南极恩克斯堡岛土壤中可培养细菌多样性

【背景】南极地区环境苛刻植被稀少,无冰区面积约占其总面积的0.4%,但是土壤中存在着丰富的微生物群落,对于极地微生物资源仍需要进一步挖掘。【目的】获得南极恩克斯堡岛土壤中可培养细菌多样性信息。【方法】对来源于南极恩克斯堡岛的5个土壤样品采用直接涂布、微好氧富集和有氧富集后涂布3种方法进行细菌分离培养。【结果】共获得144株可培养细菌,分布于5门30个属。这些菌株来自变形菌门(Proteobacteria,38.9%)、放线菌门(Actinobacteria,34.0%)、厚壁菌门(Firmicutes,22.2%)、异常球菌-栖热菌门(Deinococcus-Thermus,3.5%)、拟杆菌门(Bacteroidetes,1.4%)等。不同土壤样品中可培养细菌多样性存在差异。企鹅粪土中Bacillus是主要属类;Pseudomonas、Streptomyces在植被覆盖区土壤中是优势菌属;Psychrobacter在湖水边土壤中是优势属类,Flavobacterium、Chryseobacterium仅从该样品中分离得到;放线菌门类在干燥土壤样中占优势,Pseudarthrobacter、Rhodococcus、Microbacterium等属仅从这类干燥样品中分离出。经16S rRNA基因序列对比发现,有2株菌为潜在新种。【结论】南极恩克斯堡岛土壤中存在可开发利用的菌种资源,本文为研究该地细菌多样性提供一定的基础数据。     

Impact of commonly used agrochemicals on bacterial diversity in cultivated soils

The effects of three selected agrochemicals on bacterial diversity in cultivated soil have been studied. The selected agrochemicals are Cerox (an insecticide), Ceresate and Paraquat (both herbicides). The effect on bacterial population was studied by looking at the total heterotrophic bacteria presence and the effect of the agrochemicals on some selected soil microbes. The soil type used was loamy with pH of 6.0–7.0. The soil was placed in opaque pots and bambara bean (Vigna subterranean) seeds cultivated in them. The agrochemicals were applied two weeks after germination of seeds at concentrations based on manufacturer’s recommendation. Plant growth was assessed by weekly measurement of plant height, foliage appearance and number of nodules formed after one month. The results indicated that the diversity index (Di) among the bacteria populations in untreated soil and that of Cerox-treated soils were high with mean diversity index above 0.95. Mean Di for Ceresate-treated soil was 0.88, and that for Paraquattreated soil was 0.85 indicating low bacterial populations in these treatment-type soils. The study also showed that application of the agrochemicals caused reduction in the number of total heterotrophic bacteria population sizes in the soil. Ceresate caused 82.50% reduction in bacteria number from a mean of 40 × 10⁵ cfu g⁻¹ of soil sample to 70 × 10⁴ cfu g⁻¹. Paraquat-treated soil showed 92.86% reduction, from a mean of 56 × 10⁵ cfu g⁻¹ to 40 × 10⁴ cfu g⁻¹. Application of Cerox to the soil did not have any remarkable reduction in bacterial population number. Total viable cell count studies using Congo red yeast-extract mannitol agar indicated reduction in the number of Rhizobium spp. after application of the agrochemicals. Mean number of Rhizobium population numbers per gram of soil was 180 × 10⁴ for the untreated soil. Cerox-treated soil recorded mean number of 138 × 10⁴ rhizobial cfu g⁻¹ of soil, a 23.33% reduction. Ceresate- and Paraquat-treated soils recorded 20 × 10⁴ and 12 × 10⁴ cfu g⁻¹ of soil, respectively, representing 88.89% and 93.33% reduction in Rhizobium population numbers. Correspondingly, the mean number of nodules per plant was 44 for the growth in untreated soil, 30 for the plant in the Cerox-treated soil, 8 for the plant in Paraquat-treated soil and 3 for the plant in Ceresate-treated soil. The study has confirmed detrimental effect of insecticide on bacterial populations in the soil. Total heterotrophic counts, rhizobial counts as well as the number of nodules of all samples taken from the chemically treated soils were all low as compared to values obtained for the untreated soil. However, the effect of the insecticide was minimal in all cases as compared to the effects of the herbicides on the soil fauna. Indiscriminate use of agrochemicals on farms can therefore affect soil flora and subsequently food production.     

Fermentation dynamics and diversity of bacterial community in four typical woody forages

Woody forage is a new feeding resource used worldwide, and silage is the main long-term storage, mediated by micro-organisms present during their processing. The objectives of our work were to evaluate the fermentation dynamics and to characterize the bacterial community of our typical woody forages. We selected four typical woody forages: paper mulberry (Broussonetia papyrifera), mulberry (Morus alba L.), moringa tree (Moringa oleifera), and Anthocephalus chinensis (Neolamarckia cadamba). The materials were ensiled and sampled after ensiling 1, 3, 5, 7, 15, 30, and 60 days. Our results indicate that woody forages have good forage properties with relatively high crude protein content and low neutral detergent fiber and acid detergent fiber content. However, the water soluble carbohydrate content in paper mulberry was considerably low (18.67 g kg−1), which makes ensiling difficult. The lactic acid and acetic acid contents in each of the four materials were reduced after 3 days of ensiling and increased again after 30 days of ensiling, with the exception of Anthocephalus chinensis. Anthocephalus chinensis and moringa tree were well-preserved after 7 and 60 days of ensiling, respectively, with low pH and ammonia nitrogen content. Cyanobacteria was predominant in moringa tree and Anthocephalus chinensis before ensiling, and Lactobacillus became dominant after 15 days of ensiling. Enterobacter dominated the paper mulberry and mulberry during fermentation process and accelerated their poor silage quality. Therefore, the conformity of bacterial community succession with ensiling parameters guaranteed the final quality of woody forage silages, and this might aid in controlling the manufacturing process.     

Distribution and genetic diversity of bacterial thiopurine methyltransferases in soils emitting dimethyl selenide

Dimethyl selenide (DMSe) and dimethyl diselenide (DMDSe) emissions by soil samples spiked with selenite or (methyl)selenocysteine, with or without a supplement of nutrient broth and glucose were measured. DMSe was the main form of volatile Se produced, and was observed for both Se-substrates. DMDSe was only emitted from soils spiked with (methyl)selenocysteine. Two bacterial thiopurine methyltransferases (TPMTs), TPMT-I and TPMT-E, have been reported to be involved in DMSe and DMDSe emissions [J. Bacteriol. 184 (2002) 3146; Appl. Environ. Microbiol. 69 (2003) 3784]. To establish if these TPMTs or other members of their gene family could have contributed to the DMSe emissions observed, the diversity of bTPMT gene (tpm) sequences among the soils of this study was investigated. Total DNAs from these soils were extracted and screened using the tpm PTCF2-PTCR2 consensus primers defined to PCR amplify this gene family. The PCR products obtained from two soils were cloned, analysed by PCR-RFLP, and sequenced. Their analysis showed an important diversity of tpm lineages (around 12) in soils. Phylogenetic analysis of the deduced TPMT sequences of these soils revealed lineages not previously recorded in the databases, sequences closely related or identical to freshwater TPMTs, or sequences encoding TPMTs closely related to those of Pseudomonas fragi TPMT-K, Pseudomonas Hsa.28 TPMT-I, or Colwellia psychrerythraea TPMT-Z. Nested PCRs, allowing detection of about 13 distinct tpm soil and freshwater lineages by PTCF2-PTCR2 PCR screenings, were performed on the soil total DNAs. These PCRs confirmed the sequencing data, and allowed to recover lineages not detected by the cloning strategy. These results indicate that soils, like the freshwater samples, harbour TPMT-I gene sequences but may also have distinct tpm lineages. This study further supports our hypothesis that TPMTs contribute to DMSe soil emissions.     

Seasonal and altitudinal changes of culturable bacterial and yeast diversity in Alpine forest soils

The effect of altitude and season on abundance and diversity of the culturable heterotrophic bacterial and yeast community was examined at four forest sites in the Italian Alps along an altitude gradient (545–2000 m). Independently of altitude, bacteria isolated at 0 °C (psychrophiles) were less numerous than those recovered at 20 °C. In autumn, psychrophilic bacterial population increased with altitude. The 1194 bacterial strains were primarily affiliated with the classes Alpha-, Beta-, Gammaproteobacteria, Spingobacteriia and Flavobacteriia. Fifty-seven of 112 operational taxonomic units represented potential novel species. Strains isolated at 20 °C had a higher diversity and showed similarities in taxa composition and abundance, regardless of altitude or season, while strains isolated at 0 °C showed differences in community composition at lower and higher altitudes. In contrast to bacteria, yeast diversity was season-dependent: site- and altitude-specific effects on yeast diversity were only detected in spring. Isolation temperature affected the relative proportions of yeast genera. Isolations recovered 719 strains, belonging to the classes Dothideomycetes, Saccharomycetes, Tremellomycetes and Mycrobotryomycetes. The presence of few dominant bacterial OTUs and yeast species indicated a resilient microbial population that is not affected by season or altitude. Soil nutrient contents influenced significantly abundance and diversity of culturable bacteria, but not of culturable yeasts.     

Microbial community succession and bacterial diversity in soils during 77,000 years of ecosystem development

The origins of the biological complexity and the factors that regulate the development of community composition, diversity and richness in soil remain largely unknown. To gain a better understanding of how bacterial communities change during soil ecosystem development, their composition and diversity in soils that developed over c. 77 000 years of intermittent aeolian deposition were studied. 16S rRNA gene clone libraries and fatty acid methyl ester (FAME) analyses were used to assess the diversity and composition of the communities. The bacterial community composition changed with soil age, and the overall diversity, richness and evenness of the communities increased as the soil habitat matured. When analysed using a multivariate Bray-Curtis ordination technique, the distribution of ribotypes showed an orderly pattern of bacterial community development that was clearly associated with soil and ecosystem development. Similarly, changes in the composition of the FAMEs across the chronosequence were associated with biomarkers for fungi, actinomycetes and Gram-positive bacteria. The development of the soil ecosystem promoted the development of distinctive microbial communities that were reminiscent of successional processes often evoked to describe change during the development of plant communities in terrestrial ecosystems.     

A meta-analysis of the publicly available bacterial and archaeal sequence diversity in saline soils

An integrated view of bacterial and archaeal diversity in saline soil habitats is essential for understanding the biological and ecological processes and exploiting potential of microbial resources from such environments. This study examined the collective bacterial and archaeal diversity in saline soils using a meta-analysis approach. All available 16S rDNA sequences recovered from saline soils were retrieved from publicly available databases and subjected to phylogenetic and statistical analyses. A total of 9,043 bacterial and 1,039 archaeal sequences, each longer than 250 bp, were examined. The bacterial sequences were assigned into 5,784 operational taxonomic units (OTUs, based on ≥97 % sequence identity), representing 24 known bacterial phyla, with Proteobacteria (44.9 %), Actinobacteria (12.3 %), Firmicutes (10.4 %), Acidobacteria (9.0 %), Bacteroidetes (6.8 %), and Chloroflexi (5.9 %) being predominant. Lysobacter (12.8 %) was the dominant bacterial genus in saline soils, followed by Sphingomonas (4.5 %), Halomonas (2.5 %), and Gemmatimonas (2.5 %). Archaeal sequences were assigned to 602 OTUs, primarily from the phyla Euryarchaeota (88.7 %) and Crenarchaeota (11.3 %). Halorubrum and Thermofilum were the dominant archaeal genera in saline soils. Rarefaction analysis indicated that less than 25 % of bacterial diversity, and approximately 50 % of archaeal diversity, in saline soil habitats has been sampled. This analysis of the global bacterial and archaeal diversity in saline soil habitats can guide future studies to further examine the microbial diversity of saline soils.     

盐度对滨海土壤细菌多样性和群落构建过程的影响

滨海盐土是重要的农业土地后备资源.微生物是土壤中物质循环的关键动力,然而盐度对土壤微生物群落特征影响的研究还很缺乏.本研究采集滨海地区的土壤样品,研究非盐、轻盐和高盐3组不同盐度对土壤细菌数量、多样性和群落构建的影响.结果表明:与非盐和轻盐土壤相比,高盐土壤的脱氢酶活性和细菌数量显著降低,而细菌α多样性没有变化,细菌群...     

Environmental influences on bacterial diversity of soils on Signy Island,maritime Antarctic

Soil bacterial diversity at environmentally distinct locations on Signy Island, South Orkney Islands was examined using the denaturing gradient gel profiling approach. A range of chemical variables in soils at each site was determined in order to describe variation between locations. No apparent differences in Shannon Diversity Index (H′) were observed. However, as revealed in an analysis of similarity (ANOSIM), the dominant bacterial communities of all eight studied locations were significantly different. Within this, higher levels of similarity were observed between penguin rookeries, seal wallows and vegetated soils, all of which share varying levels of impact from vertebrate activity, in contrast with more barren soil. In addition, the lowest H′ value was detected from the latter soil which also has the most extreme environmental conditions, and its bacterial community has the greatest genetic distance from the other locations. DGGE analyses indicated that the majority of the excised and sequenced bands were attributable to the Bacteroidetes. Across a range of ten environmental variables, multivariate correlation analysis suggested that a combination of pH, conductivity, copper and lead content potentially contributed explanatory value to the measured soil bacterial diversity.     

Cultivation-dependent characterization of bacterial diversity from British Columbia forest soils subjected to disturbance

Bacteria from forest surface organic matter and mineral soil horizons were cultivated using four methods and characterized by fatty acid methyl ester (FAME) analysis. Soil samples from a British Columbia Ministry of Forests Long-Term Soil Productivity (LTSP) installation were collected during winter and summer from two disturbance treatments (whole-tree harvesting with no soil compaction (plot N) and whole-tree harvesting plus complete surface organic matter removal with heavy soil compaction (plot S)) and from an unlogged reference plot (REF). Seventy-five percent of 1795 bacterial isolates were affiliated with 42 genera representing beta- and gamma-Proteobacteria, Actinobacteria, the Bacillus/Clostridium group, and the Cytophaga-Flexibacter-Bacteroides group. Approximately half of the culture collection represented genetic diversity confined to four bacterial genera: Pseudomonas, Bacillus, Paenibacillus, and Arthrobacter. A significantly higher proportion of bacterial isolates belonging to Actinobacteria, and the member genus Arthrobacter, were isolated from plot S soil samples compared with soil samples from plots N and REF. Twenty-five percent of bacterial isolates were not conclusively identified to genus with FAME analysis. Sherlock Tracker cluster analysis and partial 16S rRNA gene sequence analysis enabled classification of a subset of these isolates.     

Molecular characterization of bacterial diversity from British Columbia forest soils subjected to disturbance

Bacteria from forest soils were characterized by DNA sequence analysis of cloned 16S rRNA gene fragments (16S clones). Surface organic matter and mineral soil samples from a British Columbia Ministry of Forests Long-Term Soil Productivity (LTSP) installation were collected during winter and summer from two disturbance treatments: whole-tree harvesting with no soil compaction (plot N) and whole-tree harvesting plus complete surface organic matter removal with heavy soil compaction (plot S). Phylogenetic analyses revealed that 87% of 580 16S clones were classified as Proteobacteria, Actinobacteria, Acidobacterium, Verrucomicrobia, Bacillus/Clostridium group, Cytophaga-Flexibacter-Bacteroides group, green nonsulfur bacteria, Planctomyces, and candidate divisions TM6 and OP10. Seventy-five 16S clones could not be classified into known bacterial divisions, and five 16S clones were related to chloroplast DNA. Members of Proteobacteria represented 46% of the clone library. A higher proportion of 16S clones affiliated with y-Proteobacteria were from plot N compared with plot S. 16S rRNA gene fragments amplified with Pseudomonas-specific primers and cloned (Ps clones) were examined from mineral-soil samples from plots N and S from three LTSP installations. A significantly greater proportion of sequenced Ps clones from plot N contained Pseudomonas 16S rRNA gene fragments compared with Ps clones from plot S.     

Organic amendments and land management affect bacterial community composition, diversity and biomass in avocado crop soils

Background and aims

The avocado-producing area of southern Spain includes conventional orchards and organic orchards that use different organic amendments. To gain insight into the effects of these amendments, physicochemical properties and microbial communities of the soil were analysed in a representative set of commercial and experimental orchards.

Methods

The population size of several groups of culturable microorganisms was determined by plating on different selective media. Bacterial community structure was studied by denaturing gradient gel electrophoresis (DGGE)

Results

Commercial composts showed the largest effects, especially the animal compost, enhancing the population sizes of some microbial groups and affecting bacterial community structure in superficial and deep soil layers. Moreover, animal and vegetal compost, manure and blood meal addition are related to high bacterial diversity in the superficial soil layer.

Conclusions

All of the organic amendments used in this study affect soil properties in one or more of the characteristics that were analysed. Culturable microbial population data revealed the most evident effects of some of the organic treatments. However, molecular analysis of soil bacterial communities by DGGE allowed the detection of the influence of all of the analysed amendments on bacterial community composition. This effect was stronger in the superficial layer of the avocado soil.     

Moderate grazing increased alpine meadow soils bacterial abundance and diversity index on the Tibetan Plateau

The response of grassland soil bacterial community characteristics to different grazing intensities is central ecological topics. However, the underlying mechanisms between bacterial abundance, diversity index, and grazing intensity remain unclear. We measured alpine meadow soil bacterial gene richness and diversity index under four grazing intensities using 16S rDNA sequence analysis on the Tibetan Plateau. The results suggest that extreme grazing significantly decreased alpine meadow both bacterial gene abundance and diversity index (p < .05). The lowest operational taxonomic unit numbers were 3,012 ± 447 copies under heavy grazing in the growing season. It was significantly lower than heavy grazing with approximately 3,958 ± 119 copies (p < .05). The Shannon index for medium and high grazing grassland bacterial diversity was slightly higher than for light grazing in the growing season. Furthermore, the lowest index was approximately 9.20 ± 0.50 for extreme grazing of grassland in the growing season. The average bacterial gene abundance and diversity index in the dormancy period were slightly higher than that in the growing season. Soil bulk density, pH, ammonium, and nitrate nitrogen were the main positive factors driving grazed grassland bacterial communities. Our study provides insight into the response of alpine meadows to grazing intensity, demonstrating that moderate grazing increases bacterial community diversity in grazed grasslands.     

Multifunctionality and diversity in bacterial biofilms

Bacteria are highly diverse and drive a bulk of ecosystem processes. Analysis of relationships between diversity and single specific ecosystem processes neglects the possibility that different species perform multiple functions at the same time. The degradation of dissolved organic carbon (DOC) followed by respiration is a key bacterial function that is modulated by the availability of DOC and the capability to produce extracellular enzymes. In freshwater ecosystems, biofilms are metabolic hotspots and major sites of DOC degradation. We manipulated the diversity of biofilm forming communities which were fed with DOC differing in availability. We characterized community composition using molecular fingerprinting (T-RFLP) and measured functioning as oxygen consumption rates, the conversion of DOC in the medium, bacterial abundance and the activities of five specific enzymes. Based on assays of the extracellular enzyme activity, we calculated how the likelihood of sustaining multiple functions was affected by reduced diversity. Carbon source and biofilm age were strong drivers of community functioning, and we demonstrate how the likelihood of sustaining multifunctionality decreases with decreasing diversity.     

长期施肥对稻田土壤细菌、古菌多样性和群落结构的影响

稻田土壤是“迷失碳”的重要吸纳场所之一,也是温室气体(CH4和N2O等)的重要排放源.大气温室气体的动态变化与土壤碳氮转化的微生物过程紧密相关.以湖南桃江国家级稻田肥力变化长期定位试验点为平台,采用PCR-克隆测序和实时荧光定量PCR技术,研究不施肥(CK)、施氮磷钾肥(NPK)和氮磷钾肥+秸秆还田(NPKS)3种长期施肥制度(＞25 a)对稻田土壤细菌和古菌群落结构及数量的影响.细菌和古菌16S rRNA基因文库分析结果表明:稻田土壤细菌主要类群为变形菌、酸杆菌、绿弯菌,而古菌主要为泉古菌和广古菌.长期施肥导致土壤细菌和古菌种群结构产生明显差异,与CK相比,NPK和NPKS处理稻田土壤的变形菌、酸杆菌和泉古菌相对丰度增加.LIBSHUFF软件分析结果也表明,16S rRNA基因文库在CK、NPK及NPKS处理间存在显著差异.3种施肥处理的稻田土壤细菌16S rRNA基因拷贝数为每克干土0.58× 1010～1.06×1010个,古菌为每克干土1.16×106 ～ 1.72×106个.施肥(NPK和NPKS)后,细菌和古菌的多样性和数量增加,且NPKS＞NPK.说明长期施肥显著影响土壤细菌和古菌群落结构、多样性及数量.     

海南西海岸四种真红树根系土壤放线菌物种多样性及其延缓衰老活性初筛

为更好地开发利用海洋微生物资源积累丰富的放线菌菌种,该文以海南西海岸潮间带区域四种真红树根系土壤为研究对象,分析了红树林根系放线菌物种多样性组成及其代谢产物活性。该研究以9种不同培养基为分离介质,采用纯培养方法和三区划线法分离纯化菌株,结合放线菌形态学特征及其16S rRNA基因序列结果开展多样性分析。放线菌发酵液经乙酸乙酯萃取,利用秀丽隐杆线虫(Caenorhabditis elegans)测试其延缓衰老活性。结果表明:(1)共分离获得22株放线菌,隶属于4目7科9属,其中链霉菌属(Streptomyces)为优势菌群,并初步确认IMDGX 6012、IMDGX 6028、IMDGX 6118、IMDGX 6326、IMDGX 6119 5株放线菌可能为潜在的新物种。(2)发酵产物延缓衰老研究结果表明,8株放线菌的代谢产物可显著延长线虫寿命(P0.05);其中,IMDGX 6028和IMDGX 6118作为拟无枝酸菌属(Amycolatopsis)和短小杆菌属(Curtobacterium)的潜在新物种,具有极显著延缓衰老活性(P 0. 01),与空白组比较,可分别延长线虫寿命的22.2%和26.6%。综上结果表明海南西海岸真红树根系土壤具有丰富的可培养放线菌资源,具有发现放线菌新物种和延缓衰老活性菌株的潜力。