A família Baculoviridae é um grande grupo de vírus com cerca de 700 espécies de insetos hospedeiros, com dois fenótipos: o ODV (occlusion derived virion), que faz a infecção primária do intestino médio; e o BV (budded virus), responsável pela infecção sistêmica. No Brasil, o nucleopoliedrovírus Anticarsia gemmatalis (AgMNPV) é utilizado como controle biológico da lagarta-da-soja Anticarsia gemmatalis. O genoma do AgMNPV-2D contém 152 ORFs, 26 das quais codificam proteínas estruturais. Entre elas, a glicoproteína GP64 é fundamental para infecção secundária. Este estudo visa identificar proteínas estruturais do AgMNPV-2D por duas abordagens de espectrometria de massas. Também comparar a variabilidade da gp64 de isolados geográficos por sequenciamento por Sanger e de alta cobertura. Assim, identificamos as substituições de gp64 e vimos que ela não suporta a separação geográfica dos isolados. Também identificamos 44 e 33 proteínas em ODV e BV, respectivamente. Seis novas proteínas foram identificadas no ODV e sete delas no BV. Além disso, 11 proteínas celulares foram identificadas no AgMNPV-2D, possivelmente necessárias para infecção. Este achado contribui para o entendimento da morfogênese do AgMNPV e fatores associados à multiplicação viral. / Baculoviridae are arthropod-specific viruses with more than 700 host insects, which produce two phenotypes: the budded virus (BV) and, the occlusion-derived virus (ODV) for intra and across host spread, respectively. Brazil uses the Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) as a biological control agent of the velvet bean caterpillar (A. gemmatalis). The genome of the AgMNPV-2D carries 152 ORFs, 26 of which code for structural proteins. Herein, the structural proteins of AgMNPV-2D were analyzed by two mass spectrometry techniques. The additional objective was to compare the gene gp64. of different geographical populations by Sanger and next generation sequencing. This analysis allowed us to observe the substitutions of gp64 and refuted the notion of a geographical isolation of the samples. We also observed a total of 44 proteins of the ODV and 33 of the BV. Six new proteins were found in the ODV and seven in the BV. Furthermore, 11 cellular proteins were also identified, which are possibly assorted during viral morphogenesis. These findings may provide novel insights into AgMNPV biology and its host interaction, leading us to a better understanding about morphogenesis and also the associated factors of the viral multiplication.

Thesis (M.Sc. ( Physiology )) --University of Limpopo, 2011 / Refer to document / the National Research Foundation

Isolated populations have potential to become new species that should have less genetic variation than their ancestors. Small populations are more likely to lose genetic variation, which is, thus, expected to be greater in ancestors. Aliciella caespitosa and A. tenuis, two endemic species, may be derived from small populations of A. subnuda, a widespread species. Chloroplast DNA sequences were used to test this hypothesis. Allozyme data were used to compare genetic variation and numbers of alleles. Chloroplast data do not support the proposed relationships between A. subnuda and the other two species. Allozyme data were not more variable in A. subnuda. The data suggest that A. tenuis is derived from A. caespitosa, although the former did not show lower allozyme diversity. I detected fewer alleles in A. tenuis. These data suggest that the original population of A. tenuis was not small enough to lose genetic variation relative to its progenitor.

La réponse des arbres à l'augmentation des sécheresses liées au réchauffement climatique dépend de leur capacité d'adaptation, c’est-à-dire de la variation génétique et de l’héritabilité de caractères adaptatifs impliqués dans la résistance à la sécheresse. Dans le chapitre I, nous identifions des proxys facilement mesurables de caractères adaptatifs impliqués dans la résistance à la sécheresse en comparant la microdensité du bois d’individus morts et survivants après la sécheresse de 2003. Les variables les plus discriminantes sont les densités moyennes de segments de haute et basse densité, la proportion du segment de haute densité et le coefficient de variation du segment de haute densité. Les arbres survivants ont toujours une densité plus élevée et des profils généralement plus hétérogènes. Si ces traits sont suffisamment variables génétiquement et héritables, alors il est possible de sélectionner des arbres plus résistants à la sécheresse dans les populations d'amélioration et dans les peuplements forestiers destinés à être régénérées naturellement. Nos résultats suggèrent qu’une sélection naturelle directionnelle pour la densité du bois du douglas se produit dans des environnements plus ou moins limités en eau. Cette sélection agit dans des directions différentes selon les caractéristiques des pressions de sélections subies dans les régions étudiées. Dans le chapitre II, nous évaluons le potentiel d’adaptation à la sécheresse du douglas introduit en France. Ce potentiel d’adaptation dépend de la variation génétique et de l'héritabilité des caractères adaptatifs mis en évidence dans le chapitre I. Nous trouvons une grande variabilité des estimations d’héritabilité et d’AGCV entre variables, provenances, sites et, dans une faible mesure, entre cernes annuels. La plupart des variables possèdent des héritabilités élevées à assez élevées au moins pour certaines provenances dans certains sites. Certaines variables tendent à avoir des héritabilités et AGCV généralement plus élevées : ce sont plutôt des variables de la partie de faible densité du cerne. Certaines variables de résistance à la sécheresse possèdent à la fois une héritabilité et une AGCV élevées : ce sont de bons critères de sélection pour la résistance à la sécheresse en amélioration génétique ou en régénération naturelle. Les estimations différentes entre sites suggèrent que les estimations d’héritabilités augmentent avec le caractère favorable du milieu. Les estimations sont significativement différentes entre provenances, avec de forts effets d’interaction avec les sites. En revanche il y a peu ou pas de différences significatives entre cernes. Dans le chapitre III, nous avons tenu compte de la variation associée aux cernes de croissance pour étudier les relations entre les estimations annuelles de paramètres génétiques et des variables climatiques et édaphiques. Les estimations d'héritabilité et de variation génétique de la plupart des variables de microdensité sont corrélées significativement avec la plupart des variables environnementales testées. De rares variables n’ont montré aucune corrélation significative dans aucun cas. Les caractéristiques des relations significatives sont variables entre les caractères, les sites et les provenances. Les prédicteurs climatiques les plus importants sont la température, l'évapotranspiration, la réserve en eau du sol et le déficit en eau. Les précipitations affectent peu l´estimation des paramètres génétiques. De façon générale, meilleures sont les conditions de croissance, plus élevées sont les estimations. Toutes les composantes des essais expérimentaux modifient les estimations des paramètres génétiques. Certaines peuvent être fortement déterminées par le choix du matériel végétal et du site. D'autres, comme la variation climatique temporelle, sont moins contrôlées et peuvent affecter de façon plus ou moins aléatoire les estimations. / Forest response to the drought increase associated to the climatic warming relies on tree adaptive potential, i.e. the genetic variation and the heritability of adaptive traits involved in resistance to drought. In the first chapter, we identify easy-to-measure proxies of adaptive traits for resistance to drought. We compare the wood microdensity of dead and surviving trees after the 2003 heat wave in France. The most discriminating variables are the mean density of high and lowdensity segments, high-density proportion and coefficient of variation of the lowdensity segment. The wood of the surviving trees is always denser and more heterogeneous. If these adaptive traits are variable and heritable, then it is possible to select for improved resistance to drought in the breeding population as well as in natural regeneration. Our results also suggest that directional selection is going on in more or less water-stressed environments. The direction is variable according to the nature of the selection pressure in the different regions. In the Chapter II, we estimate the evolutionary potential to drought of the introduced Douglas-fir in France. This evolutionary potential relies on the magnitude of the genetic variation and of the heritability of the adaptive traits found in the first chapter. The heritability and the genetic variation are highly variable between provenances, sites and, to a much lower extent, between annual rings. Most variables have moderate to high heritability estimates for at least some provenances in some sites. Some traits tend to have generally higher heritability and genetic variation estimates. These are mostly variables of the density part of the annual ring. The variables having at the same time relatively high estimates of heritability and genetic variation are good candidates for becoming efficient selection traits for resistance to drought in tree breeding as well as in natural regeneration. The significant between-site variation suggests that the heritability estimates increase with site quality. The estimates are also significantly different between provenances with a strong provenance × site interaction. Conversely there is little significant between annual-ring variation. The chapter III takes advantage of the annual-ring variation to study the relationships between the genetic parameter estimates and climatic and soil variables. The heritability and genetic variation estimates of most variables significantly relates with most tested environmental variables. Very few variables never correlates with any environmental variable. The significant relationships are very variable between traits, provenances and sites. The most important predictors are temperature, evapotranspiration, and soil water reserve and water deficit. Rainfall marginally influences the genetic parameter estimates. Generally, the better the growing conditions, the higher the estimates. All components of the experimental trials affect the genetic parameters estimates. Thus, the choice of the plant material and of the experimental site strongly determines the genetic parameter estimates. The uncontrolled climatic variation may randomly affect the estimates.

A number of morphological, floral, rhizome, root and herbage characteristics were studied in order to describe genetic variation and environmental responses in T. ambiguum. From each of six lines, 30 genotypes were clonally progagated into five diverse edaphic and altitudinal sites and grown for one season. Comparisons of growth and form were also made between T. ambiguum, T. repens cv. 'Grasslands Huia' and Lotus pedunculatus cv. 'Grasslands Maku'. The polyploid lines were found to have larger leaves than the diploids while all the lines of T. ambiguum had larger leaves than did white clover. Tetraploid lines had longer petioles and were taller and more erect than the diploids or hexaploids. Floral initiation was found to become later as ploidy level increased but the tetraploid lines exhibited a very large variation in flowering date. The cultivar Treeline was found to produce the most herbage under all conditions although not significantly more than cultivar Prairie or C.P.I. 57353. However, as nodulation was not studied it was not possible to determine whether variety differences were due to root nodulating ability or some other genetically determined parameter. None of the T. ambiguum varieties produced as much herbage as cv. Huia or cv. Maku at any site. However, all the T. ambiguum lines performed relatively better under harsher conditions. Because a large proportion of T. ambiguum was below ground the best T. ambiguum line, cultivar Treeiine, produced equivalent total plant dry weight to cv. Huia and cv. Maku at 1200 m. a.s.l., the high altitude site. Cultivar Prairie was found to have the highest proportion of rhizomes to total plant mass bu because cv. Treeline had higher total plant dry weight both cultivars produced equivalent mean rhizome dry weight. The number of rhizomes, number of daughter plants and rhizome dry weight were all highly correlated and these three characteristics showed similar trends among varieties. Rhizome length was found to increase with ploidy level, as did rhizome internode length. However, the number of nodes was found to be higher in the diploids than in the polyploids. Cultivar Treeline was found to have a high proportion of branching nodes on its rhizomes while C.P.I. 57353 and cv. Summit had the least. Rhizome production was restricted in the Wakanui silt loam soil of high bulk density. However, herbage growth and rhizome branching was increased, probably because of the higher fertility. It was shown using factor analysis on genotypic correlations, that rhizome characteristics and herbage yields were relatively independently inherited. However, morphological characters tended to be related to herbage yields. The polyploid varieties were found to be more genetically variable than the diploids . Broad sense heritabilities were calculated for all parameters measured, and in general, morphological characteristics had higher values than agronomic characteristics. As the genetic variation within each line was higher than the variation among lines, it is apparent that selection within lines should result in the largest gains. Therefore, comparison of the mean performance of the presently highly variable lines is relatively uninformative. By sacrificing some genetic diversity, large gains could be made in performance. It is suggested that cv. Prairie be used as the basis for selecting a highly rhizomatous cultivar while cv. Treeline could be used in the selection of a cultivar with higher herbage production suitable for high country conditions. In a second trial, an established stand of cv. Treeline produced up to 13250 kg per ha for one season under good growing conditions. The management required to produce this amount of herbage was to irrigate and cut to ground level every two months. The growth rate was considerably depressed when cut monthly. It was also shown that root and rhizome yield reached 12600 kg per ha, indicating a massive reserve of assimilates, particularly useful for surviving periods of stress. Seed yields were found to be adequate, reaching levels equivalent to 500-700 kg per ha. These results were discussed in relation to earlier observations on T. ambiguum by workers in Russia, Australia, U.S.A. and New Zealand. Suggestions were made for further genetic and agronomic testing.

<p>Although the practice of releasing fish into the wild is common in the management and conservation of fish populations, the success of release programmes and the potential harmful genetic and ecological effects that may follow are rarely considered. This thesis focuses on genetic and ecological consequences of fish releases, exemplified by supportive breeding of brown trout (<i>Salmo trutta</i>) and translocation of European eel (<i>Anguilla anguilla</i>). Specific questions addressed include: What is the relative performance of hatchery produced fish released to support wild populations, and do released hatchery fish contribute to the natural productivity? What is the variation in reproductive success in the wild, and how does it affect the genetic consequences of a supportive breeding programme? Is there a spatial genetic structure in the European eel that must be considered in the management of this rapidly declining species?</p><p>Experiments conducted under natural and near-natural conditions in the River Dalälven, Sweden, suggest that hatchery produced trout can reproduce in the wild. In fact, when the pronounced variation between individual breeders was accounted for, there were no detectable differences between hatchery produced and wild born trout in reproductive success or offspring survival. These results were supported by molecular genetic data suggesting a pronounced gene flow from hatchery to wild trout in the river. Hatchery reared trout were, however, found to exhibit reduced survival rates immediately following release into the wild, an effect that was most likely due to phenotypic responses to the hatchery environment during ontogeny and a lack of experience of the wild.</p><p>In sharp contrast to recently published studies, the present genetic analyses of European eels sampled across the whole distribution range suggest no spatial genetic structure but a subtle temporal genetic heterogeneity within sampled locations. These results emphasise the need to consider temporal replication when assessing population structure of marine species.</p><p>The results obtained have general implications for the management and conservation of fish populations. First, supportive breeding of threatened salmonid populations might be successful, not only for boosting the census size and thereby reducing the short-term probability of extinction, but also for reducing the risks of inbreeding depression and loss of adaptive potential in future generations. However, the results also highlight the need to restore the natural productivity of a population under supportive breeding to avoid a potential reduction in fitness due to hatchery selection. Further, the lack of a detectable spatial genetic structure in the European eel suggests that the management strategy of translocating juvenile eels from locations were they are overabundant to other suitable freshwater habitats does not necessarily have to include genetic considerations with respect to the geographical origin of the translocated eels.</p>

<p>Genetic and epigenetic variation is a key determinant of human diversity and has an impact on disease predisposition. Single nucleotide polymorphisms (SNPs) and copy number polymorphisms (CNPs) are the main forms of genetic variation. The challenge is to distinguish normal variations from disease-associated changes. Combination of genetic and epigenetic alterations, often together with an environmental component, can cause cancer. In paper I, we investigated possible alterations affecting the transcriptional regulation of PDGFRα in patients affected with central nervous system tumors by characterizing the haplotype combinations in the <i>PDGFRA</i> gene promoter. A specific over-representation of one haplotype (H2δ) in primitive neuroectodermal tumors and ependymomas was observed, suggesting a functional role for the ZNF148/PDGFRα pathway in the tumor pathogenesis. In paper II, 50 glioblastomas were analyzed for DNA copy number variation with a chromosome 22 tiling genomic array. While 20% of tumors displayed monosomy 22, copy number variations affecting a portion of chromosome 22 were found in 14% of cases. This implies the presence of genes involved in glioblastoma development on 22q. Paper III described the analysis of copy number variation of 37 ependymomas using the same array. We detected monosomy in 51.5% of the samples. In addition, we identified two overlapping germline deletions of 2.2 Mb and 320 kb (the latter designated as Ep CNP). In order to investigate whether Ep CNP was a common polymorphism in the normal population or had an association with ependymoma development, we constructed a high-resolution PCR product-based microarray covering this locus (paper IV). For this purpose, we developed a program called <i>Sequence Allocator</i>, which automates the process of array design. This approach allowed assessment of copy number variation within regions of segmental duplications. Our results revealed that gains or deletions were identical in size and encompassed 290 kb. Therefore, papers I-IV suggest that some SNPs and CNPs can be regarded as tumor-associated polymorphisms. Finally, paper V describes variation of DNA methylation among fully differentiated tissues by using an array covering ~9% of the human genome. Major changes in the overall methylation were also found in colorectal cancer cell lines lacking one or two DNA methyltransferases.</p>

Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (leaves 150-153).

Abiotic and biotic variation has been shown to be important in regulating nutrient cycling and belowground communities in natural systems. However, genetic variation in dominant plants as a driver of rates of nutrient cycling is still poorly understood and few studies have looked at genotype interactions across multiple environments. Using Populus angustifolia and a common garden approach, we hypothesized that all three factors: tree genetic variation, environmental conditions and genetic by environment (G x E) interactions would affect soil carbon (C) storage and nitrogen (N) cycling. Replicated copies of five different reciprocally planted Populus genotypes were studied in three separate 18-21 year old common gardens at different elevations (1300m, 1384m and 1587m) in northern Utah, to measure the genotype and environmental effects on pools of soil C and N as well as rates of soil net N nitrification and net mineralization. Our results indicate that genotypes influence pools of soil C, total N and C:N, but genotype did not influence net rates of nitrogen mineralization. Environmental variation significantly influenced pools of soil C, total N, soil C:N and rates of net nitrification and net N mineralization. As predicted, G x E interactions significantly influenced both pools and processes of soil C and N cycling. Overall, we found that genetic variation in plant traits (tree diameter and leaf/root chemistry) as well as soil texture across gardens were significant predictors of soil C and N pools and fluxes across seasons. These data help us understand the relative role of genotypic variation on above- and belowground interactions in different environments and the consequences of these interactions on ecosystem processes. The results from this study show that across an environmental gradient Populus angustifolia genotypes can influence nitrogen mineralization through feedbacks between environmental variation, tree phenotype and soils.

Although the practice of releasing fish into the wild is common in the management and conservation of fish populations, the success of release programmes and the potential harmful genetic and ecological effects that may follow are rarely considered. This thesis focuses on genetic and ecological consequences of fish releases, exemplified by supportive breeding of brown trout (Salmo trutta) and translocation of European eel (Anguilla anguilla). Specific questions addressed include: What is the relative performance of hatchery produced fish released to support wild populations, and do released hatchery fish contribute to the natural productivity? What is the variation in reproductive success in the wild, and how does it affect the genetic consequences of a supportive breeding programme? Is there a spatial genetic structure in the European eel that must be considered in the management of this rapidly declining species? Experiments conducted under natural and near-natural conditions in the River Dalälven, Sweden, suggest that hatchery produced trout can reproduce in the wild. In fact, when the pronounced variation between individual breeders was accounted for, there were no detectable differences between hatchery produced and wild born trout in reproductive success or offspring survival. These results were supported by molecular genetic data suggesting a pronounced gene flow from hatchery to wild trout in the river. Hatchery reared trout were, however, found to exhibit reduced survival rates immediately following release into the wild, an effect that was most likely due to phenotypic responses to the hatchery environment during ontogeny and a lack of experience of the wild. In sharp contrast to recently published studies, the present genetic analyses of European eels sampled across the whole distribution range suggest no spatial genetic structure but a subtle temporal genetic heterogeneity within sampled locations. These results emphasise the need to consider temporal replication when assessing population structure of marine species. The results obtained have general implications for the management and conservation of fish populations. First, supportive breeding of threatened salmonid populations might be successful, not only for boosting the census size and thereby reducing the short-term probability of extinction, but also for reducing the risks of inbreeding depression and loss of adaptive potential in future generations. However, the results also highlight the need to restore the natural productivity of a population under supportive breeding to avoid a potential reduction in fitness due to hatchery selection. Further, the lack of a detectable spatial genetic structure in the European eel suggests that the management strategy of translocating juvenile eels from locations were they are overabundant to other suitable freshwater habitats does not necessarily have to include genetic considerations with respect to the geographical origin of the translocated eels.