Dissertations / Theses on the topic 'Germplasm conservation'

The development of regional germplasm pools to conserve genetic resources from a specific region has been suggested to increase germplasm utilization and to reduce maintenance costs. However, the effects of selection on genetic variability within germplasm pools have not been documented. This research attempted to study effects of germplasm pooling and multiple-trait selection on phenotypic and genetic variability within nondormant alfalfa (Medicago sativa L.) germplasm pools. Five germplasm pools differing in geographical representation were formed from 12 Middle Eastern ecotypes based on agronomical and morphological similarities. These germplasm pools included three restricted pools, representing variability among relatively similar ecotypes, and two broader-based pools. Syn-1 seed of germplasm pools were evaluated for blue aphid (Acyrthosiphon kondoi Shinji) resistance and forage yield in saline and non-saline environments in the greenhouse. Fifty six plants (p = 12.5%) were selected using Simple Weighted Index in each pool and in the 'Hejazi' ecotype, and interpollinated to form six Cycl-1 selected populations. These populations and six randomly selected populations were evaluated both in the greenhouse, to measure response to selection, and in the field, to measure effect of selection on phenotypic variability. The initial screening study indicated that all germplasm pools had low aphid resistance and good potential for increased yield in saline and non-saline environments. No significant differences were observed in the field between selected and random populations for any of the agronomical or morphological traits evaluated. This indicates that multiple-trait selection did not affect variability for traits not targeted by selection. Response to selection for aphid resistance was significant only in the restricted pools. Response to selection for forage yield in saline and non-saline environments was highest in the most broad-based pool. Half-sib analysis among 25 families indicated that genetic variability in selected populations was dependent on the level of variability present in the base population. The results of this study indicate that development of single Arabian alfalfa germplasm pool may be adequate to conserve the genetic variability among the Arabian alfalfa ecotypes since most of the desired traits among the Arabian ecotypes are common to most ecotypes.

Thesis (M.S.)--West Virginia University, 2007.
Title from document title page. Document formatted into pages; contains vii, 48 p. : col. ill. Includes abstract. Includes bibliographical references (p. 40-42).

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.

Genetic resources are important for the development of every country and for humanity. Collection, conservation and reasonable utilization of genetic resource is required mission. Understanding the importance of genetic resource, especially rice germplasm, since 2001, Center for conservation and development of crop genetic resources (CCD-CGR) of Hanoi University of Agriculture (Vietnam National University of Agriculture) has been collected, conserved and evaluated rice germplasm from different provinces of Vietnam for breeding programs. So far, 1090 accessions of local rice of Vietnam have been collected. Evaluation of agronomic properties and screening of some important genes using DNA molecular markers have revealed that Vietnamese rice germplasm has high level diversity and containing important genes for quality and resistance for disease and pests. These genetic resources are potential materials for national breeding programs. Based on the collected germplasm, 3 new glutinous rice varieties have been successfully created with high yield and good quality. In addition, the degradation of local rice varieties is also a matter of concern. So far, 4 specialty rice varieties Deo Dang, Ble chau, Pu de and Khau dao have been successfully restored for the north provinces of Vietnam. The main results of this study are germplasms for rice breeding programs and new improved varieties that bring economic benefits to farmers and the country
Nguồn gene là tài nguyên sống còn của mỗi quốc gia và của toàn nhân loại. Vì vậy thu thập, bảo tồn, đánh giá và khai thác hợp lý nguồn tài nguyên này có ý nghĩa rất lớn. Nhận thức được tầm quan trọng của nguồn gen nhất là nguồn gen cây lúa, ngay từ đầu những năm 2000, Trung tâm bảo tồn và phát triển nguồn gene cây trồng thuộc Trường Đại học nông nghiệp, nay là Học Viện nông nghiệp Việt Nam đã tiến hành thu thập, lưu giữ, đánh giá và khai thác nguồn gene lúa. Kết quả đã thu thập, lưu giữ được 1090 mẫu giống lúa địa phương Việt Nam. Đánh giá đặc điểm nông sinh học và phát hiện một số gene quy định các tính trạng chất lượng và kháng sâu bệnh bằng chỉ thị phân tử DNA. Đây là nguồn gene quan trọng cho chọn tạo giống. Dựa vào nguồn gene thu thập được, cho đến nay, Trung tâm bảo tồn và phát triển nguồn gene cây trồng đã lai và chọn tạo được thành công 03 giống lúa nếp chất lượng cao. Ngoài ra, thoái hóa giống cũng là vấn đề đang được quan tâm. Cho đến nay 4 giống lúa đặc sản Đèo đàng, Ble châu, Pu đe và Khẩu dao đã được phục tráng và đưa vào sản xuất. Kết quả của những nghiên cứu này là ngân hàng các giống lúa làm nguồn gene để chọn tạo giống mới đem lại lợi ích kinh tế cho người nông dân và đất nước

Genetic resources are important for the development of every country and for humanity. Collection, conservation and reasonable utilization of genetic resource is required mission. Understanding the importance of genetic resource, especially rice germplasm, since 2001, Center for conservation and development of crop genetic resources (CCD-CGR) of Hanoi University of Agriculture (Vietnam National University of Agriculture) has been collected, conserved and evaluated rice germplasm from different provinces of Vietnam for breeding programs. So far, 1090 accessions of local rice of Vietnam have been collected. Evaluation of agronomic properties and screening of some important genes using DNA molecular markers have revealed that Vietnamese rice germplasm has high level diversity and containing important genes for quality and resistance for disease and pests. These genetic resources are potential materials for national breeding programs. Based on the collected germplasm, 3 new glutinous rice varieties have been successfully created with high yield and good quality. In addition, the degradation of local rice varieties is also a matter of concern. So far, 4 specialty rice varieties Deo Dang, Ble chau, Pu de and Khau dao have been successfully restored for the north provinces of Vietnam. The main results of this study are germplasms for rice breeding programs and new improved varieties that bring economic benefits to farmers and the country.
Nguồn gene là tài nguyên sống còn của mỗi quốc gia và của toàn nhân loại. Vì vậy thu thập, bảo tồn, đánh giá và khai thác hợp lý nguồn tài nguyên này có ý nghĩa rất lớn. Nhận thức được tầm quan trọng của nguồn gen nhất là nguồn gen cây lúa, ngay từ đầu những năm 2000, Trung tâm bảo tồn và phát triển nguồn gene cây trồng thuộc Trường Đại học nông nghiệp, nay là Học Viện nông nghiệp Việt Nam đã tiến hành thu thập, lưu giữ, đánh giá và khai thác nguồn gene lúa. Kết quả đã thu thập, lưu giữ được 1090 mẫu giống lúa địa phương Việt Nam. Đánh giá đặc điểm nông sinh học và phát hiện một số gene quy định các tính trạng chất lượng và kháng sâu bệnh bằng chỉ thị phân tử DNA. Đây là nguồn gene quan trọng cho chọn tạo giống. Dựa vào nguồn gene thu thập được, cho đến nay, Trung tâm bảo tồn và phát triển nguồn gene cây trồng đã lai và chọn tạo được thành công 03 giống lúa nếp chất lượng cao. Ngoài ra, thoái hóa giống cũng là vấn đề đang được quan tâm. Cho đến nay 4 giống lúa đặc sản Đèo đàng, Ble châu, Pu đe và Khẩu dao đã được phục tráng và đưa vào sản xuất. Kết quả của những nghiên cứu này là ngân hàng các giống lúa làm nguồn gene để chọn tạo giống mới đem lại lợi ích kinh tế cho người nông dân và đất nước.

Agrobiodiversity constitutes an essential resource for traditional rural populations. Home gardens are "hotspots" of agrobiodiversity and important loci of in situ conservation efforts. This study seeks to understand the factors affecting gardeners' choices and to assess the accessibility of planting material in rural communities of the Peruvian Amazon. Household surveys and garden inventories conducted in 15 villages of the Corrientes river (n = 300), and case studies in three of these villages (n = 89), allowed to describe the local and regional patterns of garden agrobiodiversity and the structure of planting material exchange networks. Analyses reveal a strong link between species diversity and both household cultural and socioeconomic characteristics, and village ethnicity and size. Planting material flows primarily through matrilineal bonds, from advice-givers to advice-seekers, from old to young and from rich to poor. Farmers with exceptional species diversity, propensity to give and/or expertise are identified and their role in the conservation of cultivated plants is assessed. Expertise is not found to be as closely related to high species diversity as expected, but knowledge and planting stock dissemination go hand-in-hand.

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Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico
Brazil is one of the major centers of diversity for polyploid cotton plants; these plants belong to the genus Gossypium, which has three known species: G. hirsutum, G. barbadense and G. mustelinum. The Northeast is the only region where the three species occur, the last group being endemic. Northeast s cotton plants can be important sources of variability for genetic breeding. It is believed that great part of local diversity is being lost, due to economic, political, cultural and agricultural problems. In an attempt to mitigate this loss and delineate conservation strategies it is necessary to know how the species are found where they occur. The objective was to characterize and determine how plants are maintained in situ in the states of Maranh?o, Piau?, Cear?, Rio Grande do Norte and Para?ba at the beginning of the XXI century. The in situ characterization of G. hirsutum and G. barbadense was conducted through structured interviews with the cotton plants owners and through the analysis of the environment. The data were collected during expeditions undertaken between the years 2004 to 2005. Twenty-two plants were collected in the state of Para?ba, forty-four in the state of Rio Grande do Norte, one hundred and forty-six in the state of Cear?, forty in the state of Maranh?o and ninety-one plants in the state of Piau?. All plants collected in the states of Para?ba and Rio Grande do Norte belonged to moco type. Moco cotton plants also predominated in the other states, representing 92%, 62% and 78% of plants collected in Cear?, Piau? and Maranh?o, respectively. The other cotton plants collected belong to the species G. barbadense. The cotton plants were found in situ as dooryard plants, roads side, feral populations, cultivation or local varieties. Great part were dooryard plants (45.2%), being major in Piau? and Maranh?o. Cultivation predominated in Cear?; in Rio Grande do Norte feral populations were the most frequent and, in Para?ba, local varieties. The maintenance of moco plants is related, mainly, to the phytotherapic domestic use (20.9%) and to confection of lamp wicks (29.7%). Few inhabitants in Para?ba, Rio Grande do Norte, Piau? and none in Maranh?o used harvest the plants, storage the seeds or gin; however, in Cear?, 40.5% of owners affirmed that they harvested and commercialized the fiber. It was found that the maintenance of species is dependent of the fragile cultural habits of local inhabitants, therefore the maintenance in situ is not a suitable way to conservation of genetic resources. The efforts must be directed to the continuity of collections, maintenance and characterization ex situ
O Brasil ? um dos importantes centros de diversidade de algodoeiros polipl?ides pertencente ao g?nero Gossypium, com 3 esp?cies conhecidas: G. hirsutum, G. barbadense e G. mustelinum. O Nordeste ? a ?nica regi?o com ocorr?ncia das tr?s esp?cies, sendo a ?ltima end?mica. Os algodoeiros desta regi?o podem ser fontes importantes de variabilidade para o melhoramento gen?tico. Acredita-se que grande parte da diversidade local esteja sendo perdida, devido a problemas econ?micos, pol?ticos, culturais e agr?colas. Na tentativa de mitigar tal perda e delinear estrat?gias de conserva??o ? necess?rio conhecer como as esp?cies se encontram no local em que ocorrem. Objetivou-se caracterizar e determinar o modo com que as plantas s?o mantidas in situ nos estados do Maranh?o, Piau?, Cear?, Rio Grande do Norte e Para?ba no inicio do s?culo XXI. A caracteriza??o in situ de G. hirsutum e G. barbadense foi realizada por meio de entrevista estruturada com o propriet?rio e pela an?lise do ambiente. Os dados foram tomados durante expedi??es empreendidas entre os anos de 2004 a 2005. Foram coletadas 22 plantas no estado da Para?ba, 44 no estado do Rio Grande do Norte, 146 no estado do Cear?, 40 no estado do Maranh?o e 91 plantas no estado do Piau?. Todas as plantas coletadas nos estados da Para?ba e Rio Grande do Norte eram do tipo moc?. O algodoeiro moc? tamb?m predominou nos demais estados, representando 92%, 62% e 78% das plantas coletadas no Cear?, Piau? e Maranh?o, respectivamente. Os demais algodoeiros coletados pertencem a esp?cie G. barbadense. Os algodoeiros moc?s foram encontrados in situ na forma de planta de fundo de quintal, beira de estrada, feral, lavoura, variedade local. Em grande parte eram do tipo fundo de quintal (45,2%), sendo maioria no Piau? e Maranh?o. No Cear? predominou o tipo lavoura, no Rio Grande do Norte tipo feral e na Para?ba variedades locais. A manuten??o das plantas do tipo moc? est? ligada, principalmente, ao uso dom?stico fitoter?pico (20,9%) e confec??o de pavios para candeeiro (29,7%). Poucos moradores na Para?ba, Rio Grande do Norte, Piau? e nenhum no Maranh?o apresentaram o h?bito de realizar a colheita, armazenamento e beneficiamento das sementes, entretanto no Cear? 40,5% dos propriet?rios afirmaram realizar a colheita e comercializar a fibra. Verificou-se que a manuten??o da esp?cie ? dependente dos fr?geis h?bitos culturais da popula??o local, portanto a manuten??o in situ n?o ? um meio adequado ? conserva??o dos recursos gen?ticos. Os esfor?os devem ser direcionados para a continuidade das coletas, caracteriza??o e manuten??o ex situ

Resumo: A construção da Usina Hidrelétrica Engenheiro Sergio Motta, na região do Pontal do Paranapanema, promoveu severos impactos aos ambientes naturais, especialmente sobre as comunidades florestais remanescentes. Com a finalidade de compensar a perda das espécies florestais na área de inundação do reservatório, desenvolveu-se por meio do convênio CESP - ESALQ/USP-IPEF, o Projeto Banco Ativo de Germoplasma (BAG), implantando-se em forma de teste de progênies, delineamento de blocos ao acaso, constituído por três repetições, contendo parcelas lineares de oito plantas de trinta progênies de cada uma das espécies eleitas para conservação. O presente estudo objetivou verificar o comportamento, variação genética e propor estratégia adequada de seleção para caracteres silviculturais de 32 espécies do Banco Ativo de Germoplasma de espécies de Floresta Estacional Semidecidual (BAG - SP), implantado em Rosana - SP e de 31 espécies conservadas no Banco Ativo de Germoplasma de espécies do Cerrado (BAG - MS), implantado em Anaurilândia - MS. Foram avaliados os caracteres DAP (diâmetro a altura do peito), altura e sobrevivência. Os resultados, obtidos por meio do programa estatístico SELEGEN-REML/BLUP, possibilitaram observar que o sistema de plantio adotado na implantação do Banco Ativo de Germoplasma da Floresta Estacional Semidecidual (BAG - SP) e do Banco Ativo de Germoplasma do Cerrado (BAG - MS), mostra-se promissor para as espécies que se encontram conservadas nestes. Aos 9,3 anos após o início da implantação o BAG - SP apresenta, em média, 67,5% de sobrevivência, DAP de 5,42 cm e 5,73 m de altura. Aos 8,3 anos após o início da implantação o BAG - MS apresenta, em média, 65% de sobrevivência, DAP de 7,51 cm e 5,79 m de altura. A variação genética presente nas populações que compõe o... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The construction of the Hydroeletric Power Plant Engenheiro Sergio Motta; in the region of Pontal do Paranapanema, caused severe impacts to natural environments, especially on the remaining forest communities. To compensate the loss of forest species in the floodplain of the reservoir, was developed, through the partnership CESP - ESALQ/ USP-IPEF, the Project Active Germplasm Bank (AGB), implemented in the form of a progeny test, designed random blocks, consisting of three replicates, containing linear parcels of eight plants from thirty progenies of each one of the elected species for conservation. This work aimed to study the genectic variation for the silvicultural characters of 32 species from the Active Germplasm Bank of species of Semideciduous Seasonal Forest (AGB - SP), implemented in Rosana - SP and 31 species conserved in the Active Germplasm Bank of Cerrado species (AGB-MS), implemented in Anaurilândia - MS. Were evaluated the DBH (diameter at breast height), height and survival. The results obtained through the statistical program SELEGEN-REML/BLUP, allowed to observe that the plantation system adopted in the implementation of the Active Germplasm Bank of the Semideciduous Seasonal Forest (AGB-SP) and the Active Germplasm Bank of Cerrado (AGB-MS) shows promise for the species that are conserved in these. At 9.3 years after the beginning of the implementation, the AGB - SP showed 67.5% of survival, DBH of 5.42 cm and 5.73 m of height and the AGB-MS, at 8.3 years after the beginning of the implementation showed 65% of survival, DBH of 7.51cm and 5.79 m of height. The genetic variation present in populations that make up the AGBSP and AGB-MS validates the ex situ conservation program that was proposed and enables the development of an improving program for these species, in order to produce improved seeds. The strategy... (Complete abstract click electronic access below)
Orientador: Mario Luiz Teixeira de Moraes
Coorientador: Marcos Deon Vilela de Resende
Banca: Marco Eustaquio de Sa
Banca: João Antonio da Costa Andrade
Banca: Flávio Bertin Gandara
Banca: Reginaldo Brito da Costa
Doutor

Petróleo Brasileiro S/A
Heart of palm (Euterpe edulis Martius) is a characteristic species of Atlantic Forest. Presently, natural populations are being drastically reduced due to human action. This system of exploitation resulted in substantial genetic erosion in the populations and did not permitted the maintenance of the demographic structure through natural regeneration. The only way of heart of palm propagation was through seeds that do not tolerate storage for a long time. Then, the development of strategies for germplasm conservation has become necessary. Heart of palm does not present a natural system of vegetative propagation, then the technique of culture of tissues showed to be adequated for the conservation and multiplication of germplasm in large scale. The information of geographic distribution of genetic resources is fundamental in the formulation of strategies of in situ conservation and also to help in the collection of vegetal material and to establish ex situ germplasm banks. The main goal of this study was to elucidate some aspects of somatic embryogenesis and analyze the in vitro germination of immature zygotic embryos of heart of palm, aiming the ex situ conservation of germplasm of this species. In the first study, zygotic embryos and leaf sheaths were the explant sources. Zygotic embryos were inoculated in MS culture medium (MURASHIGE and SKOOG, 1962) supplemented with Morel vitamins (MOREL and WETMORE, 1951) 2,4-D (0, 30, 35, 40 mg.L-1), 3 mg.L-1 of 2iP, 0.5 g.L-1 of glutamine, 0.5 g.L-1 of activated charcoal, 30 g.L-1 of glucose and 5 mg.L-1. The delineate used in this study was in Random Blocks, in that the treatments consisted of four concentrations of 2,4-D, combined with two sources of carbohydrate in factorial scheme (4x2), in six repetitions. Each parcel was constituted by two test tubes, each one with one embryo. Leaf sheaths extracted of in vitro germinated plants were inoculated in MS medium supplemented with Picloram (72.3 mg.L-1) or 2,4-D (66.3 mg.L-1), 3 mg.L-1 of 2iP, glutamine (0; 0.29; 0.58; 1.17 g.L-1), 1.5 g.L-1 de activated charcoal and 2.5 g.L-1 of Phytagel. The delineate was in Random Blocks with eight treatments and three repetitions. Each parcel was constituted by three Petri plates in ten transversal sections of leaf sheaths. Indirect somatic embryos were induced from zygotic embryos in a medium supplemented with 40 mg.L-1 of 2,4-D and 30 g.L-1 of sucrose. It was verified a high formation of leaf sheaths by using MS medium supplemented with 1.17 g.L-1 of glutamine and 66.3 mg.L-1 of 2,4-D. In the second study, the objective was to elucidate the influence of saline concentration and of sucrose in the culture medium during the germination of zygotic embryos and to study the influence of the addition of different concentration of Calcium chloride to the MS medium of culture in the induction of somatic embryogenesis from immature zygotic embryos of E. edulis. The evaluation of germination in vitro of zygotic embryos of E. edulis occurred through the inoculation in medium of culture added by Morel vitamins, 7 g.L-1 of agar and 1.5 g.L-1 of activated charcoal. The treatments were different concentrations of the mediums MS (MS and MS/2) combined with different concentrations of sucrose (20, 30 and 40 g.L-1). It was used the Random Blocks Delineate, with six repetitions, each one constituted by five test tubes with one embryo per tube. For the induction of somatic embryogenesis, immature zygotic embryos were extracted and inoculated in a culture medium MS supplemented with Morel Vitamins, 7 g.L-1 of agar, 0.5 g.L-1 of glutamine, 3 mg.L-1 of 2iP, 100 mg.L-1 of 2,4-D and 1.5 g.L-1 of activated charcoal and different concentrations of calcium chloride (0, 2, 4, 8, 12 mM). It was used the statistical delineate in Random Blocks with six repetitions. Each parcel was constituted by one flask containing four immature zygotic embryos. For the conversion of the somatic embryos in seedlings it was used the complete medium of culture MS and the medium with the half of original saline concentration (MS/2). It was verified that the concentration was not affected by the saline concentration of the medium culture and of sucrose. But the growing in height and the production of fresh mass of the seedlings were affected by the treatments. The increasing of concentration of sucrose of 20 to 40 g.L-1 in the medium culture resulted in an increasing in the fresh mass of the seedlings. The composition of the different mediums of culture did not influence the mean number of roots per seedling of heart of palm. The induction of somatic embryogenesis in immature zygotic embryos was not significantly influenced by the addition of different concentrations of calcium chloride to the medium, after 60 days. But, they were observed significant differences among the concentrations of Calcium chloride for the number of somatic embryos formed, after 150 days. The increasing on the concentration of Calcium chloride in the medium of culture resulted in a decrease in the mean number of somatic embryos produced per experimental unit. Both medium MS and MS/2 did not differ significantly in the capacity of germination of the somatic embryos. The process of induction of somatic embryogenesis in the immature zygotic embryos of E. edulis occurred directly from the cotyledonary node of the embryo. The present study evidenced that the supplementation of the medium of culture (MS or MS/2) with sucrose (30 or 40 g.L-1) was necessary for the growing of the seedlings of heart of palm originated from immature zygotic embryos. The results confirmed the possibility to propagate the palm E. edulis through direct somatic embryogenesis, because they were produced complete seedlings, and the importance of supplementation of the medium of culture with a source of organic nitrogen in the in vitro morphogenesis of leaf sheaths of heart of palm.
O palmiteiro (Euterpe edulis Mart.), uma espécie característica da Mata Atlântica, foi drasticamente reduzido, devido à ação antrópica em diversas populações. A exploração indiscriminada resultou em erosão genética nas populações, não permitindo mais a manutenção da estrutura demográfica, através da regeneração natural. A única forma de propagação do palmiteiro é através de produção de sementes, as quais não toleraram armazenamento por longos períodos. Desta maneira, o desenvolvimento de estratégias para a conservação de germoplasma tornou-se necessário. Como o palmiteiro não apresenta um sistema de propagação vegetativa natural, a técnica de cultura de tecidos mostrou-se adequada tanto para a conservação, quanto para a multiplicação em larga escala de germoplasma. A informação da distribuição geográfica dos recursos genéticos tem sido fundamental na formulação de estratégias de conservação in situ, bem como no auxílio para a coleta de material vegetal e estabelecimento de bancos de germoplasma ex situ. O objetivo geral do presente estudo foi elucidar alguns aspectos da embriogênese somática, e analisar a germinação in vitro de embriões zigóticos imaturos de palmiteiro, visando à conservação ex situ de germoplasma de palmiteiro. No primeiro estudo, embriões zigóticos e bainhas foliares serviram como fonte de explantes. Embriões zigóticos foram inoculados em meio MS (MURASHIGE e SKOOG, 1962), suplementado com as vitaminas de Morel (MOREL e WETMORE, 1951), 2,4-D (0, 30, 35, 40 mg.L-1), 3 mg.L-1 de 2iP, 0,5 g.L-1 de glutamina, 0,5 g.L-1 de carvão ativado, 30 g.L-1 de glicose ou sacarose e, geleificado com 5 g.L-1 de ágar. O delineamento experimental foi em Blocos ao acaso, em que os tratamentos constaram de quatro concentrações de 2,4-D, combinados com duas fontes de carboidrato em esquema fatorial (4x2), em 6 repetições. Cada parcela foi constituída por dois tubos de ensaio, com um embrião cada. Bainhas foliares, extraídas de plântulas germinadas in vitro, foram inoculadas em meio MS, suplementado com Picloram (72,3 mg.L-1) ou 2,4-D (66,3 mg.L-1), 3 mg.L-1 de 2iP, glutamina (0; 0,29; 0,58; 1,17 g.L-1), 1,5 g.L-1 de carvão ativado, sendo geleificado com 2,5 g.L-1 de Phytagel. O delineamento foi em Blocos ao acaso, com 8 tratamentos, em 3 repetições. Cada parcela foi constituída por três placas de Petri, com 10 secções transversais das bainhas foliares. Embriões somáticos indiretos foram induzidos a partir de embriões zigóticos, em meio suplementado com 40 mg.L-1 de 2,4-D e 30 g.L-1 de sacarose. Verificou-se elevada formação de calos em bainhas foliares, utilizando o meio MS suplementado com 1,17 g.L-1 de glutamina e 66,3 mg.L-1 de 2,4-D. No segundo estudo, o objetivo foi elucidar a influência da concentração salina e da sacarose no meio de cultura, durante a germinação de embriões zigóticos, e estudar a influência da adição de diferentes concentrações de Cloreto de cálcio ao meio de cultura MS, na indução de embriogênese somática, em embriões zigóticos imaturos de E. edulis. A avaliação da germinação in vitro de embriões zigóticos de E. edulis ocorreu através da inoculação, em meio de cultura adicionado das vitaminas de Morel, 7 g.L-1 de ágar e 1,5 g.L-1 de carvão ativado. Os tratamentos foram diferentes concentrações do meio MS (MS e MS/2), combinados com diferentes concentrações de sacarose (20, 30 e 40 g.L-1). Foi utilizado o delineamento em Blocos ao acaso, em 6 repetições, cada repetição foi constituída por cinco tubos de ensaio, contendo um embrião cada. Para a indução de embriogênese somática, embriões zigóticos imaturos foram extraídos e inoculados em meio de cultura MS, suplementado com as vitaminas de Morel, 7 g.L-1 de ágar, 0,5 g.L-1 de glutamina, 3 mg.L-1 de 2iP, 100 mg.L-1 de 2,4-D e 1,5 g.L-1 de carvão ativado, além de diferentes concentrações de Cloreto de cálcio (0, 2, 4, 8, 12 mM). Utilizou-se o delineamento estatístico em Blocos ao acaso, em 6 repetições. Cada parcela foi constituída por um frasco, contendo quatro embriões zigóticos imaturos. Para a conversão dos embriões somáticos em plântulas, foram empregados o meio de cultura MS completo, e o com a metade da concentração salina original (MS/2). Verificou-se que a germinação de embriões zigóticos de palmiteiro não foi afetada pela concentração salina do meio de cultura e de sacarose. Entretanto, o crescimento em altura e a produção de massa fresca das plântulas foram afetados pelos tratamentos. O aumento da concentração de sacarose, de 20 para 40 g.L-1, no meio de cultura, resultou em acréscimo na massa fresca das plântulas. A composição dos diferentes meios de cultura não influenciou o número médio de raízes por plântula de palmiteiro. A indução de embriogênese somática em embriões zigóticos imaturos de E. edulis não foi influenciada significativamente pela adição de diferentes concentrações de Cloreto de cálcio ao meio de cultura, aos 60 dias. Entretanto, foram observadas diferenças significativas entre as diversas concentrações de Cloreto de cálcio, quanto ao número de embriões somáticos formados, aos 150 dias. O aumento na concentração de Cloreto de cálcio, no meio de cultura, resultou em decréscimo no número médio de embriões somáticos de palmiteiro produzidos. Os meios de cultura MS e MS/2 não diferiram significativamente, quanto à capacidade de germinação dos embriões somáticos. O processo de indução de embriogênese somática, em embriões zigóticos imaturos de E. edulis, ocorreu diretamente a partir do nó cotiledonar. O presente estudo evidenciou que a suplementação do meio de cultura (MS ou MS/2) com sacarose (30 ou 40 g.L-1) foi necessária para o desenvolvimento das plântulas de palmiteiro, oriundas de embriões zigóticos imaturos. Resultados confirmaram a possibilidade de propagar a palmeira E. edulis através da embriogênese somática direta, pois foram produzidas plântulas completas, além de demonstrarem a importância da suplementação do meio de cultura com uma fonte de nitrogênio orgânico, na morfogênese in vitro de bainhas foliares de palmiteiro.

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A construção da Usina Hidrelétrica Engenheiro Sergio Motta, na região do Pontal do Paranapanema, promoveu severos impactos aos ambientes naturais, especialmente sobre as comunidades florestais remanescentes. Com a finalidade de compensar a perda das espécies florestais na área de inundação do reservatório, desenvolveu-se por meio do convênio CESP – ESALQ/USP-IPEF, o Projeto Banco Ativo de Germoplasma (BAG), implantando-se em forma de teste de progênies, delineamento de blocos ao acaso, constituído por três repetições, contendo parcelas lineares de oito plantas de trinta progênies de cada uma das espécies eleitas para conservação. O presente estudo objetivou verificar o comportamento, variação genética e propor estratégia adequada de seleção para caracteres silviculturais de 32 espécies do Banco Ativo de Germoplasma de espécies de Floresta Estacional Semidecidual (BAG – SP), implantado em Rosana – SP e de 31 espécies conservadas no Banco Ativo de Germoplasma de espécies do Cerrado (BAG - MS), implantado em Anaurilândia – MS. Foram avaliados os caracteres DAP (diâmetro a altura do peito), altura e sobrevivência. Os resultados, obtidos por meio do programa estatístico SELEGEN-REML/BLUP, possibilitaram observar que o sistema de plantio adotado na implantação do Banco Ativo de Germoplasma da Floresta Estacional Semidecidual (BAG – SP) e do Banco Ativo de Germoplasma do Cerrado (BAG - MS), mostra-se promissor para as espécies que se encontram conservadas nestes. Aos 9,3 anos após o início da implantação o BAG – SP apresenta, em média, 67,5% de sobrevivência, DAP de 5,42 cm e 5,73 m de altura. Aos 8,3 anos após o início da implantação o BAG – MS apresenta, em média, 65% de sobrevivência, DAP de 7,51 cm e 5,79 m de altura. A variação genética presente nas populações que compõe o...
The construction of the Hydroeletric Power Plant Engenheiro Sergio Motta; in the region of Pontal do Paranapanema, caused severe impacts to natural environments, especially on the remaining forest communities. To compensate the loss of forest species in the floodplain of the reservoir, was developed, through the partnership CESP – ESALQ/ USP-IPEF, the Project Active Germplasm Bank (AGB), implemented in the form of a progeny test, designed random blocks, consisting of three replicates, containing linear parcels of eight plants from thirty progenies of each one of the elected species for conservation. This work aimed to study the genectic variation for the silvicultural characters of 32 species from the Active Germplasm Bank of species of Semideciduous Seasonal Forest (AGB – SP), implemented in Rosana – SP and 31 species conserved in the Active Germplasm Bank of Cerrado species (AGB-MS), implemented in Anaurilândia – MS. Were evaluated the DBH (diameter at breast height), height and survival. The results obtained through the statistical program SELEGEN-REML/BLUP, allowed to observe that the plantation system adopted in the implementation of the Active Germplasm Bank of the Semideciduous Seasonal Forest (AGB-SP) and the Active Germplasm Bank of Cerrado (AGB-MS) shows promise for the species that are conserved in these. At 9.3 years after the beginning of the implementation, the AGB – SP showed 67.5% of survival, DBH of 5.42 cm and 5.73 m of height and the AGB-MS, at 8.3 years after the beginning of the implementation showed 65% of survival, DBH of 7.51cm and 5.79 m of height. The genetic variation present in populations that make up the AGBSP and AGB-MS validates the ex situ conservation program that was proposed and enables the development of an improving program for these species, in order to produce improved seeds. The strategy... (Complete abstract click electronic access below)

Corn is one of the most cultivated cereal in Brazil, making the country's third largest producer of culture. Due to their genetic diversity, which allows its adaptation to different environments. Among the cultivars, are local, traditional or creole called (CLTCs), which are an alternative for small farms and also constitute an important source of genetic resistance and tolerance to different types of stress and adaptation to different environments and local managements. This study aimed to evaluate the adaptability and phenotypic stability of 15 Land varieties conserved in Family Seed Banks in Ibarama - RS as well, estimate genetic parameters in sib progenies derived from a field of three recombination Creole cultivars corn. Evaluation of adaptability and stability was performed in two consecutive years and in three areas, totaling six environments, using the randomized block design with three replications of two lines of 5m spacing of 0.9 m between rows and 0.2 m between plants. Productivity data were evaluated by the proposed Eberhart and Russell (1966) method. Among the cultivars stood out Cinquentinha with adaptability to harsh environments and to Sertanejo favorable and the others were considered wide adaptability. As for stability, all landraces cultivars showed high stability and predictability of production. For estimation of genetic parameters, three cultivars of yellow endosperm in order to reduce plant height were recombined. The progenies were recombined into three environments in randomized block design, with rows of 5m length and spacing of 0.9 m between rows and 0.2 m between plants. Heritability, phenotypic variance, genotypic and environmental and Cve / CVG: characteristics such as plant height, plant height and ear height and productivity for each of these characteristics were estimated using the GENES (Cruz, 1997) the following genetic parameters were evaluated program . The selection in sib progenies of creole maize in order to reduce the size of the plant is related to the selection of plants with lower ear height and these progenies have potential and wide genetic variability for the improvement of the characters plant height, height of spike and grain yield in the present study, by genetic selection gains can be expected.
O milho é um dos cereais mais cultivados no Brasil, fazendo com que o país seja o terceiro maior produtor mundial da cultura. Devido a sua diversidade genética, que permite sua adaptação a diferentes ambientes. Dentre as cultivares utilizadas, estão as denominadas locais, tradicionais ou crioulas (CLTCs), que constituem uma alternativa para pequenas propriedades rurais e constituem, também, uma importante fonte genética de tolerância e resistência para diferentes tipos de estresse e de adaptação aos variados ambientes e manejos locais. O presente trabalho teve como objetivo avaliar a adaptabilidade e a estabilidade fenotípica de 15 cultivares crioulas conservadas em Bancos Familiares de Sementes em Ibarama - RS, bem como, estimar parâmetros genéticos em progênies de meios irmãos oriundas de um campo de recombinação entre três cultivares crioulas de milho. A avaliação da adaptabilidade e estabilidade foi realizada em duas safras consecutivas e em três áreas, totalizando seis ambientes, utilizando-se o delineamento blocos ao acaso com três repetições de duas linhas de 5m com espaçamento de 0,9m entre fileiras e 0,2m entre plantas. Os dados de produtividade foram avaliados pelo método proposto por Eberhart e Russel (1966). Dentre as cultivares destacaram-se Cinquentinha com adaptabilidade a ambientes desfavoráveis e Sertanejo para favoráveis e as demais foram consideradas de adaptabilidade ampla. Já para estabilidade, todas as cultivares crioulas apresentaram alta estabilidade e previsibilidade de produção. Para a estimação de parâmetros genéticos, foram recombinadas três cultivares de endosperma amarelo com objetivo de reduzir a altura de planta. As progênies foram recombinadas em três ambientes no delineamento blocos ao acaso, com linhas de 5m de comprimento e espaçamento de 0,9m entre fileiras e 0,2m entre plantas. Foram avaliadas características como altura de planta, altura de inserção da espiga e produtividade e para cada uma dessas características foram estimadas com auxílio do programa GENES (CRUZ, 1997) os seguintes parâmetros genéticos: herdabilidade, variâncias fenotípicas, genotípicas e ambientais e Cve/Cvg. A seleção em progênies de meios irmãos de milho crioulo com o objetivo de reduzir o porte de planta está relacionada à seleção de plantas com menor altura de espiga e essas progênies apresentam potencial e ampla variabilidade genética para o melhoramento dos caracteres altura de planta, altura de espiga e produtividade de grãos, avaliados no presente estudo, podendo ser esperados ganhos genéticos mediante seleção.

Dissertação apresentada à Escola de Ciências Agrárias e Veterinárias - Departamento de Agronomia - da Universidade de Trás-os-Montes e Alto Douro, como requisito para a obtenção do grau de Mestre em Engenharia Agronómica.
Cryopreservation is a recent tools that has enormous potential in all aspects of human life. With this tool, the limitations for preserving genetic material become almost impossible to grasp. With this dissertation there in an attempt to understand better how this new technology works and how to put it to good use in practical manners regarding honey bee conservation. This dissertation has two parts, the theoretical part and the practical part. The practical part was only possible to do in Länderinstitut für Bienenkunde, Berlin. When it comes to results found in this dissertation, data suggests that there is a better time for the collection of semen samples that obtain better results when put through cryopreservation protocols. We can also affirm that there is a possibility of sending semen samples by mail. The last conclusion regarding sperm analyses is that, there is no relationship between inbreeding factors and semen quality. Regarding the RNA experiments, even though they present to be a solid foundation for later experiments, by their preliminary nature, there is no definitive conclusion to be taken from them.
A criopreservação é uma ferramenta relativamente recente com um enorme potencial em todos os aspetos da vida humana. Com esta ferramenta, as limitações no que toca a preservação de material genético tornam-se impossíveis de visualizar. Com esta dissertação á uma tentativa de melhor entender como esta tecnologia funciona e como a por em pratica no que toca a preservação da abelha. Esta dissertação é constituída por duas partes, a parte teórica e a parte pratica. A parte pratica teve de ser realizada Länderinstitut für Bienenkunde em Berlim. No que toca aos resultados desta experiencia, os dados recolhidos sugerem existe uma altura em que a recolha de amostra de sémen se traduz em melhores resultados quando exposta a protocolos de criopreservação. Também podemos afirmar que existe a possibilidade de transporte do sémen através do correio. Por último, podemos concluir não existe uma ligação entre factor de consanguinidade e qualidade do sémen. Relativamente aos ensaios com recurso a RNA, podemos afirmar que estes constituem uma base sólida para experiências futuras embora, não seja possível retirar conclusões definitas devido á sua natureza extremamente preliminar do estudo.

Dissertation submitted in compliance with the requirements for the Master's Degree in Technology: Biotechnology, Technikon Natal, 2002.
Lack of suitable techniques for field collection of the germplasm of different species, and spoilage of samples in transit, hinder efforts to collect, conserve, distribute and regenerate most plant germplasm in vitro. The aims of this investigation, therefore, were to address problems encountered in collection of field germplasm from species and hybrids of Eucalyptus (TAG5, TAGI4, ZG14, GC550 and GU2IO) that are propagated vegetatively and Trichilia dregeana, which has recalcitrant seeds. Simple in vitro culture-based protocols were developed to minimise contamination and maintain viability of plant material for sufficient time for it to be transported from the field to the tissue culture laboratory. From the two simulations of 48 h 'transportation' conditions for explants of Eucalyptus species investigated, those in bottles containing sterile vermiculite exhibited no contamination and greater than 50% bud break, regardless of whether or not field surface sterilization with alcohol had been done. In contrast, when explants were enclosed in cling wrap, contamination was high and bud break levels low. For selection of the more suitable Eucalyptus explant, two types were investigated: nodal explants each with one half leaf (type 1) and stem segments with three nodes (type 2). As type 2 showed considerably better shoot yields (up to 1624 shoots per 100 explant), and were more practical to use with respect to space, such trinodal stem segments were deemed best for collection. Of the sterilization procedures investigated, treatment with 70% (v/v) alcohol prior to storage was found to be most suitable in almost all cases. For plant material with high endogenous microbial contamination, the bud break medium was supplemented with Benomyl and calcium hypochlorite, each at 0.5 and 1.0 g.r). Alcoholtreated, stored explants cultured on bud break medium with 1.0 g.r) calcium hypochlorite exhibited low levels of contamination and an increased final yield (up to a maximum of 930 shoots per 100 explants). Thus, this protocol was employed for field material of E. grandis clones TAG5, TAGI4 and ZGI4. For these clones, stored type 2 explants
M

The trees belonging to the genus Podocarpus, of which only four species are native to South Africa, are renowned for their superior quality timber. Prior to 1880, Podocarpus henkelii, together with P. falcatus and P. latifolius, played a significant role in the development of the country as they were heavily utilised as timber trees for the building of dwellings, furniture and other necessary items. Due to this over-exploitation in the timber trade, all the Podocarpus species in South Africa have been afforded a ‘Protected’ status on the IUCN red data list of species that are either threatened or in danger of extinction. However, despite the obvious need to conserve the threatened genetic diversity of these species, few attempts (aside from in vitro micropropagation) have been made to explore ex situ Podocarpus germplasm conservation in the long-term. Consequently, the primary aim of this study was to establish a protocol for the long-term conservation of germplasm of Podocarpus henkelii Stapf ex Dallim. Jacks. The seeds of Podocarpus henkelii exhibit recalcitrant behaviour and can therefore not be stored in conventional seed banks. This has necessitated the investigation of alternative methods of germplasm conservation with a focus on cryopreservation which is presently considered the most reliable, efficient and cost-effective means of storing the genetic resources of recalcitrant-seeded species for prolonged periods. The first objective of this study was to investigate the effect of slow (two-step) and ultra-rapid cooling on the post-thaw survival of variously treated P. henkelii embryos. The results of this investigation revealed that the rate of cooling employed had a significant effect on explant viability as none of the precultured, cryoprotected embryos that were slowly cooled survived cryostorage while some of the preconditioned embryos responded to ultra-rapid cooling (i.e. 36% shoot production and 88% callus formation). For ultra-rapid cooling, it was found that flash-drying prior to cooling was a prerequisite for survival as osmotic dehydration alone did not effectively prepare the tissues for the stresses imposed during cryostorage. Furthermore, for those flash drying intervals that yielded positive results, preconditioning explants with 10% glycerol proved the most effective pre-cooling treatment. However, due to the low recovery numbers after ultra-rapid cooling, a third cryopreservation technique i.e. cryogenic vitrification, was investigated. For cooling by vitrification, data obtained from preliminary experiments showed that precultured explants needed to be initially loaded with 18% sucrose (w/v) + 14% glycerol (v/v) for 20 min and subsequently immersed in Plant Vitrification Solution 3 (PVS3) at 0°C for 10 min prior to cooling. However, relatively low success was achieved for P. henkelii embryos cooled by vitrification as the highest post-cooling survival obtained was only 20% germination, 27% shoot formation and 37% callus formation. Due to the low post-thaw survival obtained despite the rigorous manipulations employed in the development of the slow cooling, ultra-rapid cooling and vitrification protocols, it was decided that an alternative explant should be investigated for the conservation of P. henkelii germplasm. The explant of choice was adventitious buds induced to form on, and subsequently excised from, mature P. henkelii embryos. The first objective was to develop a suitable protocol for the induction of adventitious buds on P. henkelii embryos. The medium that induced in the highest percentage of embryos (85%) to form adventitious buds consisted of Douglas-fir cotyledon revised (DCR) basal medium supplemented with 30 g L-1 sucrose, 0.05 mg L-1 NAA, 0.5 mg L-1 BA and 6 g L-1 agar. This medium also resulted in the highest average number of buds formed per embryo (i.e. 35 ± 3 buds per embryo). Once the adventitious bud induction medium was developed, it was necessary to optimise the size of adventitious bud clumps to be used as explants for cryopreservation. Three bud clump sizes were investigated: ca 3, 5 and 10 buds per clump. However, none of the bud clumps survived excision from the mother-tissue despite the investigation of three different types of bud-break media. The resultant tissue mortality is suggested to have occurred because the adventitious bud clumps were excised prior to bud break and shoot development which could have exacerbated excision-related cellular and sub-cellular damage. It was therefore decided that attempts should be made to induce adventitious buds directly on P. henkelii embryos post-cooling, thereby eliminating the possibility of potentially lethal excision-related damage. The protocols that yielded the best results after ultra-rapid cooling and cooling by vitrification were used in this experiment. For ultra-rapid cooling, embryos were first cryoprotected with 5% followed by 10% glycerol for 1 h in each and subsequently flash dried for 30 min prior to immersion in nitrogen slush. For cooling by vitrification, embryos that were first precultured on 0.3 M sucrose for 1 d were loaded with 10% glycerol + 14% sucrose (LS4). The loaded explants were then immersed in ice-cold PVS3 and maintained on ice for 10 min prior to cryostorage. The effect of each pretreatment (either independently or in combination) on adventitious bud production pre-cooling was also investigated. For both protocols the various pretreatments decreased not only the capacity of the embryos to form buds but also the average number of buds formed per embryo (i.e. 7 ± 2 buds per embryo and 14 ± 2 buds per embryo were formed on treated embryos prior to ultra-rapid cooling and cooling by vitrification, respectively). Thus, it was predicted that even if the percentage of cryopreserved embryos forming buds was minimal, the number of possible plantlets that could be regenerated from adventitious buds per cryopreserved explant would compensate for the low recovery of embryos post-cooling. However, none of the embryos that were cryopreserved by either ultra-rapid cooling or by vitrification formed adventitious buds after eight weeks in culture. The very restricted success achieved in this study despite the investigation of three cryopreservation techniques and two different explants only serves to reinforce the difficulties associated with the conservation of recalcitrant germplasm. The large size and structural complexity of P. henkelii embryos, coupled with their high water content post-shedding, are just some of the characteristics to which their intractability to the manipulations involved in the development of a successful cryopreservation protocol could be attributed. For future investigations, development of adventitious buds produced on cryopreserved root segments (as opposed to entire roots), and/or use of seedling meristems as explants which might be amenable to cryopreservation are suggested as possible avenues for the long-term conservation of P. henkelii genetic diversity.
Thesis (M.Sc.)-University of KwaZulu-Natal, Westville, 2012.

The zygotic germplasm of plant species producing desiccation-sensitive seeds can be conserved in the long-term only by cryopreservation. Usually the embryonic axis is excised from the cotyledons and is used as the explant for cryopreservation as it is small and provides a large surface area:volume ratio. However the shoot of the axis of most species studied does not develop after excision, with the result that survival after cryopreservation is often recorded as callus production or simply explant enlargement and/or greening. Thus, besides explant size, factors such as in vitro regeneration techniques, physical injury induced upon excision and developmental status of the seed could compromise the success of cryopreservation. This study investigated the effect of the factors mentioned above, with particular attention to the developmental status of the seeds on explant in vitro development (section 3.1), response to dehydration (section 3.2) and cryopreservation of the desiccation-sensitive embryonic axes (section 3.3) of two species: Trichilia dregeana, T. emetica and embryos of a third, Strychnos gerrardii. For all three species, investigations were conducted on the embryonic axes/embryos excised from mature seeds immediately after fruit harvesting and from mature seeds stored under hydrated conditions for different periods (in order to achieve different degrees of development). In addition, preliminary studies were carried out on axes of T. dregeana to assess whether generation of reactive oxygen species (ROS) occurs in response to wounding upon axis excision (section 3.4). Excised embryonic axes of T. dregeana and T. emetica did not develop shoots in vitro unless the explants included attached cotyledonary segments. Following the development associated with short-term storage, however, the excised axes could develop shoots after complete cotyledon excision. The embryos from the (endospermous) seeds of S. gerrardii which included the paper-thin cotyledons, developed normally in vitro, with percentage germination increasing with seed storage time. For all three species, in vitro axis germination was promoted when activated charcoal was included in the germination medium, regardless of the developmental stage of the seeds. When dehydrated to approximately 0.3 g H2O g-1 dry mass (g g-1), embryonic axes from all three species failed to develop shoots even though a minimum of 50% produced roots in all cases. Hence, shoot production was shown to be more sensitive to desiccation than was root production. Furthermore, the sensitivity of the shoot apical meristem to desiccation was not ameliorated with seed storage for T. dregeana and T. emetica, but did decrease for S. gerrardii when seeds were stored for 6 – 8 weeks. The application of certain cryoprotectants did facilitate production of shoots after dehydration by a few axes of both Trichilia spp. For T. dregeana explants, combination of glycerol and sucrose allowed for 10% of the axes to retain the ability for shoot production after dehydration while for T. emetica explants, the combination of DMSO and glycerol (10 - 20% shoot production after dehydration) was best. The efficacy of the cryoprotectants was not influenced by storage period. The provision of cryoprotectants still needs to be tested for S. gerrardii. Survival of subsequent cryopreservation of T. dregeana and S. gerrardii explants was best achieved with rapid cooling in nitrogen slush, with the cooling procedure for T. emetica explants still to be optimized. The highest post-cryopreservation survival of T. dregeana axes was achieved when seeds had been stored for three months, while the seed storage period did not affect post-thaw survival of the axes of T. emetica or S. gerrardii. A small proportion of S. gerrardii explants only, produced shoots after cryopreservation, whereas the surviving embryonic axes of T. dregeana and T. emetica regenerated only as non-embryogenic callus. Although callus production is less desirable than successful seedling establishment, it has the potential for micropropagation if embryogenicity can be induced. Ultrastructural examination of the shoot apical meristem of T. dregeana after a 3-d recovery period, following excision, revealed considerable cellular derangement, although damage of individual organelles could not be resolved microscopically. Preliminary studies on T. dregeana involving a colorimetric assay using epinephrine, confirmed the generation of ROS in response to wounding associated with axis excision. Reactive oxygen species generated appeared to persist over prolonged periods rather than occurring only as a single oxidative burst. Hence, ROS production at the wound site could be the primary factor contributing to lack of shoot development. Axes immersed in the anti-oxidant, ascorbic acid (AsA) immediately after excision, showed lower ROS production and 10% shoot development when cultured in vitro, indicating that the oxidative burst coincident with, and after excision might be counteracted if immediate ROS production can be adequately quenched. Future investigations should aim to identify the specific ROS associated with wounding and optimize an anti-oxidant treatment(s) that will facilitate shoot development. Thus, the successful cryopreservation of the germplasm of the species tested, and others producing recalcitrant seeds, depends on a spectrum of species-specific factors, some still to be elucidated.
Thesis (M.Sc.)-University of KwaZulu-Natal, Westville, 2007.

Indigenous communities have developed a wealth of knowledge, which plays a crucial role in providing leads for the use of genetic resources and bioprospecting. However, such knowledge is under increasing threat due to the misappropriation of the biological resources and associated traditional knowledge of indigenous communities, through both bioprospecting, as well as the inappropriate exercise of intellectual property rights. The internationally agreed Convention on Biological Diversity (CBD) attempts to provide a bulwark against biopiracy and although it assists indigenous communities to regain some control, the CBD has proven inadequate in the protection of the traditional knowledge of indigenous communities. The subsequent Nagoya Protocol on Access to Genetic Resources and the Fair and Equitable Sharing of Benefits Arising from their Utilization to the Convention on Biological Diversity (Nagoya Protocol) attempts to address some of these limitations but unfortunately has its own shortcomings, as it was largely concluded on the basis of a compromise between developed and developing countries. This dissertation will undertake a critical analysis of the provisions of the CBD and Nagoya Protocol, with a view to establishing the level of protection these instruments afford indigenous communities. It will be shown that notwithstanding the drawbacks of both the CBD and Nagoya Protocol, they nevertheless represent major achievements in the journey to protect the genetic resources and associated traditional knowledge of indigenous communities. It is in this context that this dissertation will analyse South Africa’s Access and Benefit Sharing (ABS) regime in relation to the protection it affords indigenous communities and in the light of the implementation challenges that such legislation presents. A particular focus will be on whether South Africa’s ABS legislation complies with the country’s international obligations relating to the protection of indigenous communities and whether South Africa’s approach to the protection of the genetic resources and associated traditional knowledge of indigenous communities, in the context of bioprospecting, is adequate or whether there exists potential for its enhancement.
Thesis (LL.M.)-University of KwaZulu-Natal, Durban, 2013.

M.Sc.
Climate change, loss of habitat and over-exploitation of natural resources as well as the introduction of invasive alien species through human activities are resulting in an ever increasing risk of extinction of many plant and animal species. There are two major approaches to conserving threatened and endangered species. Firstly the large scale preservation of natural habitat and ecological processes, thereby protecting the species inhabiting the habitat. The second approach involves the ex-situ breeding of rare and endangered species. It is estimated that in the next 200 years approximately 800 mammalian species will require the assistance of breeding programs to ensure long term genetic viability. Biological Resource Banks (BRB) can potentially contribute to this challenge by providing a source of genes that can be used to counter the effects of external selection pressures, genetic drift and inbreeding depression in small or fragmented populations. These banks commonly contain biological materials such as cryopreserved sperm, embryos and cell cultures mainly as genetic and research resources. . Biological resource banks can potentially use these cryopreserved gametes together with assisted reproductive technologies (ART), such as artificial insemination (AI), in vitro fertilisation (IVF), embryo transfer (ET), intracytoplasmic sperm injection (ICSI) and nuclear transfer (NT) to maintain genetic heterogeneity in ex-situ and wild populations. Ascertaining the appropriate protocols for developing the ARTs necessary for non-domestic species is one of the major challenges faced by reproductive physiologists. Typically, there is very little available information about the processing of semen, the effects of diluents, concentration and type of cryoprotectants and freeze-thaw methods for sperm samples of non-domestic species. Procedures proven to be highly effective in humans and laboratory or domestic species, are frequently adopted and modified for use in related wildlife species. It is thus necessary to gain knowledge of the reproductive physiology of wildlife species in order to define effective protocols for the cryopreservation of biomaterials which assists in the conservation of South Africa‘s diverse wildlife species. Sperm quality assessment is a useful tool for assessing the reproductive health of free-ranging populations as well as for selecting individuals for future assisted reproduction programs.