyanobacteria, which accounted for about half of the total, followed by the Alphaproteobacteria and Firmicutes. The moraine lake and stream libraries were dominated by the Bacteroidetes and Betaproteobacteria, followed by the Actinobacteria. The results indicated that snow and water were highly diverse systems even in the same glacier. Microbial communities in the snow on the Yala Glacier were distinctly different from those in the East Rongbuk Glacier on the northern slope of Himalayas. However, microbes in the moraine lakes at two glaciers had similar community features. The snow habitat was easily affected by various environmental factors, while the aquatic habitats were comparatively stable in different glaciers.     

Differential bacterial endophytome in Foc-resistant banana cultivar displays enhanced antagonistic activity against Fusarium oxysporum f.sp. cubense (Foc)

Diverse endophytes with multiple functions exist in different banana cultivars. However, the diversity of cultivable bacterial endophytome that contributes to antifungal activity against Fusarium oxysporum f.sp. cubense (Foc) in resistant and susceptible banana cultivars is mostly unknown. In the present study, we isolated bacterial endophytes from resistant Yengambi KM5 (AAA) and susceptible banana cultivar Ney Poovan (AB) to determine the diversity of cultivable bacterial endophytes. Our study revealed the presence of 56 cultivable bacterial endophytes and 6 nectar-associated bacteria in YKM5 and 31 cultivable bacterial endophytes in Ney Poovan. The identified cultivable bacterial genera in YKM5 included Alcaligenes, Arthrobacter, Azotobacter, Acinetobacter, Agrobacterium, Bacillus, Brucella, Brevundimonas, Brachybacterium, Beijerinckia, Klebsiella, Leclercia, Lysinibacillus, Myroides, Ochrobactrum, Pseudomonas, Rhizobium, Stenotrophomonas, Serratia, and Verticiella. In Ney Poovan, the cultivable endophytic bacterial genera present were Agrobacterium, Bacillus, Bradyrhizobium, Enterobacter, Klebsiella, Lysinibacillus, Micrococcus, Ochrobactrum, Pseudomonas, Rhizobium, and Sphingobium. Thus, the composition and diversity of cultivable endophytic bacterial genera were higher in Foc-resistant YKM5. The antifungal efficacy of bacterial endophytes Brachybacterium paraconglomeratum YEBPT2 (65.5%), Brucella melitensis YEBPS3 (63.3%), Bacillus velezensis YEBBR6 (63.3%), and nectar-associated Bacillus albus YEBN2 (61.1%) from YKM5 showed the highest antifungal activity against Foc, compared with the antifungal activity of endophytes from the susceptible cultivar.     

Bacterial Population Association with Phytoplankton Cultured in a Bivalve Hatchery

Bacterial populations association with phytoplankton cultures used as food for bivalve larvae were enumerated and identified from their partial 16S rDNA gene sequences. Microalgae were provided from different European hatcheries during the larval production season. Average concentration (direct counts) of bacteria ranged from 1.3 × 10⁵ to 5.3 × 10⁸ mL^–1 while culturable bacteria represented from 10% to >60% of total bacteria. In most cases, three to six representatives of each type of colony were collected on solid medium. The identity of isolates from the same colony type was checked by two different randomly amplified polymorphic DNA (RAPD) typing methods, after which the 16S rDNA gene of one to three isolates by colony type were partially sequenced. Algae harbored a large spectrum of bacteria belonging to the -Proteobacteria, -Proteobacteria, -Proteobacteria, Cytophaga– Flavobacterium– Bacteroides (CFB) group, Actinobacteria, and Bacillus. Members of the Roseobacter clade and CFB group were the most abundant. In the majority of cases one strain constituted 50% or more of the culturable bacterial flora. About half of the isolates were common to two hatcheries or at least two microalgal cultures. Several isolates were closely related to bacteria associated with harmful dinoflagellates in culture. Thus, the algal cultures seemed to favor certain bacterial species which belonged to distantly separated groups. As some of them could disturb the development of bivalve larvae, the control of bacterial populations would undoubtedly make it possible to reduce larval losses in bivalve rearing.     

Microorganisms Associated with Feathers of Barn Swallows in Radioactively Contaminated Areas Around Chernobyl

The Chernobyl catastrophe provides a rare opportunity to study the ecological and evolutionary consequences of low-level, environmental radiation on living organisms. Despite some recent studies about negative effects of environmental radiation on macroorganisms, there is little knowledge about the effect of radioactive contamination on diversity and abundance of microorganisms. We examined abundance patterns of total cultivable bacteria and fungi and the abundance of feather-degrading bacterial subset present on feathers of barn swallows (Hirundo rustica), a colonial migratory passerine, around Chernobyl in relation to levels of ground level environmental radiation. After controlling for confounding variables, total cultivable bacterial loads were negatively correlated with environmental radioactivity, whereas abundance of fungi and feather-degrading bacteria was not significantly related to contamination levels. Abundance of both total and feather-degrading bacteria increased with barn swallow colony size, showing a potential cost of sociality. Males had lower abundance of feather-degrading bacteria than females. Our results show the detrimental effects of low-level environmental radiation on total cultivable bacterial assemblage on feathers, while the abundance of other microorganism groups living on barn swallow feathers, such as feather-degrading bacteria, are shaped by other factors like host sociality or host sex. These data lead us to conclude that the ecological effects of Chernobyl may be more general than previously assumed and may have long-term implications for host–microbe interactions and overall ecosystem functioning.     

Culturable bacteria isolated from snow cores along the 1300?km traverse from Zhongshan Station to Dome A, East Antarctica

The abundance and community composition of culturable bacteria in four snow cores along the 1300 km traverse from Zhongshan Station to Dome A, East Antarctica, were investigated through the combination of liquid and solid media and small subunit 16S rRNA sequences. Under aerobic cultivation conditions, the average concentrations of bacterial colonies from each snow core varied from 0.008 to 0.32 CFU mL⁻¹. A total of 37 and 15 isolates with different morphologic characteristics were recovered from solid and liquid media PYGV, respectively. The phylogenetic analysis of 14 representatives with different ARDRA patterns from RFLP showed that all the isolates were affiliated with five phylogenetic groups: Firmicutes, Actinobacteria, Alphaproteobacteria, Gammaproteobacteria and Bacteroidetes. Actinobacteria represented the largest cluster with 43% of strains, and these strains exhibited unique phenotypic properties. The community compositions of culturable bacteria in the four snow cores were distinctly different from each other and the concentrations and community sizes of culturable bacteria along the traverse decreased with increases of latitude, altitude and distance from coast, which likely reflected the different bacterial sources and biogeographies under the different regional climate conditions in the snow cover of East Antarctica.     

Abundance and Diversity of Glacial Bacteria on the Tibetan Plateau with Environment

The relation of glacial microorganism and their living environment is concerned but less understood for both glaciologists and microbiologists. Here we present the results about glacial snow bacteria from 3 glaciers on the Tibetan Plateau. The concentrations and diversity of bacteria collected from snow pit samples of East Rongbuk, Laohugou and Hailuogou glaciers on the Tibetan Plateau were investigated by epifluorescence microscope, denaturing gradient gel electrophoresis and Shannon-Weaver index. Concentrations and community diversity of bacteria in the East Rongbuk glacier with lower concentration and smaller size of microparticle were lower than in the Laohugou with higher concentration and larger size of microparticle. Bacterial concentration in East Rongbuk Glacier was close to that in polar regions, suggesting that Mt. Everest is a bacterial background for remote regions away from direct influence of anthropogenic sources. In addition, altitude difference was another factor for higher concentrations and community diversity of bacteria in Hailuogou and Laohugou glaciers than in East Rongbuk Glacier. The highest concentrations and community diversity of bacteria in Hailuogou Glacier were attributed to its most diverse atmospheric circulations and highest temperature among the 3 glaciers. We suggest a complicated correlation between glacial bacteria and their regional living environments.     

Prokaryotic diversity in sediments beneath two polar glaciers with contrasting organic carbon substrates

Microbial ecosystems beneath glaciers and ice sheets are thought to play an active role in regional and global carbon cycling. Subglacial sediments are assumed to be largely anoxic, and thus various pathways of organic carbon metabolism may occur here. We examine the abundance and diversity of prokaryotes in sediment beneath two glaciers (Lower Wright Glacier in Antarctica and Russell Glacier in Greenland) with different glaciation histories and thus with different organic carbon substrates. The total microbial abundance in the Lower Wright Glacier sediment, originating from young lacustrine sediment, was an order of magnitude higher (~8 × 10⁶ cells per gram of wet sediment) than in Russell Glacier sediment (~9 × 10⁵ cells g⁻¹) that is of Holocene-aged soil origin. 4% of the microbes from the Russell Glacier sediment and 0.04–0.35% from Lower Wright Glacier were culturable at 10°C. The Lower Wright Glacier subglacial community was dominated by Proteobacteria, followed by Firmicutes. The Russell Glacier library was much less diverse and also dominated by Proteobacteria. Low numbers and diversity of both Euryarchaeota and Crenarchaeota were found in both sediments. The identified clones were related to bacteria with both aerobic and anaerobic metabolisms, indicating the presence of both oxic and anoxic conditions in the sediments.     

胡杨叶片及韧皮部内生细菌多样性及生物学功能分析

[目的]通过分析胡杨叶片及树干内生细菌群落多样性、结构特征及生物学功能,探究内生细菌与宿主胡杨的互作机制.[方法]利用Illumina MiSeq高通量测序技术,对采集自新疆喀什地区胡杨林的3组胡杨树叶和3组树干样本的内生细菌进行Alpha、Beta多样性分析、群落组成分析以及通过比对代谢数据库进行群落功能预测,并对样...     

生姜根系不同生态位细菌群落多样性特征、组成及结构差异

生姜作为常见的调味品和传统中药材,是我国重要的经济作物之一。作为取食部分的生姜块茎与根系直接相连,其产量、品质与根相关细菌群落密切相关。然而,关于生姜根系微环境中细菌群落的特点仍鲜有报道,土壤环境能否衍生出宿主特异性内生菌群落尚不清楚。以生姜根系不同生态位细菌群落为研究对象,采用高通量测序技术,对非根际、根际及根内细菌进行16S rRNA基因测序。结果表明,不同生态位细菌群落多样性存在显著差异,其中非根际及根际细菌群落多样性(Shannon index, Observed species, Faith′s PD)显著高于内生菌群落。同时,各生态位共现网络稳定性和复杂度表现为非根际>根际>根内细菌群落。而在组成上,细菌群落在不同生态位差异显著(R²=0.57,P=0.001)。其中变形菌门(Proteobacteria)是根内的优势门,该门类下假单胞菌属(Pseudomonas)、短波单胞菌属(Brevundimonas)、寡养单胞菌属(Stenotrophomonas)及泛菌属(Pantoea)在根内显著富集。在根际细菌中,拟杆菌门(Bacteroid...     

Diversity of Both the Cultivable Protease-Producing Bacteria and Their Extracellular Proteases in the Sediments of the South China Sea

Protease-producing bacteria are known to play an important role in degrading sedimentary particular organic nitrogen, and yet, their diversity and extracellular proteases remain largely unknown. In this paper, the diversity of the cultivable protease-producing bacteria and their extracellular proteases in the sediments of the South China Sea was investigated. The richness of the cultivable protease-producing bacteria reached 10⁶ cells/g in all sediment samples. Analysis of the 16S rRNA gene sequences revealed that the predominant cultivated protease-producing bacteria are Gammaproteobacteria affiliated with the genera Pseudoalteromonas, Alteromonas, Marinobacter, Idiomarina, Halomonas, Vibrio, Shewanella, Pseudomonas, and Rheinheimera, with Alteromonas (34.6%) and Pseudoalteromonas (28.2%) as the predominant groups. Inhibitor analysis showed that nearly all the extracellular proteases from the bacteria are serine proteases or metalloproteases. Moreover, these proteases have different hydrolytic ability to different proteins, reflecting they may belong to different kinds of serine proteases or metalloproteases. To our knowledge, this study represents the first report of the diversity of bacterial proteases in deep-sea sediments.     

Remarkable impact of PAHs and TPHs on the richness and diversity of bacterial species in surface soils exposed to long-term hydrocarbon pollution

Nowadays, because of substantial use of petroleum-derived fuels the number and extension of hydrocarbon polluted terrestrial ecosystems is in growth worldwide. In remediation of aforementioned sites bioremediation still tends to be an innovative, environmentally attractive technology. Although huge amount of information is available concerning the hydrocarbon degradation potential of cultivable hydrocarbonoclastic bacteria little is known about the in situ long-term effects of petroleum derived compounds on the structure of soil microbiota. Therefore, in this study our aim was to determine the long-term impact of total petroleum hydrocarbons (TPHs), volatile petroleum hydrocarbons (VPHs), total alkyl benzenes (TABs) as well as of polycyclic aromatic hydrocarbons (PAHs) on the structure of bacterial communities of four different contaminated soil samples. Our results indicated that a very high amount of TPH affected positively the diversity of hydrocarbonoclastic bacteria. This finding was supported by the occurrence of representatives of the α-, β-, γ-Proteobacteria, Actinobacteria, Flavobacteriia and Bacilli classes. High concentration of VPHs and TABs contributed to the predominance of actinobacterial isolates. In PAH impacted samples the concentration of PAHs negatively correlated with the diversity of bacterial species. Heavily PAH polluted soil samples were mainly inhabited by the representatives of the β-, γ-Proteobacteria (overwhelming dominance of Pseudomonas sp.) and Actinobacteria.