<p>&nbsp / </p> <p align="left">The domesticated apple (<i><font face="TimesNewRomanPS-ItalicMT">Malus </font><font face="TimesNewRomanPSMT">x </font><i><font face="TimesNewRomanPS-ItalicMT">domestica </font><font face="TimesNewRomanPSMT">Borkh.), belonging to the </font><i><font face="TimesNewRomanPS-ItalicMT">Malus </font><font face="TimesNewRomanPSMT">genus of the Rosaceae family, is one of the edible pomaceous fruits. Since it is one of the important commercial fruit crops worldwide, the quality of the fruit is crucial to breeders and farmers as it ultimately determines acceptance of a cultivar for consumption. Fruit quality is also a critical determinant factor that is used to estimate the potential of apples to have a long shelf life. The introduction of marker-assisted selection (MAS) has allowed hastening of traditional breeding and selection of high-quality apple cultivars. The availability of genetic linkage maps, constructed by positioning molecular markers throughout the apple genome, enables the detection and analysis of major genes and quantitative trait loci (QTLs) contributing to the quality traits of a given genotype.&nbsp / herefore, the primary aim of this study was to construct a genetic linkage map of the &lsquo / Golden Delicious&rsquo / x &lsquo / Dietrich&rsquo / population for the identification of QTLs associated with fruit quality traits and then to examine the apple fruit pulp proteome with a specific focus on fruit firmness. In this regard, genomic DNA was extracted from leaves of the &lsquo / Golden Delicious&rsquo / x Dietrich&rsquo / population and used in megaplex PCR reactions. The PCR products were analysed prior to scoring of alleles. Polymorphic markers were then used to construct genetic linkage maps. The genetic linkage maps constructed in this study comprise of 167 simple sequence repeats (SSR) markers, 33 of these were newly developed markers. The 17 linkage groups of apple were constructed and aligned to existing apple genetic maps. The maps span 1,437.8 cM and 1,491.5 cM for &lsquo / Golden Delicious&rsquo / and &lsquo / Dietrich&rsquo / , respectively.</font></i></i></i></p>

<p>&nbsp / </p> <p align="left">The domesticated apple (<i><font face="TimesNewRomanPS-ItalicMT">Malus </font><font face="TimesNewRomanPSMT">x </font><i><font face="TimesNewRomanPS-ItalicMT">domestica </font><font face="TimesNewRomanPSMT">Borkh.), belonging to the </font><i><font face="TimesNewRomanPS-ItalicMT">Malus </font><font face="TimesNewRomanPSMT">genus of the Rosaceae family, is one of the edible pomaceous fruits. Since it is one of the important commercial fruit crops worldwide, the quality of the fruit is crucial to breeders and farmers as it ultimately determines acceptance of a cultivar for consumption. Fruit quality is also a critical determinant factor that is used to estimate the potential of apples to have a long shelf life. The introduction of marker-assisted selection (MAS) has allowed hastening of traditional breeding and selection of high-quality apple cultivars. The availability of genetic linkage maps, constructed by positioning molecular markers throughout the apple genome, enables the detection and analysis of major genes and quantitative trait loci (QTLs) contributing to the quality traits of a given genotype.&nbsp / herefore, the primary aim of this study was to construct a genetic linkage map of the &lsquo / Golden Delicious&rsquo / x &lsquo / Dietrich&rsquo / population for the identification of QTLs associated with fruit quality traits and then to examine the apple fruit pulp proteome with a specific focus on fruit firmness. In this regard, genomic DNA was extracted from leaves of the &lsquo / Golden Delicious&rsquo / x Dietrich&rsquo / population and used in megaplex PCR reactions. The PCR products were analysed prior to scoring of alleles. Polymorphic markers were then used to construct genetic linkage maps. The genetic linkage maps constructed in this study comprise of 167 simple sequence repeats (SSR) markers, 33 of these were newly developed markers. The 17 linkage groups of apple were constructed and aligned to existing apple genetic maps. The maps span 1,437.8 cM and 1,491.5 cM for &lsquo / Golden Delicious&rsquo / and &lsquo / Dietrich&rsquo / , respectively.</font></i></i></i></p>

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2003. / Härtill 5 uppsatser.

Esta tese alia dois enfoques principais dentro da Química Farmacêutica. Por um lado, busca explorar uma nova fonte de insumos naturais, cascas de Malus  domestica, com vistas a obtenção dos triterpenos ácidos ursólico (AU) e betulínico (AB); e por outro lado, estuda a relação dos derivados triterpênicos semissintéticos obtidos com a proteína-alvo (PfATP6) destacada atualmente na literatura da terapia da malária com vistas ao planejamento de novos antimaláricos. O primeiro Capítulo inclui o desenvolvimento de um método eficiente, fácil e extremamente rápido onde são combinadas as técnicas de extração por ultrassom (UAE) e análise por Cromatografia Líquida de Alta Eficiência Acoplada a Detector de Espectroscopia de Massas (LC-MS), para identificação e doseamento dos ácidos ursólico (AU) e betulínico (BA) em extratos de cascas frescas de maçã de cinco clones das cultivares Gala e Fuji (“Baigent”, “Fuji Mishima”, “Fuji Suprema”, “Fuji Select” and “Maxi Gala”) oriundas da Região Sul do Brasil. Os parâmetros cromatográficos do método analítico incluem: ionização por eletro spray em modo positivo (ESI+), fluxo de 1,0 mL/min, em modo de eluição isocrático, consistindo de 80% acetonitrila e 20% acetato de amônio 10 mM em pH 6,0 e temperatura ambiente. O método desenvolvido foi validado e mostrou ser seletivo, sensível (LOD e LOQ de 0,087 e 0,266 μg/mL para BA, e 0,398 e 2,117 μg/ mL para UA), coeficiente de regressão linear (r >0.99), preciso, exato e robusto para os analitos de interesse. A otimização do método combinado de UAE com LC-MS permitiu concluir os procedimentos de extração e análise em tempo inferior a 4 h, uma vez que o método não requer a secagem da amostra, etapa que demanda longos tempos de processamento. Este método foi aplicado e forneceu a primeira caracterização fitoquímica dos cinco clones de maçã estudados. Os resultados demonstraram que o método combinado de UAE-LC-MS é adequado às práticas do controle de qualidade. O segundo Capítulo apresenta o estudo da interação dos ligantes semissintéticos derivados dos AU e AB, sintetizados pelo nosso grupo de pesquisa, com a proteína Ca2+-ATPase do Plasmodium falciparum (PfATP6), através do emprego da técnica de Docking Molecular. A PfATP6 é descrita como um importante alvo para novos antimaláricos como Artemisinina (ART), cujo mecanismo de ação inclui, dentre outros, a modulação da homeostasia do cálcio intracelular. Investigações conduziram à hipótese do extravasamento do cálcio, do interior do retículo sarco-endoplasmático como mecanismo plausível para ação dos derivados triterpênicos do AU e AB. Os escores de energia determinados no Docking, de cada um dos nove ligantes (derivados triterpênicos) e quatro compostos de controle (ácidos ursólico e betulínico, artemisinina (ART) e tapsigargina (TPG) foram determinados (análises realizadas em triplicata) e correlacionados com os valores de IC50, para a atividade antimalárica. Os resultados mostraram excelente correlação entre os escores de energia (com a PfATP6) e os valores de IC50, superior a 80% (r > 0,83360). O estudo fornece fortes evidências de que a PfATP6 pode constituir um alvo para os derivados pentacíclicos do estudo, bem como permitiu identificar o perfil conformacional dos ligantes e os principais resíduos do sítio de ligação (SL) da PfATP6 envolvidos nas interações; bem como, contribuiu para a melhor compreensão das propriedades envolvidas na interação do ligante com o receptor e mecanismo de ação. O terceiro Capítulo faz uso de Métodos Clássicos e Quânticos para descrever as mudanças conformacionais da proteína PfATP6. As simulações de Dinâmica Molecular do receptor na forma isolada e complexada com os ligantes foram realizadas durante 10 ns. As conformações finais obtidas para os receptores foram avaliadas em termos RMSD, efeito da presença dos ligantes no sítio de ligação e tipos de interações estabelecidas entre o ligante e o receptor. Os resultados mostraram que as proteínas PfATP6 e SERCA tendem a manter sua conformação nativa e que o modelo utilizado é adequado aos propósitos do estudo. O monitoramento dos resíduos da região citoplasmática das proteínas permitiu evidenciar o efeito alostérico da presença dos ligantes AU e ART no SL, sobre os domínios A e N, da PfATP6. Esse efeito reproduz as conformações E1 e E2, bem estabelecidas para PfATP6, na presença e ausência de Ca2+. As análises de Dinâmica Molecular corroboram os achados do Capítulo II ao evidenciarem o estabelecimento de interações de hidrogênio com os resíduos importantes do SL da PfATP6. Esses resultados fundamentam os indícios de que as bombas de Ca2+-ATPase (SERCA), possam ser de fato, um alvo para os derivados triterpênicos dos AU e AB. / This thesis combines two main focuses within Pharmaceutical Chemistry. On the one hand, it seeks to explore a new source of natural inputs, bark of Malus domestica, in order to obtain ursolic (UA) and betulinic acid (BA) triterpenes. On the other hand, it studies the relation of the semi-synthetic triterpenic derivatives obtained with the target proteins (PfATP6), currently highlighted in the literature on malaria therapy with a view to planning new antimalarial drugs. The first chapter includes the development of an efficient, easy and extremely fast method where ultrasonic extraction techniques (UAE) and high-performance liquid chromatography coupled to mass spectroscopy (LC-MS) are combined for identification and assay of ursolic acid (UA) and betulinic acid (BA) in fresh apple peel extracts from five clones of the Gala and Fuji cultivars (Baigent, Fuji Mishima, Fuji Suprema, Fuji Select and Maxi Gala ") in the Southern Region of Brazil. Chromatographic parameters of the analytical method include: electrospray ionization (ESI +), flow rate of 1.0 mL/min in isocratic elution mode, consisting of 80% acetonitrile and 20% 10 mM ammonium acetate at pH 6.0 and room temperature. The method was validated and proved to be selective, sensitive (LOD and LOQ of 0.087 and 0.266 μg/mL for BA, and 0.398 and 2.117 μg/mL for UA), linear regression coefficient (r> 0.99), accurate, robust for analytes of interest. The optimization of the combined method of UAE with LC-MS allowed to complete the procedures of extraction and analysis in less than 4 h, since the method does not require drying the sample, a stage that demands long processing times. This method was applied and provided the first phytochemical characterization of the five apple clones studied. The results demonstrated that the combined UAE-LC-MS method is suitable for quality control practices. The second chapter presents the study of the interaction of the semi-synthetic ligands derived from the UA and BA, synthesized by our research group, with the Plasmodium falciparum Ca2+-ATPase protein (PfATP6), using the Molecular Docking technique. PfATP6 is described as an important target for new antimalarials such as Artemisinin (ART), whose action mechanism includes, among others, the modulation of intracellular calcium homeostasis. Investigations led to the hypothesis of extravasation of calcium from the interior of the sarco-endoplasmic reticulum as a plausible mechanism for the action of the triterpenic derivatives of UA and BA. The Docking energy scores (binding energy) of each of the nine ligands (triterpenic derivatives) and four control compounds (ursolic and betulinic acids, artemisinin (ART) and tapsigargine (TPG)) with PfATP6, were calculated (analyses performed in triplicate) and correlated with its antimalarial IC50 value. The results showed an excellent correlation between energy scores (with PfATP6) and IC50 values, higher than 80% (r > 0.83360). The study supplies strong evidence that PfATP6 may be a target for the pentacyclic derivatives of the study, and also allowed identifying the conformational profile of the ligands and the main residues of the PfATP6 binding site (BS) involved in the interactions. It further contributed to a better understanding of the properties involved in the interaction of the ligand with the receptor and its action mechanism. The third chapter uses Classical and Quantum Methods to describe the conformational changes of the PfATP6 protein. The Molecular Dynamics simulations of the receptor in the isolated and complexed form with the ligands were performed for 10 ns. The final conformations obtained for the receptors were evaluated in RMSD terms, effect of the presence of ligands at the binding site and types of interactions established between the ligand and the receptor. The results showed that the PfATP6 and SERCA proteins tend to maintain their native conformations and that the model used is adequate for the purposes of the study. The monitoring of the residues of the cytoplasmic region of the proteins allowed evidencing the allosteric effect of the presence of UA and ART ligands in BS, on the A- and N-domains of PfATP6. This effect reproduces the well-established E1 and E2 conformations for PfATP6, in the presence and absence of Ca2+, respectively. Molecular Dynamics analyses corroborate the findings of Chapter II, by showing the possibility of establishing hydrogen interactions with the important residues of PfATP6 BS. These results support the evidence that Ca2+-ATPase (SERCA) pumps may be a target for the triterpenic derivatives of UA and BA.

In 2013 and 2014 the occurrence of originator of Monilia Heat Stroke and Monilia Rot Fungi of the genus Monilia was observed in the Botanical Gardens and Arboretum, Mendel University and at other locations, e.g. Hranice na Moravě, Lednice, Namest nad Oslavou The following ones have been included in the observations: genera Cotoneaster, Malus, Prunus Persica, Pyrus, Sorbus, Cydonia, Crataegus, Chaenomeles and Mespilus. Based on morphological data, determination of Monilia species have been performed. Most commonly encountered species in the area BZA were Monilia laxa and M. fructigena. According to themorphological observation, it was determined thath kind M. polystroma appeared to be the most abundant species to the genus. The occurrence of lesser known species M. linhartiana on fruits Cydonia oblonga and Sorbus mougeotii has been confirmed.

Esta tese alia dois enfoques principais dentro da Química Farmacêutica. Por um lado, busca explorar uma nova fonte de insumos naturais, cascas de Malus  domestica, com vistas a obtenção dos triterpenos ácidos ursólico (AU) e betulínico (AB); e por outro lado, estuda a relação dos derivados triterpênicos semissintéticos obtidos com a proteína-alvo (PfATP6) destacada atualmente na literatura da terapia da malária com vistas ao planejamento de novos antimaláricos. O primeiro Capítulo inclui o desenvolvimento de um método eficiente, fácil e extremamente rápido onde são combinadas as técnicas de extração por ultrassom (UAE) e análise por Cromatografia Líquida de Alta Eficiência Acoplada a Detector de Espectroscopia de Massas (LC-MS), para identificação e doseamento dos ácidos ursólico (AU) e betulínico (BA) em extratos de cascas frescas de maçã de cinco clones das cultivares Gala e Fuji (“Baigent”, “Fuji Mishima”, “Fuji Suprema”, “Fuji Select” and “Maxi Gala”) oriundas da Região Sul do Brasil. Os parâmetros cromatográficos do método analítico incluem: ionização por eletro spray em modo positivo (ESI+), fluxo de 1,0 mL/min, em modo de eluição isocrático, consistindo de 80% acetonitrila e 20% acetato de amônio 10 mM em pH 6,0 e temperatura ambiente. O método desenvolvido foi validado e mostrou ser seletivo, sensível (LOD e LOQ de 0,087 e 0,266 μg/mL para BA, e 0,398 e 2,117 μg/ mL para UA), coeficiente de regressão linear (r >0.99), preciso, exato e robusto para os analitos de interesse. A otimização do método combinado de UAE com LC-MS permitiu concluir os procedimentos de extração e análise em tempo inferior a 4 h, uma vez que o método não requer a secagem da amostra, etapa que demanda longos tempos de processamento. Este método foi aplicado e forneceu a primeira caracterização fitoquímica dos cinco clones de maçã estudados. Os resultados demonstraram que o método combinado de UAE-LC-MS é adequado às práticas do controle de qualidade. O segundo Capítulo apresenta o estudo da interação dos ligantes semissintéticos derivados dos AU e AB, sintetizados pelo nosso grupo de pesquisa, com a proteína Ca2+-ATPase do Plasmodium falciparum (PfATP6), através do emprego da técnica de Docking Molecular. A PfATP6 é descrita como um importante alvo para novos antimaláricos como Artemisinina (ART), cujo mecanismo de ação inclui, dentre outros, a modulação da homeostasia do cálcio intracelular. Investigações conduziram à hipótese do extravasamento do cálcio, do interior do retículo sarco-endoplasmático como mecanismo plausível para ação dos derivados triterpênicos do AU e AB. Os escores de energia determinados no Docking, de cada um dos nove ligantes (derivados triterpênicos) e quatro compostos de controle (ácidos ursólico e betulínico, artemisinina (ART) e tapsigargina (TPG) foram determinados (análises realizadas em triplicata) e correlacionados com os valores de IC50, para a atividade antimalárica. Os resultados mostraram excelente correlação entre os escores de energia (com a PfATP6) e os valores de IC50, superior a 80% (r > 0,83360). O estudo fornece fortes evidências de que a PfATP6 pode constituir um alvo para os derivados pentacíclicos do estudo, bem como permitiu identificar o perfil conformacional dos ligantes e os principais resíduos do sítio de ligação (SL) da PfATP6 envolvidos nas interações; bem como, contribuiu para a melhor compreensão das propriedades envolvidas na interação do ligante com o receptor e mecanismo de ação. O terceiro Capítulo faz uso de Métodos Clássicos e Quânticos para descrever as mudanças conformacionais da proteína PfATP6. As simulações de Dinâmica Molecular do receptor na forma isolada e complexada com os ligantes foram realizadas durante 10 ns. As conformações finais obtidas para os receptores foram avaliadas em termos RMSD, efeito da presença dos ligantes no sítio de ligação e tipos de interações estabelecidas entre o ligante e o receptor. Os resultados mostraram que as proteínas PfATP6 e SERCA tendem a manter sua conformação nativa e que o modelo utilizado é adequado aos propósitos do estudo. O monitoramento dos resíduos da região citoplasmática das proteínas permitiu evidenciar o efeito alostérico da presença dos ligantes AU e ART no SL, sobre os domínios A e N, da PfATP6. Esse efeito reproduz as conformações E1 e E2, bem estabelecidas para PfATP6, na presença e ausência de Ca2+. As análises de Dinâmica Molecular corroboram os achados do Capítulo II ao evidenciarem o estabelecimento de interações de hidrogênio com os resíduos importantes do SL da PfATP6. Esses resultados fundamentam os indícios de que as bombas de Ca2+-ATPase (SERCA), possam ser de fato, um alvo para os derivados triterpênicos dos AU e AB. / This thesis combines two main focuses within Pharmaceutical Chemistry. On the one hand, it seeks to explore a new source of natural inputs, bark of Malus domestica, in order to obtain ursolic (UA) and betulinic acid (BA) triterpenes. On the other hand, it studies the relation of the semi-synthetic triterpenic derivatives obtained with the target proteins (PfATP6), currently highlighted in the literature on malaria therapy with a view to planning new antimalarial drugs. The first chapter includes the development of an efficient, easy and extremely fast method where ultrasonic extraction techniques (UAE) and high-performance liquid chromatography coupled to mass spectroscopy (LC-MS) are combined for identification and assay of ursolic acid (UA) and betulinic acid (BA) in fresh apple peel extracts from five clones of the Gala and Fuji cultivars (Baigent, Fuji Mishima, Fuji Suprema, Fuji Select and Maxi Gala ") in the Southern Region of Brazil. Chromatographic parameters of the analytical method include: electrospray ionization (ESI +), flow rate of 1.0 mL/min in isocratic elution mode, consisting of 80% acetonitrile and 20% 10 mM ammonium acetate at pH 6.0 and room temperature. The method was validated and proved to be selective, sensitive (LOD and LOQ of 0.087 and 0.266 μg/mL for BA, and 0.398 and 2.117 μg/mL for UA), linear regression coefficient (r> 0.99), accurate, robust for analytes of interest. The optimization of the combined method of UAE with LC-MS allowed to complete the procedures of extraction and analysis in less than 4 h, since the method does not require drying the sample, a stage that demands long processing times. This method was applied and provided the first phytochemical characterization of the five apple clones studied. The results demonstrated that the combined UAE-LC-MS method is suitable for quality control practices. The second chapter presents the study of the interaction of the semi-synthetic ligands derived from the UA and BA, synthesized by our research group, with the Plasmodium falciparum Ca2+-ATPase protein (PfATP6), using the Molecular Docking technique. PfATP6 is described as an important target for new antimalarials such as Artemisinin (ART), whose action mechanism includes, among others, the modulation of intracellular calcium homeostasis. Investigations led to the hypothesis of extravasation of calcium from the interior of the sarco-endoplasmic reticulum as a plausible mechanism for the action of the triterpenic derivatives of UA and BA. The Docking energy scores (binding energy) of each of the nine ligands (triterpenic derivatives) and four control compounds (ursolic and betulinic acids, artemisinin (ART) and tapsigargine (TPG)) with PfATP6, were calculated (analyses performed in triplicate) and correlated with its antimalarial IC50 value. The results showed an excellent correlation between energy scores (with PfATP6) and IC50 values, higher than 80% (r > 0.83360). The study supplies strong evidence that PfATP6 may be a target for the pentacyclic derivatives of the study, and also allowed identifying the conformational profile of the ligands and the main residues of the PfATP6 binding site (BS) involved in the interactions. It further contributed to a better understanding of the properties involved in the interaction of the ligand with the receptor and its action mechanism. The third chapter uses Classical and Quantum Methods to describe the conformational changes of the PfATP6 protein. The Molecular Dynamics simulations of the receptor in the isolated and complexed form with the ligands were performed for 10 ns. The final conformations obtained for the receptors were evaluated in RMSD terms, effect of the presence of ligands at the binding site and types of interactions established between the ligand and the receptor. The results showed that the PfATP6 and SERCA proteins tend to maintain their native conformations and that the model used is adequate for the purposes of the study. The monitoring of the residues of the cytoplasmic region of the proteins allowed evidencing the allosteric effect of the presence of UA and ART ligands in BS, on the A- and N-domains of PfATP6. This effect reproduces the well-established E1 and E2 conformations for PfATP6, in the presence and absence of Ca2+, respectively. Molecular Dynamics analyses corroborate the findings of Chapter II, by showing the possibility of establishing hydrogen interactions with the important residues of PfATP6 BS. These results support the evidence that Ca2+-ATPase (SERCA) pumps may be a target for the triterpenic derivatives of UA and BA.

Made available in DSpace on 2014-06-11T19:26:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2005-06-20Bitstream added on 2014-06-13T20:15:27Z : No. of bitstreams: 1 moreira_gc_me_botfca.pdf: 522936 bytes, checksum: 59249a8e43961d1234678ad95d42c9af (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O presente trabalho teve por objetivo verificar os efeitos do ácido ascórbico e da radiação gama na conservação pós-colheita de maçã Royal Gala minimamente processada, submetida à refrigeração, visando estender a sua vida útil, retardando seu amadurecimento e reduzindo a incidência de patógenos, sem prejuízo das suas características físicas, químicas e organolépticas. Foram realizados 4 experimentos: Experimento 1 - os frutos processados em cubos de 2 x 2 cm; Experimento 2 - os frutos processados em rodelas de 1 cm; Experimento 3 - os frutos cortados em 4 partes. Para os experimentos 1, 2 e 3, após o processamento nos diferentes cortes, os frutos foram acondicionadas em bandejas de isopor envolvidas em filme plástico e submetidas a diferentes concentrações de ácido ascórbico (0%, 1%, 2%, 3% e 4%); Experimento 4 - os frutos processados em cubos de 2 x 2 cm, imersos em solução de 3% de ácido ascórbico, acondicionados em bandejas de isopor envolvidas em filme plástico e irradiadas com diferentes doses de radiação gama (T1 - Testemunha (0,0kGy), T2 - 0,1kGy, T3 - 0,2kGy, T4 - 0,3kGy, T5 - 0,4kGy, T6 - 0,5kGy). As frutas foram irradiadas na EMBRARAD, localizada em Cotia - SP. Foram utilizadas nessa pesquisa maçãs Royal Gala, provenientes da Empresa Agroban (Vacaria - RS). Os frutos foram transportados ao Laboratório de Frutas e Hortaliças, pertencente ao Departamento de Gestão e Tecnologia Agroindustrial, da Faculdade de Ciências Agronômicas - UNESP - Câmpus de Botucatu, SP. Para cada experimento, os frutos após receberem cada tratamento, foram armazenados em câmara fria a 5l1ºC e 85 - 90% de umidade relativa. As avaliações foram realizadas a cada dois dias, durante 10 dias de armazenamento. As alterações na qualidade pós-colheita foram detectadas por meio das análises de perda de massa fresca... / The present work had as objective to verify the effects of the ascorbic acid and the gamma radiation in the postharvest conservation of the minimally processed apple 'Royal Gala' minimally processed, submitted to the refrigeration aiming to extend its useful life, delaying its ripening and reducing the microorganisms incidence, without damage of its physical, chemistries and sensorial characteristics. Four experiments were accomplished: Experiment 1 - the fruits processed in cubes of 2 x 2 cm; Experiment 2 - the fruits processed in slices of 1 cm; Experiment 3 - the fruits cut in 4 parts. For the experiments 1, 2 and 3, after the processing in the different cuts, the fruits were conditioned in isopor trays involved in plastic film and submitted to different concentrations of ascorbic acid (0%, 1%, 2%, 3% and 4%); Experiment 4 - the fruits processed in cubes of 2 x 2 cm, immersed in solution of 3% of ascorbic acid, conditioned in isopor trays involved in plastic film and irradiated with different gamma radiation doses (T1 - control (0,0kGy), T2 - 0,1kGy, T3 - 0,2kGy, T4 - 0,3kGy, T5 - 0,4kGy, T6 - 0,5kGy). The fruits were irradiated in EMBRARAD, located in Cotia - SP. Apples ' Royal Gala' were used in this research, coming from the Company Agroban (Vacaria - RS). The fruits were transported to the Laboratory of Fruits and Vegetables, belonging to the 'Departamento de Gestão e Tecnologia Agroindustrial', of 'Faculdade de Ciências Agronômicas - UNESP - Campus of Botucatu, SP'. Each fruit after receiving treatment, was stored for each experiment in a cold camera to 5l1ºC and 85 - 90% of relative humidity. The evaluations were accomplished every other day, for 10 days of storage. The alterations in the quality postharvest were detected through the analyses of loss of fresh mass... (Complete abstract, click electronic access below)

Made available in DSpace on 2014-08-20T13:25:42Z (GMT). No. of bitstreams: 1 Dissertacao_ Fernando_Jose_ Hawerroth.pdf: 3146259 bytes, checksum: 9a1582822ee01601422d0cc7ea2ea69e (MD5) Previous issue date: 2008-03-30 / Temperature is the main climate factor related to induction, maintenance and dormancy release in apple (Malus domestica Borkh.). The inadequate chilling exposure for this species causes the occurrence of budbreak problems, resulting in decrease in yield potential. Thus, the knowledge of physiological principles and environmental factors determining the dormancy phenomenon, especially winter temperature effects, it is necessary for the cultivars efficient selection in a productive region. Similarly cultural practices can be developed and adapted to minimize the problems cause by lack chilling during winter. The budbreak promoters application is one of main management s strategies used to decrease the problems of insufficient winter chilling on temperate fruit trees. Because the influence of budbreak promoters on plant s productive behavior, detecting this effects along of productive season became important. This study aimed to evaluate the budbreak and depth dormancy of apple cultivars exposed to different chilling conditions during the winter, and evaluate the apple productive response by budbreak promoters application. For this, three experiments were carried out in this research. In the first experiment, one-yearold twigs of Castel Gala and Royal Gala cultivars, grafted on M7 rootstock, were submitted to temperatures of 5, 10 and 15 C for different exposure periods (168, 336, 672, 1008 and 1344 hours). After treatments execution, the plants were kept in a greenhouse at 25ºC. Budbreak was quantified when accumulated 3444, 6888, 10332, 13776, 17220 and 20664 GDHºC after temperature treatments. The cultivars responded differently to temperature effect during the winter period. The temperature of 15 °C showed greater effectiveness on Castel Gala budbreak, while temperatures of 5 and 10ºC showed better performance in Royal Gala apple trees. It was not possible to estimate chilling requirement of cultivars studied through autumn grafted twigs, because the low budbreak obtained, derived of high apical dominance expressed by apple tree in this evaluation system. The objective of the second experiment was to evaluate the dormancy progression of apple cultivars with different chilling requirements. One-year-old twigs of Castel Gala , Condessa , Daiane , Fuji , Imperatriz and Royal Gala cultivars were exposed to 0, 336, 672, 1008 and 1344 chilling units at 4.5 ± 0.5 º C. After chilling exposure, the twigs were segmented into four cuttings (terminal, axillary superior, axillary median and axillary basal) and placed in growth chamber at 23±1ºC. The depth dormancy was measured by the mean time of budbreak (MTB). Through the method was not possible to group the cultivars according chilling requirement, indicating that correlation between the depth dormancy and chilling requirement isn t observed for all apple cultivars. In the third experiment were evaluated different hydrogen cyanamide and mineral oil combinations on phenology, budbreak and fruit production of Imperial Gala and Suprema s Fuji apple trees, during 2007/2008 growing season. Five budbreak promoters treatments were tested (1. control; 2. mineral oil 3.2%; 3. mineral oil 3.2% + hydrogen cyanamide 0.20%; 4. mineral oil 3.2% + hydrogen cyanamide 0.39%; 5. mineral oil 3.2% + hydrogen cyanamide 0.59%). The budbreak promoters application anticipated and reduced the blooming period, increasing the blooming overlap between Imperial Gala and Suprema´s Fuji cultivars. The budbreak promoters equalized and increased the budbreak in axillary and terminal buds, with the major budbreak level observed in Imperial Gala cultivar with 0.44% of hydrogen cyanamide and 3.2% of mineral oil treatment. The increase of hydrogen cyanamide concentration showed the tendency to decrease the fruit set. It was observed different treatment responses on production and mean fruit weight, probably to be related to fruit production predominance in different fructification structures. / A temperatura é o principal fator climático relacionado a indução, manutenção e superação da dormência na macieira (Malus domestica Borkh.). O inadequado suprimento em frio para esta espécie determina a ocorrência de problemas relacionados a brotação, repercutindo na diminuição do potencial produtivo. Dessa forma, o conhecimento dos princípios fisiológicos e dos fatores ambientais determinantes no fenômeno da dormência, sobretudo o efeito das temperaturas hibernais, faz-se necessário para a eficiente seleção de cultivares em determinada região produtora, assim como para eficiente elaboração e adequação de práticas culturais para minimização dos problemas oriundos do insuficiente acúmulo de frio hibernal. A aplicação de indutores de brotação é uma das principais estratégias de manejo utilizadas para minimizar os problemas da falta de frio em frutíferas de clima temperado. Em razão da influência dos indutores de brotação sobre a resposta produtiva das plantas, a mensuração de seus efeitos ao longo do ciclo produtivo torna-se importante. Este estudo objetivou avaliar a brotação e profundidade de dormência de cultivares de macieira frente a diferentes condições de exposição ao frio durante o período hibernal, e avaliar a resposta produtiva de macieiras frente à aplicação de indutores de brotação. Para tanto, foram realizados três experimentos. No primeiro experimento, ramos de um ano das cultivares Castel Gala e Royal Gala, enxertadas no porta-enxerto M7, foram submetidos as temperaturas de 5, 10 e 15ºC durante diferentes períodos de exposição (168; 336; 672; 1008 e 1344 horas). Após a efetivação dos tratamentos, as plantas foram mantidas em casa de vegetação a 25ºC. A brotação foi quantificada quando acumulada soma térmica de 3444, 6888, 10332, 13776, 17220 e 20664 GDHºC após tratamentos térmicos. As cultivares estudadas responderam diferentemente as temperaturas durante o período hibernal. A temperatura de 15ºC apresentou maior efetividade na brotação de gemas da cultivar Castel Gala, enquanto as temperaturas de 5 e 10ºC apresentaram melhor desempenho na cultivar Royal Gala. Não foi possível estimar o requerimento em frio das cultivares estudadas através do uso de ramos enxertados no outono, visto os baixos percentuais de brotação obtidos, decorrentes da alta dominância apical apresentada pela macieira neste sistema de avaliação. O objetivo do segundo experimento foi avaliar a dinâmica da dormência em cultivares com diferentes requerimentos em frio. Ramos de um ano das cultivares Castel Gala, Condessa, Daiane, Fuji, Imperatriz e Royal Gala foram expostos a 0, 336, 672, 1008 e 1344 unidades de frio a 4,5±0,5ºC. Após a exposição ao frio, os ramos foram segmentados em quatro estacas (terminal, axilar superior, axilar mediana e axilar inferior) e colocados em câmara de crescimento à 23±1ºC. A profundidade de dormência foi avaliada pelo tempo médio para brotação das gemas (TMB). Através do método utilizado não foi possível agrupar as cultivares segundo o requerimento em frio, indicando que a correlação entre a profundidade de dormência e o requerimento em frio não é valida para todas as cultivares de macieira. No terceiro experimento foram avaliadas diferentes combinações de cianamida hidrogenada e óleo mineral sobre a fenologia, brotação de gemas e produção de frutos em macieiras Imperial Gala e Fuji Suprema , durante o ciclo 2007/2008. Foram testados cinco níveis de indutor de brotação (1. testemunha; 2. óleo mineral 3,2%; 3. óleo mineral 3,2% + cianamida hidrogenada 0,20%; 4. óleo mineral 3,2% + cianamida hidrogenada 0,39%; 5. óleo mineral 3,2% + cianamida hidrogenada 0,59%). A aplicação dos indutores de brotação antecipou e reduziu o período de florescimento, aumentando a coincidência de florescimento das cultivares Imperial Gala e Fuji Suprema. Os indutores de brotação uniformizaram e aumentaram a brotação de gemas axilares e terminais, obtendo-se máxima brotação na cultivar Imperial Gala com 0,44% de cianamida hidrogenada e 3,2% de óleo mineral. O aumento da concentração de cianamida hidrogenada apresentou a tendência de diminuir a frutificação efetiva. Observaram-se respostas diferenciadas entre tratamentos quanto à produção e a massa média dos frutos, podendo ser atribuídas ao predomínio da produção de frutos em distintas estruturas de frutificação.

Philosophiae Doctor - PhD / The domesticated apple (Malus x domestica Borkh), belonging to the Malusgenus of the Rosaceae family, is one of the edible pomaceous fruits. Since it is one of the important commercial fruit crops worldwide, the quality of the fruit is crucial to breeders and farmers as it ultimately determines acceptance of a cultivar for consumption. Fruit quality is also a critical determinant factor that is used to estimate the potential of apples to have a long shelf life / South Africa

>Magister Scientiae - MSc / The Rosaceae family contains major temperate crops such as the domesticated apple(Malus x domestica Borkh.), peach (Prunus persica L. Batsch) and European pear (Pyrus communis L.). However, despite its evident economic importance, it is generally poorly studied in genomic terms, relative to the other major crop groups. Microsatellite and Diversity Array Technology (DArT) genetic markers have been exploited in this work and are essential tools in genetic map construction and marker-assisted selection (MAS) of high quality apples and other rosaceous crops. Microsatellites are advantageous in that they are co-dominant, highly polymorphic, abundant, transferable and reliably reproducible; hence their use in this study. In order for budbreak to take place in a timely and homogenous fashion, apple trees need a period of exposure to low temperatures.Within orchards the application of chemicals that induce budbreak in unsuitable environments is required to produce apples from cultivars that require high chilling levels. However, this and other practices using chemicals in orchards tend to pollute the environment. One of the solutions to this problem is to breed low chill apples such as ‘Anna’ cultivar, which was used as one of the parents in this study.This work was aimed at understanding the underlying genetic factors that determine chilling requirements for the time of initial vegetative budbreak trait in the apple cross ‘Anna’ x ‘Austin’. This was achieved through linkage map construction using SSR and DArT molecular markers followed by QTL analysis. This thesis has therefore exploited the large number of Expressed Sequence Tags (ESTs) and genome sequence data for the apple, using Tandem Repeats Finder, to design a total of 98 new SSR primers pairs. The other 369 SSR markers used in this work were from published work. JoinMap! 4.1 software was used to create an integrated genetic map with 17 linkage groups, for the domesticated apple cultivar, ‘Austin’ x ‘Anna’ mapping population with 80 individuals.The result of this process was a genetic map 1 212cM in length, and a total of 429 markers (314 DArT and 115 SSR), at an average density of a marker every 4 cM. This map was used identify the Quantitative Trait Loci (QTLs) determining chilling requirements for time of vegetative budbreak (IVB). In this process, putative IVB QTLs were identified in the ‘Anna’ x ‘Austin’ mapping population using the rMQM analysis function of MapQTL! 6.0, for both adult and seedling data collected over 3 growing seasons from 1996 to 1998. These QTLs were detected on linkage groups 2, 9 and 14,and explained 0.3 to 12.8 % of the observed phenotypic variation for the adult population,and 5.3 - 21 % for the seedling population. Seedling (LG 14) and adult (LGs 5, 7, 10) specific QTLs were also detected for the ‘Anna’ x ‘Austin’ cross. These QTLs will provide the basis for marker validation on related mapping populations in the apple breeding programme, and for the future identification of candidate genes controlling the process of budbreak.