Thesis (Ph. D.)--University of Alberta, 2009. / Title from pdf file main screen (viewed on Dec. 16, 2009). "A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Plant Science, Department of Agricultural, Food and Nutritional Sciences, University of Alberta." Includes bibliographical references.

Cryopreservation is the most promising method for the long-term conservation of germplasm of plants producing desiccation-sensitive seeds. While such seeds are generally termed recalcitrant in the context of conventional storage practices, the term ‘cryo-recalcitrant’ is used for germplasm which is not readily amenable to cryopreservation. Cryo procedures usually involve a sequential combination of steps which must be optimised to limit the stresses experienced by specimens, thus promoting their survival. The present contribution reports on the effects of some of the steps involved in cryopreservation on the survival of the embryos of the amaryllids, Ammocharis coranica, Brunsvigia grandiflora and Haemanthus albiflos, with the ultimate aim of developing a protocol(s) for the successful cryopreservation of the germplasm of these species. The main foci of the investigations were the effects of rapid (flash) drying, the use of the cryoprotectant additives, glycerol (5 & 10%) and DMSO (0.1 & 0.25%), and employment of different cooling rates on the zygotic embryos of the selected species, which are known to be recalcitrant as well as being cryo-recalcitrant. Furthermore, this study reports on attempts at improving the rapidity of dehydration during flash drying by applying a vacuum, and also of providing cathodic protection (via highly reducing cathodic water and/or direct exposure to a static {negatively-charged} cathodic field during flash drying) to the explants at various stages in the protocol. These techniques were employed in attempts to ameliorate the adverse effects of reactive oxygen species associated with stresses imposed by the procedures during the cryopreservation process. The embryos of Ammocharis coranica, Brunsvigia grandiflora and Haemanthus albiflos were initially at water contents (WCs, dry mass basis) of 3.28±0.52, 2.55±0.22, 4.48±0.92 g g-1, respectively, after harvest. These embryos proved to be tolerant to moderately rapid water loss in the short term, with >60% retaining germinability at water contents ≥0.5 g g-1. The results from this study confirmed that dehydration to water contents below 0.5 g g-1 (dry mass basis) compromised survival, and that this effect was exacerbated if the embryos were cryoprotected prior to drying. Interestingly, the rate of water loss in embryos of these species differed, with A. coranica and H. albiflos drying at a (comparably) much slower rate than those of B. grandiflora. Subsequent rapid cooling yielded promising results when compared with slow cooling, as 30% of glycerol cryoprotected, rapidly cooled A. coranica embryos that had been flash-dried to 0.36±0.10 g g-1 generated normal seedlings. It was clear, however, that the effects of these procedures were exacerbated when all the steps of the cryo procedure were applied sequentially. However, the work also showed that these adverse effects may be ameliorated if each step of the cryopreservation protocol is optimised on a species-specific basis, thus promoting the chances of survival after cryopreservation and facilitating subsequent seedling establishment. This was evident in the 30% germination obtained when embryos of A. coranica, which had been cryoprotected with glycerol prior to flash drying before exposure to rapid cooling, while those that had not been cryoprotected or were cryoprotected with DMSO before drying did not survive. The incorporation of cathodic protection during flash drying appeared promising as it promoted the survival of 10% of H. albiflos embryos dehydrated to WCs between 0.37 and 0.26 g g-1 (whereas no survival was achieved without the inclusion of this step), and 70% of A. coranica embryos that were dehydrated to 0.35±0.21. In addition, the reduction of the explant size, from a whole 6 mm embryo to a 3-4 mm excised axis, promoted survival by up to 30% for A. coranica and H. albiflos, even at higher WCs. However, survival in these cases was based on observations of abnormal development, i.e. the development of roots or shoots, or calli. No surviving embryos were obtained from B. grandiflora after cooling, regardless of the preconditioning treatment or rate of cooling, and this was accredited to the greater degree of sensitivity of these embryos to the cryo procedures than those of the other two species. The use of cathodic water to re-hydrate explants after dehydration and of applying a vacuum during flash drying did not result in any observable benefits, and require further investigation for optimisation. The very limited success towards establishing a cryopreservation protocol for the species investigated in this study reinforces the difficulties associated with the cryopreservation of recalcitrant germplasm, which informs the cryo-recalcitrance of some explants. However, the results obtained have helped to identify a number of intervention points that could be used to minimise the damage incurred during the various procedural steps involved in cryopreservation. / Thesis (M.Sc.)-University of KwaZulu-Natal, Durban, 2013.

Coming together to save the Earth. Ten years have passed since the Earth Summit in Rio created such passion and expectations. Whether the Convention on Biological Diversity has met those expectations or not vary from person to person. / Evidently, the Convention on Biological Diversity is complex, not only in its language, but also in its attempt to balance between conservation and sustainable use, and between the providers of and benefiters from biological diversity. / Subsequent to its conclusion, the Parties have strived to achieve these objectives. This thesis attempts to assist in this process by exploring the means of implementing the Convention on Biological Diversity and their consequences. / The scope of the thesis is limited to the matters of sovereignty rights and access to genetic resources, in an effort to clear up the uncertainties in the applications of these components. This thesis attempts to contribute a pragmatic perspective to these matters, which, at their core, rely on the crossing points in the implementation of the Convention on Biological Diversity and the Agreement on Trade-Related Aspects of Intellectual Property Rights. This thesis focuses on ways to reconcile property rights in genetic resources with patent rights in invention using genetic resources. It proposes one interpretation of property rights in genetic resources so as to avoid any conflict with patent rights and accordingly, avoid conflict between the requirements of the Convention on Biological Diversity and those of the Agreement on Trade-Related Aspects of Intellectual Property Rights. / I hope that the views and proposals expressed in this thesis will be considered along with other diverse approaches to the implementation of the Convention on Biological Diversity.

Cryopreservation of gametes is a valuable tool for the fast-growing aquaculture industry in New Zealand. In the present study, research was aimed to improve the cryopreservation of Pacific oyster (Crassostrea gigas) oocytes. For this, two main approaches were used: the modification of an existing published (standard) cryopreservation protocol for oyster oocytes and the modification of the oocytes themselves prior to cryopreservation. The objectives in the chapters of this thesis were: (a) determination of the cryobiological characteristics of oyster oocytes; (b) assessment and reduction of intracellular ice formation (IIF) in oocytes; and (c) modification of the lipid fraction (cholesterol and fatty acids) of oocytes prior to cryopreservation. Knowledge of the membrane permeability parameters in response to concentrations of water and ethylene glycol (EG), the influence of temperature upon these parameters, and the osmotic tolerance limits of oyster oocytes were used to develop computer models that simulated the cellular volume changes that oocytes underwent during EG addition and removal. The models predicted that when one part of EG was added in one step to one part of oocyte suspension and equilibrated for 20 min at 20 �C, similar volume changes in oocytes would be obtained, compared to a more complicated multi-step addition method. This method of addition resulted in similar post-thaw fertilization rates to those obtained by using the multi-step addition method, thus reducing oocyte handling. Cryomicroscopy was used to assess the effect of cooling rates and EG concentration on the temperature at which oocytes underwent IIF. It was found that IIF occurred at higher subzero temperatures when fast cooling rates were used (30 and 5 �C min⁻�) and at EG concentrations ranged between 0 and 15%. At a relatively slower cooling rate of 0.3 �C min⁻� and with 10% EG, which are the conditions employed in the standard cryopreservation protocol, no IIF occurred. The steps of the standard protocol that were more likely to cause oocyte damage were identified by evaluating the fertilization rate of oocytes at each step. Results showed that oocytes were most damaged by cooling them to -35 �C and followed by plunging them in liquid nitrogen. Contrary to what had been observed under the cryomicroscope, transmission electron microscopy (TEM) analysis revealed that all oocytes cryopreserved by the standard protocol contained cytoplasmic ice. In addition, it was also observed that oocytes were at two developmental stages when frozen (prophase and metaphase I). These observations prompted the development of alternative cooling programmes aimed to reduce intracellular ice. The effect of cooling rate, plunge temperature and time held at the plunge temperature were thus evaluated, based on post-thaw fertilization rate of oocytes. Overall, neither the cooling rate nor the holding time had an effect on oocyte fertility. However, the plunge temperature had an effect, where oocytes plunged at -60 �C had lower post-thaw fertilization rates than oocytes plunged at -35 �C. Through the slowing of the cooling rate, lengthening of the holding time and lowering of the plunge temperature, it was possible to reduce the amount of ice in the cytoplasm. However, the reduction of intracellular ice did not improve the post-thaw fertilization rate of the oocytes; on the contrary, post-thaw fertilization decreased notoriously. From these results, it can be suggested that oyster oocytes are more likely to be damaged by exposure to high intra and extracellular solute concentration than IIF during cryopreservation. In an effort to modify the lipid content of oyster oocytes prior to cryopreservation and thus, making them more resistant during cryopreservation, oocytes were incubated in solutions that would add or remove cholesterol or in solutions rich in long chain fatty acids (EPA or DHA). Oocytes incubated in cholesterol-rich solutions showed a positive uptake of fluorescently labelled cholesterol and this effect was dose dependent. Nevertheless, this uptake did not improve the post-thaw fertilization rate nor did it increase the total cholesterol content of the oocytes. When oocytes were incubated in non-conjugated or conjugated EPA or DHA, no increase in the proportion of these fatty acids was identified in the fatty acid profiles of whole oocytes and no improvement of the post-thaw fertilization rate was recorded. Given that there was no uptake of fatty acids from the incubation media by the oocytes, a different approach was taken. This involved the supplementation of lipid-rich diets to the oyster broodstock during gametogenesis (cold-conditioning) and vitellogenesis (warm-conditioning). Despite results showing that lipid content and, indeed, fatty acid profile was altered through the diet, the results also showed that fresh oocytes from broodstock fed during cold-conditioning did not show any improvement in their fertilization rates, nor did they benefit from a lipid-rich diet during warm-conditioning. On the other hand, cryopreserved oocytes did have higher post-thaw fertilisation rates when broodstock were fed during cold-conditioning and, although no effect was found from feeding broodstock with either of the lipid-rich diets during warm-conditioning, trends indicated that a diet consisting of fresh microalgae or the commercial supplement Algamac would yield the highest post-thaw fertilization rates. This thesis has furthered the understanding of some of the factors that determine cryosurvival in oyster oocytes and has demonstrated that both physical and biological issues must be taken into consideration for cryopreservation. Specifically, the results in this thesis helped to modify an empirically developed cryopreservation protocol for Pacific oyster oocytes. In addition, the results also showed strong evidence of the survival of oyster oocytes to intracellular ice and highlighted the importance of supplying the broodstock with lipid-rich food during the periods of gamete formation and maturation in order to obtain oocytes that are more amenable to cryopreservation. These benefits could be of significant practical importance and may be extended for the development or refinement of cryopreservation protocols for other shellfish species of commercial importance to the aquaculture industry of New Zealand.

Diss. (sammanfattning). Uppsala : Sveriges lantbruksuniv., 2003. / Härtill 7 uppsatser.

Thesis (Ph. D.)--Ohio State University, 2007.

Thesis (Ph. D.)--Wageningen Universiteit, 2005. / Thesis propositions sheet inserted. Includes bibliographical references (p. [249]-262).

O Brasil é país detentor de grande biodiversidade, no entanto pequena parcela dessa já foi estudada e catalogada. Os recentes avanços antrópicos sobre os ecossistemas naturais tem levado a sua rápida fragmentação e eliminação de alguns biótipos ainda não estudados ou catalogados. Nessa realidade de erosão genética acelerada encontra-se também espécies de jabuticabeira (Plinia sp.) que são endêmicas do Centro Sul/Sudoeste do Paraná, no ecossistema Floresta com Araucária. Medidas de conservação de germoplasma para uso atual e futuro, bem como táticas de manejo e conservação dos recursos naturais são preponderantes para reduzir ao mínimo os danos causados a biodiversidade brasileira. Para tanto é necessário entender a diversidade genética existente nos ecossistemas naturais, que é o insumo básico para a sobrevivência e evolução entre os indivíduos frente as modificações ambientais. No presente trabalho foi realizado a caracterização genética de 110 jabuticabeiras que constituem o banco ativo de germoplasma desta espécie na UTFPR – Câmpus Dois Vizinhos. Foi possível identificar a transferibilidade de 9 marcadores moleculares microssatélites (SSR) com caráter polimórfico para a população estudada, e realizar a padronização de reação de PCR para cada um deles. Da análise das jabuticabeiras do banco ativo de germoplasma chegou-se a conclusão de que o mesmo abrigou valor considerável de diversidade alélica. No entanto tal diversidade está mal distribuída, já que 11 indivíduos sozinhos já são capazes de representar 59,2% de todos alelos da coleção de 110 plantas. Os valores encontrados de heterozigosidade observada e conteúdo informativo (PIC) nessa subpopulação de 11 indivíduos foram superiores aos valores encontrados para o conjunto integral do banco ativo. O agrupamento dos indivíduos mostrou a existência de 8 diferentes grupos, sendo que 89 indivíduos ficaram no grupo 1, demonstrando o seu possível grau de parentesco e baixo nível de diversidade genética. / Brazil is a country of great biodiversity holder, however small portion of that has been studied and cataloged. Recent advances human activities on natural ecosystems has led to its rapid fragmentation and elimination of some biotypes still, not it studied or cataloged. In this reality of genetic erosion is also accelerated jabuticaba tree species (Plinia sp.). That are endemic from South Central / Southeastern Paraná State, in Araucaria forest ecosystem. Germplasm conservation measures for current and future use, as well as tactics for management and conservation of natural resources are crucial to minimize the damage caused to Brazilian biodiversity. So it is necessary to understand the genetic diversity in natural ecosystems, which is the basic input for the survival and evolution among individuals facing environmental changes. In the present study was to characterize genetic of 110 jabuticaba tree fruit, that constitute the germoplasm bank of jabuticaba tree fruit from UTFPR - Câmpus Dois Vizinhos. It was possible to identify transferability of 9 microsatellite markers (SSR) with polymorphic character for the population studied, and it carry out standardization of PCR reaction for each of them. The analysis of plant germoplasm bank came to the conclusion that houses a considerable amount of allelic diversity, but this diversity is poorly distributed, since 11 individuals alone are already able to represent 59.2% of all alleles of the collection of 110 plant. The found values of, observed heterozygosity and information content (PIC) in this subpopulation of 11 individuals were higher than the values found for the full set of the germoplasm bank. The grouping of individuals showed the existence of 8 different groups, and 89 subjects were in group 1, demonstrating a possible relationship and low level of genetic diversity.

Made available in DSpace on 2015-04-20T12:22:42Z (GMT). No. of bitstreams: 1 Lucifrancy Vilagelim Costa.pdf: 2348102 bytes, checksum: 1b6decf454525c5bf05fd0c336e86cc9 (MD5) Previous issue date: 2012-06-21 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The Capsicum peppers are used in different ways by many cultures throughout the world. In Brazil it is found wide variability of this genus and Amazon is an important center of diversity, there is wide genetic variability expressed on several characteristics of plants, flowers and fruits. Information about the diversity in germplasm collection serve to increase the efficiency of improvement works of cultivated species, through morphological characterization is possible to describe accesses that exists in collections, this is important to organize the information and let it available for use in breeding programs. To study the genetic variability of Capsicum spp. It was stablished a collection of its kind in the Faculty of Agrarian Sciences, Federal University of Amazonas, UFAM. The objective of this study was to characterize morphologically 40 accesses collected in the Amazonas. Were analyzed, with small modifications, 61 morphological descriptors recommended by the International Plant Genetic Resources Institute (IPGRI) for the genus Capsicum, including characteristics of the seedling, plant, flowers and fruits. The accesses were grown in the sector of Olericulture in the Faculty of Agrarian Sciences, Federal University of Amazonas. Using 17 descriptors, identified as essential by IPGRI, we calculated the genetic similarity among accesses using the general similarity coefficient of Gower. The accesses were grouped by the hierarchical method of distances average (UPGMA - Unweighted Pair-Group Method using an Arithmetic Average) and the graphical representation of the similarity between them was made by the method of Principal Coordinates Analysis (PCO). Of the 61 descriptors evaluated only 10 (color, hypocotyl, leaf shape, corolla length, calyx pigmentation, spots or streaks with nodal anthocyanin, peduncle length, appendix at the end of the fruit, surface of the seed, seed size and seed color) showed no variation. According to the analyzed characteristics, especially the flowers, the most important for the differentiation of species, among the 40 accesses of the collection, three species were identified, 35 of C. chinenseJacq, a genotype C. baccatum and four C. frutescens. The characteristics of the fruit showed a great variation of classes; it was identified six colors of immature fruits and six in mature stage, four shapes of fruit and great variation in size and weight of fruits. The most part of the accesses showed high pungency. It was identified two morphotypesmurupi pepper, two finger-girl, four chillies, seven eye-of-fish, thirteen sweet and twelve curabiá. The similarity coefficient ranged from 0.42 to 1.0, indicates that there are in the collection accesses totally similar and different, indicating that despite the existence of duplicates is important the maintenance of the accesses. The collection presents a high genetic variability for characteristics of plants, flowers and fruits mainly for the specie C. chinense native to the Amazon region / As pimentas do gênero Capsicum são utilizadas de diferentes formas por muitas culturas no mundo. No Brasil é encontrada ampla variabilidade desse gênero sendo a Amazônia um importante centro de diversidade, existindo uma ampla variabilidade genética expressa em diversas características de plantas, flores e frutos. Informações a respeito da diversidade em coleção de germoplasma servem para aumentar a eficiência de trabalhos de melhoramento de espécies cultivadas, por meio da caracterização morfológica é possível descrever acessos existentes nas coleções, para que as informações sejam organizadas e disponibilizadas para uso em programas de melhoramento. Para estudar a variabilidade genética de Capsicum spp. foi estabelecida uma coleção do gênero na Faculdade de Ciências Agrárias da Universidade Federal do Amazonas/UFAM. O objetivo desse trabalho foi caracterizar morfologicamente 40 acessos coletados no Amazonas. Foram analisados, com pequenas modificações, 61 descritores morfológicos recomendados pelo International Plant Genetic Resources Institute/ IPGRI para o gênero Capsicum, incluindo características da plântula, planta, flores e frutos. Os acessos foram cultivados no setor de Olericultura FCA, da UFAM. Utilizando-se 17 descritores, identificados como essenciais pelo IPGRI, foi calculada a similaridade genética entre os acessos usando o coeficiente de similaridade geral de Gower. Os acessos foram agrupados pelo método hierárquico das médias das distâncias (UPGMA Unweighted Pair Group Method Using na Arithmetic Average) e a representação gráfica da similaridade entre eles realizada pelo método da Análise de Coordenadas Principais (PCO). Dos 61 descritores avaliados apenas 10 (cor do hipocótilo, forma da folha, comprimento da corola, pigmentação do cálice, manchas ou estrias com antocianina nodal, comprimento do pedúnculo, apêndice na ponta do fruto, superfície da semente, tamanho da semente e cor da semente) não apresentaram variação. De acordo com as características analisadas, principalmente das flores, as mais importantes para a diferenciação de espécies, dentre os 40 acessos da coleção, foram identificadas três espécies, sendo 35 de C. chinense Jacq, um genótipo de C. baccatum L. e quatro de C. frutescens L. As características de fruto apresentaram grande variação de classes: foram identificadas seis cores de frutos imaturos e seis no estágio maduro, quatro formas de fruto e grande variação no tamanho e peso. A maioria dos acessos apresentou pungência alta. Foram identificadas duas pimenteiras do morfotipos murupi, duas dedo-de-moça, quatro malagueta, sete olho-de-peixe, treze de cheiro e doze curabiá. O coeficiente de similaridade variou de 0,42 a 1,0, indica que na coleção existem acessos totalmente similares e diferentes, indicando que apesar da existência de duplicatas é importante a manutenção dos acessos avaliados. A coleção apresentou uma alta variabilidade genética para características da planta, flores e de frutos principalmente para a espécie C. chinense nativas da região Amazônica

Master of Science / Department of Horticulture, Forestry, and Recreation Resources / Candice A. Shoemaker / Seed saving, whether performed by subsistence farmers, by breeders at land grant universities, or the seed industry, provide the world’s farmers with the needed supply of seeds to produce food crops annually. This thesis considers historical agricultural trends as they relate to the process of seed saving. It utilizes information gathered from primary source materials, historical monographs, and FAO and NGO publications. Chapter two discusses six universal agricultural dichotomies and how they relate to seed saving over time and globally. The dichotomies examined as they relate to seed saving include: subsistence and commercial, commons and commodity, public and private, basic and applied, global north and south, and urban and rural food production. Subsistence and commons oriented agricultural systems have historically traveled towards commercial and commodity forms in developed countries like the United States. As a result, seed saving moved from something farmers did to either public or private institutions that performed basic and applied research for genetic improvement of agricultural crops. As breeding programs looked outward for better breeding materials (germplasm) the importance of the Global North and South and Urban and Rural locales became important. Based on the results of this examination, it is clear that prioritizing subsistence practices and understanding seeds as a commonly held resource play important roles in maintaining agricultural diversity, particularly for more commercialized and commodity oriented agricultures. This shift from subsistence to commercial agriculture in the Global South jeopardizes subsistence agriculture’s ability to maintain agricultural diversity. Chapter three utilized a case study framework and focused on American seed saving within the Corn Belt from 1890 to 1950. The Corn Show, a common annual showcase of corn seed savers in the Midwest, supported both subsistence and commercial agricultural ideals. It also set the stage for the introduction of hybrid corn and suggested an alternative to the commodification of seeds by the industry. These results suggest that seed saving programs today could benefit from a culture that values subsistence practices while still utilizing the benefits of contemporary methods that are common to commodification. The added benefit of community and diversity that are realized by seed saving could develop a culture of seed production that is capable of contributing to rural development goals. This thesis concludes by tying together its discussion of dichotomies, reinforcing the connectedness between different agricultural production systems, and thus, the need for many different types of seed saving.