No description available.

The wild origin, early breeding history, and diversity of the China Rose group, including R. chinensis and its varieties, cultivars, and hybrids, are largely unknown. The aims of this study were to investigate the genetic diversity and relationships of the China Roses with related species and hybrids, including information in support of, or refuting, the hypothesis that these roses are the hybrid result of the wild R. chinensis var. spontanea and R. odorata var. gigantea. Ninety Rosa accessions, including China Roses, a Miscellaneous Old Garden Rose, Noisettes, early Polyanthas, Bourbons, Teas, and species from Sections Indicae and Synstylae were surveyed using 23 microsatellite primer pairs. The trnH-psbA chloroplast intergenic spacer was also sequenced for the China Roses, Misc. Old Garden Rose, and the species to look specifically at maternal relationships. A total of 291 alleles were scored for the 23 microsatellites, with alleles per locus ranging from 6-22 and averaging 12.65. A dendrogram based on Dice similarity and a three-dimensional Principle Coordinate Analysis (PCoorA) graph were plotted with the data. In the cluster analysis, the similarity coefficients ranged from ~0.15-0.99, with the cultivated roses forming well-defined groups at about 0.45 similarity. These groups generally reflected the American Rose Society horticultural classifications. A large number of sports and synonyms in the China Rose group were identified through this analysis as well. The PCoorA gave a better graphical representation of the relationships of the species and cultivars, and with the inclusion of the chloroplast sequence haplotypes, some maternal relationships could also be identified. This study shows that the cultivated China Roses are a closely related group and identified which accessions were likely Hybrid China Roses. The results also suggest that the China Roses were maternally derived from R. chinensis var. spontanea. Based on the microsatellites and chloroplast sequence haplotypes, the identity of the R. odorata var. gigantea accessions in this study are suspect, but the China Roses may also have this species in their background as the result of natural or artificial hybridization.

Le microbiote pulmonaire complexe de patients atteints de mucoviscidose (MV) peut être modifié par des facteurs de l'hôte (état clinique), xénobiotiques (antibiothérapie, habitat) ainsi que bactériens (pathogènes, commensaux). Notre étude a porté sur ces facteurs bactériens et plus particulièrement sur l'effet de Pseudomonas aeruginosa sur la structuration des microbiotes MV. Le but de notre travail a été d'étudier la structure et la composition des microbiotes pulmonaires chez des patients MV en fonction de la présence ou non de P. aeruginosa, ainsi que l'effet de ces microbiotes sur ce pathogène. Différentes approches nous ont permis de mettre en évidence ces résultats : i) certains genres bactérien (Stenotrophomonas, Prevotella) contenus dans un microbiote particulier plus pauvre et moins diverse sont associés préférentiellement à la présence de P. aeruginosa, ii) aucune diminution de richesse et de diversité n'est observée en amont d'une primo-colonisation par P. aeruginosa mais celle-ci entraine ultérieurement une diminution de richesse du microbiote, iii) le microbiote MV in vitro s'appauvrit et diminue en diversité plus rapidement lors d'une colonisation par une souche ayant un phénotype de « primo-colonisation », iv) les souches de primo-colonisation sont plus virulentes que les souches issues de colonisation chronique, v) ces souches sécrètent des molécules appartenant à la famille des 4-hydroxy-2-heptylquinoline (HAQs) bactéricides vis-à-vis de S. aureus. Une meilleure compréhension de cette modulation des microbiotes MV par P. aeruginosa offriraient de nouvelles stratégies thérapeutiques afin notamment de contrer l'implantation de ce pathogène au niveau pulmonaire et de limiter le développement d'infections chroniques chez les patients MV / Cystic fibrosis microbiota could be modified by different factors such as host factor (clinical state), xenobiotic factor (antibiotic, environment) or bacterial factors (pathogen, commensal). Our study focused on bacterial factors and more particularly on the impact of Pseudomonas aeruginosa on the CF microbiota structure. The objectives were to study the structure and the composition of CF microbiota whether the presence or not of P. aeruginosa and the impact of the CF microbiota on this pathogen. Different experimentations were used to get results : i) bacterial genus (Stenotrophomonas, Prevotella) were preferentially associated with P. aeruginosa in a poor and no diverse microbiota, ii) no previous decrease of microbiota richness was observed when the initial P. aeruginosa pulmonary colonization took place but this event was associated with a later decrease of microbiota richness, iii) In vitro model of CF microbiota showed a more important richness and diversity decrease with early colonization strains, iv) early colonization strains were more virulent than chronic colonization strains, v) these strains secreted bactericid molecules toward Staaphylococcus aureus belonging to 4-hydroxy-2-heptylquinoline (HAQs). Improve our understanding on how P. aeruginosa could modulate CF microbiota could permit to find new therapeutic strategies to prevent the early colonization and then to limit the chronic infections in CF patients

Hybridization is an important mechanism in diversification. It often makes taxonomy difficult. Lack of strong supported intrageneric classification in genus Pteris (Pteridaceae) could be caused by natural hybridization. Most hybridization documnted in Pteris was based on limited evidence. This study focuses on Pteris cadieri complex, the taxon with putative hybridization. The species complex displayed significant morphological variation and was associated with hybrid origin. Reproductive biology revealed variation in spore number per sporangium, spore size, spore shape and apogamous reproduction, which imply its hybrid origin. Cytology analysis using chromosome counting and flow cytometry identified diploids, triploids, and tetraploids. CpDNA and nuclear DNA supported that Pteris cadieri complex is hybrid origin: paternal and maternal lineages were inferred and 11 taxa were identified. Furthermore, comparing materials form Hainan and Taiwan, , it is clear that the species complex is composed by taxa arisen from multiple hybridization. Systematic inconsistency existed between chloroplast and nuclear phylogenies in Pteris impled that other taxa might have involved in hybridization events, in addition to the Pteris cadieri complex. Hybridization may be very common in Pteris. To infer intrageneric taxonomy of Pteris, effect of reticulate evolution should never be neglected. Finally, based on morphological and evolutionary traits, the taxonomy of Pteris cadieri complex is revised. There are Pteris cadieri Christ, Pteris dimorpha Copel. var. dimorpha, Pteris dimorpha var. plumbea (Christ) Y.-S. Chao, H.-Y. Liu & W.-L. Chiou, Pteris grevilleana Wall. ex Agardh var. grevilleanan, Pteris grevilleana Wall. ex Agardh var. ornata Alderw., and Pteris hainanensis Ching.

The 16S, 23S, and 5S ribosomal RNA (rRNA) genes are highly conserved sequences in bacteria. For this reason, rRNA genes are often used for phylogenetic classification. On the other hand, the regions between the structural sequences, known as intergenic spacer regions (ITS), are under less evolutionary pressure to be conserved. Because they are not as highly conserved, they can be used to differentiate strains of the same bacterial specie. The purpose of this study was to evaluate the 16S-23S ITS of Flavobacterium columnare, an important pathogen of cultured fish, by comparing the 16S-23S ITS sequences from 70 isolates. We developed two PCR assays that amplify overlapping regions of one large previously identified ITS. The primers targeted the 16S sequence and isoleucine tRNA encoding sequences and the 23S sequence and alanine tRNA encoding sequences. The PCR products were cloned and sequenced. We also targeted I-CeuI restriction fragments from the ATCC type strain that were separated by pulse field gel electrophoresis and analyzed the 16S-23S ITS regions. We found that the genome of this species harbors at least 6 intergenic spacer regions that are very similar and contain the same tRNA encoding sequences. This suggests that earlier studies that used the ITS for distinguishing between strains of Flavobacterium columnare may be comparing sequences from different structural RNA operons and thus have misleading data.

Flera olika arter av mjölksyrabakterier som ingår i släktena Lactobacillus och Bifidobacterium har hittats hos bin och i deras honung. Idag finns ingen effektiv metod för identifiering av bakterierna. Syftet med detta projekt är att utveckla en metod för snabb identifiering genom att hitta lämpliga primers till olika mjölksyrabakterier och därmed få fram en Polymeraskedjereaktion (PCR) metod. Ribosomal ribonukleinsyra (rRNA) generna eller 16S-23S rRNA intergenic spacer region (ISR) används ofta vid design av primers, som därefter används i PCR för att identifiera olika bakterier. Deoxiribonukleinsyra (DNA) visualiseras i agarosgelen med hjälp av SYBRgreen I som fluorescens på ultraviolett (UV)-ljusbord. I detta projekt har 16S rRNA och 16S-23S rRNA ISR amplifierats i enkel PCR och multiplex PCR och visualiserats i agarosgel i försök att identifiera mjölksyrabakterierna. 16S rRNA har visat sig ha mycket liten variation mellan bakterierna och ansågs därför inte lämplig att använda för identifiering av närbesläktade arter. 16S-23S rRNA ISR visade större variation, fram för allt mellan lactobacillerna och bifidobakterierna. Gruppering av bakterierna med hjälp av multiplex PCR gjordes med viss framgång, med undantag av några bakterier som inte hamnade i den förväntade gruppen. Dock behövs fler försök för att stödja dessa resultat. / Several different lactic acid bacterium (LAB) species from the genera Lactobacillus and Bifidobacterium was discovered in bees and in their honey. Today there is no rapid and reliable method to identify these LAB. Therefore a rapid polymerase chain reaction (PCR) method to identify the LAB is needed. The aim of this project is to find primers suitable for the different LAB. Ribosomal ribonucleic acid (rRNA) genes or 16S-23S rRNA intergenic spacer region (ISR) are often used to designing of primers followed by PCR assays, for identification of different bacteria. To visualize deoxyribonucleic acid (DNA) in agarose gels, SYBRgreen I was used as fluorescence and then viewed under ultraviolet (UV) light. In this project the 16S rRNA and 16S-23S rRNA ISR was used as a target in a PCR and a multiplex PCR amplification. The PCR product was analyzed in agarose gel in an attempt to identify the LAB. 16S rRNA sequence have to little variation and is not suitable to identify closely related species. 16S-23S rRNA ISR sequence exhibits greater variations, especially between Lactobacillus and Bifidobacterium. Differentiation of the bacteria into groups by multiplex PCR was done with good result, except for some of the bacteria that did not end up in the expected group. More studys is needed to support these results.

No description available.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Land-use change and soil type can have significant impact on microbial communities of soil. The Pampa biome in recent decades has undergone severe changes in landscape due to landuse change, mainly for the introduction of exotic tree plantation and croplands. Different landuse in Pampa biome were evaluated to determine the effect on the structure of soil microbial communities. Furthermore, due to the presence of various soil types present in this biome, we investigated whether different soil type harbor different microbial communities. Soil samples were collected at two sites with different land-uses (native grassland, native forest, exotic tree plantation and cropland) and in a typical toposequence in Pampa biome formed by Paleudult, Albaqualf and alluvial soils. The structure of soil microbial community (archaeal and fungal) was evaluated by RISA and soil functional capabilities were measured by microbial biomass carbon and metabolic quotient. We detected different patterns in fungal and archaeal community driven by land-use change and soil type showing that both factors are significant drivers of microbial community structure and activity. Acacia and Eucalyptus afforestation presented the most dissimilar communities when compared with natural vegetation. Although differences in the communities were detected, the soils tested shared most of the taxonomic unities and only a proportion of the community suffers changes caused by human interference. / A mudança do uso da terra e o tipo de solo podem exercer impactos significantes sobre a comunidade microbiana do solo. O bioma Pampa Brasileiro, nas últimas décadas, tem sofrido severas mudanças na paisagem devido à mudança no uso da terra, principalmente pela introdução de plantações de árvores exóticas e pelos cultivos agrícolas. Diferentes usos do solo no bioma Pampa foram avaliados para determinar o efeito sobre a estrutura das comunidades microbianas do solo. Além disso, devido à presença de vários tipos de solo presentes neste bioma, foi investigado se diferentes tipos de solos abrigam diferentes comunidades microbianas. Amostras de solo foram coletadas em duas áreas com diferentes usos do solo (pastagem nativa, mata nativa, plantações de árvores exóticas e cultivo agrícola) e em uma topossequência típica no bioma Pampa formado por Argissolo, Planossolo e solos aluviais. A estrutura das comunidades microbianas do solo (arqueas e fungos) foi avaliada por RISA e capacidades funcionais do solo foram mensuradas através de carbono da biomassa microbiana e quociente metabólico. Diferentes padrões foram detectados nas comunidades de fungos e arqueas influenciados pela mudança no uso da terra e pelo tipo de solo, mostrando que ambos são importantes fatores da estrutura e atividade da comunidade microbiana. Florestamentos de acácia e eucalipto apresentaram as comunidades mais diferentes quando comparados com a vegetação natural. Embora diferenças nas comunidades foram detectadas, os diferentes usos e tipos de solos avaliados compartilham grande parte das unidades taxonômicas e mostram que apenas uma parte da comunidade sofre alterações causadas pela interferência humana.

A protein, B0238.11, was identified in a yeast one-hybrid screen to bind to the ribosomal intergenic spacer region (IGS) of Caenorhabditis elegans. Proteins interacting with this region of the DNA have been implicated in ribosome biogenesis in other model organisms, so it is also possible that B0238.11 plays a role in RNA transcription by interacting with RNA polymerase I or other transcription machinery. Thus, the goal of this study was to further characterize the structure and function of B0238.11. I used yeast two-hybrid experiments to identify proteins that interact with B0238.11 within the nucleus. RPS-0, K04G2.2, DPY-4, EFT-3, PAL-1, and B0238.11, itself, were found to bind to B0238.11. Additionally, I analysed the amino acid sequence of B0238.11 using in silico bioinformatics methods to determine its structure and putative function and also to identify and characterize the other interacting proteins. I found that B0238.11 contains a high-mobility group box domain, which is also found in HMO1P in yeast and UBF in vertebrates. These other proteins also bind to the IGS, are known to form homodimers and have been implicated in the initiation of ribosomal RNA transcription. Here I scrutinize the validity of the interaction between each protein and B0238.11. I conclude that B0238.11 is likely to be a C. elegans homolog of UBF and present an updated interactome map for B0238.11. / Synopsis: I carried out yeast two-hybrid assay to find proteins interacting with B0238.11 (O16487_CAEEL). I found that this protein's DNA-binding profile and protein interaction profile mimic other HMG-box containing proteins UBF and HMO1P which are involved in ribosomal RNA transcription initiation. Acknowledgements: I would like to thank my supervisor, Dr. Teresa J. Crease, for not only giving me the opportunity to investigate an interesting topic in Molecular Biology, but also for her patient guidance, encouragement and sound advice. I feel extremely lucky to have a supervisor who cared so much about my work, who responded to my questions and queries so promptly, and was always available to discuss project and career related matters. I would also like to thank Dr. Todd Gillis and Dr. Terry Van Raay for their careful consideration of this project and timely constructive criticisms that helped shape my project. I would like to thank all the members of my committee for helping me see things from different perspectives and helping me develop and critical and mature understanding of the scientific process. I must also express my gratitude to Dr. Robin Floyd for allowing me to build upon his work and Dr. Marian Walhout, at the University of Massachusetts, for providing the Caenorhabditis elegans complimentary DNA library. A large part of this project would not have been possible without the people at the genomics facility in the Department of Integrative Biology, I commend their professionalism and punctuality in delivering results. Completing this work would have been all the more difficult were it not for the support and friendship provided by my peers Shannon Eagle, Tyler Elliott, Nick Jeffery, Joao Lima, Sabina Stanescu, Fatima Mitterboeck and Paola Pierossi. And finally, I would like to thank my parents and siblings Sara Omar and Ali Omar for their continued support through good times and bad, and letting me use their laptops when mine broke down.