Nous avons étudié les phénomènes de polyploïdisation chez l'igname Dioscorea alata qui comprend trois cytotypes diploïde (2n=40), triploïde (2n=60) et tétraploïde (2n=80) en vue d'optimiser les stratégies de production d'hybrides polyploïdes. Chez ce complexe polyploïde l'augmentation de la ploïdie est corrélée avec une augmentation de la vigueur et des rendements plus élevés et plus stables. Dans un premier temps nous avons démontré l'autotétraploïdie des variétés tétraploïdes à l'aide de trois approches différentes: des analyses d'hérédité de marqueurs microsatellites, l'observation de phénomènes de double réduction et l'étude des méioses des cellules mères de pollen. Nous avons ensuite déterminé quels sont les mécanismes les plus probables à l'origine des polyploïdes naturels via l'étude de la transmission de l'hétérozygotie parentale à l'aide de microsatellites et par l'étude des incompatibilités au niveau de l'albumen lors de différents croisements intracytotypes et intercytotypes en utilisant la cytométrie en flux. Les résultats obtenus montrent que les polyploïdes de D. alata seraient apparus via la formation de gamètes non réduits de clones diploïdes. Le pool triploïde se serait édifié et diversifié uniquement par voie femelle, du fait de la non viabilité des croisements intercytotypes et de la stérilité des femelles et mâles triploïdes. Le pool tétraploïde serait apparu par union de deux gamètes non réduits de clones diploïdes (polyploidisation sexuelle bilatérale). Par la suite ce pool aurait été diversifié via des croisements intercy / We studied polyploidisation phenomena in the yam Dioscorea alata that includes three cytotypes -diploid (2n=40), triploid (2n=60) and tetraploid (2n=80) -in order to optimise polyploid hybrid production strategies. In this complex polyploid, the increase in ploidy is correlated with an increase in vigour and higher and more stable yields. We first showed the autotetraploidy of tetraploid varieties using three different approaches: heredity analyses of microsatellite markers, the observation of double reduction phenomena, and the study of meiosis of pollen mother cells. We then determined the mechanisms most likely to be at the origin of natural polyploids through the study of the transmission of parental heterozygoty using microsatellites and the study of incompatibilities at the endosperm level at the time of different intracytotypic and intercytotypic crosses using flow cytometry. The results obtained reveal that the polyploids of D. alata probably appeared through the formation of non-reduced gametes of diploid clones. The triploid pool would then have been constituted and diversified through the female pathway as a result of the non-viability of intercytotypic crosses and the sterility of female and male triploids. The tetraploid pool would have appeared as a result of the union of two non-reduced gametes of diploid clones (bilateral sexual polyploidisation). This pool would then have diversified through intercytotypic crosses with the formation of 2n gamètes through both the female and the male pathway, as well as by intracytotypic crosses within the 4X pool.

Submitted by (lucia.rodrigues@ufrpe.br) on 2017-03-17T14:50:17Z No. of bitstreams: 1 Joyce Silva Lima.pdf: 2222741 bytes, checksum: 8a6c7b004633c1014edd4a6cdb6ce33f (MD5) / Made available in DSpace on 2017-03-17T14:50:17Z (GMT). No. of bitstreams: 1 Joyce Silva Lima.pdf: 2222741 bytes, checksum: 8a6c7b004633c1014edd4a6cdb6ce33f (MD5) Previous issue date: 2012-02-17 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The yam crop (Dioscorea spp.) has a great socioeconomic importance for the Northeast region of Brazil, due to the excellent nutritional quality and energy of its tubers, with potential for export, especially for Europe. However information about the occurrence and distribution of yam viruses are limited in Brazil, the major constraints to yam production, are the viral pathogens. In this study, surveys were conducted to detect the occurrence of viruses yam in produtive areas in northeastern Brazil. The analysis of sequences of virus in yam leaf samples from Alagoas, Pernambuco and Paraiba states were evaluated using the molecular techniques polymerase chain reaction (PCR) and rolling circle amplification (RCA) which revealed the occurrence of badnavirus and geminivirus. Analyzing the partial nucleotide sequence in the RT/RNaseH encoding region, was observed that the isolates of badnavirus correspond to the species Dioscorea alata bacilliform virus (DaBV) and presented nucleotide sequences identity with 50 isolates of DaBV varying between 78-97%. It was also observed that a group with four isolates unrelated to any species of Badnavirus genus, indicating the possibility of being a new phytovirus genus or corresponding to endogenous sequence. Some samples showed the presence of a new begomovirus closely related to species that infect tomato and weeds. A curtovirus was also detected in yam samples and it was related to the described specie Beet mild curly top virus, not reported in Brazil in any host. These results suggest that DaBV is a widespread virus found in northeastern Brazil, the possible presence of endogenous sequences in yam and records the first report of geminivirus in yam and monocotyledonous in Brazil. / A cultura do inhame (Dioscorea spp.) apresenta grande importância socioeconômica para a região Nordeste do Brasil, devido à excelente qualidade nutritiva e energética de suas túberas, apresentando potencial de expansão via exportação de túberas, especialmente para a Europa. Patógenos virais estão entre os principais fatores que ameaçam a produção e produtividade de inhame e circulação segura de germoplasma, contudo informações sobre a ocorrência e distribuição de vírus de inhame são limitadas no Brasil. Neste estudo, as pesquisas foram conduzidas para detectar a ocorrência de viroses nas zonas produtoras do Nordeste do Brasil. A análise de um total de 425 amostras de folhas de inhame obtidas em áreas de cultivo, localizadas nos estados de Alagoas, Pernambuco e Paraíba foram avaliadas através das técnicas moleculares de reação em cadeia da polimerase (PCR) e amplificação em circulo rolante (RCA) que revelaram a ocorrência de badnavírus e geminivírus. Analisando a sequência parcial de nucleotídeos da região codificante RT/RNaseH observou-se que os isolados de badnavirus correspondem à espécie Dioscorea alata baciliforme virus (DaBV) e apresentam identidade de seqüências de nucleotídeos de 50 isolados de DaBV variando entre 78-97%. Constatou-se ainda um grupo de quatro isolados que não se relacionaram com nenhuma espécie do gênero Badnavirus, podendo constituir um novo gênero de fitovírus ou corresponder a sequências endógenas integradas. Em algumas amostras foi constatada a presença de um novo begomovirus estreitamente relacionado com espécies que infectam tomate e plantas daninhas. Um curtovirus também foi detectado nas amostras de inhame e relacionado com uma espécie já descrita Beet mild curly top vírus, ainda não relatada no Brasil em nenhum hospedeiro. Estes resultados sugerem a ampla disseminação do DaBV no Nordeste brasileiro e a possível presença de sequências endógenas em plantas de inhame e registra o primeiro relato de geminivírus na cultura do inhame e em monocotiledôneas em geral no Brasil.

Submitted by Reginaldo Soares de Freitas (reginaldo.freitas@ufv.br) on 2016-11-03T17:18:19Z No. of bitstreams: 1 texto completo.pdf: 669306 bytes, checksum: 9b386dbcaba82f69fd43166410e1d916 (MD5) / Made available in DSpace on 2016-11-03T17:18:19Z (GMT). No. of bitstreams: 1 texto completo.pdf: 669306 bytes, checksum: 9b386dbcaba82f69fd43166410e1d916 (MD5) Previous issue date: 2004-12-08 / O objetivo deste trabalho foi desenvolver um revestimento comestível a base de amido de inhame “caramujo" adicionado de quitosana, como antimicrobiano, e avaliar seu efeito na conservação de cenoura minimamente processada. O amido foi extraído dos tubérculos e sua composição físico-química e propriedades reológicas foram determinadas. Raizes de cenoura “Brasília”, foram fatiadas em rodelas e minimamente processadas. Foram preparados filmes e revestimentos a partir de soluções aquosas de 4% de amido de inhame (p/p) + 2% de glicerol (p/p), 4% de amido de inhame (p/p) + 2% de glicerol (p/p) + quitosana nas concentrações de 0,5, 0,75, 1,0 e 1,5%. As amostras de cenouras foram imersas nos revestimentos durante 3 minutos e secas a 20°C por 3 horas. As amostras do grupo controle foram imersas em água destilada esterilizada. Todas as amostras foram acondicionadas em bandejas de poliestireno expandido, envoltas em filme de poli (vinil cloreto) e estocadas a 8 ± 2°C por 15 dias. Foram realizadas análises microbiológicas, perda de massa, índice de esbranquiçamento, aceitabilidade de cor e sabor com 0, 5, 10 e 15 dias de estocagem. Os filmes foram formados pelo método “casting” e sua permeabilidade ao vapor de água foi determinada. O rendimento da extração do amido a partir dos tubérculos foi de 9,3%. Os componentes físico- químicos, em base seca, foram de 11,57% de umidade, 0,64% de proteínas, 0,052% de matéria graxa e 0,17% de cinzas. Os conteúdos de amido e de amilose foram de 99,17% e 22%, respectivamente. A temperatura inicial de pasta foi de 60,3oC e a máxima temperatura de viscosidade foi de 98,5oC, apresentando baixa tendência a retrogradação (100 UA). Os filmes produzidos foram visualmente transparentes com espessura de 30 ± 5 μm com valores de permeabilidade ao vapor de água entre 4,58 g mm kPa-1 d-1 m-2, para o filme de amido e glicerol, e 3,57 g mm kPa-1 d-1 m-2, para o filme de amido+glicerol+1,5% de quitosana, respectivamente. permeabilidade Os valores de foram estatisticamente diferentes (P<0,05) entre eles. A permeabilidade ao vapor de água dos filmes diminuiu com o aumento do teor de quitosana. Para a sanitização da cenoura em rodelas foi utilizada a concentração de 200 mg/L de cloro residual total, que se apresentou como a mais eficiente para o controle de coliformes totais e fecais e aeróbios mesófilos. O tempo ideal de centrifugação para amostras contendo 1500 g de cenoura em rodelas foi de 6 minutos, quando se utilizo centrífuga doméstica (ARNO) e 3,5 minutos para amostras contendo 3000 g quando foi usada centrífuga industrial. Nas fotomicrografias foi observado que os revestimentos apresentaram distribuição homogênea sobre a superfície da cenoura com uma espessura entre 15 a 20 μm. Os revestimentos apresentaram efeito significativo (P<0,05) para o controle de coliformes totais, psicrotróficos, fungos filamentosos e leveduras e nos atributos cor, sabor e índice de esbranquiçamento. Entretanto, não apresentaram diferença significativa (P>0,05) para perda de massa. Durante o armazenamento, todas as amostras apresentaram contagens <3 NMP/g para coliformes fecais e Escherichia coli e ausência de Salmonella sp em 25/g. Os revestimentos que continham quitosana apresentaram efeito antimicrobiano para coliformes totais, psicrotróficos, fungos e leveduras. O revestimento com 1,5% de quitosana apresentou a melhor eficiência sobre esses grupos microbianos com reduções de 2,1, 1,83 e 3,64 ciclos log, respectivamente, em relação ao controle. O grupo controle e as amostras revestidas somente com amido e glicerol apresentaram contagens mais altas dos grupos de microrganismos avaliados durante a estocagem. No tempo zero de estocagem, os revestimentos que continham quitosana mantiveram melhor a cor da cenoura, obtendo as notas hedônicas mais altas. Após 15 dias de estocagem, o revestimento de amido e glicerol, obteve a nota mais alta (7,4) para o atributo cor, enquanto que o grupo controle (sem revestimento) apresentou nota 4,7. Quanto ao sabor, as amostras revestidas com amido e glicerol também obtiveram a maior nota (6,5). Notas em torno de 5 foram obtidas para o controle e amostras revestidas com amido e quitosana. Para o índice de esbranquiçamento, as amostras revestidas não difereram entre elas e difereram do controle. O menor índice de esbranquiçamento foi de 37,6, apresentado pela amostra revestida com amido e 1,5% de quitosana, em comparação a 44,2, apresentado pelo grupo controle. O amido de inhame apresentou-se como uma boa fonte para a produção de filmes e revestimentos comestíveis, com grande potencial de aplicação na indústria de alimentos, com a vantagem de serem biodegradáveis. O uso do revestimento comestível antimicrobiano, constituído de amido de inhame e quitosana é uma alternativa viável para controlar o crescimento microbiano, manter a cor, retardar o aparecimento do esbranquiçamento na cenoura minimamente processada e estender a vida de prateleira do produto. / This work aimed to develop an edible coating from yam starch incorporated with chitosan, as an antimicrobial compound, and to evaluate its effect on minimally processed carrots conservation. The starch was extracted from yam and had characterized its physical, chemical and reological properties. Brazilian variety carrots were sliced and minimally processed. Films and coatings were prepared as the following composition: 4% yam starch (p/p) + 2% glycerol (p/p), 4% yam starch (p/p) + 2% glycerol (p/p) added with 0,5, 0,75, 1,0 and 1,5 % of chitosan. Samples of sliced carrots were immersed into these coatings for 3 minutes and dried at 20oC/ 3 hours. The control samples were immersed into pure water. All the samples were acondicionated in polystyrene trays, wrapped in polyvinyl cloride and stored at 10oC during 15 days. Microbiological analyses, mass loss, whitish index, sensorial evaluation of color and flavor were realized at 0, 5, 10 and 15 days of storage. The casted films had its vapor permeabilities determined. The starch presented the following characteristics: yield of 9,3%, humidity content of 11,57%, proteins content of 0,64%, lipids:0,052%, ash:0,17%, starch:99,17% and amylose:22%. The gel initial temperature was 60,3oC and the temperature of maximum viscosity was of 98,50C, showing a low tendency for retrogradation (100 UA). The films with thickness of 30± 5μm showed transparency and presented a vapor permeability ranging from 4,58 g.mm.dia-1.m-2.KPa-1, for starch-glycerol film, to 3,57 g.mm.dia-1.m-2.KPa-1, for the film containing 1,5% of chitosan + starch +glycerol. The permeabilities values showed significant (p<0,05) difference among them. The film permeability decreased as the chitosan level in the film increased. The concentration of 200 mg/L of total residual clorine was used for carrots sanification treatment and, among the others treatments, presented the highest efficiency on coliforms and mesophilics aerobes inhibition. The centrifugation time for 1500 g of sliced carrots was 6 minutes for houseware centrifuge and 3.5 minutes for industrial centrifuge. Photomicrografies from the coated carrots showed a homogeneous distribution of the coating on the carrots and a thickness ranging from 15 to 20 μm . The coatings showed a significant (P<0,05) effect on microbiological control of coliforms, psychrotrophic and fungi as well as in the sensorial evaluation of color and flavor and whitiness index, but did not showed significant difference for mass loss ( P<0,05) During storage period, all the samples presented countings <3NMP/g for coliforms fecal and E. coli and no presence of Salmonella in 25 g. The edible coatings with chitosan inhibited the microrganisms growth. Coating containing 1,5% of chitosan reduced 2,1, 1,83 and 3,64 log cycles for coliforms, psychrotrophic and fungi and yeast, respectively, compared with the control. The control group and the samples coated with starch and glycerol showed the highest microorganisms countings evaluated during the storage period. Chitosan-coatings protected carrots against color loss at the beginning of the storage, but at the end the treatment with starch and glycerol obtained the highest score for color attribute. For flavor sensorial analyses, this treatment also was evaluated by the painelist and obtained the highest score (6,4) compared with the control and the chitosan-starch treatments (5,0). The whitish index was lower for the chitosan 1,5% (37,6) compared with the value of 44,2 showed in the control treatment. The yam starch presents as a good source for production of edible coating and film with a potential availability for food industry. The edible coating incorporated with chitosan demonstrated to be a barrier hurdle for food preservation, inhibiting microorganism growth, decreasing color loss and whitish index of the minimally processed carrots and, therefore, increase its shelf-life.

Le taro (Colocasia esculenta L.), la grande igname (Dioscorea alata L.) et le manioc (Manihot esculenta Crantz) représentent trois cultures amylacées d'importance dans les régions tropicales et subtropicales du monde. Afin d'évaluer la diversité chimique en molécules bioactives de ces espèces, nous avons développé des méthodologies analytiques CLHP/DBD et CPG/SM à moyen débit permettant de doser les teneurs de 129 caroténoïdes et composés phénoliques/indoliques et de 16 acides organiques/gras dans les tissus souterrains consommés. Ces substances ont ensuite été dosées pour la première fois sur un large échantillonnage représentatif de la diversité agro-morphologique de ces trois espèces (respectivement 173, 113 et 79 cultivars de taro, grande igname et manioc cultivés sur un même site). Les résultats montrent l'existence d'une énorme diversité qualitative et quantitative des marqueurs chimiques et une absence de chimiotype marqué. De nombreux métabolites secondaires n'ont été détectés que dans peu de variétés en accord avec les résultats d'études antérieures sur la diversité, et la distribution, d'allèles génétiques neutres. Bien qu'appréciées, les substances colorées ou perceptibles en bouche n'ont pas fait l'objet d'amélioration variétale poussée sur leurs teneurs et ne peuvent être perçues que dans un petit nombre de cultivars. Dans l'ensemble, les résultats soutiennent l'importance de la sélection participative de petits agriculteurs pour la création et le maintien d'une large biodiversité chimique chez ces plantes. Ils offrent également des perspectives et des outils nouveaux pour améliorer la qualité nutritionnelle de ces espèces / Taro (Colocasia esculenta L.), the greater yam (Dioscorea alata L.) and cassava (Manihot esculenta Crantz) are three important staple crop species from tropical and subtropical regions. In order to evaluate the diversity in bioactive molecules of these three species, we have developed medium throughput HPLC/DAD and GC/MSbased analytical methodologies to estimate the contents of 129 carotenoids and phenolic/indolic compounds and 16 organic/fatty acids in the consumed underground tissues. The contents of these substances have then been quantified for the first time in a large sampling representative of the agro-morphological biodiversity of these three species (respectively 173, 113 and 79 landraces of taro, greater yam and cassava that were cultivated on a common site). Results demonstrate the existence of a large qualitative and quantitative diversity of chemical traits and the absence of clear-cut chemotype. Most secondary metabolites have only been detected in few landraces in agreement with results from previous studies on the diversity, and distribution, of neutral genetic alleles in these plants. Though well appreciated by consumers, colored and mouth-perceivable substances have not been subjected to major content improvement through targeted selection and are in fact only detectable in a limited number of landraces. As a whole, these results support the importance of participatory selection by small farmers in the creation, and maintaining, of a large chemical diversity in these species. They also offer novel tools and perspectives for the improvement of the nutritional value of these species by plant breeders