Made available in DSpace on 2017-07-25T19:29:39Z (GMT). No. of bitstreams: 1 Geisa Barbosa.pdf: 1077715 bytes, checksum: 67df48b716e73523f8fbdca95d4ff28e (MD5) Previous issue date: 2008-09-03 / Coffee is one of the most valuable agricultural commodities, ranking second on international trade exchanges. There is much agronomic research on coffee, but molecular knowledge of its fruit development and ripening is limited. The Brazilian Coffee Genome Project provides the genomic resources required to study the proteomics of Coffea arabica, to add to results derived from DNA microarray and EST studies. This work reports a comparative proteomic investigation of immature coffee fruit in two developmental stages: stage 1, intense cell division, and stage 2, intense cell expansion. Proteins were extracted using a modified SDSPhenol method and two-dimensional electrophoresis gels stained with Coomassie blue revealed about 300 well-resolved polypeptide spots in the pH range 3-10 and 500 wellresolved polypeptide spots in the pH range 4-7. Nineteen variable polypeptides were excised, trypsin-digested and analyzed by MALDI-TOF mass spectrometry. Peptide MS data were searched against the coffee EST database and six protein spots were identified, according to their putative and assigned functions in known biosynthetic pathways: Fructose bifosfato aldolase, glyceraldehyde-3-phosphate dehydrogenase and malate dehydrogenase enzymes are part of glycolytic pathway, 11S storage globulin protein has the reserve function, the thaumatin-like protein signals a response to stress, and chloroplast precursor is involved in pathway photosynthesis. These proteins, except taumatina-like, had increased their expression at the stage of expansion of the fruit indicating that they are more required with the fruit development. / O café é um dos mais importantes produtos agrícolas, ocupando o segundo lugar no mercado internacional de ‘commodities’. Existem muitas pesquisas em café, mas o conhecimento molecular sobre desenvolvimento e amadurecimento de fruto ainda é limitado. O Projeto Genoma Café fornece recursos requeridos para o estudo proteômico de café, para adicionar resultados derivados do estudo de microarranjos de DNA e ESTs. Este trabalho relata uma investigação proteômica comparativa do fruto imaturo de Coffea arabica em dois estádios de desenvolvimento: chumbinho, intensa divisão celular, e expansão do fruto, intensa expansão celular. As proteínas foram extraídas usando o método SDS-Fenol modificado e separadas por eletroforese bidimensional. Os géis corados com Coomassie blue revelaram aproximadamente 300 spots bem resolvidos na faixa de pH entre 3-10 e 500 spots na faixa de pH 4-7. O método de identificação de proteínas por padrão de massas peptídicas foi aplicado nos 16 spots que apresentaram maior diferença de expressão na faixa de pH 3-10, utilizando-se um espectrômetro de massa (MS) do tipo MALDI-TOF e comparando os espectros de digestão tríptica contra bancos de dados baseados em ESTs de Coffea. Foram identificadas seis prováveis proteínas que foram classificadas de acordo com sua função e vias biossintéticas conhecidas: as enzimas frutose bifosfato aldolase, gliceraldeído-3-fosfato desidrogenase e malato desidrogenase fazem parte da via glicolítica; a proteína 11S possui função de reserva, a taumatina-like sinaliza resposta a um estresse, e precursor de cloroplasto, encontra-se envolvida na via fotossintetizante. Essas proteínas, exceto a taumatina-like, tiveram sua expressão aumentada no estádio de expansão do fruto indicando que são mais requeridas à medida que o fruto se desenvolve.

A perda da vitalidade pulpar, em consequência de injurias traumáticas, lesões cariosas profundas e malformações dentárias, pode interromper o desenvolvimento radicular em dentes permanentes jovens. O tratamento endodôntico em dentes com rizogênese incompleta constituise num desafio ao clínico para a realização de tratamentos convencionais, devido principalmente à dificuldade de debridamento das paredes dentinárias e do adequado selamento apical. Além disso, em função das paredes dentinárias delgadas e uma proporção coroa-raiz inadequada, a ocorrência de fraturas no trans e pós-operatório não é um evento raro. Existem diferentes estratégias terapêuticas para o manejo destes dentes com rizogênese incompleta, incluindo os procedimentos de apicificação e revascularização da polpa. O objetivo desta tese foi revisar sistematicamente a literatura para avaliar a efetividade dos procedimentos de apicificação e revascularização da polpa em relação aos desfechos clínicos, radiográficos e de retenção dentária. Dois revisores de maneira independente revisaram a literatura e avaliaram os artigos. A busca foi realizada na literatura via PubMed/MEDLINE e Embase até junho de 2017 para selecionar estudos de caso-controle e coorte. Um total 231 artigos foram localizados nas bases de dados, dos quais 3 permaneceram após a aplicação dos critérios de elegibilidade. Após a seleção dos artigos, os dados de cada estudo foram coletados e o risco de viés avaliado. Foram obtidas estimativas de efeito comparando o sucesso clínico e radiográfico e retenção dentária entre o plug apical de agregado de trióxido mineral (MTA) versus revascularização da polpa (BC) e hidróxido de cálcio (CH). As duas meta-análises comparando tanto o MTA vs BC para os desfechos clínicos e radiográficos (Z=0.164, p=0.869, RR=1.010, 95%CI:0.895-1.140, I=0%) e retenção dentária (Z=0.855, p=0.393, RR=1.050, 95%CI:0.939-1.173, I=0%) ou MTA vs CH para os desfechos clínicos e radiográficos e retenção dentária (Z=-1.596, p=0.110, RR=0.816, 95%CI:0.635-1.048, I=0%) não demonstraram diferenças estatisticamente significativas. Todos estudos incluídos na revisão sistemática foram considerados de alta qualidade metodológica. Em conclusão, a técnica de revascularização da polpa, por possibilitar o desenvolvimento radicular adicional, deve ser a primeira opção de tratamento em dentes permanentes com rizogênese incompleta que apresentem proporção coroa-raiz inadequada, entretanto, o plug apical de MTA parece apresentar resultados mais fidedignos em relação ao sucesso em geral, e assim, nos casos de dentes permanentes que apresentem proporção coroaraiz satisfatória, esta técnica de ser a primeira opção de tratamento. / The loss of pulp vitality, as a consequence of traumatic injuries, deep carious lesions and dental malformations, may interrupt the root development in young permanent teeth. The endodontic treatment in immature teeth constitutes a challenge to the practitioner to perform conventional treatments, mainly due to difficulty of dentinal walls debridement and adequate apical sealing. In addition, due to the thin dentinal walls and inadequate crown-root proportion, occurrence of trans and postoperative fractures is not a rare event. There are different therapeutic strategies to manage these immature teeth including apexification and pulp revascularization procedures. The aim of this thesis was to systematically review the literature to evaluate the effectiveness of apexification and pulp revascularization procedures regarding the clinical, radiographic and tooth retention outcomes. Two reviewers independently performed the screening and evaluation of articles. The literature was screened via PubMed/MEDLINE and Embase databases until June 2017 to select case-control and cohort studies. A total of 231 articles were retrieved from the databases, in which only 3 remained after the eligibility criteria. After the selection of the articles, the data of each study was extracted and bias risk assessed. Pooled-effect estimates were obtained comparing the clinical and radiographic success and retention rates among the mineral trioxide aggregate (MTA) apical plug vs pulp revascularization (BC) and calcium hydroxide (CH). The two meta-analysis comparing either MTA vs BC for clinical and radiographic outcomes (Z=0.164, p=0.869, RR=1.010, 95%CI:0.895-1.140, I=0%) and tooth retention rates (Z=0.855, p=0.393, RR=1.050, 95%CI:0.939-1.173, I=0%) or MTA vs CH for clinical and radiographic outcomes and tooth retention rates (Z=-1.596, p=0.110, RR=0.816, 95%CI:0.635-1.048, I=0%) showed no statistically significant differences. All studies included in the systematic review were classified as high quality. In conclusion, the revascularization treatment, as it allows additional root development may be the first treatment option in immature necrotic permanent teeth that have inadequate crown-root ratio, however, the MTA apical plug seems to provide more predictable outcomes regarding the overall success, and thus, in cases of permanent teeth having adequate crown-root proportion, it should be the first treatment option.

RESUMO Urbinatti PR. Observações ecológicas de Ae. albopictus (Diptera: Culicidae) em áreas de proteção ambiental e urbana na periferia da Grande São Paulo. São Paulo; 2004. [Tese de Doutorado – Faculdade de Saúde Pública da Universidade de São Paulo]. Objetivos. Contribuir para o conhecimento da ecologia de uma população de Ae. albopictus em área de proteção ambiental e urbanizada. Métodos. Para avaliar aspectos da infestação desse culicídeo, escolheram-se duas áreas próximas, o Parque Ecológico do Tietê (PET), em área de proteção ambiental, (ambientes 1 e 2) e outra antrópica, Jardins São Francisco e Piratininga (ambientes 1 e 2), na Zona Leste da Capital. A primeira etapa da pesquisa foi desenvolvida no PET, de abril de 2001 a março de 2002 e a segunda foi nos Jardins São Francisco e Piratininga, de abril de 2002 a março de 2003. Com a finalidade de medir e comparar a infestação de Ae. albopictus entre os dois ambientes do PET e Jardins, utilizaram-se, 10 larvitrampas e 10 ovitrampas, sendo instaladas no peridomicílio e com monitoramento quinzenal no PET e a cada dez dias nos Jardins. Foram efetuadas coletas de amostras de água para análise de parâmetros físicos e químicos e de fauna associada. Resultados. Ae. albopictus foi à espécie mais freqüente em relação as demais coletadas, nos criadouros naturais e artificiais, correspondendo a 97,70 % e 92,50 % do total de imaturos, respectivamente no PET, Jardins São Francisco e Piratininga. Observou-se que os índices de densidade e positividade das ovitrampas e larvitrampas, no PET apresentaram valores elevados quando comparados aos Jardins São Francisco e Piratininga. Verificou-se correlação positiva significante entre a densidade de larvas e pupas de Ae. albopictus coletada em larvitrampas e as temperaturas ambiente e da água nos pneus nas duas áreas. Observou-se diferença estatisticamente significante entre a densidade de larvas de Ae. albopictus nas larvitrampas dos ambientes 1 e 2 do PET e Jardins, na presença e ausência de fauna associada.Conclusões. A presença freqüente de Ae. albopictus tanto em áreas de proteção ambiental como em alterada, reforça a evidência de sua ampla valência ecológica em colonizar ecótopos naturais e artificiais. Diante deste comportamento e aliado a competência e capacidade vetora, espera-se que os resultados dessa pesquisa possam fornecer informações que possibilitem subsidiar ações de controle caso esse mosquito represente ameaças à Saúde Pública. Descritores: Aedes albopictus, vetor, imaturos, ovitrampa, larvitrampa, urbanização / SUMMARY Urbinatti PR. Ecological observations of Aedes albopictus (Diptera: Culicidae) in urban and preserved areas in the outskirts of Greater Sao Paulo. São Paulo; 2004. [Tese de Doutorado – Faculdade de Saúde Pública da Universidade de São Paulo]. Objective. To contribute to the knowledge of the ecology of the Ae. albopictus population both in a protected and in an urban environment. Methods. Two different environments were chosen to evaluate aspects of the infestation of the East End of the city: a protected environment - the Parque Ecológico do Tietê (PET) divided into environments 1 and 2, and another anthropogenically altered one - the Jardins São Francisco and Piratininga, divided in the same way. The first step of the investigation was undertaken in PET between April 2001 and March 2002 and the second step in the Jardins, between April 2002 and March 2003. To measure and compare Ae. Albopictus infestation as between PET and Jardins, 10 larvitraps and 10 ovitraps were installed around a domiciliary area with monitoring every 15 days in PET and every 10 days in the Jardins. The analysis of water samples for their chemical and physical parameters and associated fauna was undertaken. Results. Ae. albopictus was the most frequently collected species, both in artificial and natural breeding sites, accounting for 97.70% and 92.50% of the total of immature specimens collected in PET and in the Jardins, respectively. As regards the positive rates of ovitraps and larvitraps, PET presented higher values than the Jardins. There was a significant correlation between larval and pupal densities in the larvitraps and the temperature of water in tires and sites and a significant difference between larval densities in the larvitraps of environments 1 and 2 in both PET and Jardins, as regards the presence and absence of associated fauna. Conclusions. The frequent presence of Ae. albopictus in protected areas and in altered ones reinforces the evidence of its ecological adaptability in the colonization of both natural and artificial environments. This behavior associated with its competence and vectorial capacity should contribute with information that will be useful for control actions if these insects come to represent a public health threat. Key words: Aedes albopictus, vector, immature, ovitrap, larvitrap, urbanization

São estimadas 90 mil espécies de insetos para o Cerrado, o segundo maior bioma brasileiro, com enorme heterogeneidade de habitats e rica fauna. Dentre esses insetos, os Lepidoptera representam cerca de 10% do total de espécies. Entretanto, estudos sobre seus imaturos (ovo, larva e pupa) ainda são incipientes para o bioma, principalmente sobre as mariposas, cujas histórias naturais de muitas espécies ainda são desconhecidas. A falta desses estudos faz com que sejam ignoradas as interações multitróficas em que esses organismos estão envolvidos e, portanto, inviabiliza futuros estudos relacionados ao seu papel em comunidades e redes ecológicas. Esse é o caso de muitas espécies da família Geometridae, incluindo Oospila pallidaria, que é uma mariposa esmeralda (devido à coloração verde esmeralda das suas asas). Oospila pallidaria é uma espécie herbívora que não possuía até o momento nenhuma informação publicada sobre a sua biologia e fatores que condicionam sua ocorrência, tais como: a fenologia da planta hospedeira, que pode determinar o período de melhores condições para crescimento e reprodução; a qualidade nutricional dos recursos alimentares (e. g. quantidade de água e nitrogênio), que pode determinar quais recursos conferem melhor desenvolvimento/performance a um herbívoro; os inimigos naturais, que podem restringir a ocorrência dos herbívoros e a temperatura e pluviosidade, que podem alterar a qualidade e a distribuição geográfica dos recursos alimentares utilizados pelos herbívoros. Nesse sentido, o objetivo principal deste estudo foi descrever, pela primeira vez, aspectos da biologia e história natural de O. pallidaria (Capítulo 1) e avaliar a sua performance larval, a partir de diferentes dietas (Capítulo 2). Os resultados apresentados no Capítulo 1 mostraram que os ovos de O. pallidaria, verdes e com duração média de sete dias, foram solitários e ovipostos principalmente nos tricomas das folhas maduras. As larvas se alimentaram de folhas maduras predominantemente, mas utilizaram folhas jovens e botões florais oportunisticamente. As larvas tiveram coloração críptica, se camuflaram em meio às folhas de Mimosa setosa (Leguminosae: Mimosoideae), sua única hospedeira, e desenvolveram o comportamento de auto-limpeza. Há cinco ínstares larvais, com coloração que variou de amarelo a verde. As larvas apresentaram também um par de projeções no protórax e linha mediana marrom na região dorsal. O comprimento máximo do corpo das larvas foi de 28 mm. As pupas foram predominantemente verdes, com no máximo 10 mm de comprimento. O desenvolvimento do ovo ao adulto durou cerca de 50 dias ( = 42; S = 6; n = 11). Seu único inimigo natural registrado foi o microhimenóptero Cotesia sp. (Braconidae, Microgastrinae). Fêmeas de O. pallidaria produziram 65 ovos em média (S = 7,07; n = 2). A ocorrência de O. pallidaria foi sazonal e sobreposta ao pico de presença de folhas maduras. Larvas foram negativamente relacionadas à temperatura e pluviosidade, com significância estatística somente para a primeira (r = - 0.5889, P < 0.05). No Capítulo 2, foi mostrado que as folhas maduras de M. setosa foram o único recurso alimentar disponível durante todo o ano. Os botões florais foram o recurso com maior qualidade nutricional (conteúdo relativo de água e nitrogênio total), seguidos por folhas jovens e maduras respectivamente. Entretanto, a sobrevivência de O. pallidaria foi maior com folhas maduras. Os resultados indicam que para o herbívoro especialista O. pallidaria, a fenologia da planta hospedeira é crucial para sua sobrevivência, em especial a presença de folhas maduras de M. setosa. Por outro lado, os botões florais são recursos efêmeros, porém importantes para a sobrevivência das larvas de últimos ínstares no final da estação seca, quando as folhas maduras estão ressecadas e/ou senescentes. / It is estimated 90.000 species of insects for the Cerrado, the second largest Brazilian biome, with great diversity of habitats and rich fauna. Among these insects, Lepidoptera represents about 10% of all species. However, studies on their immature stages (egg, larvae and pupae) are incipient for the biome, mostly on moths, whose natural histories of many species are unknown. The lack of these studies makes the multitrophic interactions in that these organisms are involved ignored and, therefore, prevent future studies related to their role in communities and ecological networks. This is the case of many species of Geometridae, including Oospila pallidaria, which is an emerald moth (due to emerald green color of their wings). Oospila pallidaria is a herbivorous species that had not published any information on their biology and factors that influence their occurrence, such as: the phenology of the host plant, which can determine the time of better conditions for growth and reproduction; the nutritional quality of food resources (e. g. amount of water and nitrogen), which can determine what resources provide better development/performance to a herbivore; natural enemies, that could restrict the occurrence of herbivores and the temperature and rainfall, which can change the quality and geographical distribution of food resources used by herbivores. Accordingly, the aim of this study was to describe, for the first time, aspects of the biology and natural history of O. pallidaria (Chapter 1) and to assess their larval performance from different diets (Chapter 2). The results presented in Chapter 1 showed that eggs of O. pallidaria, green and lasting an average of seven days, were lonely and laid especially in trichomes of mature leaves. The larvae fed on mature leaves predominantly, but used young leaves and flower buds opportunistically. Larvae had cryptic coloration, they camouflaged among the leaves of Mimosa setosa (Leguminosae: Mimosoideae), a single host, and developed the self-cleaning behavior. There are five larval instars, with color ranging from yellow to green. The larvae also had a pair of projections on prothorax and brown midline in the dorsal region. The maximum length of the larvaes body was 28 mm. The pupae were predominantly green, with a maximum of 10 mm in length. The development from egg to adult lasted about 50 days ( = 42, S = 6, n = 11). Their only natural enemy recorded was the microhymenopteran Cotesia sp. (Braconidae, Microgastrinae). Oospila pallidaria females produced 65 eggs on average (S = 7.07; n = 2). The occurrence of O. pallidaria was seasonal and overlapped on the peak presence of mature leaves. Larvae were negatively related to temperature and rainfall, with statistical significance only for the first (r = - 0.5889, P < 0.05). In Chapter 2, it was shown that mature leaves of M. setosa were the only food source available throughout the year. The flower buds were the resource with higher nutritional quality (relative water content and total nitrogen), followed by young and mature leaves respectively. However, the survival of O. pallidaria was greater with mature leaves. The results indicate that for the specialist herbivore O. pallidaria, the host plant phenology is critical for its survival, especially the presence of mature leaves of M. setosa. On the other hand, the flower buds are ephemeral resources, but important for the survival of the last instar larvae in the dry season, when mature leaves are withered and/or senescent.

Orbital ATK tests horizontally restrained solid rocket motors (SRMs) in Promontory, Utah, USA. During SRMs tests, an estimated 1.5 million kg (~3.3 million lbs.) of entrained soil and combustion products (mainly aluminum oxide, gaseous hydrogen chloride, and water) are deposited over a wide area. Local farmers have expressed concerns regarding the deposition of this material, referred to as Test Fire Soil (TFS), on crops surrounding the test site. Greenhouse studies of mature corn and alfalfa (54 days to harvest) exposed to a worse case TFS deposition density to soil (3200-g/m2) resulted in reductions in germination and plant growth. Application of TFS only on leaves caused visual changes in leaf appearance (e.g. “burning”) but did not impact on growth. The visual damage to the leaves was thought to be caused by the high chloride in the TFS. The pH (measured from 2-10 in the field) of the TFS was also thought to be a potential cause of the visual damage. Follow-up studies were conducted using younger corn, alfalfa and winter wheat (12-54 days from emergence), to evaluate the impact of a more typical TFS deposition density and application scenario. The relationship between deposition amount and the crop impact was also examined. Impact was evaluated by comparing the growth, tissue composition (focusing on chloride and aluminum) and chlorophyll content of controls relative to the treatments. The individual effects of chloride and pH on corn leaves were determined by monitoring visual appearance and chlorophyll content. Leaves exposed to a typical TFS deposition loading (70-g/m2) did not contain elevated chloride or aluminum levels relative to the controls. Higher TFS loadings (700-g/m2 and 3200-g/m2) resulted in significantly elevated chloride content in corn leaves at 29 days from emergence. Corn leaves treated with TFS at the 700-g/m2 loading 12 days from emergence had comparable chloride levels to controls. High and low pH and chloride solutions visually impacted corn leaves and reduced leaf chlorophyll. This study indicated SRMs testing can be conducted under typical conditions during the juvenile growth stages of field corn, alfalfa, and winter wheat without adverse impact on crop yield and plant tissue composition.

During starch synthesis, knowledge of how the supra molecular organization of starch granules occur is still unknown. This dissertation begins exploring the changes to wheat starch morphology, structure and organization during granule development. Starches from wheat seeds harvested at different days after anthesis were isolated and studied for their molecular organization at different scales. The interaction of granular starch to iodine vapor was used to reveal information about the physical state of the polymers in the granules. Both large and small granule populations initiated as spherical shaped granules, but large granules changed into lenticular shape, while small granules remained spherical. Blocklet, lamellae, and crystalline structures were well developed already at seven days of maturity, suggesting their presence at the center of mature granules. Furthermore, (i) the size of blocklets decreased from center to the periphery in both granule populations, but at all developmental stages, they were larger in small granules than in large granules; (ii) Lamellae repeat distance was shorter close to the center of granules; (iii) B-type crystallites were mixed with A-type crystallites at the center of granules from both populations, but the proportion of B-type crystallites was higher in large granules, which correlated with a higher proportion of amylopectin long chains in these granules; (iv) The relative crystallinity (RC) of starches from both populations was higher at the center of the granule than that at the periphery. RC was lower in large granules than in small granules suggesting influence of the higher amylose content in large granules, which introduces more defects reducing RC. AFM visualized surfaces with features different from the regular type of surface among immature starch granules, which was interpreted as the amorphous growth ring of the starch granule. Exposure of starches to iodine vapor demonstrated different levels of molecular organization between them and revealed possible interaction of inter-cluster chain segments and spacer segments of amylopectin with iodine. In-situ AFM imaging of the granule surface exposed to iodine vapor under humid conditions revealed that glucan polymers or polymer segments in between and on top of blocklets have the ability to interact with iodine.

No description available.

Although cottonseed is an important source of oil and fiber, the development of cotton embryos has not been investigated as well as development of cotton fiber. The development of cotton embryos in late heart-stage and early cotyledonary stage is less well investigated than the first 10-14 days after anthesis, or the late stages of embryo development during seed-fill and desiccation. This analysis focused on cotton embryos in the late heart-stage and early cotyledonary stage of development (1.5-4.0 mm or about 13-18 DPA). In vitro analyses are important tools for studying embryos in isolation from the endosperm and fiber and when it is necessary to monitor the developing embryo continuously. The original goal of this work was to develop an in vitro culture method that would support continued development of excised zygotic embryos from the early cotyledonary stage into complete plants with true shoots, i.e. true leaves or visible buds and then to use this method to study aspects of developmental regulation during cotyledonary stage and the transition to later stages. Not all embryos were competent to develop true shoots (an apical bud or a leaf plus a bud) in culture. A number of cultural variables were tested and eliminated. Embryo maturity at the time embryos were excised and the presence or absence of light during the first 14 days of culture affected the competence of immature embryos to developed true shoots. The effect of light was verified in several large replicated experiments. Morphological changes occurring during in vivo development were examined microscopically. The transition from heart-stage to early cotyledonary stage and the development of the first leaf from initials to a large structure were identified. Embryonic shoot apices continued to grow in cultured 1-3 mm embryos. The size and shape of light-treated and dark-treated embryonic apices was compared. A germination test of mature seeds identified seedlings with a similar phenotype occurring at similar rates in seedlings and light-cultured embryos and possible causes were discussed.

The objective of this study was to optimize regeneration parameters of immature inflorescence culture of Triticum aestivum cv. Y&uuml / regir-89. The effects of dark incubation period and explant region on regeneration success were tested. Immature inflorescences were cut into 3 pieces as tip, mid, base and put onto 2mg /L 2,4-dichlorophenoxyacetic acid containing callus induction medium. These explants were taken to regeneration after 6, 9, 13 weeks of dark incubation period. The regeneration capacities of calli were determined as rooting and shooting percentages. Shooting percentages were found to be 72.0 % for 6 weeks of dark incubation and 64.1 % for 9 weeks of dark incubation while it decreases to 26.1 % in 13 weeks of dark incubation period. This showed that prolonged dark incubation period decreased regeneration capacity of the callus. There was no significant difference in regeneration capacities of tip, mid and base regions of immature inflorescences, which reveals the potential of every region of inflorescence to be used as explant source in further transformation studies. Besides regeneration studies, optimization of transformation parameters for Turkish wheat cultivar Y&uuml / regir by using Agrobacterium tumefaciens AGLI containing binary vector pALl56 was performed. Transformation efficiencies were determined by monitoring the transient expression of uidA gene via histochemical GUS assay. Three to four weeks old calli were found to be more responsive to Agrobacterium-mediated transformation. Different media were tested for utilization during co-cultivation period. It was found that including phenolic compound acetosyringone along with ascorbic acid as an antioxidant was essential for succesful transformation.

iv The objective of this study was to optimize tissue culture and regeneration parameters of immature inflorescence culture of Triticum aestivum cv. Bezostaja- 01 and Triticum durum cv. Kiziltan-91. The effects of callus age and vernalisation time of explants on regeneration success were evaluated. For determination of optimum vernalisation time of immature inflorescence, plants subjected to 4 &deg / C for 1, 2, 3, 4, and 5 weeks, respectively. Tillers containing immature inflorescences were collected at the same time. Percentage of inflorescence formed tillers over total explants were reached the highest value, 79 %, at 4 weeks cold treated Kiziltan cultivar and, 73 %, at 5 weeks cold treated Bezostaja cultivar. Isolated immature inflorescences were put onto 2mg /L 2,4-dichlorophenoxyacetic acid and picloram containing callus induction medium for Kiziltan and Bezostaja cultures, respectively. Callus induction rate were found to be 100 % for Kiziltan and Bezostaja. These explants were taken to regeneration after 6, 9, 12 and 15 weeks of dark incubation period. The regeneration capacities of calli were determined as shooting percentage and data were collected after 4, 8, 12, and 15 week regeneration period. The highest shooting percentage of 69 %, were obtained from 6 weeks old calli produced from 4 weeks vernalised explants in Kiziltan cultures at the end of 15 weeks regeneration period. However, shooting percentage was 57.2 % for 9 weeks old calli while it decreases to 37.6 % in 12 weeks old calli and 44.2 % in 15 weeks old calli at the end of 15 weeks regeneration period. This showed that prolonged dark incubation period decreased regeneration capacity of the callus. However, there was no significant difference in regeneration capacities of calli produced from Bezostaja immature inflorescence and the highest shooting percentage was obtained from 9 weeks old calli produced from 5 weeks vernalised explants, 27.4 %. Besides regeneration studies, optimization of transformation parameters for winter wheat cultivars Kiziltan and Bezostaja by using Agrobacterium tumefaciens AGLI containing binary vector pALl56 was performed. Transformation efficiencies were determined by monitoring the transient expression of uidA gene via histochemical GUS assay. Three to four weeks old calli were found to be more responsive to Agrobacterium-mediated transformation in Kiziltan cultures. However, four to five weeks old calli were found to be more responsive to Agrobacterium-mediated transformation in Bezostaja cultures. Different transformation protocols were used. It was found that MGL based and MMA based protocols could be used for Bezostaja and Kiziltan transformation, respectively. The highest GUS expression, 84%, was obtained from 28 weeks old calli produced from 5 weeks vernalised explants in Bezostaja cultures.