旨在了解秀丽白虾线粒体16S rRNA基因序列及线粒体16S r RNA基因在虾类物种鉴定及系统分类中的作用,采用PCR特异扩增测序的方法,对秀丽白虾(Exopalaemon modestus)的16S rRNA基因片段进行了分析,并探讨了长臂虾亚科虾类的系统发育关系...旨在了解秀丽白虾线粒体16S rRNA基因序列及线粒体16S r RNA基因在虾类物种鉴定及系统分类中的作用,采用PCR特异扩增测序的方法,对秀丽白虾(Exopalaemon modestus)的16S rRNA基因片段进行了分析,并探讨了长臂虾亚科虾类的系统发育关系。结果表明:54尾秀丽白虾16S r RNA基因共发现5种单倍型和8个变异位点。13种长臂虾间的遗传距离在0~0.2614之间,7个属间的遗传距离为0.1344~0.2769。NJ法和ME法构建的系统进化树显示,除洁白长臂虾外,同属的不同种先聚成一支,其中东方白虾和脊尾白虾亲缘关系最近。长臂虾属和小长臂虾属亲缘关系很近,长臂虾属为多起源;推测洁白长臂虾与宽额拟瘦虾之间的分化时间距今约22.34百万年。本研究结果表明,线粒体16S rRNA基因序列不能单独对长臂虾亚科虾类进行物种鉴定,但进行系统分类和亲缘关系分析是可行的。展开更多
Metagenomic approach to the characterization of uncultured and mixed bacterial consortium has been used in this study for a few arctic micro-organisms. Arctic soil, collected from one spot at Ny Alesund in Svalbard wa...Metagenomic approach to the characterization of uncultured and mixed bacterial consortium has been used in this study for a few arctic micro-organisms. Arctic soil, collected from one spot at Ny Alesund in Svalbard was used as the source of such organisms and enriched in culture medium. Only psychrotrophic microbes were chosen which could grow from 5°C to 20°C. Total genomic DNA isolated from the consortium was used as template for amplifying the 16S rRNA genes using the conserved forward and reverse primers. The amplified mixture of 1.4 kb DNA was cloned in pGEMT Easy vector and individual 16S rDNA sequences were determined by automated sequencing. A phylogenetic tree was constructed using MEGA 5 software which enlists 19 isolates in the consortium. Most of these arctic bacteria were Bacillus species as these were isolated from a single spot.展开更多
Weaning of piglets is generally considered as a stressor which changes intestinal ecosystem and leads to clinical implications. Microbiota inhabiting in small intestine (especially ileum) are assumed to promote heal...Weaning of piglets is generally considered as a stressor which changes intestinal ecosystem and leads to clinical implications. Microbiota inhabiting in small intestine (especially ileum) are assumed to promote health, but their functional properties are yet poody dascdbed. As indicated by the 16S rRNA gene sequences of ileal micrebiota in nursing piglets (at the age of 21 and 28 d) and 28-day-old weaned piglets (weaned at 21 d of age), the microbiota were mainly comprised of gram-positive bacteria. There were 40 operational taxonomic units (OTUs) (from 171 clones) in the ileum of nursing piglets aged 21 d, 61 OTUs (from 194 clones) in the ileum of nursing piglets aged 28 d, and 56 OTUs (from 171 clones) in the ileum of weaned piglets aged 28 d. The flea of nursing piglets aged 21 d were dominantly occupied by Lactobacilli (87.7%) as well as Streptococ cus ( 3.5 % ). Lactobacillus amy/ovorus (41.5 % ), Lactobaci/lus sp. ( 19.3 % ), Lactobaci/lus reuteri ( 12.3 % ), Lactobacillus salivarius ( 9.4 % ) and L. mucosae (4.7%) were the predominant species among Lactobacil/L Similar results were obtained in the nursing piglets at 28 d of age ex- cept that Lactobaci/li decreased to 71.1% and Streptococcus increased to 21.1% significantly. Lactobacillus (52.0%) and Streptococcus (26.3%) were the two major groups in the ileum of weaned piglets aged 28 d. Lactobacillus amylovorus (31.6%) and Lactobaci/lus reuteri ( 16.4% ) was the two most important species in Lactobacillus. Therefore, Lactobacilli were predominant in the ileum of nursing and weaned piglets, and they had the highest diversity, followed by Streptococcus. The diversity of ileal microbiota was not different remarkably between the nursing piglets and the weaned piglets, but the composition changed significantly. These findings are helpful to understand ileal bacterial ecophysiology and further develop nutritional regimes to prevent or counteract complications during the weaning transition.展开更多
The sequences of the mitochondrial 16S rRNA gene of 16 Oedipodidae species were amplified and sequenced. All sequences were aligned and analyzed and the phylogenetic relationships were inferred. The properties of 16S ...The sequences of the mitochondrial 16S rRNA gene of 16 Oedipodidae species were amplified and sequenced. All sequences were aligned and analyzed and the phylogenetic relationships were inferred. The properties of 16S gene in Oedipodidae showed typical patterns of many insects such as a high A+T content and variable distance-dependent transition/transversion ratios. The 0.2 weight for sites of loops may be advisable for phylogeny reconstruction using the maximum parsimony method. The phylogenetic analysis results do not support the current subfamily classification systems of Oedipodidae. Bryodemellinae and Bryodeminae are closely related and should be merged as one subfamily. The status of Oedipodinae and Locustinae is also problematic.展开更多
Molecular diversity and development of the Lactobacillus community in the intestinal tract, as influenced by age and intestinal compartment, were studied in one litter of 12 conventionally raised piglets. Piglets were...Molecular diversity and development of the Lactobacillus community in the intestinal tract, as influenced by age and intestinal compartment, were studied in one litter of 12 conventionally raised piglets. Piglets were euthanized at each week (3 animals per time). Digesta and tissue samples from stomach, duodenum, jejunum, ileum, caecum, colon, and rectum were collected and analysed by using 16S ribosomal RNA-based methods. DGGE (denaturing gradient gel electrophoresis) profiles revealed that the Lactobacillus communities throughout the GI tract from duodenum to rectum showed good stability at same age. This indicates that fecal Lactobacillus communities can effectively represent the intestinal community. Two dominant bands were found in tissue samples of the small intestine, suggesting that the lactobacilli can adhere to the small intestinal wall. The Lactobacillus communities in different GI tract compartments developed over time. A successional change of Lactobacillus communities was observed from birth, through creep feeding to one week after weaning, showing a trend from simple to complex and back to simple. Furthermore, a clone library of Lactobacillus spp. 16S rRNA gene sequences were generated from jejunal and colonic chymes. Six dominant DGGE bands generated from jejunal chymes were matched with sequences that show 94-98% similarity to the bands derived from L. reuteri, L. delbrueckii, and L. crispatus. Seven dominant DGGE bands generated from colon chymes were matched with sequences that show 88-99% similarity to those derived from L. reuteri, L. delbrueckii, L. amylovorus/L. sobrius, and L. acidophilus. Amplicons related to L. reuteri were found in all DGGE fingerprints from jejunal digesta of age of weeks 1, 3, and 4. Amplicons related to L. amylovorus/L. sobrius were present in all DGGE fingerprints from colonic digesta of age of week 1, 3, and 4. Amplicons related to L. delbrueckii were found before weaning, L. crispatus after creep feeding before weaning and L. acidophilus after weaning. This indicates that L. reuteri and L. amylovorus/L. sobrius probably belong to the permanent composition, while L. delbruckii, L. acidophilus, and L. crispatus probably belong to the temporal groups of Lactobacillus communities in the GI tract of piglets.展开更多
A study was undertaken to investigate the heterotrophic bacterial flora associated with the sea anemones. Samples of the sea anemones Anthopleura midori were collected from the coast of Weihai and bacteria were isolat...A study was undertaken to investigate the heterotrophic bacterial flora associated with the sea anemones. Samples of the sea anemones Anthopleura midori were collected from the coast of Weihai and bacteria were isolated from these samples. Additionally, high numbers of viable bacteria were obtained from the celom wall and surface of anemone, the community of cultivable bacteria was very diverse. As a result of this isolation, 60 strains were obtained, 56 of them were selected for identification and characterization by 16S rRNA gene sequence analysis and limited phenotypic testing. Among these isolates, 16 strains were phylogenetically related to members of the genus Pseudoalteromonas and neighboring taxa. Other isolates included members of the genera Colwcllia, Vibrio, Acinetobactcr, Pseudomonas, Endozoicomonas, Roseovarius, Paraeoceus, Loktanella, Leisingcra, Sulfitobacter, Bacillus, Staphylococcus, Plantibacter, Microbaeterium, Micro- coccus, Joostella, Psychroserpens, Cellulophaga, Krokinobaeter, Polaribacter and Psychrobaeter. Seven potential novel species were found. Among 60 strains, 17 of them can produce proteolytic exoenzyme, 20 can produce lipolytic exoenzyme. Strain NQ8 has strong antagonistic effects on some Vibrio strains. This study demonstrates that the culturable fraction of bacteria from the sea anemones Anthopleura midori is diverse and appears to possess much potential as a source for the discovery of novel bioactive materials.展开更多
This study deals with the biogeography of Antarctic microflora (Antarctica acts as best model to study microbial biogeography) such as cyanobacteria and selected halophiles with special emphasis on Halomonas variabi...This study deals with the biogeography of Antarctic microflora (Antarctica acts as best model to study microbial biogeography) such as cyanobacteria and selected halophiles with special emphasis on Halomonas variabilis and Bacillus licheniformis.Halophiles are known to be resistant not only to salt stress,but also to extreme temperature,pressure,and aridity and they are capable of surviving in harsh environments such as polar regions,deep-sea habitats,and deserts.Many microbes are known to be resistant to hostile environmental conditions,and are capable of surviving in harsh environments.Our group has isolated 444 strains belonging to 28 genera of halophiles from various environments around the world.The 16S rRNA gene sequences revealed that many of the isolated strains from geographically distant habitats having different environmental conditions,were closely related to each other,with some strains possessing 100% identical sequences.Organisms possessing survival mechanism such as spore formation are usually ubiquitous.The genus Halomonas is represented by potentially endemic strains and the ubiquitous H.variabilis,while spore-forming B.licheniformis showed cosmopolitan distribution.One potentially endemic (moderate endemicity that is regional and/or continental distribution) strain was reported from Syowa station,East Antarctica,and Mario Zucchelli station,West Antarctica,which are geographically separated by 3000 km.Moreover,15 strains having 100% similarity with B.licheniformis were considered cosmopolitans.The results of this work provide support for the middle-ground model that some microbes have moderate endemicity and others have cosmopolitan distribution.These results will contribute to a greater understanding of microbial biogeography with special emphasis on Antarctica.展开更多
The abundance and diversity of archaeal assemblages were analyzed in soils collected from Gyorae Gotjawal forest, Jeju, Korea. Gotjawal soil refers to soil derived from a lava-formed forest, characterized by high orga...The abundance and diversity of archaeal assemblages were analyzed in soils collected from Gyorae Gotjawal forest, Jeju, Korea. Gotjawal soil refers to soil derived from a lava-formed forest, characterized by high organic matter content, fertility, and poor rocky soil. Using domain-specific primers, archaeal 16S rRNA gene sequences were PCR amplified for clone library construction, and a total of 185 archaeal clones were examined. The archaeal clones were affiliated with the phyla Thaumarchaeota (96.2%) and Euryarchaeota (3.8%). The most abundant thaumarchaeal group (90.3% of the clones) was the group I.1b clade, which includes soil ammonia-oxidizing archaea. The unique characteristics of Gotjawal soil, including basalt morphology, vegetation, and groundwater aquifer penetration, may be reflected in the archaeal community composition. Further study is necessary to understand the unique factors of Gotjawal soils that influence archaeal abundance, composition, and diversity.展开更多
文摘旨在了解秀丽白虾线粒体16S rRNA基因序列及线粒体16S r RNA基因在虾类物种鉴定及系统分类中的作用,采用PCR特异扩增测序的方法,对秀丽白虾(Exopalaemon modestus)的16S rRNA基因片段进行了分析,并探讨了长臂虾亚科虾类的系统发育关系。结果表明:54尾秀丽白虾16S r RNA基因共发现5种单倍型和8个变异位点。13种长臂虾间的遗传距离在0~0.2614之间,7个属间的遗传距离为0.1344~0.2769。NJ法和ME法构建的系统进化树显示,除洁白长臂虾外,同属的不同种先聚成一支,其中东方白虾和脊尾白虾亲缘关系最近。长臂虾属和小长臂虾属亲缘关系很近,长臂虾属为多起源;推测洁白长臂虾与宽额拟瘦虾之间的分化时间距今约22.34百万年。本研究结果表明,线粒体16S rRNA基因序列不能单独对长臂虾亚科虾类进行物种鉴定,但进行系统分类和亲缘关系分析是可行的。
文摘Metagenomic approach to the characterization of uncultured and mixed bacterial consortium has been used in this study for a few arctic micro-organisms. Arctic soil, collected from one spot at Ny Alesund in Svalbard was used as the source of such organisms and enriched in culture medium. Only psychrotrophic microbes were chosen which could grow from 5°C to 20°C. Total genomic DNA isolated from the consortium was used as template for amplifying the 16S rRNA genes using the conserved forward and reverse primers. The amplified mixture of 1.4 kb DNA was cloned in pGEMT Easy vector and individual 16S rDNA sequences were determined by automated sequencing. A phylogenetic tree was constructed using MEGA 5 software which enlists 19 isolates in the consortium. Most of these arctic bacteria were Bacillus species as these were isolated from a single spot.
基金Supported by grants from Knowledge Innovation Project of Chinese Academy of Sciences (KSCX2-YW-N-051 and SW-323)NSFC(30901040, 30901041, 30928018, 30828025, 30700581, and 30771558)+2 种基金National Basic Research Program of China(2009CB118800)National 863 project (2008AA10Z316)National Scientific and Technological Supporting Project(2007BAQ01047 and 2006BAD12B07)~~
文摘Weaning of piglets is generally considered as a stressor which changes intestinal ecosystem and leads to clinical implications. Microbiota inhabiting in small intestine (especially ileum) are assumed to promote health, but their functional properties are yet poody dascdbed. As indicated by the 16S rRNA gene sequences of ileal micrebiota in nursing piglets (at the age of 21 and 28 d) and 28-day-old weaned piglets (weaned at 21 d of age), the microbiota were mainly comprised of gram-positive bacteria. There were 40 operational taxonomic units (OTUs) (from 171 clones) in the ileum of nursing piglets aged 21 d, 61 OTUs (from 194 clones) in the ileum of nursing piglets aged 28 d, and 56 OTUs (from 171 clones) in the ileum of weaned piglets aged 28 d. The flea of nursing piglets aged 21 d were dominantly occupied by Lactobacilli (87.7%) as well as Streptococ cus ( 3.5 % ). Lactobacillus amy/ovorus (41.5 % ), Lactobaci/lus sp. ( 19.3 % ), Lactobaci/lus reuteri ( 12.3 % ), Lactobacillus salivarius ( 9.4 % ) and L. mucosae (4.7%) were the predominant species among Lactobacil/L Similar results were obtained in the nursing piglets at 28 d of age ex- cept that Lactobaci/li decreased to 71.1% and Streptococcus increased to 21.1% significantly. Lactobacillus (52.0%) and Streptococcus (26.3%) were the two major groups in the ileum of weaned piglets aged 28 d. Lactobacillus amylovorus (31.6%) and Lactobaci/lus reuteri ( 16.4% ) was the two most important species in Lactobacillus. Therefore, Lactobacilli were predominant in the ileum of nursing and weaned piglets, and they had the highest diversity, followed by Streptococcus. The diversity of ileal microbiota was not different remarkably between the nursing piglets and the weaned piglets, but the composition changed significantly. These findings are helpful to understand ileal bacterial ecophysiology and further develop nutritional regimes to prevent or counteract complications during the weaning transition.
基金This study was supported by the National Natural Science Foundation of China (NSFC grants No. 30070114, 30470238).
文摘The sequences of the mitochondrial 16S rRNA gene of 16 Oedipodidae species were amplified and sequenced. All sequences were aligned and analyzed and the phylogenetic relationships were inferred. The properties of 16S gene in Oedipodidae showed typical patterns of many insects such as a high A+T content and variable distance-dependent transition/transversion ratios. The 0.2 weight for sites of loops may be advisable for phylogeny reconstruction using the maximum parsimony method. The phylogenetic analysis results do not support the current subfamily classification systems of Oedipodidae. Bryodemellinae and Bryodeminae are closely related and should be merged as one subfamily. The status of Oedipodinae and Locustinae is also problematic.
基金supported by the National Natural Science Foundation of China (30771570)the Natural Science Foundation of Jiangsu Province, China(BK2006143)
文摘Molecular diversity and development of the Lactobacillus community in the intestinal tract, as influenced by age and intestinal compartment, were studied in one litter of 12 conventionally raised piglets. Piglets were euthanized at each week (3 animals per time). Digesta and tissue samples from stomach, duodenum, jejunum, ileum, caecum, colon, and rectum were collected and analysed by using 16S ribosomal RNA-based methods. DGGE (denaturing gradient gel electrophoresis) profiles revealed that the Lactobacillus communities throughout the GI tract from duodenum to rectum showed good stability at same age. This indicates that fecal Lactobacillus communities can effectively represent the intestinal community. Two dominant bands were found in tissue samples of the small intestine, suggesting that the lactobacilli can adhere to the small intestinal wall. The Lactobacillus communities in different GI tract compartments developed over time. A successional change of Lactobacillus communities was observed from birth, through creep feeding to one week after weaning, showing a trend from simple to complex and back to simple. Furthermore, a clone library of Lactobacillus spp. 16S rRNA gene sequences were generated from jejunal and colonic chymes. Six dominant DGGE bands generated from jejunal chymes were matched with sequences that show 94-98% similarity to the bands derived from L. reuteri, L. delbrueckii, and L. crispatus. Seven dominant DGGE bands generated from colon chymes were matched with sequences that show 88-99% similarity to those derived from L. reuteri, L. delbrueckii, L. amylovorus/L. sobrius, and L. acidophilus. Amplicons related to L. reuteri were found in all DGGE fingerprints from jejunal digesta of age of weeks 1, 3, and 4. Amplicons related to L. amylovorus/L. sobrius were present in all DGGE fingerprints from colonic digesta of age of week 1, 3, and 4. Amplicons related to L. delbrueckii were found before weaning, L. crispatus after creep feeding before weaning and L. acidophilus after weaning. This indicates that L. reuteri and L. amylovorus/L. sobrius probably belong to the permanent composition, while L. delbruckii, L. acidophilus, and L. crispatus probably belong to the temporal groups of Lactobacillus communities in the GI tract of piglets.
基金The State Key Laboratory of Microbial Technology under contract No. M2010-04Independent Innovation Foundation of Shandong University (IIFSDU)+1 种基金the National Natural Science Foundation of China under contract No. 40730847Special Non-profit Research Project from Ministry of Agriculture of the Peoples Republic of China under contract No. nyhyzx07-046
文摘A study was undertaken to investigate the heterotrophic bacterial flora associated with the sea anemones. Samples of the sea anemones Anthopleura midori were collected from the coast of Weihai and bacteria were isolated from these samples. Additionally, high numbers of viable bacteria were obtained from the celom wall and surface of anemone, the community of cultivable bacteria was very diverse. As a result of this isolation, 60 strains were obtained, 56 of them were selected for identification and characterization by 16S rRNA gene sequence analysis and limited phenotypic testing. Among these isolates, 16 strains were phylogenetically related to members of the genus Pseudoalteromonas and neighboring taxa. Other isolates included members of the genera Colwcllia, Vibrio, Acinetobactcr, Pseudomonas, Endozoicomonas, Roseovarius, Paraeoceus, Loktanella, Leisingcra, Sulfitobacter, Bacillus, Staphylococcus, Plantibacter, Microbaeterium, Micro- coccus, Joostella, Psychroserpens, Cellulophaga, Krokinobaeter, Polaribacter and Psychrobaeter. Seven potential novel species were found. Among 60 strains, 17 of them can produce proteolytic exoenzyme, 20 can produce lipolytic exoenzyme. Strain NQ8 has strong antagonistic effects on some Vibrio strains. This study demonstrates that the culturable fraction of bacteria from the sea anemones Anthopleura midori is diverse and appears to possess much potential as a source for the discovery of novel bioactive materials.
基金partly supported by the Special Coordination Fund"Archaean Park Project:International research project on interaction between sub-vent biosphere and geo-environments"of the Ministry of Education,Culture,Sports,Science and Technology of Japanthe NIPR collaborative research program No.99(2002-2004)+2 种基金the Fund"Construct the Genetic Resource Library of Unidentifed Microbes Based on Genome Information"from the New Energy and Industrial Technology Development Organization,JapanGrants-in-Aid for Scientifc Research(11833012,14340268,and 18255005)from the Japan Society for the Promotion of Scienceand Antarctic samples from Livingston Island were collected during the Spanish program LIMNOPOLAR in the International Polar Year(IPY)2007-2008 campaign
文摘This study deals with the biogeography of Antarctic microflora (Antarctica acts as best model to study microbial biogeography) such as cyanobacteria and selected halophiles with special emphasis on Halomonas variabilis and Bacillus licheniformis.Halophiles are known to be resistant not only to salt stress,but also to extreme temperature,pressure,and aridity and they are capable of surviving in harsh environments such as polar regions,deep-sea habitats,and deserts.Many microbes are known to be resistant to hostile environmental conditions,and are capable of surviving in harsh environments.Our group has isolated 444 strains belonging to 28 genera of halophiles from various environments around the world.The 16S rRNA gene sequences revealed that many of the isolated strains from geographically distant habitats having different environmental conditions,were closely related to each other,with some strains possessing 100% identical sequences.Organisms possessing survival mechanism such as spore formation are usually ubiquitous.The genus Halomonas is represented by potentially endemic strains and the ubiquitous H.variabilis,while spore-forming B.licheniformis showed cosmopolitan distribution.One potentially endemic (moderate endemicity that is regional and/or continental distribution) strain was reported from Syowa station,East Antarctica,and Mario Zucchelli station,West Antarctica,which are geographically separated by 3000 km.Moreover,15 strains having 100% similarity with B.licheniformis were considered cosmopolitans.The results of this work provide support for the middle-ground model that some microbes have moderate endemicity and others have cosmopolitan distribution.These results will contribute to a greater understanding of microbial biogeography with special emphasis on Antarctica.
文摘The abundance and diversity of archaeal assemblages were analyzed in soils collected from Gyorae Gotjawal forest, Jeju, Korea. Gotjawal soil refers to soil derived from a lava-formed forest, characterized by high organic matter content, fertility, and poor rocky soil. Using domain-specific primers, archaeal 16S rRNA gene sequences were PCR amplified for clone library construction, and a total of 185 archaeal clones were examined. The archaeal clones were affiliated with the phyla Thaumarchaeota (96.2%) and Euryarchaeota (3.8%). The most abundant thaumarchaeal group (90.3% of the clones) was the group I.1b clade, which includes soil ammonia-oxidizing archaea. The unique characteristics of Gotjawal soil, including basalt morphology, vegetation, and groundwater aquifer penetration, may be reflected in the archaeal community composition. Further study is necessary to understand the unique factors of Gotjawal soils that influence archaeal abundance, composition, and diversity.