Made available in DSpace on 2016-04-28T14:58:35Z (GMT). No. of bitstreams: 1 2005-Cleiton Mateus Sousa.pdf: 885284 bytes, checksum: 42cf8890c05e973310a56f9182c0e1e0 (MD5) Previous issue date: 2005-01-20 / Gerberas are cut flowers with large color diversity and good durability. The use of growth regulators on in vitro propagation is related to the capacity to induce proliferation of sprouts and differentiation of roots. However, citokinins in excess can induce the formation of abnormal plant, with reduced acclimatization potential. The mineral and organic composition of media culture plays an important role during the process. The objective of this dissertation was to adjust protocols for in vitro propagation of Gerbera jamesonii. The following aspects were evaluated: (i) the effect of the inflorescence developmental stage on the performance of in vitro culture initiated from chapters (ii) gerbera varieties (iii) growth regulator balance and (iv) the effect of salt concentration of MS medium on the proliferation of in vitro sprouting and rooting and acclimatization of three varieties of gerbera. For variety Phanter, the regeneration of explants prepared from inflorescences depended on its stage of development. Explants prepared from young chapters showed better results. The concentration and source of citokinin, and the concentration of salts in MS medium influenced the proliferation of sprouts in explants of variety Ornela. BAP was more efficient than kinetin on proliferation of sprouts. Concentrations above 2.0 mg.L-1 of BAP induced the formation of plants with undesirable characteristics during in vitro multiplication, while this was not observed in the presence of the same kinetin in larger concentration (4.0 mg.L-1). The combinations of 0.5 mg.L-1 BAP with the different levels of IBA showed the best results for multiplication and production of sprouts, with potential for acclimatizing variety Ornela. The presence of BAP inhibited the differentiation of roots, while the same was not observed in largest concentration of kinetin. The concentration of salts below 50% limited the proliferation of sprouts in variety Ornela. The varieties Capuccino, Mirage and Ornela, when maintained in the same balance of BAP and IBA, showed divergences in results for number of sprouts; when acclimatized, they presented high survival rates. Difficulties on in vitro gerbera propagation were mostly found on the regeneration phase and on the formation of sprouts in explants prepared from chapters. For the other phases, no major difficulties were detected. / As g?rberas s?o flores de corte que apresentam diversidade na colora??o e boa durabilidade. O uso de fitorreguladores na propaga??o de plantas in vitro esta relacionado com a capacidade de induzir a prolifera??o de brotos e a diferencia??o de ra?zes. No entanto, citocininas em excesso podem induzir a forma??o de pl?ntulas anormais e com baixo potencial para serem aclimatadas. Al?m disso, a composi??o mineral e org?nica do meio de cultura apresenta importante papel durante o processo. O objetivo deste trabalho foi ajustar protocolos para a propaga??o massal in vitro de variedades de Gerbera jamesonii. Avaliou-se: o efeito do est?dio de desenvolvimento de infloresc?ncias na implanta??o de culturas in vitro a partir de cap?tulos; variedades; balan?os de fitorreguladores; concentra??o dos sais do meio MS na prolifera??o de brotos e enraizamento in vitro e a aclimatiza??o de tr?s variedades de g?rbera. A regenera??o de explantes preparados a partir de infloresc?ncias depende do est?dio de desenvolvimento das mesmas. Explantes preparados a partir de cap?tulos jovens apresentaram melhores resultados. A prolifera??o de brotos em explantes de g?rbera, variedade Ornela foi influenciada tanto pela concentra??o quanto pela fonte de citocinina, assim como pela concentra??o dos sais do meio MS. A mol?cula BAP mostrou ser mais eficiente que kinetina para induzir a prolifera??o de brotos. BAP acima de 2,0 mg.L-1 induziu a forma??o de plantas com caracter?sticas indesej?veis durante a multiplica??o in vitro, enquanto isto n?o foi observado mesmo na maior concentra??o de kinetina (4,0 mg.L-1). As combina??es de 0,5 mg.L-1 de BAP com os diferentes n?veis de AIB (0; 0,05 e 0,5 mg.L-1) apresentaram os melhores resultados para a multiplica??o e produ??o de brotos in vitro com potencial para serem aclimatados da variedade Ornela. Somente o BAP inibiu a diferencia??o de ra?zes. A concentra??o dos sais abaixo de 50% limitou a prolifera??o de brotos na variedade Ornela. As variedades Capuccino Mirage e Ornela, quando mantidas no mesmo balan?o de BAP e AIB apresentaram resultados divergentes quanto ? prolifera??o de brotos, e na aclimatiza??o apresentaram alta taxa de sobreviv?ncia. Na propaga??o in vitro de g?rbera, somente na fase de implanta??o das culturas in vitro que verificou-se dificuldade de regenera??o e na forma??o de brotos em explantes preparados a partir de cap?tulos. Nas demais fases, as variedades estudadas apresentaram facilidade e resultados bastante promissores.

Made available in DSpace on 2016-04-28T14:59:00Z (GMT). No. of bitstreams: 1 Cleiton Mateus Sousa.pdf: 6410770 bytes, checksum: 26a93d90d24ec7420524c2e0478aec38 (MD5) Previous issue date: 2008-07-25 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The determination of plant hormones can be an excellent tool for to study the plant development. Today, the complexity of methods, equipment and reagents of high cost limit the use with practice of routine. Some advantages of methods immunoassay can exceed those limitations, showing a potential of practical use in determination of plant hormones. The test Enzyme-Linked Immunosorbent Assay (ELISA) is utilized in detection of molecules with low molecular weight. However, the ELISA requires specific antibodies against molecules of interest, in the case, plant hormones. Today in the market there is not available antibodies for the determination of hormonal molecules. Faced with that, proposed be produced and characterize antibodies against molecules of AIA, 2ip and zeatin and subsequently, for use the detection those molecules. For that, the hormonal molecules were conjugated with protein (BSA) for subsequent immunization of chicken or rabbits. From the serum of rabbits or yolks eggs the antibodies were purified and characterized through the test of precipitation of double diffusion, SDS-PAGE and test ELISA. The antibodies that presented better results were utilized in detection of plant hormones molecules in samples of plantlets in vitro of gerbera. The test of precipitation of double diffusion revealed that the antibodies productized were capable of will detect the molecule in study. Through in the SDS-PAGE was verified that the antibodies obtained in eggs yolks of chicken presented superior purity than them obtained in serum of rabbits. From the results of the test ELISA, observed itself that the antibodies against AIA did not present potential of practical use. The detection of plant hormones in samples of tissue stayed restricted to 2ip and zeatin. The detection of those two molecules in crude extract, obtained from plantlets in vitro of gerbera, revealed that the level endogenous of zeatin was higher of level 2ip. Plantlets with six weeks after multiplication presented higher level of zeatin than plantlets with one week after multiplication. For the 2ip, that difference was not evident. The use of antibodies obtained in eggs yolks of chicken permitted the determination of zeatin and 2ip in samples plant utilizing the test ELISA. / A determina??o do n?vel hormonal end?geno pode ser uma excelente ferramenta para estudar o desenvolvimento vegetal. Atualmente, fica limitada devido a complexidade das metodologias adotadas, uma vez que demandam equipamentos e reagentes de alto custo e ainda apresentam baixo rendimento. Por outro lado, os ensaios imunoenzim?ticos possuem algumas vantagens que superam essas limita??es, demonstrando um potencial de uso pr?tico na dosagem de horm?nios vegetais. Entre os ensaios imunoenzim?ticos, destaca-se o teste Enzyme-Linked Immunosorbent Assay (ELISA), o qual vem sendo utilizado na detec??o de mol?culas com baixo peso molecular. No entanto, o teste ELISA exige anticorpos espec?ficos e que sejam capazes de reconhecer as mol?culas de interesse, no caso os horm?nios vegetais. Hoje no mercado n?o h? anticorpos dispon?veis para a determina??o das mol?culas hormonais. Diante disso, prop?s-se produzir e caracterizar anticorpos contra mol?culas de AIA, 2ip e zeatina e posteriormente, us?-los na detec??o dessas mol?culas. Para isso, as mol?culas hormonais foram conjugadas com prote?na (BSA) para posterior imuniza??o de galinhas poedeiras ou coelhos. A partir do soro de coelhos ou de gemas de ovos de galinhas os anticorpos foram purificados e caracterizados atrav?s do teste de imunodifus?o, SDSPAGE e teste ELISA. Os anticorpos que apresentaram melhores resultados foram utilizados na detec??o de mol?culas hormonais em amostras de tecidos de pl?ntulas mantidas in vitro. O teste de imunodifus?o revelou que os anticorpos obtidos foram capazes de detectarem a mol?cula em estudo. Atrav?s do SDS-PAGE verificou-se que os anticorpos obtidos em gemas de ovos de galinhas apresentaram maior pureza que os obtidos em coelhos, sugerindo que os mesmos possuem maior potencial de uso pr?tico. A partir dos resultados do teste ELISA, observou-se que os anticorpos contra AIA n?o apresentaram potencial de uso pr?tico na determina??o dessa mol?cula em amostras vegetais. Sendo assim, a detec??o de mol?culas hormonais em amostras de tecidos vegetais ficou restrita a 2ip e a zeatina. A detec??o dessas duas mol?culas em extrato bruto, obtidos a partir de pl?ntulas de g?rbera mantidas in vitro, revelou que o n?vel end?geno de zeatina foi superior ao n?vel de 2ip. Pl?ntulas com seis semanas ap?s a repicagem apresentaram maior n?vel de zeatina do que pl?ntulas rec?m repicadas, enquanto para o 2ip, essa diferen?a n?o foi evidente. O uso de anticorpos obtidos em gemas de ovos de galinhas permitiu a detec??o e quantifica??o de zeatina e 2ip em amostras vegetais utilizando o teste ELISA.