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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
81

MAPEAMENTO DE QTLs PARA TOLERÂNCIA À MURCHA BACTERIANA (Ralstonia solanacearum Smith) EM TABACO

Souza, Adenilson Mroginski 27 September 2018 (has links)
Submitted by Angela Maria de Oliveira (amolivei@uepg.br) on 2018-12-04T18:41:29Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Dissertação_final_Adenilson.pdf: 1632453 bytes, checksum: ca3bab925a332585e719109a07462e72 (MD5) / Made available in DSpace on 2018-12-04T18:41:29Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Dissertação_final_Adenilson.pdf: 1632453 bytes, checksum: ca3bab925a332585e719109a07462e72 (MD5) Previous issue date: 2018-09-27 / Os objetivos deste trabalho foram a obtenção de um mapa genético de alta densidade utilizando marcadores SNP (Single Nucleotide Polymorphism) obtidos através de genotipagem por sequenciamento (GBS) e identificar as regiões genômicas ligadas à tolerância à murcha bacteriana (Ralstonia solanacearum Smith) em uma população de duplo-haploides (DH) de tabaco. As linhagens endogâmicas NC95 (tolerante) e NC2326 (suscetível) foram cruzadas entre si gerando a população F1, anteras foram coletadas destas plantas para produção de haploides e posterior duplicação cromossômica através da cultura de anteras, gerando 180 famílias duplo-haploides que foram avaliadas em ambiente controlado quanto à tolerância à murcha bacteriana, após inoculação com R. solanacearum, através de uma escala de notas com amplitude de 0 a 4. As famílias DH foram genotipadas utilizando a metodologia de GBS e os dados resultantes desta genotipagem foram alinhados com o genoma de referência do tabaco para posterior obtenção dos marcadores SNP utilizados na construção do mapa de ligação. O mapa de ligação juntamente com os dados de fenotipagem foram utilizados para realizar o mapeamento de QTLs através do mapeamento por intervalo composto. Foram identificados 6.842 SNPs, utilizados para construção de um mapa de ligação com 70.583 cM, sendo este o maior mapa de ligação utilizando marcadores SNP disponível para tabaco e com o maior número de marcadores. Utilizando este mapa de ligação foram mapeados 13 QTLs para tolerância à murcha bacteriana em oito grupos de ligação, dos quais oito QTLs ainda não tinham sido identificados na literatura especializada. Os locos presentes nos grupos de ligação 3, 17 e 22 apresentaram os maiores efeitos na variação fenotípica. O elevado número de QTLs mapeados nesta população confirma o padrão de herança quantitativa da tolerância de tabaco à murcha bacteriana causada por R. solanacearum. / The objectives of this work were to obtain a high-density genetic map using SNP (Single Nucleotide Polymorphism) markers obtained through Genotyping-by-Sequencing (GBS) and to identify the genomic regions linked to bacterial wilt tolerance (Ralstonia solanacearum Smith) in a tobacco double-haploid (DH) population. The inbred lines NC95 (tolerant) and NC2326 (susceptible) were crossed generating the F1 population, anthers were collected from these plants for haploid production and subsequent chromosomic duplication using anthers culture, generating 180 double-haploid families that were evaluated in a controlled environment for tolerance to bacterial wilt after inoculation with R. solanacearum, using an assessment scale from 0 to 4. The DH families were genotyped using the GBS methodology and the resulting data from this genotyping were aligned with the reference genome and then to obtain the SNP markers used to construct the genetic linkage map. The linkage map jointly with the phenotyping data were used to QTL mapping through the composite interval mapping method. A total of 6,842 SNPs was identified and used to construct a linkage map with 70,583 cM, being the largest SNP-based genetic linkage map available for tobacco and presenting the highest number of markers. Using this linkage map, 13 QTLs were mapped for bacterial wilt tolerance in eight linkage groups, from those eight QTLs had not yet been identified in the specialized literature. The loci present in linkage groups 3, 17 and 22 had the highest effects on phenotypic variation. The high number of QTLs mapped in this population confirms the quantitative genetic control of tobacco tolerance to bacterial wilt caused by R. solanacearum.
82

?Fernandes, Ana Rosa de Figueiredo. Patogenicidade de Pratylenchus brachyurus e severidade da murcha bacteriana quando associada a este nemat?ide em fumo. 2009. / ?Fernandes, Ana Rosa de Figueiredo. Pathogenicity of Pratylenchus brachyurus and severity of bacterial wilt in association with this nematode in tobacco. 2009.

Fernandes, Ana Rosa de Figueredo 29 October 2009 (has links)
Made available in DSpace on 2016-04-28T14:57:35Z (GMT). No. of bitstreams: 1 Ana Rosa de Figueiredo Fernandes.pdf: 5291957 bytes, checksum: e9ba6c76361eb4a2e901846a5bb5c2dd (MD5) Previous issue date: 2009-10-29 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / ?The pathogenicity of Pratylenchus brachyurus on tobacco (Nicotiana tabacum) cultivar K326 was evaluated in experiments testing different initial population densities of the nematode (Pi = 0; 1,000; 3,000; 9,000 and 27,000 eggs and larvae per plant), in eight replications. The isolate was monoxenically multiplied on carrot discs. At the end of the experiment (70 days after inoculation), the above ground part of each plant was weighed and measured and, the respective root system was processed. The reproduction rate of Pratylenchus brachyurus was estimated by extracting the nematodes of the roots and of the soil. Necroses on the roots of the tobacco plants were observed. Pratylenchus brachyurus caused well-delimited root lesions on tobacco root. None of the analyzed plants died; however plant growth was reduced. Also, it was observed a decrease of the fresh mass of roots and of the dry top weight, mainly under the highest population density (27,000 nematodes per plant). The reproduction rate was low. Besides the interaction between Pratylenchus brachyurus and Ralstonia solanacearum in two tobacco varieties, Virg?nia K326 and Burley By21 was evaluated. Test plants were inoculated with the nematode in the initial population density of 3,000 individuals (eggs and larvae) by plant and, 15 days later, the plants were inoculated with a sample of soil infested by the bacterium, in estimated population density of 106 CFU.g-1 of soil. Besides the mixed inoculation with Pratylenchus brachyurus and Ralstonia solanacearum, tobacco plants were single inoculated. The plants were monitored daily. Dates of the beginning of symptom expression were registered, as well as symptom evolution. When the plants were inoculated with Pratylenchus brachyurus and Ralstonia solanacearum, symptom of wilt that is caused by the bacterium Ralstonia solanacearum began earlier than in single inoculated plants, and they also evolved quicker, inclusive in the variety Virg?nia K326 that was considered more tolerant. In this case, the presence of Pratylenchus brachyurus, in the root system of these plants, probably favored the infection by Ralstonia solanacearum, even before the symptoms caused by the nematodes could be detected. This study was done in greenhouse and under laboratory conditions. / ?A patogenicidade de Pratylenchus brachyurus em fumo Virg?nia K326 foi avaliada em experimento no qual foram testadas diferentes densidades populacionais iniciais do nemat?ide (Pi = 0; 1.000; 3.000; 9.000 e 27.000 ovos e larvas por planta), em oito repeti??es. O isolado foi multiplicado axenicamente em discos de cenoura. Ao final do experimento (85 dias), as plantas tiveram sua parte a?rea pesada e medida e foi processado o sistema radicular de cada uma. O fator de reprodu??o foi estimado por extra??o dos nemat?ides das ra?zes e do solo. Necroses nas ra?zes das plantas foram observadas. Pratylenchus brachyurus causou les?es radiculares delimitadas nas ra?zes de fumo. N?o houve morte de nenhuma planta analisada, por?m a altura das plantas foi afetada e tamb?m a massa seca da parte a?rea, principalmente sob a densidade populacional mais alta, de 27.000 nemat?ides por planta. O fator de reprodu??o foi baixo. Al?m disto, avaliou-se a intera??o entre Pratylenchus brachyurus e Ralstonia solanacearum em fumo Virg?nia variedade K326 e fumo Burley variedade BY21. As plantas foram inicialmente inoculadas com o nemat?ide na densidade populacional de 3.000 ovos e larvas por planta e 15 dias ap?s, foram inoculadas com uma amostra de solo infestado pela bact?ria, na densidade populacional estimada de 106 UFC.g-1 de solo. Al?m da inocula??o mista com Pratylenchus brachyurus e Ralstonia solanacearum, testemunhas foram inoculadas isoladamente. As plantas foram monitoradas diariamente, anotando-se a data de in?cio e da evolu??o dos sintomas. Quando as plantas foram inoculadas com os Pratylenchus brachyurus e Ralstonia solanacearum, os sintomas de murcha, que s?o causados pela bact?ria se manifestaram mais cedo e tamb?m evolu?ram mais r?pido, inclusive na variedade K326 que foi considerada mais tolerante. Neste caso, provavelmente a presen?a de Pratylenchus brachyurus, no sistema radicular dessas plantas, favoreceu a infec??o por Ralstonia solanacearum, antes mesmo que os sintomas causados pelo nemat?ide pudessem ser observados. Este trabalho foi realizado em casa de vegeta??o e em laborat?rio.
83

Análise molecular da interação entre Tospovirus e o gene resistência Sw-5 / Molecular analysis of the tospovirus Sw-5 resistance gene interaction

Lau, Douglas 31 March 2004 (has links)
Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2017-04-27T13:57:34Z No. of bitstreams: 1 texto completo.pdf: 2630530 bytes, checksum: 112de5671b9d3e521f0446f6d6d7237e (MD5) / Made available in DSpace on 2017-04-27T13:57:34Z (GMT). No. of bitstreams: 1 texto completo.pdf: 2630530 bytes, checksum: 112de5671b9d3e521f0446f6d6d7237e (MD5) Previous issue date: 2004-03-31 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Alguns aspectos da interação entre o gene de resistência Sw-5 e os tospovírus, foram analisados neste trabalho. A capacidade de Sw-5 conferir resistência em Nicotiana benthamiana foi avaliada em plantas transformadas com uma construção na qual a ORF e a região 3' do gene estavam sob controle do promotor 35S. As plantas transformadas foram resistentes à infecção por tospovírus. A comparação do espectro da resistência destas plantas com o observado para outras solanáceas , indica que as vias de sinalização e as respostas de defesa ativadas por Sw-5 estão conservadas nesta família, e que o polimorfismo genético nos componentes das vias de transdução de sinais pode resultar em diferentes níveis de resistência. A fim de identificar o gene de avirulência dos tospovírus, os genes N, NSm e NSs , foram expressos isoladamente ou em combinações, por meio do vetor viral PVX, em plantas com Sw-5. A expressão destes genes não foi capaz de desencadear a resposta de hipersensibilidade e tampouco interferiu na infecção da planta por PVX. Portanto, outro componente dos tospovírus deve ser responsável pelo desencadeamento da reação de resistência. Independentemente da presença do gene Sw-5, a expressão do gene NSs por meio do PVX , agravou os sintomas provocados por este vírus em algumas solanáceas, o que pode ter relação com a capacidade desta proteína de suprimir silenciamento gênico. Em plantas com Sw-5, a co-expressão da região 5' deste gene por meio do vetor PVX , favoreceu a infecção sistêmica por tospovírus. Uma vez que o efeito foi observado tanto para expressão senso quanto anti-senso, a redução dos níveis de mRNA de Sw-5 provocada por silenciamento gênico , poderia ser a causa desta interferência na resistência, embora a análise de Northern blot não tenha demonstrado tal redução. Uma seqüência homóloga a Sw-5 contendo uma ORF sem deleções ou interrupções prematuras foi clonada a partir do acesso LA371-20 de Lycopersicon peruvianum. Análises moleculares demonstraram que esta seqüência é originada do loco Sw-5 ou de região próxima a este e que segrega com a resistência a tospovírus. A capacidade deste e de outros três homólogos oriundos de distintos acessos de tomateiro de conferir resistência a tospovírus, foi avaliada em plantas transgênicas de tomateiro e tabaco. As plantas transformadas foram suscetíveis ao vírus. A não-funcionalidade destes homólogos pode ser devido à estrutura das construções utilizadas na transformação. Alternativamente, outros homólogos presentes nestes acessos de tomateiro podem ser os responsáveis pela resistência. / In this work some aspects of the tospovirus - Sw-5 interaction was analyzed. The capacity of the Sw-5 to confer resistance in Nicotiana benthamiana was evaluated in transgenic plants transformed with Sw-5 ORF containing its own 3 ́ UTR region under 35S promoter control. Transgenic plants were resistant to tospovirus infection. Comparisons of the resistance spectrum with other members of the Solanaceae suggest that the signal transduction pathways and resistance responses triggered by Sw-5 are conservated in solanaceae and that the genetic polymorphism in the signal transduction components may result in different resistance levels. The N, NSm and NSs genes isolated or in combination were expressed by PVX vector in plants harboring Sw-5 in order to detect the tospovirus avirulence gene. These genes were not able to trigger the hypersensitive response and to affect PVX infection in Sw-5 plants, which suggest that another tospovirus component is the elicitor of the resistance response. Independently of the Sw-5 gene, PVX clones harboring NSs gene induced more severe symptoms in some solanaceae plants. Gene silencing may be the cause of this symptoms. In transgenic plants harboring Sw-5 gene, the co-expression of the 5 ́ region of this gene by PVX favored tospovirus infection. As this effect was observed both for 5 ́ sense and anti- sense constructions it is possible that it has been caused by reduction on mRNA levels by gene silencing, although Northern blot analysis did not agree with this hypothesis. An Sw-5 homolog was cloned from LA371-20 accession. This homolog is localized in or near of Sw-5 locus and segregated with tospovirus resistance. The capacity of this and other three homologs originated from others accessions to confer tospovirus resistance was evaluated in tobacco and tomato transgenic plants. All the transformants were susceptible to the virus. Other homologs presents in the different accessions evaluated may be responsible for the resistance, although problems in the structure of the constructions can not be discarded. / Tese importada do Alexandria
84

Rendimento e Qualidade do Tabaco Virgínia Afetados Pela Adubação Nitrogenada e Potássica / Yield and Quality of Virgínia Tobacco Affected by Nitrogen and Potassium Fertilization

Jesus, Juliano de 13 December 2016 (has links)
Submitted by Claudia Rocha (claudia.rocha@udesc.br) on 2018-03-07T14:51:34Z No. of bitstreams: 1 PGCS16MA153.pdf: 966411 bytes, checksum: 76076bab2e5f299ad53df33d518a0ef9 (MD5) / Made available in DSpace on 2018-03-07T14:51:34Z (GMT). No. of bitstreams: 1 PGCS16MA153.pdf: 966411 bytes, checksum: 76076bab2e5f299ad53df33d518a0ef9 (MD5) Previous issue date: 2016-12-13 / Virgínia tobacco is a demanding crop in nitrogen (N) and potassium (K) making necessary fertilization to supply these nutrients as a way to guarantee them in available amounts throughout the cycle. The demand of the consumer market requires a high quality product with balanced chemical contents. The objective of this work was to evaluate the use of N and K fertilizers of controlled release in the productivity and quality of the Virgínia tobacco. Two experiments were carried out, in the field, in the 2014/2015 and 2015/2016 growing seasons, in Ituporanga, SC, Southern of Brazil. A randomized complete block design was used, with four replications. In the 2014/2015 season, nine treatments were used; in the 2015/2016 season, ten. Two fertilizer sources were used in pre-planting fertilization and seven sources in sidedressing fertilization. In the experiment of the 2015/2016 season, two control treatments were used, being one with pre-planting fertilization and another with no fertilization. The productivity (dry mass), the economic yield and the amounts of N and K in the soil and the plant were evaluated in two agricultural years with contrasting climatic conditions. The results showed that the limitation of N and K in the soil reduces the yield potential (kg ha-1) and qualitative (IQS) of Virgínia tobacco, reducing the level of nicotine. The use of controlled release fertilizers such as Sulfammo and Agrodiza Fuerza provided tobacco plants with lower productive and qualitative potential in a period with rainfall above average when compared to fertilizers NKalcio (14-00-15) NIP, NKalcio (14-00-15) SOP, Unifertil (03-15-15), Salitre of Chile (15-00-14) ACF and Salitre of Chile (15-00-14) produced By SQM. The efficacy of the fertilizers NKalcio (14-00-15) NIP, NKalcio (14-00-15) SOP, Unifertil (15-03-15) and Salitre of Chile (15-00-14) ACF was similar of that presented by Chilean Salitre (15-00-14), traditional from SQM / O tabaco Virgínia é uma cultura exigente em nitrogênio (N) e potássio (K) fazendo-se necessária adubação de cobertura para repor esses nutrientes, como forma de garanti-los em quantidades necessárias ao longo do ciclo. A demanda do mercado consumidor exige um produto de alta qualidade que ofereça teores químicos balanceados. Este trabalho teve por objetivo avaliar o uso de fertilizantes nitrogenados e potássicos minerais de liberação controlada na produtividade e qualidade da cultura do tabaco Virgínia. Foram conduzidos dois experimentos, a campo, nas safras 2014/2015 e 2015/2016, no município de Ituporanga, SC. Foi utilizado o delineamento experimental de blocos ao acaso, com quatro repetições, sendo que na safra 2014/2015 foi trabalhado com nove tratamentos e na safra 2015/2016 com dez. Foram usadas duas fontes de fertilizantes na adubação de pré-plantio e sete fontes na adubação de cobertura. No experimento da safra 2015/2016 foram utilizados dois tratamentos controle, um somente com adubação de pré-plantio e outro sem adubação. Foram avaliados a produtividade (massa seca), o rendimento econômico e as quantidades de N e K no solo e na planta, em dois anos agrícolas com condições climáticas contrastantes. Os resultados mostraram que a limitação de N e K+ no solo reduz o potencial produtivo (kg ha-1) e qualitativo (IQS) do tabaco Virgínia, reduzindo o nível de nicotina. O uso de fertilizantes de liberação controlada, como Sulfammo e Agrodiza Força, reduz a operação de aplicação de fertilizante em cobertura, porém esses dois fertilizantes proporcionaram plantas de tabaco com menor potencial produtivo e qualitativo em um período com chuvas acima da média quando comparado aos fertilizantes NKalcio (14-00-15) NIP, NKalcio (14-00-15) SOP, Unifertil (15-03-15), Salitre do Chile (15-00-14) ACF e Salitre do Chile (15-00-14) produzido pela empresa SQM. Os fertilizantes NKalcio (14-00-15) NIP, NKalcio (14-00-15) SOP, Unifertil (15-03-15) e Salitre do Chile (15-00-14) ACF apresentam potencial similar ao Salitre do Chile (15-00-14), tradicional da SQM
85

Tnt1 retrotransposon expression and ethylene phytohormone interplay mediates tobacco (Nicotiana tabacum) defense responses / A dinâmica entre a expressão do retrotransposon Tnt1 e o fitormônio etileno envolvida nas respostas de defesa em tabaco (Nicotiana tabacum)

Danielle Maluf Quintanilha 10 October 2014 (has links)
Tnt1 is a transcriptionally active LTR-retrotransposon, present in over 600 copies in the Nicotiana tabacum genome. Under normal growth conditions, Tnt1 expression is limited to basal levels, but its expression is further induced under biotic and abiotic stresses. Transgenic tobacco plants (HP plants) expressing a Tnt1 reverse transcriptase hairpin were generated. These showed pleiotropic phenotypes such as cell death spots on the leaves and callose deposition and other severe abnormal development in aerial and underground portions. RNA sequencing of leaves with cell death spots revealed a rewiring of transcriptional regulatory networks related to stress responses exclusive to HPs. Among the positively modulated genes were ethylene synthesis and response cascade genes. The objective of the present work was to unravel the relation observed between Tnt1 and ethylene, generating a model. The results obtained suggest that HP seedlings and plants have increased ethylene synthesis when compared to the wildtype. Folding prediction of Tnt1 messenger RNA allowed the identification of ethylene-responsive sequences in putative stem loop locations. Thus it is possible that Tnt1 expression can produce small RNAs targeted to sequences present in the Tnt1 retrotransposon itself as well as at the promoter region of other ethylene responsive genes. Quantification of the expression of Tnt1 and ethylene related genes revealed \"phase opposition\" expression kinetics in the HPs, which led us to hypothesize that there might be an antagonistic relationship between the expression of Tnt1 and the expression of ethylene responsive genes involved in plant defense responses. Our findings suggest that Tnt1 could generate sRNAs that exerts transcriptional control over itself as well as other genes. Our model establishes a completely new biological role for a retrotransposon: Tnt1 would provide feedback control to ethylene-mediated gene regulation in tobacco defense responses, bringing the system back to a homeostatic condition and turning the defense responses down. / Tnt1 é um retrotransposon com LTR transcricionalmente ativo, e está presente em mais de 600 cópias no genoma de Nicotiana tabacum. Em condições normais de crescimento Tnt1 é expresso em níveis basais. No entanto, sua expressão é induzida pelo estímulo de estresses bióticos e abióticos. Plantas de tabaco transgênicas (chamadas de HP) expressando um grampo da transcriptase reversa de Tnt1 foram geradas. Estas apresentaram fenótipos como: pontos de morte celular e deposição de calose nas folhas e severas anomalias de desenvolvimento severas nas porções aérea e radicular das plantas. Sequenciamento de RNA de folhas com os pontos de morte celular revelou uma reorganização de redes de regulação transcricional relacionadas a resposta a estresses. Essas novas redes surgiram exclusivamente nas plantas HP. Entre os genes modulados positivamente estavam genes de síntese e de resposta ao etileno. O presente trabalho teve como objetivo elucidar a relação observada entre Tnt1 e o fitormônio etileno gerando um modelo de atuação. Os resultados obtidos permitiram demonstrar que plântulas e plantas HP adultas tem um aumento na síntese de etileno quando comparadas à selvagem. A predição do dobramento do RNA mensageiro de Tnt1 permitiu a identificação de sequências responsivas ao etileno localizadas em posição potencial para formar grampos. Desta forma, é possível que a expressão de Tnt1 leve à produção de pequenos RNAs que tem como alvo sequências responsivas a etileno presentes tanto no próprio elemento quanto em regiões promotoras de outros genes. A quantificação da expressão de Tnt1 versus genes relacionados ao etileno revelou um padrão em \"oposição de fase\" nas HPs, o que nos levou a hipotetizar que talvez ocorra uma relação antagonista entre a expressão de Tnt1 e a expressão de genes responsivos ao etileno envolvidos em respostas de defesa vegetais. Nossos resultados sugerem que Tnt1 pode gerar pequenos RNAs que exercem controle transcricional sobre Tnt1 e outros genes endógenos. Nosso modelo estabelece um novo papel biológico para um retrotransposon: Tnt1 agiria como um modulador da indução de genes mediada por etileno nas respostas de defesa de tabaco, trazendo o sistema de volta à condição homeostática e encerrando as respostas de defesa.
86

Métodos para superação de dormência em sementes de tabaco / Methods to release dormancy in tobacco seeds

Gmach, Janice Regina 20 August 2009 (has links)
Made available in DSpace on 2014-08-20T13:44:42Z (GMT). No. of bitstreams: 1 dissertacao_janice_regina_gmach.pdf: 2217576 bytes, checksum: 44546cca74b285343c5577b59d2b02c1 (MD5) Previous issue date: 2009-08-20 / The growing modernization in agricultural production and the use of small and high cost seeds, usually with the waste of sowing and the disposal seedlings recommend the use of techniques such as pelleting of seeds. However, the use of pelleted seeds may present some problems because the pellet formed around the seed can affect their performance during the germination. The cultivation of tobacco is in itself one of the major agricultural activities in the South Region of Brazil, due to the system of production that allows the cultivation in small areas, with profit to small farmers, giving it relevant socioeconomic role, besides providing raw material for a major industrial complex. Its planting is done through seeds, whose small size hinders its handling, these seeds have difficulties in germination, showing dormancy varying levels in pelleted seeds, a fact that is not seen in raw seeds. This study aimed to verify the efficiency of the use of GA3 and KNO3 as a method of overcoming dormancy in tobacco pelleted seeds, as well as use of two methods for evaluation of tobacco pelleted seeds germination. The results show that the potassium nitrate showed higher percentage of germination in pelleted seeds in both varieties and that the methodology UC (moisture continues) is more sensitive to responses of tobacco pelleted seeds. / A crescente modernização na produção agrícola, assim como a utilização de sementes pequenas e de alto custo, geralmente com desperdícios na semeadura e no descarte de mudas, recomendam o uso de técnicas como a peletização de sementes. No entanto, o uso de sementes peletizadas pode apresentar também alguns problemas, pois o pélete formado ao redor da semente pode afetar seu desempenho durante a germinação. O cultivo do tabaco constitui-se em uma das principais atividades agrícolas da região Sul do Brasil, devido ao sistema de produção que possibilita o cultivo em pequenas áreas, com boa remuneração para os pequenos agricultores, o que lhe confere relevante papel sócio-econômico, além de se constituir em matéria prima para um importante complexo industrial. Seu plantio é feito por meio de sementes, cujo tamanho reduzido dificulta o seu manuseio, essas sementes apresentam dificuldades na germinação, apresentando níveis variáveis de dormência em sementes peletizadas, fato este não observado em sementes nuas. O presente trabalho teve por objetivo verificar a eficiência do uso de AG3 e KNO3 como método de superação de dormência em sementes peletizadas de tabaco, bem como a utilização de duas metodologias para avaliação de germinação de sementes peletizadas de tabaco. Os resultados demonstram que o nitrato de potássio apresentou maiores percentuais de germinação em sementes peletizadas em ambos os cultivares e que a metodologia do UC (Umedecimento Contínuo) é mais sensível ás respostas de sementes de tabaco peletizadas.
87

Etude de l’implication des deux voies de biosynthèse des isoprénoïdes pour la spécificité et la régulation de la prénylation des protéines chez les plantes / Implication of two isoprenoid biosynthesis pathways in the specificity and regulation of protein prenylation in plants

Huchelmann, Alexandre 26 November 2013 (has links)
La prénylation de type I des protéines correspond à une modification post-traductionnelle faisant intervenir une liaison thioéther entre une cystéine localisée dans un motif CaaX en position C terminale et un groupement prényle en C 15 (farnésyle) ou C 20 (géranylgéranyle). Ces réactions sont catalysées par des protéine prényltransférases (PPTs) appartenant à la même famille fonctionnelle et comprenant la protéine farnésyltransférase (PFT) et géranylgéranyltransférase de type I (PGGT-I). Les plantes se distinguent par une double origine des substrats prényle (farnésyle diphosphate et géranylgéranyle diphosphate) utilisés comme précurseurs pour la biosynthèse des isoprénoïdes. Ces derniers sont biosynthétisés par l'intermédiaire de deux voies métaboliques distinctes, la voie cytosolique du mévalonate (MVA) et la voie plastidiale du méthylérythritol phosphate (MEP). Il est maintenant clair que la géranylgéranylation des protéines végétales dépend de la voie du MEP. Durant ce travail de thèse doctorale une étude comparative des spécificités de substrat a été réalisée. Elle a permis de montrer que la PFT est spécifique de son substrat protéique alors que la PGGT-I est spécifique de son substrat prényle. Ces spécificités peuvent néanmoins être modifiées in vivo, par exemple lors d’une augmentation de la concentration en MVA, suggérant que cette flexibilité des propriétés enzymatiques a un rôle régulateur dans certaines conditions physiologiques. Pour cette raison, nous avons entrepris une caractérisation de la prénylation des protéines dans des plantes de tabac élicitées, qui induisent la synthèse de MVA pour produire le capsidiol, une phytoalexine sesquiterpénique. La biosynthèse de ce métabolite secondaire capsidiol dérivant de la voie du MVA, est dépendante de la prénylation des protéines, notamment de protéines géranylgéranylées d’origine plastidiale. Le monoterpène S-carvone a été identifié comme un inhibiteur de la biosynthèse de capsidiol en interférant avec l’activité des PPTs in vivo. Les travaux ont également permis d’envisager l’existence d’un nouveau mode de prénylation des protéines spécifique aux feuilles. / Type-I protein prenylation is a post-translational modification of a protein bearing a CaaX motif with a prenyl moiety, this by a thioether linkage. The enzymes catalyzing those reactions are called protein prenyltransferase (PPTs). Two enzymes are involved, the protein farnesyltransferase (PFT) and the protein geranylgeranyltransferase type I (PGGT-I). They respectively use farnesyl diphosphate and geranylgeranyl diphosphate as substrate. Those precursors are synthetized in plants by two differentbiosynthetic pathways: the cytosolic mevalonate (MVA) and the plastidial methylerythritol phosphate (MEP) pathways. Protein geranylgeranylation is dependent of the MEP pathway. Those specificities can be modified A comparative analysis of PPTs specificity was done during this PhD thesis, revealing that PFT is specific for its protein substrate, while PGGT-I is specific for its prenyl substrate. But those specificities can be modulated in vivo, for instance by increasing the concentration of MVA. This suggests that the regulation of protein prenylation specificities can become functionally important during physiological processes. For that reason we characterized protein prenylation in elicited tobacco plants, which produce the sesquiterpene phytoalexin capsidiol. This metabolite is synthesized via the MVA pathway, and this process depends of protein prenylation, in particular geranylgeranylation, with the substrate coming from plastids. S-Carvone, a monoterpene, was identified as an inhibitor of PPTS, resulting in a lack of capsidiol production. This work also suggests that a new mechanism of prenylation might exist, specifically in leaves.
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Bioatividade de inseticidas botânicos sobre Ascia monuste orseis (lepidoptera: pieridae) / Bioactivity of botanical insecticides on Ascia monuste orseis (lepidoptera: pieridae)

Biermann, Ana Cristina Sapper 29 April 2009 (has links)
Given the great importance of Ascia monuste orseis as insect-pest of Brassicaceae, this work was carried out to evaluate, under laboratory conditions, the action of botanical insecticides on larvae and egg masses, as well as the food consumption of larvae and test, in field conditions, the efficiency of control, of botanical insecticides which showed better results in laboratory tests, on larvae of A. monuste orseis in kale plants (Brassica oleracea L. var. acephala DC). In laboratory tests were evaluated, at a temperature of 25 ± 2ºC, relative humidity of 60 ± 10% and photophase of 14 hours, the effects of aqueous extracts of 10% w/v of tobacco powder (Nicotiana tabacum L.), of leaves of Melia azedarach L., Eucalyptus tereticornis Sm., Cedrella fissilis Vell. and Trichilia claussenii C. DC., of leaves and twigs of Ateleia glazioveana Baill and Ruta graveolens L., as well as DalNeem®, commercial product based on ripe fruits of neem (Azadirachta indica A. Juss) at 1, 5 and 10% v/v. After being offered discs of kale leaves treated with different treatments to larvae of three days of age, the mortality, the viability of the larval stage and food consumption were evaluated. Also the action of the extracts on egg masses was tested. From topical applications of the extracts on larvae of three days the effect of contact was evaluated through observation of the viability and duration of the larval stage, pupal viability and weight of pupae, and the number of adults with defect. Finally, in experiment conducted in the field, the efficiency of control of products that had better performance on tests in laboratory (tobacco powder, DalNeem to 1% and M. azedarach), compared with the standard treatment, product-based deltamethrin (Decis 25 EC) was estimated. It was found that, in laboratory, the tobacco powder was the treatment that best showed ovicidal activity; after ingestion of the extracts tested, the tobacco powder, followed by DalNeem were better in relation to mortality caused to the larvae; DalNeem, tobacco powder and M. azedarach affected their viability; the contact of DalNeem and tobacco powder caused total mortality of larvae and extracts of M. azedarach resulted in little more than half of viable pupae, presenting a tendency to reduce the weight of pupae and causing malformation in the wings of all adults emerged; both by ingestion and contact action, the extracts tested practically did not change the duration of the larval stage and all the extracts caused fagodeterrente effect on A. monuste orseis. To control A. monuste orseis in Brassicaceae crop, it is recommended the commercial product DalNeem to 1%, respecting the product specifications, since the tobacco powder, in the field, causes dark pigmentation on kale plants. / Devido à grande importância de Ascia monuste orseis como inseto-praga de brassicáceas, este trabalho foi desenvolvido com o objetivo de avaliar, em condições de laboratório, a ação de inseticidas botânicos sobre lagartas e posturas, além do consumo alimentar de lagartas, bem como testar, em condições de campo, a eficiência de controle, dos inseticidas botânicos que apresentaram melhores resultados nos testes em laboratório, sobre lagartas de A. monuste orseis em cultivo de couve (Brassica oleracea L. var. acephala DC). Nos testes em laboratório foram avaliados, à temperatura de 25±2ºC, umidade relativa de 60±10% e fotofase de 14 horas, os efeitos dos extratos aquosos a 10% p/v de pó-de-fumo (Nicotiana tabacum L.), de folhas de Melia azedarach L., Eucalyptus tereticornis Sm., Cedrella fissilis Vell. e Trichilia claussenii C. DC., de folhas e ramos de Ateleia glazioveana Baill e Ruta graveolens L., além de DalNeem®, produto comercial à base de frutos maduros de nim (Azadirachta indica A. Juss) a 1, 5 e 10% v/v. Após serem oferecidos discos de folhas de couve tratados com os diferentes tratamentos, a lagartas com três dias de idade, foi avaliada a mortalidade, a viabilidade da fase larval e o consumo alimentar. Foi testada, também, a ação dos extratos sobre posturas. A partir de aplicações tópicas dos extratos sobre lagartas de três dias, foi avaliada a ação de contato dos mesmos através da observação da viabilidade e da duração da fase larval, da viabilidade pupal e do peso das pupas, além do número de adultos com defeito. Finalmente, em ensaio realizado no campo, foi estimada a eficiência de controle dos produtos que melhor desempenho tiveram nos testes em laboratório (pó-de-fumo, DalNeem a 1% e M. azedarach), comparados com tratamento padrão, produto à base de deltametrina (Decis 25 CE). Verificou-se que, em laboratório, o pó-de-fumo foi o extrato que apresentou melhor atividade ovicida; após ingestão dos extratos testados, destacou-se o pó-de-fumo, seguido de DalNeem, em relação à mortalidade causada a lagartas, além do DalNeem, pó-de-fumo e M. azedarach afetarem a viabilidade das mesmas; sob ação de contato, DalNeem e pó-de-fumo ocasionaram total mortalidade às lagartas e o extrato de M. azedarach destacou-se por resultar em pouco mais da metade de pupas viáveis, por apresentar uma tendência na diminuição do peso das pupas e por ocasionar malformação nas asas da totalidade de adutos emergidos; tanto sob ação de ingestão como de contato, os extratos testados praticamente não acarretaram alteração na duração da fase larval e todos os extratos ocasionaram efeito fagodeterrente sobre o curuquerê-da-couve. Para o controle de A. monuste orseis em cultivos de brassicáceas, recomenda-se o produto comercial DalNeem a 1%, respeitando-se as especificações do produto, já que o pó-de-fumo, no campo, ocasiona pigmentação escura em plantas de couve.
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Identification et étude du rôle des protéines cibles du monoxyde d'azote (NO) dans les réponses de défense chez le tabac / Identification an characterization of nitric oxyde (No) target proteins in tabacco defense responses

Astier, Jérémy 30 May 2011 (has links)
Les études entreprises depuis une douzaine d'années indiquent que le monoxyde d'azote (NO) est un médiateur physiologique impliqué dans de nombreux processus chez les plantes, incluant la germination, le développement des racines, la fermeture des stomates ou encore la réponse adaptative aux stress biotiques et abiotiques. Malgré cet important panel de fonctions, les mécanismes sous-jacents aux effets du NO ont été peu appréhendés et restent pour l'essentiel énigmatiques. Le travail présenté dans ce manuscrit s'inscrit dans cette problématique et a consisté en l’identification et la caractérisation de protéines cibles du NO chez le tabac dans le contexte de stress biotiques et abiotiques. Nous avons démontré que la cryptogéine, un éliciteur des réactions de défense, induit la S-nitrosylation rapide et transitoire de plusieurs protéines dans des suspensions cellulaires de tabac. Après purification, une douzaine de ces protéines ont été identifiées via une analyse par spectrométrie de masse. Celles-ci incluent notamment une protéine chaperonne de la famille des AAA-ATPase nommée CDC48 (Cell Division Cycle 48). Cette dernière a fait l'objet d'une étude structure/fonction approfondie afin d'appréhender l'impact de sa S-nitrosylation. Après avoir vérifié que la protéine recombinante était S-nitrosylable in vitro, nous avons démontré que ce processus n'affecte pas la structure secondaire de la protéine mais induit des modifications locales de sa structure tertiaire et une inhibition de son activité ATPasique. Le résidu cystéine 526, localisé dans le second domaine ATPasique de la protéine, a été identifié comme site probable de S-nitrosylation. Cette localisation stratégique pourrait expliquer l'effet inhibiteur du NO sur l'activité enzymatique de CDC48. La dernière partie de ce travail a été centrée sur l'analyse des mécanismes par lesquels le NO active la protéine kinase NtOSAK (Nicotiana tabacum stress activated protein kinase) chez le tabac. Nous avons démontré que NtOSAK forme un complexe constitutif avec la glycéraldéhyde 3 phosphate deshydrogénase (GAPDH). En réponse à un stress salin, le NO promeut l'activation de NtOSAK via la phosphorylation de deux résidus serine localisés dans la boucle d'activation de l'enzyme. De plus, il induit une S-nitrosylation rapide de la GAPDH, ce processus n'affectant pas la formation du complexe. Notre hypothèse est que ce complexe constituerait une plateforme de signalisation régulée par le NO et pouvant recruter les protéines cibles de NtOSAK lors de la réponse au stress salin. / Studies conducted over the past ten years indicate that nitric oxide (NO) is a physiological mediator involved in many physiological processes in plants, including germination, root development, stomatal closure or responses against biotic or abiotic stresses. Despite this important range of functions, the mechanisms underlying the effects of NO in plants remain largely unknown. The present work aims at identifying and functionally characterizing NO target proteins in tobacco in the context of biotic and abiotic stresses. We demonstrated that cryptogein, an elicitor of defense responses, induces a rapid and transient S-nitrosylation of several proteins in tobacco cell suspensions. After purification, a dozen of these proteins have been identified through mass spectrometry analysis. These proteins include CDC48 (Cell Division Cycle 48), a chaperone-like protein belonging to the AAA-ATPase family. The regulation of CDC48 by NO was deeply investigated using a combination of structural and biochemical analyses. Once the in vitro S-nitrosylation of CDC48 was confirmed, we next demonstrated that this process does not affect the secondary structure of the protein but induces local changes in its tertiary structure together with an inhibition of its ATPase activity. The cysteine residue 526, located in the second ATPase domain of the protein, was identified as a probable S-nitrosylation site. This crucial localization may explain the inhibitory effect of NO on CDC48 enzymatic activity. The last part of this work was focused on the analysis of the mechanisms underlying the NO-dependent activation of the protein kinase NtOSAK (Nicotiana tabacum stress activated protein kinase) in tobacco. We demonstrated that NtOSAK forms a constitutive complex with glyceraldehyde 3-phosphate dehydrogenase (GAPDH). In response to salt stress, NO promotes the activation of NtOSAK via the phosphorylation of two serine residues located in the activation loop of the enzyme. Moreover, it induces a rapid S-nitrosylation of GAPDH. Interestingly, this latter process does not affect the complex formation. Our hypothesis is that once S-nitrosylated, GAPDH might act as a phosphorelay recruiting protein substrates for NtOSAK.
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Stress photooxydatif et mécanismes de tolérance en réponse à un herbicide, le clomazone, chez deux variétés de tabac (Nicotiana tabacum L.) / Photooxidative stress and tolerance mechanisms in the response to clomazone herbicide in two varities of tobacco (Nicotianan tabacum L.)

Darwish, Majd 11 December 2014 (has links)
Les effets de faibles doses de clomazone 2-(2-chlorobenzyl)-4,4-dimethyl-1,2-oxazolidin-3-one, un herbicide non-sélectif de la famille des isoxazolidinones, ont été étudiés chez Nicotiana tabacum L. De faibles concentrations de clomazone (1, 0.1, 0.01, 0.001, 0.0001 et 0.00001 µM), ont été appliquées chez deux variétés de tabac (Nicotiana tabacum L. cv. Xanthi; Nicotiana tabacum L. cv.Virginie vk51) cultivées en hydroponie. Le transport d'électrons photosynthétiques a été étudié par l'analyse de la cinétique rapide de fluorescence chlorophyllienne et par la mesure de l'évolution de l'oxygène chloroplastique. Les résultats ont montré que le clomazone à des concentrations de 1, 0.1, 0.01, 0.001et 0.0001 µM, affecte les paramètres JIP (Fv/Fm, PIabs, 1–VJ, ABS/RC, TR0/RC, DI0/RC, ET0/RC et ET0/ABS). Le clomazone à une concentration de 1 µM, conduit à une importante accumulation de H2O2 chez Virginie induisant la mort cellulaire programmée (PCD). Nos résultats montrent que la variété Xanthi présente une tolérance plus importante au clomazone que la variété Virginie, ce qui peut s'expliquer par une meilleure efficacité du transport d'électrons au delà de QA, ainsi qu'une amélioration du fonctionnement du système antioxydant enzymatique (APX et MDHAR). Le prétraitement des plantules de la variété Virginie par une alternance de cycles courts de lumière/obscurité (16 min/ 8min) pendant trois jours lorsque les plantules sont au stade de trois feuilles, a amélioré leur tolérance face à la toxicité du clomazone. Ce prétraitement induit l'activation des systèmes de détoxification des ERO via l'augmentation des activités des enzymes antioxydants et la biosynthèse de composés antioxydants tels que l'ascorbate et les composés phénoliques et réduit le niveau de mort cellulaire programmée, induit par le stress photooxydatif. Il est supposé que la stimulation des voies alternatives de dissipation des électrons, telle que la photorespiration, ainsi que les modifications du cycle circadien, soient prioritairement impliquées dans l'acquisition de la tolérance. Cet effet dit de "priming" se maintient tout au long de la croissance du tabac et se manifeste par une augmentation de la croissance et de la photosynthèse nette qui perdure jusqu'au stade de la floraison. / The effect of low doses of clomazone 2-(2-chlorobenzyl)-4,4-dimethyl-1,2-oxazolidin-3-one, a non-selective herbicide of the isoxazolidinone family, has been already determined in Nicotiana tabacum L. The plantlets of two varieties of tobacco (Nicotiana tabacum L. cv. Xanthi; Nicotiana tabacum L. cv. Virginie vk51) were subjected to low concentrations of clomazone (1, 10−1, 10−2, 10−3, 10−4 and 10−5) µM in a hydroponic system. The photosynthetic electron transport has been studied by analyzing the rapid kinetics of chlorophyll a fluorescence and measuring changes in the chloroplast oxygen evolution. The results showed that clomazone at concentrations of 1, 10−1, 10−2, 10−3 and 10−4 µM affected several parameters of chlorophyll a fluorescence and OJIP test (Fv/Fm, PIabs, 1-VJ, ABS/RC, TR0/RC, DI0/RC, ET0/RC and ET0/ABS). Clomazone at 1 µM led to a large accumulation of H2O2 in Virginie variety inducing programmed cell death. Our results show that Xanthi variety has a higher tolerance to clomazone as compared to Virginie, this can be explained by increasing the efficiency of electron transport beyond QA− (1–VJ) as well as improving the function of antioxidant enzymes (APX and MDHAR). The pretreatment of Virginie plantlets (three leaf stage) with alternation of short cycles (16 min/ 8min) of light/dark (AL) for three days, has improved its tolerance to clomazone toxicity. This pretreatment stimulated ROS-detoxification systems by increasing the activity of antioxidant enzymes and the biosynthesis of antioxidant compounds such as ascorbate acid and phenolic compounds, and consequently reduced the level of programmed cell death (PCD) provoked by the stress photooxydatif. It is suggested that the stimulation of alternative pathways of electron dissipation, such as the photorespiration, as well as changes in the circadian cycle, are primarily involved in the acquisition of the tolerance. This effect so-called "priming" has maintained during the growth of tobacco and has been revealed by increases in the growth and net photosynthesis until the flowering stage.

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