No description available.

The topic of this work was the use of variable DNA regions for clarification of phylogenic relations of closely related South African species of Crassula L. genus. Based on preliminary dendrogram which was constructed according to chloroplast regions, 69 members of Crassula genus were used for analysis. Variable regions of chloroplast DNA (matK, rps16) as well as nuclear regions of DNA (ITS, ETS) of these members were analyzed. This work was focused on variable regions of nuclear DNA mainly to ETS region which was found as the most suita-ble for phylogenic study of this group of plants. These four regions of DNA were amplified and sequenced for all 69 analyzed members of Crassula genus. The multiple alignments of all DNA sequences were evaluated. The results confirm hypothesis that chosen noncoding nucle-ar DNA regions can be more suitable for phylogenetic study of these closely related species than chloroplast DNA regions because of their much higher variability. The ETS region shoved highest variability of all regions used in this work, so it appears as suitable for detailed phylogenetic analysis of the Crassula genus.

Intelligent Transportation Systems (ITS) can include multiple technologies and applications combined to improve the overall efficiency and effectiveness of the transportation system or network. These applications are deployed with the anticipation that the desired project goals and objectives established by multiple stakeholders will be achieved. Once a system is deployed, the project goals and objectives should be evaluated. The evaluation can provide both quantitative and qualitative feedback to assess the impacts associated with the investment in building, designing and implementing these systems. This research includes a methodology to evaluate large scale ITS infrastructure projects using the Interstate 91 (I-91) ITS Project as a case study. The methodology developed includes a review of literature, a clear definition of project goals, objectives and intended outcomes, the development of hypotheses for project outcomes, specific measures of effectiveness, pre and post-data collection methods and criterion to measure the success rate of achieving the intended objective. The following recommendations should be considered by the I-91 ITS Project Team as next steps in conducting an ITS evaluation; identify and prioritize the goals and objective areas, develop a multi-phase evaluation approach, identify existing data sources of pre-deployment data, identify missing data requirements and document the existing communication protocol prior to deployment. Such a large scale evaluation requires an extensive level of effort, and priority should be given to developing a multi-phase approach. This research may be also used towards the development of an Evaluation Plan which is recommended as a component of the six step process outlined in the Evaluation Guidelines, from the United States Department of Transportation.

This study focused on isolating important wood decay fungi from two field sites located in Harrison County, MS, and Oktibbeha County, MS. Southern Yellow Pine samples of various types and treatments including: Cu8, CuOm, ACQ, PCP, proprietary organic biocide, and un-treated were collected, and fungal isolates were cultured. DNA was extracted from isolated fungal cultures and amplified by polymerase chain reaction (PCR). The internal transcribe spacer (ITS) region was sequenced, and fungal cultures were identified by comparison to sequences on GenBank using BLAST. A total of 68 fungal isolates were recovered and successfully identified from 196 samples. Thirteen basidiomycete isolates were identified, with Veluticeps fimbriata occurring most frequently. The white-rot ascomycete, Daldinia fissa was also common. Two sequence-specific oligonucleotide probes (SSOP) were designed using Lasergene® PrimerSelect software. Unsuccessful attempts were made to attach poly (dT) tails to the probes in order to cross link the probes to nylon membranes.

No description available.

Leptosphaerulina spp. are common fungi that have been reported to colonize several turfgrass species. Controversy exists regarding the relationship of Leptosphaerulina spp. and their turfgrass hosts. The fungus has been classified as a saprophyte, senectophyte, weak pathogen, and pathogen of turfgrasses. There has also been conflicting reports regarding the delineation of species within the genus Leptosphaerulina. Because of the uncertainty regarding the ecology and taxonomy of the genus in relation to turfgrasses the present study was undertaken. The ITS and EF-1á gene regions were sequenced and analyzed to compare to the multiple taxonomic schemes reported in the literature. The ITS region offered no resolution of species; however, the phylogeny of the EF-1á gene was consistent with the six-species model of Graham and Luttrell. Inoculation experiments were performed on unstressed and artificially stressed plants to determine whether the fungi are pathogens, senectophytes, or saprophytes of turfgrasses. Perennial ryegrass and creeping bentgrass plants were stressed by placing them in a dew chamber set at 38ºC, 100% R.H., and no light for two and one days respectively. Plants were inoculated with cultures of Leptosphaerulina isolated from turfgrasses, and maintained at optimum conditions reported for infection and colonization. There was no visible difference between inoculated and uninoculated plants, and examination of cleared and stained leaves with a light microscope revealed spores that germinated and produced appressoria, but failed to penetrate the epidermal cells. The lack of infection and colonization suggests that Leptosphaerulina spp. are saprophytes of turfgrasses. / Master of Science

This research investigates alternate methods of implementing a redundant communications ring for polled network applications. FDDI and a unique solution (the tone method) designed specifically for this application are compared. In polled networks one of the most critical parameters is the time required to interrogate all of the nodes in the network. This parameter, called the poll time, is compared for the two solutions under various operating conditions, including the instances when the ring is intact and when the ring has experienced a failure, causing it to operate in the redundant mode. As expected, the solution designed specifically for this application performed better than FDDI. The tone method, which requires very little overhead and is not limited by distance or number of nodes, is described in detail, as are the results of poll time comparisons. / Master of Science

Intelligent Transportation Systems (ITS) have emerged to use different technologies to promote safety, convenience, and efficiency of transportation networks. Many applications of ITS depend on the availability of the real-time positioning of the vehicles in the network. In this research, the two open challenges in the field of vehicle localization for ITS are introduced and addressed. First, in order to have safe and efficient transportation systems, the locations of the vehicles need to be available everywhere in a network. Conventional localization techniques mostly rely on Global Positioning System (GPS) technology which cannot meet the accuracy requirements for all applications in all situations. This work advances the study of vehicle positioning in ITS by introducing an integrated positioning framework which uses several resources including GPS, vehicle-to-infrastructure and vehicle-to-vehicle communications, radio-frequency identification, and dead reckoning. These technologies are used to provide more reliable and accurate location information. The suggested framework fills the gap between the accuracy of the current vehicle localization techniques and the required one for many ITS applications. Second, different ITS applications have different localization accuracy and latency requirements. A smart positioning algorithm is proposed which enable us to change the positioning accuracy delivered by the algorithm based on different applications. The algorithm utilizes only the most effective resources to achieve the required accuracy, even if more resources are available. In this way, the complexity of the system and the running time decrease while the desired accuracy is obtained. The adjective Smart is selected because the algorithm smartly selects the most effective connection which has the most contribution to vehicle positioning when a connection needs to be added. On the other hand, when a connection should be removed, the algorithm smartly selects the least effective one which has the least contribution to the position estimation. This study also provides an overview about the positioning requirements for different ITS applications. A close-to-real-world scenario has been developed and simulated in MATLAB to evaluate the performance of the proposed algorithms. The simulation results show that the vehicle can acquire accurate location in different environments using the suggested Integrated framework. Moreover, the advantages of the proposed Smart algorithm in terms of accuracy and running time are presented through a series of comprehensive simulations. / Master of Science

Os Cubozoa são animais de águas tropicais e subtropicais ao redor do mundo. A cubomedusa Chiropsalmus cf. quadrumanus (Cubozoa: Chiropsalmidae) habita na costa Atlântica americana entre os 35°N e 27°S, uma área com barreiras putativas que poderiam impedir a distribuição contínua de C. cf. quadrumanus, tornando-se um modelo interessante para estudos de estrutura populacional e filogeografia. O objetivo deste trabalho foi analisar a estrutura haplotípica de quatro populações da C. cf. quadrumanus da costa brasileira. Nossas análises foram baseadas nos marcadores mitocondriais COI e 16S e a região nuclear ITS de 40 indivíduos provenientes de Ceará (03°43S 038°29W); Rio de Janeiro (22°37S 041°54W); São Paulo (23°26S 45°04 W) e Paraná (25°41\'S 048°25\'W), assim como por estudos morfométricos de 5 indivíduos de cada população. Análises filogenéticas e de redes de haplótipos de cada marcador revelaram uma profunda divergência gênica entre duas linhagens, Ceará - São Paulo e Rio de Janeiro - Paraná, dados estes corroborados por alguns caracteres das analises morfométricas. Nossos resultados sugerem que a espécie Chiropsalmus cf. quadrumanus é um complexo de espécies resultado da combinação entre taxonomia conservadora e especiação críptica. Estudos mais abrangentes que contenham espécimes de outros pontos da costa brasileira, incluída a localidade tipo (Santa Catarina, Müller, 1859), poderão verificar a existência de outras possíveis espécies crípticas e ampliar o conhecimento sobre a distribuição das duas linhagens encontradas neste estudo. As conseqüências taxonômicas do presente trabalho são importantes e ainda deverão ser trabalhadas. A falta de informação sobre a espécie, em conjunto com atitudes taxonômicas conservadoras e eventos de especiação críptica, não nos permitem delimitar as novas espécies com clareza. De qualquer maneira, nossos resultados revelam que há um complexo de espécies com o nome Chiropsalmus quadrumanus no Atlântico Americano. O presente trabalho constitui o primeiro estudo filogeográfico para qualquer espécie de cubozoário. / Cubozoans live in tropical and subtropical waters around the world. Chiropsalmus cf. quadrumanus (Cubozoa: Chiropsalmidae) inhabits the Atlantic coast of America between 35°N and 27°S, an area with putative barriers that could prevent the continuous distribution of C. cf. quadrumanus, turning the species an interesting model to study population structure and phylogeography. The aim of this work was to analyze the haplotype structure of four populations of C. cf. quadrumanus from the Brazilian coast. We carried out our analyses based on the COI and 16S mitochondrial markers and the nuclear ITS region of 40 individuals from Ceará (03°43S 038°29W); Rio de Janeiro (22°37S 041°54W); São Paulo (23°26S 45°04 W) and Paraná (25°41\'S 048°25\'W), as well by morphometric surveys of 5 individuals from each population. Phylogenetic and haplotype network analyses of each marker reveled a deep genetic divergence between two lineages, Ceará - São Paulo and Rio de Janeiro - Paraná, this data were corroborated by some characters of the morphometrical analyses. Our results suggest that the species Chiropsalmus cf. quadrumanus is a complex of species resulting of the mixture of over-conservative taxonomy and cryptic speciation. More inclusive studies containing specimens from other points of the Brazilian coast, including the type locality (Santa Catarina, Müller, 1859),could verify the existence of other possible cryptic species and increase the knowledge about the distribution of the two lineages found in this study. The taxonomic consequences of this survey are important and should be overwrought. Lack of information about the species, conservative taxonomic attitudes and cryptic speciation events prevent us to delimit new species clearly. However, our results revealed a species complex named Chiropsalmus quadrumanus in the Atlantic coast of America. The present work represents the first phylogeographic approach of any cubozoan species.

Currently, we observe a growing number of researches that seek to unravel the causes, effects and possible biotechnological uses of the rhizosphere microbiota communities modulation in the complex interactions between plants and soil. We also know that the attack of herbivorous insects is a factor of considerable damage to agriculture and that has well established evolutionary relationships in natural systems. The present work tried to test some hypotheses about the direct connection between the rhizosphere microbiota and the insect pest attack. Beginning from the point that plants have well- established defense mechanisms against insects, it was verified that the rhizosphere microbiota seems to contribute actively to this system and thus to establish holobionte relationships. We had broad access to communities of the fungi and bacterial domain, through the new generation sequencing for rRNA 16S gene, region V3, and intergenic region ITS amplicons on soil, semi-soil and, insect gut samples from pest insects with general behavior (Order: Lepidoptera). Our results from the data analysis to Illumina Miseq sequencing outputs and, additional experiments, resulted in three articles presented here in chapters. In the first chapter, we discuss the modulating effect from the pest insect attack (Spodoptera frugiperda), on the Arabidopsis thaliana microbiota rhizosphere, for different physiological plant\'s stages. As a result, it was possible to discuss the differences between the modulation in the structure of bacterial communities and the modulation in the structure of the fungal communities after the attack of herbivorous insects. In the second chapter, we highlight the difference in the modulation of the bacterial community structure for different plant families. We used seedlings of Arabidopsis thaliana, Zea mays Sh2, Faseolus vulgaris, Solano lycopersicum and, Beta vulgaris exposed to the attack of Trichoplusia ni for one week. The rhizosphere microbiota analysis for each host plant groups, suggests that the influence of the plant species should be considered on the bacteria rhizosphere communities modulation after the insect attack. Besides, specific plant species may be less susceptible to rhizosphere modulation by insect attack. Another highlight was the microbiota rhizosphere effect in the biomass loss for plants sown on transplanted semi-soil. Based on the phenotypic data, we suggest the rhizosphere microbiota modulation after the herbivore may be involved in the plant biomass inhibition on the next seedlings generation. Finally, in the third chapter, we explore the Trichoplusia ni gut microbiota modulation through the microbial load obtained in the restricted feeding. The T. ni larvae from the same original population were divided into three populations. Each population was fed individually and restrictively with leaves of A. thaliana, S. lycopersicum or artificial caloric diet. We accessed the gut microbiota in T. ni after three generations of restricted feeding, and we verified that the gut microbiota in caterpillars of general behavior, could be altered due the obtaining of microbial load through alimentary diet. This modulation may be related to the degradation of metabolites that may be harmful to insect homeostasis. The gut microbiota of each population can also directly influence the food preferences of successive generations. In summary, all our results presented in each one of the chapters are important points that can help to clarify the complex relationships between plants/insects/microorganisms and, contributing to a better understanding of this holobiont system. / Atualmente observamos um crescente número de pesquisas que buscam desvendar as causas, os efeitos e as possíveis utilizações biotecnológicas da modulação de comunidades da microbiota de rizosfera nas interações complexas entre plantas e solo. Sabemos também que o ataque de insetos herbívoros é um fator de considerável prejuízo para a agricultura e que tem relações evolutivas bem estabelecidas em sistemas naturais. O presente trabalho procurou testar algumas hipóteses a cerca da relação direta entre a microbiota de rizosfera e o ataque de insetos praga. Partindo do ponto de que plantas possuem mecanismos de defesa contra insetos, bem conhecidos, foi verificado que a microbiota de rizosfera parece contribuir ativamente para esse sistema, e assim estabelecer relações holobiontes. Tivemos um profundo acesso á comunidades do domínio bactéria e fungi, através da tecnologia de sequenciamento de nova geração para amplicons do gene RNAr 16S, região V3 e região intergênica ITS em amostras de solo, semi- solo e intestino de insetos praga (Ordem: Lepidoptera) de comportamento generalista. Nossos resultados, resultaram em três artigos aqui apresentados em capítulos. No primeiro capítulo é discutido o efeito modulador da herbívora da praga agrícola Spodoptera frugiperda na microbiota de rizosfera de Arabidopsis thaliana em diferentes estágios fisiológicos da planta. Como resultados foi possível perceber que o efeito na modulação da estrutura de comunidades de bactérias é diferente do efeito na modulação de comunidades de fungos após o ataque de insetos herbívoros. Os efeitos são diferentes tanto em abundância relativa quando na diversidade para cada um dos domínios de microrganismos estudados. No segundo capítulo destacamos a diferença na modulação da estrutura de comunidades de bactérias para diferentes famílias de plantas. Utilizamos mudas de A. thaliana, Zea mays Sh2, Phaseolus vulgaris, Solanum lycopersicum e Beta vulgaris, expostas ao ataque de Trichoplusia ni durante uma semana. As análises da microbiota de rizosfera de cada um dos grupos de plantas hospedeiras, sugere que a influência da espécie vegetal deve ser considerada na modulação das comunidades de bactérias da rizosfera após a herbívora. Adicionalmente, determinadas espécies de plantas podem ser menos susceptíveis a modulação da rizosfera pela herbívora. Outro destaque foi o efeito da modulação da microbiota de rizosfera, na perda de biomassa de plantas semeadas em semi-solo transplantado. Com base nos dados fenotípicos das diferentes espécies de plantas avaliadas, sugerimos que a modulação da microbiota de rizosfera após a herbívora, pode estar envolvida na inibição da produção de biomassa vegetal na geração seguinte de plântulas. Por fim, no terceiro capítulo exploramos a modulação na microbiota no intestino de larvas de Trichoplusia ni através da carga microbiana obtida na alimentação restrita. Larvas T. ni de mesma origem foram divididas em três populações. Cada população foi alimentada de forma específica e restrita com folhas de A. thaliana ou S. lycopersicum ou dieta artificial calórica. Acessamos a microbiota do intestino das larvas, após três gerações de alimentação restrita e verificamos que a microbiota intestinal em lagartas de comportamento generalista, pode ser alterada devido à obtenção de carga microbiana por via alimentar. Essa modulação pode estar relacionada a degradação de metabólitos que podem ser prejudiciais à homeostase dos insetos. A microbiota intestinal de cada população também pode influenciar diretamente as preferências alimentares de gerações sucessivas. Em resumo, todos os nossos resultados, apresentados em cada um dos capítulos a seguir, são chaves no conhecimento e podem ajudar a clarificar as complexas relações entre plantas, insetos e microrganismos. Contribuindo assim para um maior entendimento desse tipo de sistema holobionte.