Abstract available in The FASEB Journal.

The development of the science of micropropagation from the very initial concept of totipotency to the modern day advancement and sophistication has been affected by a wide range of problems such as hyperhydricity, shoot-tip necrosis and somaclonal variation. These problems are largely the result of the obvious fact of trying to grow plants in an environment that is different from the one plants are used to naturally. The extent of these problems ranges from minor technical inconvenience to significant economic loss. Characterization and control of micropropagation problems has been one of the priorities of plant tissue culture research due to the enormous contribution of this discipline for plant production, improvement and conservation. The prevalence and severity of these tissue culture problems varies widely among plant species. The rationale of this research project was therefore, to identify plant species most affected by the problems studied, characterize the problem and find mechanism(s) to control or minimize the damage caused by the problem. The literatures reviewed provide sufficient background information for the experimental chapters. Due to the different nature of the problems and variation in the plant species they affect, the model plant, the methodologies used and parameters analysed were also different. The findings of these investigations, in their own different way, addressed certain problems that individually and collectively pose difficulties to the micropropagation industry. The difference in the content of the experimental chapters is therefore the result of the broader objective of the research project to tackle such difficulties. The success and failure of tissue culture system greatly depends on the choice of PGR’s. This choice can be made based on comparative study of their biological activity. Some promising reports on the role of topolins in micropropagation led to the idea of testing these cytokinins for their potential in tissue culture. As a prerequisite to subsequent investigations, the biological activity of some selected topolins and BA derivatives was tested using the soybean callus bioassay. The activity of the cytokinins tested varied significantly. The results demonstrated that the structure of a cytokinin dictates its activity. Modifications of side-chain improved the activity of oT but had no effect on pT. The presence of the methyl group had an enhancing effect on cytokinin activity of topolins or at least it did not reduce it. BA derivatives BA9THP (conjugated at N9 position), 3FBA and 2Cl6(3OHBA)R (halogenated derivatives) also showed good cytokinin activity and hold good promise for future research. In an attempt to alleviate hyperhydricity in Aloe polyphylla and optimize the micropropagation protocol, meta-topolin and its derivatives were tested at various concentrations together with BA and zeatin. Of all the cytokinins tested mT produced the best results in terms of shoot and root growth. Five μM was found to be the optimum concentration at which complete control of hyperhydricity was achieved without compromising shoot and root growth. Plantlets rooted in a multiplication media. BA generally had a negative effect on growth and development both in vitro and ex vitro. Acclimatization of plantlets was achieved easily by initially transferring plantlets to a mist house (for three weeks) followed by transfer to the greenhouse. The type of cytokinin also had an effect on ex vitro growth with BA-treated plants producing the lowest shoot and root biomass. Various experiments were conducted to characterize and control factors affecting STN in Harpagophytum procumbens. Media type and strength, PGR, carbon sources, sub-culturing, calcium and boron were tested. Results indicated that all of the tissue culture components tested affected STN. From the different media types tested, half strength was MS found to be the preferred medium. Increasing cytokinin concentration increased the incidence of STN and the problem was aggravated by the addition of auxin to the multiplication medium. Optimum shoot multiplication was achieved by omitting auxin and using the cytokinin mTR. Plantlets produced basal callus which interfered with rooting. The quantity of this basal callus was minimum when mTR was used. Sub-culturing plantlets onto fresh medium every two weeks helped minimize STN. Off all the sugars tested 3% sucrose was optimum. Other sugars either aggravated STN or inhibited growth when compared at equi-molar concentration. Increasing the concentration of either Ca or B prevented the development of necrotic shoots. When the concentration of both elements is increased simultaneously negative effects on both growth and STN were observed. Using 6 mM Ca in half strength MS medium was optimum. B was toxic at higher concentrations. Plantlets rooted readily in half strength cytokinin-free MS media supplemented with 2.5 μM IAA. Rooted plantlets produced using the optimized protocol were acclimatized successfully by transferring directly to a greenhouse in a 1:1 ratio of sand and soil mixture. The effect of meta-toplins on micropropagation and somaclonal variation of banana was investigated. Tissue cultured explants of cultivars ‘Williams’ and ‘Grand Naine’ were cultured in MS media containing the cytokinins BA, mT, MemT, MemTR and mTR at various concentrations. Results of the investigation revealed that superior multiplication and lower abnormality index was recorded from the mTR and mT treatments at 22.2 μM concentration. These treatments, however, had an inhibitory effect on rooting. The effect of these treatments (22.2 μM mT and mTR) in comparison with equi-molar concentration of BA on somaclonal variation of ‘Williams’ banana was tested using RAPD-PCR at the 7th multiplication cycle. No significant difference was found between the treatments. It should however be highlighted that cultures were initially maintained for three multiplication cycles in media containing BA. The inherent stability and initial effect of BA could have influenced the results. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.

Inhibition of Fusarium oxysporum f. sp. cubense race 4 by Burkholderia cepacia was evident when grown on various media (TSA, PDA, PSA, YM, KMB, PPM, NYGA, LA) with different carbon sources and under various pH and temperature conditions. In addition, B. cepacia was able to inhibit several fungal pathogens in vitro. Antagonism of B. cepacia against F. oxysporum f. sp. cubense occured at high levels of Fe³+, which may suggest that antagonism by B. cepacia did not involve siderophore production. Thin layer chromatogram (TLC) examination showed that B. cepacia produced several substances, one of which had similar R[f] value to that described for pyrrolnitrin. Cell-free supernatant of a 4-day culture of 6. cepacia was applied to an Amberlite XAD-2 column and inhibitory activity co-eluted with the 95% methanol (pH 9.5) fraction. The concentrated activated fractions showed inhibitory activity against F. oxysporum f. sp. cubense. A GC-MS chromatogram indicated numerous components in the antifungal extracts. The only compound identified in the Wiley 138 library, was 1,2- Benzenedicarboxylic acid, bis (2-Ethylhexyl) ester. Observations by light microscopy indicated that B. cepacia inhibited spore germination in F. oxysporum f. sp. cubense race 4 and retarded the mycelial growth. The interaction between the endophytic bacterium, B. cepacia and F. oxysporum f. sp. cubense race 4 was investigated with aid of scanning and transmission electron microscopy. This demonstrated that the bacterium was able to colonize the surface of hypha and macrospore of F. oxysporum f. sp. cubense. Mycelial deformation, terminal and/or intercalary swelling were evident. At later stages, hyphae of F. oxysporum f.sp. cubense, colonized by B. cepacia, were found to have collapsed. Further studies in vivo confirmed that B. cepacia colonized the hypha of F. oxysporum f. sp. cubense which had invaded banana roots. TEM observation showed that in the banana plant B. cepacia was closely associated with the healthy banana roots and a matrix was frequently found to be present between the bacterium and the plant surface. In addition, B. cepacia exists mainly in the intercellular space of the banana roots. UV irradiation treatment of B. cepacia resulted in a mutant that had lost inhibitory activity against F. oxysporum f. sp. cubense on TSA agar. Transposon mutagenesis of B. cepacia was performed by Tn5 insertion. Six mutants which had lost or had reduced inhibitory activity against F. oxysporum f. sp. cubense were generated. These mutants showed no inhibitory zones on TSA medium in the presence of the fungus. It was observed that one mutants. cepacia :: Tn5-188 appeared to lose the ability to colonize the fungal hypha, whilst a different mutant B. cepacia ::Tn5 - 217 was still able to colonize the fungal hyphae. TLC analyses showed that there was a decrease in antibiotic production in mutants B. cepacia :: Tn5 - 217 and B. cepacia - UV - 34, compared with the wild type. GC- MS analyses showed that there was no evidence of the peaks at 14.62 minutes, 20.0 minutes and 20.46 minutes in both chromatograms of mutants B. cepacia :: Tn5 -217 and 8. cepacia -UV - 34, compared with the wild type B. cepacia. No PCR products were detected using primers that were developed from sequences within the biosynthetic loci for Phi of P.fluorescens Q2-87(GenBank accession no. U41818) and PCA of P. fluorescens 2-79 (GeneBank no. L48616). Colony hybridization suggested that genomic DNA from B. cepacia could contain both Phi- and PCA probes. It was found that hybridization of genomic DNA digested with Cla-I of B. cepaca with Phl2a probe only occurred at low stringency. A hybridization signal was detected from a Cla-l fragment of approximately 2800bp. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 1997.

Made available in DSpace on 2014-10-09T12:54:34Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:07:20Z (GMT). No. of bitstreams: 0 / Dissertação (Mestrado) / IPEN/D / Instituto de Pesquisas Energéticas e Nucleares - IPEN/CNEN-SP

Made available in DSpace on 2014-10-09T12:54:38Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:07:47Z (GMT). No. of bitstreams: 0 / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energéticas e Nucleares - IPEN/CNEN-SP

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Orientador: Sarita Leonel / Coorientador: Emi Rainildes Lorenzetti / Banca: Jackson Mirellys Azevêdo Souza / Banca: Luiz Fernando Rolim de Almeida / Resumo: A bananeira é a segunda fruta com maior volume comercializado no mundo. Pelo fato de ser uma fruta muito apreciada pelo consumidor brasileiro, nos últimos anos surgiram diversas tecnologias que auxiliam no cultivo da bananeira, principalmente relacionadas à qualidade dos frutos. Nesse contexto, o trabalho teve como objetivo avaliar a influência da coloração dos sacos plásticos empregados no ensacamento de cachos de bananeira 'BRS Conquista'. Os tratamentos consistiram no uso de sacos de polietileno comerciais, nas cores branco, preto, vermelho e azul, além da testemunha (sem ensacamento). O delineamento experimental foi em blocos casualizados, com cinco tratamentos, quatro repetições e quatro plantas por parcela, totalizando 80 plantas. Avaliou-se a qualidade fitossanitária dos frutos, sendo o número de pontuações de danos causados por tripes da erupção e a porcentagem de área lesionada causada por antracnose, além das características físicas dos frutos, como massa do cacho, massa da ráquis, massa dos frutos, número de frutos por cacho, número de pencas por cacho, massa das pencas, número, comprimento e diâmetro de frutos da segunda penca, e as características físico-químicas dos frutos, como sólidos solúveis, acidez titulável, relação polpa/casca, índice de maturação e período de maturação. Verificou-se que os cachos que foram ensacados com sacos de coloração branca e preto apresentaram menor número de pontuações provocadas por tripes. Enquanto que para antracnose, os cachos... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Banana is the second most commercialized fruit crop in the world. Since it is very appreciated by the Brazilian consumer, in the last years, several technologies have come out to help in the development of the crop, mainly related to the quality of fruits. In this context, the objective of this work was to use banana bagging techniques for improvements in the banana cultivation process. Polyethylene bags of white, black, red, blue colors and the control (no bagging) were used in this technique. A randomized complete block design with five treatments, four replications, two plants per plot, totaling 80 plants was used. The evaluated characteristics in the post-harvest were the bunch mass, fruit mass, number of fruits per bunch, bunch number, second bunch mass, number of fruits of second batch, length, diameter, pH, soluble solids, titratable acidity, pulp/peel ratio, maturation index and maturation days. The phytosanitary quality of the fruits was evaluated, and the symptoms of thrips and anthracnose were assessed. In addition, the physical characteristics of the fruit, which is represented by the mass of the bunch, mass of the rachis, mass of the fruit, number of fruit per bunch, number of penca by bunch, productivity, mass of the second bunch, number of fruit in the second bunch, length and diameter of the fruit in the second bunch and physical chemical characteristics of the fruit, soluble solids, acidity, relation pulp/peel, index of maturation and period of maturation were evaluated. It was verified that for the variable mass of the second bunch, the white and black bags presented better results, whereas for pH variable the blue staining was the one which showed better results, and for the soluble solids it was the control group. This work showed that the efficiency of bagging is associated with the type and color of the bag which is used, since the coverings assign a thermal role inside ... / Mestre

Made available in DSpace on 2014-10-09T12:54:34Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:07:20Z (GMT). No. of bitstreams: 0 / Cascas de banana foram caracterizadas por espectroscopia de infravermelho por transformada de Fourier e microscopia eletrônica de varredura, e investigadas como biossorvente de baixo custo para a remoção de íons de urânio provenientes de soluções nítricas. A influência das seguintes variáveis foi estudada: tamanho das partículas do adsorvente, tempo de contato, pH e temperatura. O percentual de remoção aumentou de 13 para 57% quando o tamanho da partícula foi reduzido de 6,000 para 0,074mm. O tempo de contato determinado foi de 40 minutos com remoção de 60% em média. A remoção aumentou de 40 para 55% quando o pH aumentou de 2 para 5. Os modelos de isotermas lineares de Langmuir e Freundlich foram aplicados para descrever o equilíbrio de adsorção. A cinética do processo foi analisada a partir dos modelos de pseudo-primeira e segunda ordens. Parâmetros termodinâmicos como variações da entalpia (H), entropia (S) e energia livre de Gibbs (G) foram calculados. No intervalo de concentração de 50 - 500mg.L-1 , o processo de adsorção foi melhor descrito pela equação de Freundlich. A capacidade de adsorção no equilíbrio foi determinada pela equação de Langmuir, e o valor encontrado foi 11,50 mg.g-1 a 25 ± 2ºC. A cinética foi melhor representada pelo modelo pseudo-segunda ordem. O processo de biossorção para a remoção de urânio das soluções foi considerado exotérmico, e os valores de H e S obtidos foram - 9,61 kJ.mol-1 e 1,37J.mol-1, respectivamente. Os valores da energia livre de Gibbs variaram de -10,03 para -10,06 kJ.mol-1 quando a temperatura aumentou de 30 para 50ºC. Este estudo mostrou o potencial de aplicação das cascas de banana como biossorvente de íons de urânio. / Dissertação (Mestrado) / IPEN/D / Instituto de Pesquisas Energéticas e Nucleares - IPEN/CNEN-SP

Made available in DSpace on 2014-10-09T12:54:38Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:07:47Z (GMT). No. of bitstreams: 0 / Este trabalho apresenta pesquisa relativa à gestão tecnológica aplicada ao estudo de viabilidade técnica e econômica da instalação de um irradiador multipropósito no Vale do Ribeira (SP). De um lado o trabalho apresenta as características regionais que indicam que o Vale do Ribeira se mostra a região menos desenvolvida economicamente do estado de São Paulo; de outro lado, a emergência da tecnologia da irradiação de alimentos, com resultados aceitos e recomendados pelas autoridades internacionais. A gestão tecnológica, a partir dos conceitos apresentados, procura unir essa realidade regional com a tecnologia disponível, na busca do desenvolvimento regional. Os procedimentos metodológicos aplicados incluem, além da pesquisa bibliográfica, a aplicação de pesquisas qualitativa e quantitativa, bem como a utilização de métodos estatísticos e econômicos. A principal conclusão deste estudo indica a viabilidade técnica e econômica do empreendimento, embora sejam recomendados mais estudos específicos sobre a irradiação de bananas e sobre a aceitação do consumidor brasileiro a alimentos irradiados. / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energéticas e Nucleares - IPEN/CNEN-SP

Made available in DSpace on 2014-10-09T12:43:54Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:10:34Z (GMT). No. of bitstreams: 1 06784.pdf: 2785176 bytes, checksum: f2b021da60503c5d9617c8d3ec337b80 (MD5) / Dissertacao (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP