Tocopherols, collectively known as vitamin E, are lipophilic antioxidants, which are synthesized only by photosynthetic organisms. Due to their enormous potential to protect cells from oxidative damage, tocopherols are used e.g. as nutraceuticals and additives in pharmaceuticals. The most biologically active form of vitamin E is α-tocopherol. Most tocopherols are currently produced via chemical synthesis. Nevertheless, this always results in a racemic mixture of different and less effective stereoisomers because the natural isomer has the highest biological activity. Therefore, tocopherols synthesized in natural sources are preferred for medical purposes. The annual sunflower (Helianthus annuus L.) is a well-known source for α-tocopherol. Within the presented work, sunflower callus and suspension cultures were established growing under photomixotrophic conditions to enhance α-tocopherol yield. The most efficient callus induction was achieved with sunflower stems cultivated on solid Murashige and Skoog medium supplemented with 30 g l-1 sucrose, 0.5 mg l-1 of the auxin 1-naphthalene acetic acid and 0.5 mg l-1 of the cytokinin 6-benzylaminopurine. Photomixotrophic sunflower suspension cultures were induced by transferring previously established callus into liquid medium. The effects of light intensity, sugar concentration and culture age on growth rate and α-tocopherol synthesis rate were characterized. A considerable increase (max. 230 %) of α-tocopherol production in the cells was obtained within the photomixotrophic cell culture compared to a heterotrophic cell culture. These results will be useful for improving α-tocopherol yields of plant in vitro cultures.

Submitted by Franciele Moreira (francielemoreyra@gmail.com) on 2018-11-05T12:40:45Z No. of bitstreams: 2 Tese - Andreia Alves da Costa Silveira - 2018.pdf: 3591961 bytes, checksum: 1441f67b3dd43252aec552b063b6081a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-11-05T14:44:38Z (GMT) No. of bitstreams: 2 Tese - Andreia Alves da Costa Silveira - 2018.pdf: 3591961 bytes, checksum: 1441f67b3dd43252aec552b063b6081a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-11-05T14:44:38Z (GMT). No. of bitstreams: 2 Tese - Andreia Alves da Costa Silveira - 2018.pdf: 3591961 bytes, checksum: 1441f67b3dd43252aec552b063b6081a (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-07-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / In micropropagation, seedlings are produced in large scale with homogeneity, which can be potentiated with photomixotrophic systems, which improve the physiological quality of the seedlings. Endophytic fungi of bamboo have the potential of producing substances that can promote biofertilization and antagonism against pathogens. An example of application is antagonism against Magnaporhe oryzae, considered the main pathogen of rice. The objective of this work was to establish the micropropagation protocol of Bambusa oldhamii; besides characterizing endophytic fungi of this species, verify the biofertilizer potential in rice and potential antagonist against M. oryzae. Buds were inoculated in MS medium supplemented with 0.45 μM Tidiazuron-TDZ, 0.2% (w / v) PPM (Plant Preservative Mixture) and 50 mg L-1 kanamycin. The treatments consisted of: different months of collection (March-December 2016) x types of cap (heterotrophic and photomixotrophic system) x luminous conditions (100% blue (455nm), 100% red (630nm), 30% blue + 70% For the multiplication, shoots were inoculated in MS medium + 2.27 μM TDZ or 3.40 μM Paclobutrazol - PBZ. The fungal isolates that were observed in the in vitro culture were identified and characterized biochemically, besides antagonism to M. oryzae and microbiolization of rice seeds, June and July were constituted in the best months of collection, and the climatic variables that most interfered in the morphogenesis were minimum temperature and compensated mean. Photomixotrophic system was superior in the multiplication with TDZ, with increase of 59.51% in 30% blue + 70% red, 70.80% in 30% red + 7 0% blue, and 50.49% in white fluorescent lights. PBZ was higher in carotenoid production, with a mean of 128.02 μg / mL in blue light and conventional lids. TDZ was higher than PBZ when blue was not present. Five potential fungal isolates were identified, 29 (Acrocalymma sp.), 122 (Botryobambusa fusicoccum) 711 (Phoma sp.), 712 (Phoma sp.) And 27 (Arthrinium marii) isolates 711, 712, 27 and 29 produced PPO. The isolate 29 produced higher amounts of AIA, with 31.55 mg / mL on the fourth day. It was observed a reduction of mycelial growth of M. oryzae by all isolates, with emphasis on isolate 122. Isolate 711 presented phosphate solubilization, and higher mean shoot length, fresh and dry mass in rice microbiolization. An efficient micropropagation protocol of B. oldhamii using a photomixotrophic system with 30% blue + 70% red LEDs could be established, as well as the identification of a fungal isolate promising for biofertilization, which could be the basis for new studies with rooting of B. oldhamii and promotion of growth in newly acclimatized species. / Na micropropagação, mudas são produzidas em larga escala com homogeneidade, o que pode ser potencializado com sistemas fotomixotróficos, os quais melhoram a qualidade fisiológica das mudas. Fungos endofíticos de bambu possuem potencial de produção de substâncias que podem promover biofertilização e antagonismo contra patógenos. Exemplo de aplicação se dá com o antagonismo contra Magnaporhe oryzae, considerado o principal patógeno do arroz. O objetivo deste trabalho foi o estabelecimento de protocolo de micropropagação de Bambusa oldhamii; além de caracterizar fungos endofíticos desta espécie, verificar o potencial biofertilizante em arroz e potencial antagonista contra M. oryzae. Brotos foram inoculados em meio MS suplementado com 0,45 µM de Tidiazuron – TDZ, 0,2% (p/v) de PPM (Plant Preservative Mixture) e 50 mg L -1 de canamicina. Os tratamentos consistiram de: diferentes meses de coleta (março-dezembro de 2016) x tipos de tampa (sistema heterotrófico e fotomixotrófico) x condições luminosas [100% azul (455nm), 100% vermelho (630nm), 30% azul + 70% vermelho, 30% vermelho + 70% azul ou branco fluorescente]. Para a multiplicação, brotos foram inoculados em meio MS + 2,27 µM de TDZ ou 3,40 µM de Paclobutrazol - PBZ. Os isolados fúngicos que foram observados no cultivo in vitro foram identificados e caracterizados bioquimicamente, além de antagonismo à M. oryzae e microbiolização de sementes de arroz. Junho e Julho se constituíram nos melhores meses de coleta, e as variáveis climáticas que mais interferiram na morfogênese foram temperatura mínima, e média compensada. LEDs 30% azul + 70% vermelho obtiveram maiores médias, com aumento de 81,79% no número de folhas. Sistema fotomixotrófico foi superior na multiplicação com TDZ, com aumento de 59,51% em 30% azul + 70% vermelho, 70,80% em 30% vermelho + 70% azul, e 50,49% em luzes brancas fluorescentes. PBZ foi superior em produção de carotenoides, com médias de 128,02 µg/mL em luz azul e tampas convencionais. Foram identificados cinco isolados fúngicos potenciais, denominados: 29 (Acrocalymma sp.), 122 (Botryobambusa fusicoccum) 711 (Phoma sp.), 712 (Phoma sp.) e 27 (Arthrinium marii) Os isolados 711, 712, 27 e 29 produziram PPO. O isolado 29 produziu maior quantidade de AIA, com 31,55 mg/mL no quarto dia. Observou-se redução do crescimento micelial de M. oryzae por todos os isolados, com destaque para o isolado 122. O isolado 711 apresentou solubilização de fosfato, e maiores médias de comprimento da parte aérea, massa fresca e seca na microbiolização em arroz. Pôde-se estabelecer um protocolo eficiente de micropropagação de B. oldhamii utilizando sistema fotomixotrófico com LEDs 30% azul + 70% vermelho, assim como foi possível identificar um isolado fúngico promissor para a biofertilização, dados estes que podem ser o alicerce para novos estudos com enraizamento de B. oldhamii e promoção de crescimento em espécies recém-aclimatizadas.

Tocopherols, collectively known as vitamin E, are lipophilic antioxidants, which are synthesized only by photosynthetic organisms. Due to their enormous potential to protect cells from oxidative damage, tocopherols are used e.g. as nutraceuticals and additives in pharmaceuticals. The most biologically active form of vitamin E is α-tocopherol. Most tocopherols are currently produced via chemical synthesis. Nevertheless, this always results in a racemic mixture of different and less effective stereoisomers because the natural isomer has the highest biological activity. Therefore, tocopherols synthesized in natural sources are preferred for medical purposes. The annual sunflower (Helianthus annuus L.) is a well-known source for α-tocopherol. Within the presented work, sunflower callus and suspension cultures were established growing under photomixotrophic conditions to enhance α-tocopherol yield. The most efficient callus induction was achieved with sunflower stems cultivated on solid Murashige and Skoog medium supplemented with 30 g l-1 sucrose, 0.5 mg l-1 of the auxin 1-naphthalene acetic acid and 0.5 mg l-1 of the cytokinin 6-benzylaminopurine. Photomixotrophic sunflower suspension cultures were induced by transferring previously established callus into liquid medium. The effects of light intensity, sugar concentration and culture age on growth rate and α-tocopherol synthesis rate were characterized. A considerable increase (max. 230 %) of α-tocopherol production in the cells was obtained within the photomixotrophic cell culture compared to a heterotrophic cell culture. These results will be useful for improving α-tocopherol yields of plant in vitro cultures.