The banana is one of the most consumed fruits in the world and is widely consumed in Brazil, but diseases such as yellow and black Sigatoka have been reducing its production, most of ali because of lack of proper production technology. The tissue culture, specifically the micropropagation, is an alternative for the production of seedlings of banana with phitosanitarium and vegetative quality, but presents factors that difficult its application. Among which stands out contamination by fungi and bactéria, associated with oxidation of the explants by the exudation of phenolic compounds that cause rapid browning and death of apical shoots, resulting in great loss in the initial phase of the explants establishment. The objective of this work was to adapt and/or optimize the behavior of different banana cultivates, resistant to diseases, via micropropagation, adjusting the in vitro culture establishment phases, through the control of oxidation and contamination, proliferation / multiplication of shoots and rooting, for the rapid multiplication of plants with superior quality, as much in the phitosanitarium aspect as in the vegetative. The cultivates Bucaneiro (AAAA), Caipira (AAA), Fhia 18 (AAAB), BRS Garantida (AAAB), Japira (AAAB), Pacovan Ken (AAAB), PA 4244 (AAAB), Preciosa (AAAB), PV 03-76 (AAAB), Thap Maeo (AAB) and Tropical (AAAB) proceeding from the city of Baião in the state of Pará and Caipira (AAA), BRS Caprichosa (AAAB), Pacovan Ken (AAAB), Preciosa (AAAB), PV 03- 76 (AAAB), Thap Maeo (AAB) proceeding from the city of Belém in the state of Pará, were subjected to different micropropagation stages. In the study was used the streptomycin sulfate antibiotic and fungicide to reduce contamination in vitro caused by bactéria and fungi, besides the anti-oxidant PVP (polivinilpirrolidona) to control the oxidation. There was contamination reduction with the use of streptomycin sulfate in the concentration of 100 mg.L1 and of oxidation with PVP at 4 gL1. At the stage of proliferation / multiplication of shoots, the different cultivates showed means ranging from 1,67 to 7,0 shoots / explant, with greater proliferation in the initial culture (2,0 to 7,0 shoots / explant).The cultivate Caipira (AAA) stood out from the others with the highest rate of shoot multiplication after three subcultivations, 65 shoots per initial rhizome average, mostly from the rhizomes proceeding from Baião. The culture médium with half concentration of MS salts was effective in shoots rooting as much for PV 03-76 cultivate as for Pacovan to Ken cultivate. / A bananeira é uma das frutas mais consumidas no mundo, sendo amplamente consumida no Brasil, porém doenças como as Sigatokas negra e amarela vem reduzindo a sua produção, principalmente por falta de uma adequada tecnologia de produção. A cultura de tecidos, especificamente a micropropagação, é uma alternativa para a produção de mudas de bananeira com qualidade fitossanitária e vegetativa, mas apresenta fatores que dificultam sua aplicação. Dentre os quais, destaca-se a contaminação, por fungos e bactérias, associada à oxidação dos explantes pela exsudação de compostos fenólicos que provocam o rápido escurecimento e morte de ápices caulinares, resultando em grande perda na fase inicial de estabelecimento dos explantes. O objetivo desse trabalho foi adaptar e/ou otimizar o comportamento de diferentes cultivares de bananeira, resistentes a doenças, via micropropagação, ajustando as fases de estabelecimento da cultura in vitro, por meio do controle de oxidação e contaminação, proliferação/multiplicação de brotos e enraizamento, para a multiplicação rápida de plantas com qualidade superior, tanto no aspecto fitossanitário quanto vegetativo. As cultivares Bucaneiro (AAAA), Caipira (AAA), Fhia 18 (AAAB), BRS Garantida (AAAB), Japira (AAAB), Pacovan Ken (AAAB), PA 4244 (AAAB), Preciosa (AAAB), PV 03-76 (AAAB), Thap Maeo (AAB) e Tropical (AAAB) provenientes do município de Baião do estado do Pará e Caipira (AAA), BRS Caprichosa (AAAB), Pacovan Ken (AAAB), Preciosa (AAAB), PV 03-76 (AAAB), Thap Maeo (AAB), provenientes do município de Belém do Estado do Pará foram submetidas às diferentes fases do processo de micropropagação. No estudo foram utilizados o antibiótico sulfato de estreptomicina e um fungicida visando reduzir a contaminação in vitro provocada por bactérias e fungos, além do anti-oxidante PVP (polivinilpirrolidona) para controlar a oxidação. Houve redução da contaminação com uso do sulfato de estreptomicina à concentração de 100 mg.L'1 e da oxidação com PVP a 4 g.L'1. Na fase de proliferação/multiplicação de brotos, as diferentes cultivares apresentaram médias que variaram de 1,67 a 7,0 brotos/explante, com maior proliferação no cultivo inicial (2,0 a 7,0 brotos/explante). A cultivar caipira (AAA) destacou-se das demais com a maior taxa de multiplicação de brotos após os três subcultivos, média de 65 brotos por rizoma inicial, principalmente a partir dos rizomas provenientes de Baião. O meio de cultura com metade da concentração dos sais de MS foi eficiente no enraizamento de brotos tanto para a cultivar PV 03-76 quanto para a cultivar Pacovan Ken.

Vem crescendo a cada dia o interesse pelo aproveitamento biológico dos carboidratos não-disponíveis no que se refere ao amido resistente (AR) e à fibra alimentar (FA) e seus efeitos sobre a fisiologia do intestino grosso. O objetivo do presente trabalho foi avaliar o efeito trófico, decorrente da fermentação de carboidratos não-disponíveis da banana verde, no intestino grosso de ratos adultos. As amostras estudadas foram: ABV (amido isolado de plátano verde - Musa paradisíaca L.) e MBV (banana nanicão - Musa acuminata, variedade Nanicão - verde cozida com casca, descascada e seca). Foi realizado estudo de média duração (28 dias) com ratos Wistar adultos, divididos em três grupos: grupo Controle (G-C), que recebeu ração padrão (R-C), e dois grupos experimentais, que receberam rações com concentrações crescentes de AR, ou seja, G-MBV recebeu ração R-MBV, com 5% de AR, e G-ABV recebeu ração R-ABV, com 10% de AR. Foram avaliados consumo e fermentabilidade in vitro das rações; peso corpóreo; peso e umidade das fezes; pH e histologia cecais. Não foi observada diferença no consumo médio diário de ração entre os grupos. O consumo de R-ABV proporcionou menor crescimento dos animais. No G-ABV, além da queda do pH cecal, houve aumento do peso seco das fezes e do conteúdo cecal, possivelmente devido ao aumento da microbiota intestinal. Ainda nesse grupo, houve aumento do peso total do ceco, evidenciando não somente ganho de umidade (aumento do peso do conteúdo cecal) como também possível proliferação celular (aumento do peso da parede do ceco). No G-MBV, houve queda do pH cecal, devido à produção de ácidos graxos de cadeia curta pela fermentação, e ganho de umidade no conteúdo cecal, resultado coerente com a maior presença de FA solúvel nesta ração. Porém, não houve aumento do peso seco das fezes nesse grupo. Pela análise histológica do tecido cecal, foi possível evidenciar que tanto a fermentação da R-ABV como a da R-MBV exerceram efeito trófico no intestino grosso desses animais (p<0,01). Os resultados obtidos indicam que os carboidratos não-disponíveis presentes na banana verde exercem efeitos positivos sobre a fisiologia dos animais, apontando a possibilidade de utilização dessa matéria-prima na elaboração de alimentos voltados para a prevenção de determinadas doenças crônicas não-transmissíveis. / The interest in unavailable carbohydrates, mainly in the possible effects of resistant starch (RS) and dietary fiber (DF) on the physiology of the large bowel, has recently increased. The present work aimed to evaluate the trophic effect, caused by the fermentation of unavailable carbohydrates from banana, on the large bowel of adult rats. Two samples were studied: ABV (starch isolated from unripe plantain - Musa paradisíaca L.) and MBV (unripe banana -Musa acuminata, variety Nanicão - cooked with peel, peeled and dried). An assay was carried during 28 days, with adult Wistar rats, divided into three groups: Control group (G-C), fed standard diet (R-C), and two experimental groups fed crescent RS concentrations (G-MBV, fed R-MBV with 5% RS, and G-ABV, fed R-ABV with 10% RS). Consumption and in vitro fermentation of the diets; body weight; feces weight and moisture; cecum pH and histology were evaluated. No difference was observed in the average daily consumption among the groups. Rats fed R-ABV presented decreased growth. On G-ABV, besides cecum pH decrease, there was an increase in feces and cecum content dry weight, possibly due to the increase in intestinal microbiota. There was an increase in cecum total weight, evidencing not only moisture gain (increase in cecum content weight) but also possible cellular proliferation (increase in the cecum wall weight). On G-MBV, there was a decrease in the cecum pH, due to the production of short-chain fatty acids by the fermentation, and moisture gain in the cecum content, which is coherent with the greater concentration of soluble DF in this diet. However, an increase in feces dry weight was not observed in this group. Considering the histology of the cecum tissue, it was possible to evidence that the fermentation of both R-ABV and R-MBV exerted trophic effect in the large bowel of the animais (p<0,01). The results obtained indicate that unavailable carbohydrates from unripe banana exert positive effects on the physiology of the animais, pointing to the possibility of using this product on the elaboration of foods aimed to preventing certain non-transmissible chronic diseases.

No description available.

Thesis (M.Sc.)(Microbiology)--University of Pretoria, 2005. / Includes summary. Includes bibliographical references. Available on the Internet via the World Wide Web.

This dissertation undertakes a comprehensive analysis of the Costa Rican banana industry, including a case study based on fieldwork at an independent banana plantation in Costa Rica's Atlantic zone. A basic premise is that a coherent understanding of the banana industry and its consequences can only be achieved through the appreciation of the complexity of its organization. That is, the Costa Rican banana industry is a political-economic, socio-cultural, and environmental system articulated through a high degree of interaction at the micro and macro levels. Modernization and world systems theories are shown to provide partial and insufficient accounts of the dynamics at work in the Costa Rican banana industry. An embedded model of world systems theory, which includes aspects of sustainable development, is proposed to address these concerns. The analysis at various levels is intended to support the argument that consideration of environmental dynamics must be addressed in future theoretical accounts of development. / Following world systems theory, the strong role of the banana transnationals within the industry and in relation to national development is examined. Other links between the Costa Rican banana industry and the broader international political economy are also examined, including the 'banana wars', as well as environmental issues, such as DBCP and other agrochemical usage. Significant changes in the Costa Rican banana industry since the 1980's are considered. These include: (1) a sharp increase in banana production, and an increase in independent banana producers, (2) the dissolution of the banana unions, and their replacement with a new system of labour relations (solidarismo); and (3) the recent concern for issues of environmental destruction. These changes, combined with the centrality of the banana industry to Costa Rican development, have resulted in significant consequences at the micro level. These are manifested in the organization of banana production and in operations on the plantation, as seen with respect to working conditions, quality control and of transnational power. In addition, environmental degradation, and underdevelopment and marginalization beyond the plantation are examined as direct consequences of the Costa Rican banana industry. This is supported with extensive ethnographic detail.

Banana bunchy top virus (BBTV) is a circular, single-stranded (css) DNA virus that belongs to the genus Babuvirus in the family Nanoviridae. BBTV is responsible for the most devastating virus disease of banana known as "bunchy top", for which conventional control measures are generally ineffective. Genetically engineered resistance appears to be the most promising strategy to generate BBTV-resistant bananas but the success of this strategy is largely dependent upon the molecular characterisation of the target virus and knowledge of the virus life cycle, particularly the replication strategy. This PhD study was aimed at the molecular characterisation of the intergenic regions of BBTV, in order to complement the molecular information currently available and to potentially contribute to the development of transgenic resistance strategies against BBTV in banana. Three putative iterative sequences (iterons; GGGAC) previously identified in the BBTV intergenic regions were initially characterised. In order to determine their role in the binding of the master BBTV replication initiation protein (M-Rep), the putative iterons (F1 and F2 in the virion sense, and R in the complementary sense) were independently mutated in a BBTV DNA-6 greater-than-genome-length clone (1.1 mer). The DNA-6 1.1 mers (native and mutants) and the M-Rep-encoding component (DNA-1) were co-bombarded into banana (Musa spp. cv."Lady finger") embryogenic suspension cells and transient replication was evaluated by Southern hybridisation. Analysis of the DNA-6 replicative forms showed a significant decrease of approximately 41% for the F1 iteron mutant and 61% for the R iteron mutant in comparison with native levels. However, the mutation in the F2 iteron caused the most dramatic effect, decreasing replication to levels barely detectable by Southern hybridisation. These results suggest that the three iterons all play a role in BBTV replication, most likely as recognition and binding sites for the M-Rep, but that the F2 iteron appears to be the most important in replication. Following the observation that all BBTV isolates sequenced to date have identical iteron sequences, the extent to which the M-Rep would recognise, bind and initiate replication of heterologous components from geographically diverse BBTV isolates (the South Pacific and the Asian groups) was evaluated. Cross replication assays revealed that heterologous M-Reps from Fiji, Hawaii (South Pacific group) and Vietnam (Asian group) were able to initiate replication of the coat protein-encoding component (DNA-3) from the Australian BBTV isolate (South Pacific group). However, replication of DNA-3 from the Vietnamese isolate was not initiated by heterologous M-Reps from the two South Pacific isolates tested (Australia and Hawaii). These results suggest that a broad-range transgenic resistance strategy based on replication using Australian BBTV intergenic regions may be successful as this region will be recognised by the M-Reps from both Asian and South Pacific BBTV isolates. However, a Rep protein-mediated resistance strategy will more likely be specific to geographical isolates and, therefore, less suitable as a broad-range control strategy. To further characterise the BBTV intergenic regions and to gain a better understanding of the BBTV transcription process, the 5' untranslated regions (UTRs) of the major open reading frames (ORFs) associated with each of the six BBTV DNA components were mapped. In all cases, the transcription start sites were located 3' of a putative TATA box and the 5' UTRs varied in length from 23 nucleotides (DNA-6) to 5 nucleotides (DNA-3). Two potential transcription start sites (nt 84 and 87) were mapped for DNA-1, but whether these represent the transcription start sites of the two genes associated with DNA-1 remains to be determined. Two start sites were also associated with DNA-2 which is thought to be monocistronic. Whether one of these start sites is an artefact or whether they are due to natural sequence variability of BBTV is unknown. These results now enable us to define the transcribed regions of each BBTV DNA component and accurately predict their promoter regions in an attempt to gain a fundamental understanding of BBTV gene expression patterns.

Bananas are susceptible to a diverse range of biotic and abiotic stresses, many of which cause serious production constraints worldwide. One of the most destructive banana diseases is Fusarium wilt caused by the soil-borne fungus, Fusarium oxysporum f. sp. cubense (Foc). No effective control strategy currently exists for this disease which threatens global banana production. Although disease resistance exists in some wild bananas, attempts to introduce resistance into commercially acceptable bananas by conventional breeding have been hampered by low fertility, long generation times and association of poor agronomical traits with resistance genes. With the advent of reliable banana transformation protocols, molecular breeding is now regarded as a viable alternative strategy to generate disease-resistant banana plants. Recently, a novel strategy involving the expression of anti-apoptosis genes in plants was shown to result in resistance against several necrotrophic fungi. Further, the transgenic plants showed increased resistance to a range of abiotic stresses. In this thesis, the use of anti-apoptosis genes to generate transgenic banana plants with resistance to Fusarium wilt was investigated. Since water stress is an important abiotic constraint to banana production, the resistance of the transgenic plants to water stress was also examined. Embryogenic cell suspensions (ECS) of two commercially important banana cultivars, Grand Naine (GN) and Lady Finger (LF), were transformed using Agrobacterium with the anti-apoptosis genes, Bcl-xL, Bcl-xL G138A, Ced-9 and Bcl- 2 3’ UTR. An interesting, and potentially important, outcome was that the use of anti-apoptosis genes resulted in up to a 50-fold increase in Agrobacterium-mediated transformation efficiency of both LF and GN cells over vector controls. Regenerated plants were subjected to a complete molecular characterisation in order to detect the presence of the transgene (PCR), transcript (RT-PCR) and gene product (Western blot) and to determine the gene copy number (Southern blot). A total of 36 independently-transformed GN lines (8 x Bcl-xL, 5 x Bcl-xL G138A, 15 x Ced-9 and 8 x Bcl-2 3’ UTR) and 41 independently-transformed LF lines (8 x Bcl-xL, 7 x BclxL G138A, 13 x Ced-9 and 13 x Bcl-2 3’ UTR) were identified. The 41 transgenic LF lines were multiplied and clones from each line were acclimatised and grown under glasshouse conditions for 8 weeks to allow monitoring for phenotypic abnormalities. Plants derived from 3 x Bcl-xL, 2 x Ced-9 and 5 x Bcl-2 3’ UTR lines displayed a variety of aberrant phenotypes. However, all but one of these abnormalities were off-types commonly observed in tissue-cultured, non-transgenic banana plants and were therefore unlikely to be transgene-related. Prior to determining the resistance of the transgenic plants to Foc race 1, the apoptotic effects of the fungus on both wild-type and Bcl-2 3’ UTR-transgenic LF banana cells were investigated using rapid in vitro root assays. The results from these assays showed that apoptotic-like cell death was elicited in wild-type banana root cells as early as 6 hours post-exposure to fungal spores. In contrast, these effects were attenuated in the root cells of Bcl-2 3’ UTR-transgenic lines that were exposed to fungal spores. Thirty eight of the 41 transgenic LF lines were subsequently assessed for resistance to Foc race 1 in small-plant glasshouse bioassays. To overcome inconsistencies in rating the internal (vascular discolouration) disease symptoms, a MatLab-based computer program was developed to accurately and reliably assess the level of vascular discolouration in banana corms. Of the transgenic LF banana lines challenged with Foc race 1, 2 x Bcl-xL, 3 x Ced-9, 2 x Bcl-2 3’ UTR and 1 x Bcl-xL G138A-transgenic line were found to show significantly less external and internal symptoms than wild-type LF banana plants used as susceptible controls at 12 weeks post-inoculation. Of these lines, Bcl-2 3’ UTR-transgenic line #6 appeared most resistant, displaying very mild symptoms similar to the wild-type Cavendish banana plants that were included as resistant controls. This line remained resistant for up to 23 weeks post-inoculation. Since anti-apoptosis genes have been shown to confer resistance to various abiotic stresses in other crops, the ability of these genes to confer resistance against water stress in banana was also investigated. Clonal plants derived from each of the 38 transgenic LF banana plants were subjected to water stress for a total of 32 days. Several different lines of transgenic plants transformed with either Bcl-xL, Bcl-xL G138A, Ced-9 or Bcl-2 3’ UTR showed a delay in visual water stress symptoms compared with the wild-type control plants. These plants all began producing new growth from the pseudostem following daily rewatering for one month. In an attempt to determine whether the protective effect of anti-apoptosis genes in transgenic banana plants was linked with reactive oxygen species (ROS)-associated programmed cell death (PCD), the effect of the chloroplast-targeting, ROS-inducing herbicide, Paraquat, on wild-type and transgenic LF was investigated. When leaf discs from wild-type LF banana plants were exposed to 10 ìM Paraquat, complete decolourisation occurred after 48 hours which was confirmed to be associated with cell death and ROS production by trypan blue and 3,3-diaminobenzidine (DAB) staining, respectively. When leaf discs from the transgenic lines were exposed to Paraquat, those derived from some lines showed a delay in decolourisation, suggesting only a weak protective effect from the transgenes. Finally, the protective effect of anti-apoptosis genes against juglone, a ROS-inducing phytotoxin produced by the causal agent of black Sigatoka, Mycosphaerella fijiensis, was investigated. When leaf discs from wild-type LF banana plants were exposed to 25 ppm juglone, complete decolourisation occurred after 48 hours which was again confirmed to be associated with cell death and ROS production by trypan blue and DAB staining, respectively. Further, TdT-mediated dUTP nick-end labelling (TUNEL) assays on these discs suggested that the cell death was apoptotic. When leaf discs from the transgenic lines were exposed to juglone, discs from some lines showed a clear delay in decolourisation, suggesting a protective effect. Whether these plants are resistant to black Sigatoka is unknown and will require future glasshouse and field trials. The work presented in this thesis provides the first report of the use of anti-apoptosis genes as a strategy to confer resistance to Fusarium wilt and water stress in a nongraminaceous monocot, banana. Such a strategy may be exploited to generate resistance to necrotrophic pathogens and abiotic stresses in other economically important crop plants.

Thesis (Ph. D.)--University of Washington, 1998. / Vita. Includes bibliographical references (leaves [283]-294).

Thesis (Ph.D.(Plant Pathology)--University of Pretoria, 2006. / Includes bibliographical references. Available on the Internet via the World Wide Web.

Thesis (Ph. D.)--University of Washington, 1996. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.