Com o aumento da produtividade na área agropecuária, houve um aumento na busca por sistemas de produção mais eficientes e competitivos. Neste contexto, o melhoramento genético animal tem proporcionado avanços na obtenção de produtos de origem animal, principalmente na seleção de touros. A superioridade genética de touros para produção de leite não pode ser medida diretamente nos animais, assim, o valor genético é medido pelo desempenho de produção de leite de suas filhas pelo teste de progênie. A seleção de touros pelo teste de progênie pode ser considerada como método ideal quando temos características de baixa herdabilidade, populações grandes e um grande número de progênies, que hoje são obtidas pelo uso de inseminação artificial. O correto parentesco entre reprodutores é um pré-requisito para um eficiente programa de melhoramento genético animal. Muitas das questões de parentesco podem ser resolvidas pelos testes convencionais, como grupos sanguíneos e proteínas do soro. Entretanto em alguns casos, esses polimorfismos não são suficientes para esclarecer a correta paternidade e, em função disso outros métodos passaram a ser utilizados, como os marcadores moleculares microssatélites. Esta metodologia tem sido descrita para resolver com maior eficiência os casos de verificação e determinação de parentesco, inclusive em bovinos, devido ao alto polimorfismo que apresenta, aliado a resultados de clara interpretação. Para a investigação de paternidade, foram utilizados os marcadores microssatélites BM8246, BMS963, CSFM50, INRA112. Foram verificadas as paternidades de nove touros da raça Gir leiteiro em teste de progênie, calculada a taxa de erro de paternidade e o efeito que esta teria na estimativa do valor genético dos touros. Foi verificado que das 74 filhas analisadas, houve exclusão para 27 delas, correspondendo a um erro de 36% na identificação de paternidade. No entanto, a mudança observada na classificação dos touros devido à correção das paternidades incorretas e consequente mudanças nos valores genéticos dos touros, não foi significativa quando as duas avaliações foram comparadas pelo método de correlação de Spearman. / Increased production in agribusiness has intensified the search for more efficient and competitive production systems. In animal production, advances are achivied primarily through sire selection. Genetic superiority of bulls for milk production cannot be directly measured in the animal; therefore, genetic value is measured by the milk production of its daughters by a progeny test. Selection of bulls by the progeny test could be considered an ideal method when the following characteristics are present: low hereditability, large productions and a large number of progenies. However, the method presents two drawbacks that limit its use: 1) high cost and 2) increase in generation intervals. Nevertheless, these obstacles can be overcome when a large number of progenies are obtained with artificial insemination (A.I.). Correct relationship among sires is essential for an efficient animal-breeding program. Many questions about relationship can be solved by conventional tests, such as blood type and serum proteins. In some cases these polymorphisms are not enough for accurate results; consequently, other methods are used such as microsatelitte molecular markers. This methodology has been described to determine and verify parentage including bovines with high efficiency due to its greater polymorphism and easily interpreted results. Microsatelitte markers BM8246, BMS963, TEXAN15, BMS483, CSFM50, INRA112 were used to investigate paternity. Paternity of nine Gir dairy sires participating in a progeny test was verified. The paternity error rate was calculated and its effect on estimation of the genetic value of the sires determined. Of the 74 daughters analyzed 27 were excluded, which corresponds to a 36% error in paternity identification. However, the change observed in bull classification due to correction of inaccurate paternity and consequent changes of genetic value of the sires was not significant when the two evaluations were compared by the correlation method of Spearman.

The flavour or fragrance of Basmati rice is associated with the presence of 2-acetyl-1- pyrroline. This work shows that a gene with homology to betaine aldehyde dehydrogenase (BAD) has significant polymorphisms in the coding region of fragrant genotypes relative to non fragrant genotypes. Accumulation of 2-acetyl-1-pyrroline in fragrant rice genotypes may be explained by the presence of mutations resulting in loss of function of the fgr gene product. The fgr gene corresponds to the gene encoding BAD2 in rice while BAD1 is encoded by a gene on chromosome 4. Development of an allele specific amplification (ASA) based around the deletion in the gene encoding BAD2 allows, perfect, simple and low cost discrimination between fragrant and non-fragrant rice varieties and identifies homozygous fragrant, homozygous non-fragrant and heterozygous non-fragrant individuals in a population segregating for fragrance. The cDNAs transcribed from rice chromosomes 4 and 8, each encoding an enzyme with homology to betaine aldehyde dehydrogenase were cloned and expressed in E. coli. The enzyme responsible for fragrance, encoded from chromosome 8, had optimum activity at pH 10, showed low affinity towards betaine aldehyde (bet-ald) with Km value of approximately 63ìM but a higher affinity towards -aminobutyraldehyde (GABald) with a Km value of approximately 9ìM. The enzyme encoded from chromosome 4 had optimum activity at pH 9.5 and showed generally lower affinity towards most substrates compared to the enzyme encoded from chromosome 8, substrate specificities suggest that the enzymes have higher specificity to aminoaldehydes and as such both should be renamed as an aminoaldehyde dehydrogenase (AAD). The inactivation of AAD2 (BAD2) in fragrant rice varieties likely leads to accumulation of its main substrate GABald which then cyclises to 1-pyrroline the immediate precursor of 2AP.

Butternut (Juglans cinerea L.), a lesser-known relative of black walnut (Juglans nigra L.), is a native tree species beneficial for wildlife, valuable for timber, and part of the great diversity of species in the eastern forests of North America. Populations of butternut are being devastated by butternut canker disease, caused by the fungus Sirococcus clavigignenti-juglandacearum (V.M.G. Nair, Kostichka, & Kuntz), which is thought to be introduced to North America. The disease causes multiple branch and stem cankers that eventually girdle trees. Small population sizes, lack of sprouting, and shade intolerance exacerbates the disease and results in permanent losses of butternut across the native range. Fortunately, healthy, canker-free butternut trees have been found proximal to diseased trees, indicating that a breeding approach could be a feasible strategy for producing and reintroducing resistant butternuts. A successful restoration program will require an understanding of genetic variation in open-pollinated seedlings, genetic basis of disease resistance, seedling establishment procedures, site requirements, and a greater understanding of disease development over time. This dissertation is divided into six parts, with the overall goal of insight into butternut ecology and management techniques which could be used to guide restoration decisions for this important species. The first two parts are an introduction and a literature review. In the third section, butternut seedlings were propagated in nursery progeny plantings to determine the genetic and phenotypic variability among one-year-old seedlings in a controlled environment. Part four outlines the disease development of butternut seedlings across progeny in resistance screening plantings at various locations. Part five aims at aiding restoration techniques by determining the impact of phenotypic and genetic variables on establishment success across various planting sites with different silvicultural treatments. Part six describes the dynamics of a large population of healthy and diseased butternut trees including disease development across temporal scales. The information gained from this research will be directly used in gene conservation strategies, the construction of disease resistant breeding orchards, and in determining appropriate restoration techniques.

Herbaceous communities are critical to the functioning of forest ecosystems. They recycle nutrients, help prevent erosion, provide critical microhabitats and maintain biodiversity. In the eastern United States, most hardwood forests are growing on land once entirely cleared or used for some form of agriculture. Although some of these forests are nearly 150 years old, they still have depauperate native herbaceous communities when compared to remaining old-growth forests. This long-term depletion may result from dispersal limitation or environmental limitation. I tested the hypothesis that dispersal was the primary factor contributing to the absence of five spring-flowering herbaceous species in four secondary mesic hardwood forests. I transplanted adults and sowed fresh propagules into chosen forests. By establishing negative controls, I showed that propagules of experimental species were not incidentally dispersed and would not have been present at the sites had I not introduced them. In all four sites, seeds of three ant-dispersed species germinated and adults of these species survived, flowered and self-sowed viable propagules. These results strongly indicated dispersal limitation in all sites. Another ant-dispersed species showed evidence of being dispersal-limited in at least two sites. The limitations of one gravity-dispersed species were unclear. I discuss results from the first year after transplanting and offer management suggestions to facilitate the return of these species to degraded forests.

Cocoa has been useful in several ways, especially to countries that produce it and consumers all over the world. Its benefits range from improving economies to satisfying millions all over the world who desire one cocoa product or the other. The production of cocoa however is not without its own attendant problems. Aging tree stock, spread of diseases, and production capacity problems are, but a few of many challenges that seed gardens that produce cocoa face. This study investigated the pollination process on selected seed gardens in Ghana, the second largest world producer of cocoa. The purpose of this study was to investigate the artificial pollination process of cocoa on seed gardens in Ghana, and explore ways by which the Lean principle of Value Stream Mapping could be applied in improving the process. The process of pollination was observed on eighteen seed gardens, and a current value stream map depicting the existing process developed to give an indication of the current state of affairs. Subsequently, a future value stream map based on the current map was developed to show aspects of the process that could be improved to enhance the operations of the seed gardens. The future value stream map revealed that operational procedures relating to cleaning and preparation of trees, choosing male and female trees for pollination, harvesting of pollen, and manually pollinating practices presented various avenues for improving the pollination process and hence seed production output.

The flavour or fragrance of Basmati rice is associated with the presence of 2-acetyl-1- pyrroline. This work shows that a gene with homology to betaine aldehyde dehydrogenase (BAD) has significant polymorphisms in the coding region of fragrant genotypes relative to non fragrant genotypes. Accumulation of 2-acetyl-1-pyrroline in fragrant rice genotypes may be explained by the presence of mutations resulting in loss of function of the fgr gene product. The fgr gene corresponds to the gene encoding BAD2 in rice while BAD1 is encoded by a gene on chromosome 4. Development of an allele specific amplification (ASA) based around the deletion in the gene encoding BAD2 allows, perfect, simple and low cost discrimination between fragrant and non-fragrant rice varieties and identifies homozygous fragrant, homozygous non-fragrant and heterozygous non-fragrant individuals in a population segregating for fragrance. The cDNAs transcribed from rice chromosomes 4 and 8, each encoding an enzyme with homology to betaine aldehyde dehydrogenase were cloned and expressed in E. coli. The enzyme responsible for fragrance, encoded from chromosome 8, had optimum activity at pH 10, showed low affinity towards betaine aldehyde (bet-ald) with Km value of approximately 63ìM but a higher affinity towards -aminobutyraldehyde (GABald) with a Km value of approximately 9ìM. The enzyme encoded from chromosome 4 had optimum activity at pH 9.5 and showed generally lower affinity towards most substrates compared to the enzyme encoded from chromosome 8, substrate specificities suggest that the enzymes have higher specificity to aminoaldehydes and as such both should be renamed as an aminoaldehyde dehydrogenase (AAD). The inactivation of AAD2 (BAD2) in fragrant rice varieties likely leads to accumulation of its main substrate GABald which then cyclises to 1-pyrroline the immediate precursor of 2AP.

Mangos (Mangifera indica L.) are tropical/subtropical fruits belonging to the plant family Anacardiaceae. Anthracnose is the most deleterious disease of mango both in the field and during postharvest handling. It is most commonly caused by the Colletotrichum gloeosporioides complex. Mangiferin, a xanthanoid compound found in at least twelve plant families worldwide (Luo et al., 2012), is present in large amounts of the leaves and edible mangos. Even though this compound plays a pivotal role in the plant’s defense against biotic and abiotic stressors, no correlations been made between the compound and mango anthracnose resistance. Mangos were collected, grouped according to their countries of origin, and evaluated for their mangiferin concentrations at four different stages of development. Extracts of interest were then tested against different strains of C. gloeosporioides. The results demonstrated that mangiferin concentrations are significantly different at different stages in fruit development. The antifungal assays were inconclusive.

Com o aumento da produtividade na área agropecuária, houve um aumento na busca por sistemas de produção mais eficientes e competitivos. Neste contexto, o melhoramento genético animal tem proporcionado avanços na obtenção de produtos de origem animal, principalmente na seleção de touros. A superioridade genética de touros para produção de leite não pode ser medida diretamente nos animais, assim, o valor genético é medido pelo desempenho de produção de leite de suas filhas pelo teste de progênie. A seleção de touros pelo teste de progênie pode ser considerada como método ideal quando temos características de baixa herdabilidade, populações grandes e um grande número de progênies, que hoje são obtidas pelo uso de inseminação artificial. O correto parentesco entre reprodutores é um pré-requisito para um eficiente programa de melhoramento genético animal. Muitas das questões de parentesco podem ser resolvidas pelos testes convencionais, como grupos sanguíneos e proteínas do soro. Entretanto em alguns casos, esses polimorfismos não são suficientes para esclarecer a correta paternidade e, em função disso outros métodos passaram a ser utilizados, como os marcadores moleculares microssatélites. Esta metodologia tem sido descrita para resolver com maior eficiência os casos de verificação e determinação de parentesco, inclusive em bovinos, devido ao alto polimorfismo que apresenta, aliado a resultados de clara interpretação. Para a investigação de paternidade, foram utilizados os marcadores microssatélites BM8246, BMS963, CSFM50, INRA112. Foram verificadas as paternidades de nove touros da raça Gir leiteiro em teste de progênie, calculada a taxa de erro de paternidade e o efeito que esta teria na estimativa do valor genético dos touros. Foi verificado que das 74 filhas analisadas, houve exclusão para 27 delas, correspondendo a um erro de 36% na identificação de paternidade. No entanto, a mudança observada na classificação dos touros devido à correção das paternidades incorretas e consequente mudanças nos valores genéticos dos touros, não foi significativa quando as duas avaliações foram comparadas pelo método de correlação de Spearman. / Increased production in agribusiness has intensified the search for more efficient and competitive production systems. In animal production, advances are achivied primarily through sire selection. Genetic superiority of bulls for milk production cannot be directly measured in the animal; therefore, genetic value is measured by the milk production of its daughters by a progeny test. Selection of bulls by the progeny test could be considered an ideal method when the following characteristics are present: low hereditability, large productions and a large number of progenies. However, the method presents two drawbacks that limit its use: 1) high cost and 2) increase in generation intervals. Nevertheless, these obstacles can be overcome when a large number of progenies are obtained with artificial insemination (A.I.). Correct relationship among sires is essential for an efficient animal-breeding program. Many questions about relationship can be solved by conventional tests, such as blood type and serum proteins. In some cases these polymorphisms are not enough for accurate results; consequently, other methods are used such as microsatelitte molecular markers. This methodology has been described to determine and verify parentage including bovines with high efficiency due to its greater polymorphism and easily interpreted results. Microsatelitte markers BM8246, BMS963, TEXAN15, BMS483, CSFM50, INRA112 were used to investigate paternity. Paternity of nine Gir dairy sires participating in a progeny test was verified. The paternity error rate was calculated and its effect on estimation of the genetic value of the sires determined. Of the 74 daughters analyzed 27 were excluded, which corresponds to a 36% error in paternity identification. However, the change observed in bull classification due to correction of inaccurate paternity and consequent changes of genetic value of the sires was not significant when the two evaluations were compared by the correlation method of Spearman.

As a species domesticated only in the last century, agricultural cranberry plants (Vaccinium macrocarpon) remain little removed from their wild relatives. Thus, it is a potential model species for studies of the earliest stages of domestication; however, there is little available quantitative information on its wild population biology and ecology. As such information is vital to studies of the ecological changes occurring during domestication, the purpose of this study was to consolidate the relevant knowledge available and conduct a preliminary search for patterns in the invertebrate communities of wild bogs. The alpha diversity was found to be greater than the overall (gamma diversity), which is likely a result of the metric used and the fact that there was minimal overlap in rare species between bogs. In addition, alpha diversity was found to be significantly negatively correlated with bog age. Two pairs of species were found to be correlated with each other, Blunt-nosed (Limotettix vaccinia (Van Duzee)) and Sharp-nosed leafhopper (Scaphytopius sp.), along with fleabeetle (Sysena frontalis (F.)) and firebeetle (Cryptocephalus incertus (Oliv.). In addition, it was found that spiders are significantly more common in western bogs, while firebeetle is significantly more common in eastern bogs. However, this may be indicative of a correlation with bog age rather than a true correlation with geographic location. More work is needed to determine the true dynamics driving these findings, so that the information could eventually be used to improve the efficacy and decrease the environmental impact of pest management on agricultural bogs.

The biofuel market is dominated by ethanol and biodiesel derived from cellulosic and lipid-based biomass crops. This is largely due to the relatively low costs and reliability of production. At present, production of non-food plant-derived oils for biofuel production in the U.S. is minimal. A research team from the Commonwealth Scientific and Industrial Research Organization (CSIRO), an independent Australian federal government research institution, has developed an efficient transgenic system to engineer oil production in tobacco leaves. This novel system is comprised of multiple transgenes that direct the endogenous metabolic flux of oil precursors towards triacylglycerol (TAG) production. Additional genes were incorporated to store and protect the accumulated oil in vegetative tissues. Preliminary greenhouse tests by the CSIRO research group indicated an oil content of > 30% by dry weight (DW) in tobacco leaf lamina. Here we evaluated two transgenic lines against a non-transgenic control in 2017 and 2018 in greenhouse and field production systems. The 2017 pilot study showed that the high leaf-oil tobacco line was viable and will grow in the field in Kentucky. Chemical analyses revealed significantly higher oil content compared to the non-transgenic control despite several logistical setbacks. These promising discoveries prompted the deployment of additional transgenic line assessments and further data validation in 2018. Line evaluations in 2018 revealed that the LEC2:WRI1:DGAT:OLE transgenic line had the highest leaf oil content (≥ 19.3% DW-1) compared to both the WRI1:DGAT:OLE transgenic line (≤ 5.6% DW-1) and non-transgenic control (≤ 2.1% DW-1). The results of this research will contribute to the successful development of transgenic tobacco lines engineered to accumulate high concentrations of TAG in the leaves.