Propagação in vitro de antúrio (Anthurium andraeanum cv. Eidibel) via embriogênese somática

• Autor(a) principal: Pinheiro, Marcos Vinícius Marques
• Data de Publicação: 2010
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: LOCUS Repositório Institucional da UFV
• Texto Completo: http://locus.ufv.br/handle/123456789/4319
• topic: Propagação em vitro; Embriogênese somática; In vitro propagation; Somatic embryogenesis; CNPQ::CIENCIAS BIOLOGICAS::BOTANICA::FISIOLOGIA VEGETAL

In this work, the propagation in vitro of Anthurium andraeanum cv. Eidibel through somatic embryogenesis was investigated, evaluating the induction of the embryogenic cultures (Experiment I); pre-maturation of the somatic embryos (Experiment II); and maturation of the somatic embryos with subsequent regeneration of the plants (Experiment III). In Experiment I, the subdivision of plants of anturio established in vitro was used, to obtain the explant sources. Analyzed in DIC with factorial 5 x 5 x 5, with five explant types (whole leaves were cut across the midrib of leaves; petioles; stem segments with one bud; and root segment without the apex radicular; each explant was cut into pieces of about 1.0 cm); five auxins (Picloram, ANA, AIB, 2.4-D and Picloram) in five concentrations (0.0; 2.5; 5.0; 7.5 and 10.0 μM) and five additional witness, in other words, each explant source added to the treatment without auxin. The repetition was composed by five Petri dishes (90 x 15 mm), containing 25 mL of Pierik medium, and each unit experimental nine explant/placa. Cultures were maintained in growth room, at 25 ± 2 ºC, in the darkness. After 60 days of cultivation, it was evaluated the presence of embryogenic cultures, shoots, roots and mass of the produced callus. The production of the first callus was observed in petioles and stem explants, and its production was in average containing 7.5 μM ANA and 10.0 μM Picloram. In the concentrations of 10.0 μM, there was no different statistics among the treatments with the auxins ANA, 2.4-D and Picloram, being superior to the others. For the production of shoots, the explant with the largest average was the stem segments, in average without the auxins. The proliferation of roots was observed in stem segments and roots explants, mainly in average with AIB. Histochemistry analysis were determined, through the double stained with acetocarmine and Evan's blue and lugol test. However, it was observed in histological cuts that the callus produced in Pierik medium containing 10.0 μM of ANA presented well developed embryos, with protoderm and procambium, with polarization signs. For the proliferation of the embryogenic cultures, the subdivision of the selected callus was accomplished, and inoculated in Petri dishes with Pierik medium containing 10.0 μM of ANA, in five successive subcultures, with 60 days each. In the experiment II, the callus was inoculated, produced in the proliferation of the embryogenic cultures, with about 90 mg weight, in Erlenmeyers of 125 mL, containing 25 mL of average liquid , Pierik and AA2, with different concentrations of 2.4-D (0.00; 4.52; 9.05 μM) and kinetin, (0.00; 0.47; 2.32 μM), analyzed in DIC with factorial 2 x 3 x 3, maintained at growth room at 25 ± 2 ºC, in the darkness, staying under orbital agitation of 100 rpm. After 45 days of cultivation, it was evaluated a mass of the embryogenic callus; production of somatic embryos; production of secondary somatic embryos; oxidation percentage; coloration, texture of the embryogenic callus and development of the somatic embryos produced. The production of somatic embryos was better in Pierik medium with 0.47 μM of kinetin, being observed a smaller production of secondary somatic embryos, smaller oxidation percentage, with friable texture of the callus, being one of the treatments with larger development of the embryos, proven for the histological cuts, in which it was observed embryos in the globular stadium, with polarization signs, even mature embryos, with the presence of primary leaf and meristematic zone. In the Experiment III, the callus produced were inoculated in Erlenmeyers containing 25 mL of liquid and semi-solid medium, Pierik and AA2, suplemented with the kinetin concentrations (0.0; 1.16; 2.32; 4.64 μM), analyzed in DIC with factorial 2 x 2 x 4. Cultures were maintained under a 16 hours of light, photoperiod at 36 μmol m-2 s-1 provided by cool white fluorescent lamps at 25 ± 2 ºC. The liquid medium staying under orbital agitation of 100 rpm. After 45 days of cultivation, it was evaluated the number of callus with mature embryos; percentage of conversion of plants; oxidation percentage; and number of embryos with complete maturation. Pierik medium containing 2.32 μM of kinetin is recommended, in which, it was much better to the number of embryogenic callus with mature embryos, number of embryos with complete maturation and for the best conversion in plants. The plants were acclimatized ex vitro in the bench of the laboratory and transferred, in the end of two months to a greenhouse. The present study evidenced that the induction and proliferation of embryogenic cultures starting from stem segmentswere dependent on the type and concentration of the auxins. For the pre-maturation and maturation of the somatic embryos, it is necessary the retreat or the reduction of the auxin in the medium, adding ideal concentrations of cytokinins, and obtaining high conversion of the somatic embryos in plants, in the final process.