pela elaboração da composição química das soluções nutritivas utilizadas neste trabalho. À Profa. Dra. Sônia Maria de Sthephano Piedade do Departamento de Ciências Exatas, pela atenção e elaboração das análises estatísticas deste trabalho.
Curcuma zedoaria Roscoe (zedoary) is a medicinal properties-bearing Zingiberaceae from which rhizomes are commercially exploited. The objective of this work was to establish an in vitro protocol for micropropagation and callogenesis of Curcuma zedoaria Roscoe as alternative to improve plant production, turning economically feasible the exploitation of its secondary metabolites which present medicinal properties. Micropropagation by using shoot apexes produced by rhizome and from in vitro plants were carried out on Murashige & Skoog medium supplemented with 2.0 mg L -1 benzyl amino purine and 30 g L -1 sucrose. Plantlets were satisfactorily acclimated to greenhouse conditions by using plastic cover for at least 10 days. Treatment with endomycorrhiza at the ex vitro transferring time was beneficial to acclimatization, improving plant growth and development. Callus induction and growth were obtained by inoculating root segments on Murashige & Skoog medium supplemented with 1.0 mg L -1 naphtalene acetic acid and incubation in the dark at 25 ± 2ºC. Cell suspension cultures were established on liquid medium of same chemical composition and same culture conditions and a growth curve was obtained.
The objective of this work was to study the activity of sucrose metabolizing enzymes in extracts of cell suspension cultures of Bauhinia forficata Link, Curcuma zedoaria Roscoe and Phaseolus vulgaris L. Invertase pathway was identified in the three studied species. Sucrose synthase pathway was also responsible for sucrose metabolism in Curcuma zedoaria and Phaseolus vulgaris cells. Activity values higher than 300 nmol min -1 mg -1 of protein were found for acid and neutral invertases, UDPglucose pyrophosphorylase and phosphoglucomutase in the cell extract of the three plant species. Sucrose synthase showed low activity in Bauhinia forficata cells. As sucrose concentration in the culture medium decreased, sucrose synthase activity increased in C. zedoaria and P. vulgaris cells. The glycolytic enzymes activity gradually reduced at the end of the culture period, when carbohydrate was limited.Index terms: fructokinase, glycolysis, tissue culture.Enzimas do metabolismo da sacarose em cultura celular de Bauhinia forficata, Curcuma zedoaria e Phaseolus vulgaris
RESUMO: Indução de calogênese eficiente e multiplicação celular rápida são pré-requisitos fundamentais em biotecnologia de plantas. Sucesso na calogênese é dependente dos componentes do meio de cultura e da qualidade dos explantes. Neste trabalho é relatada a influência do tratamento de matrizes de Dianthus caryophyllus L. com nitrogênio na indução de calogênese in vitro. Mudas de cravo cultivadas em vasos contendo areia foram tratadas com soluções nutritivas contendo 5 níveis de nitrogênio. Explantes folha, entrenós e nó foram coletados aos 30, 45 e 60 dias após início dos tratamentos e inoculados em meio de cultura contendo os sais básicos e vitaminas de Murashige & Skoog (1962), suplementado com 1 g L -1 de caseína hidrolizada, 2 µmol L -1 de cinetina e 3 µmol L -1 de 2,4-D para indução da calogênese. Ao longo dos 60 dias de tratamento com as soluções nutritivas, as matrizes de cravo não apresentaram sintomas visíveis de deficiência ou de excesso do nutriente nitrogênio. O tratamento com nitrogênio afetou a calogênese avaliada em massa de matéria fresca e seca. A produção da massa de matéria fresca de calos foi proporcional ao tratamento com nitrogênio até concentração de 267 mg L -1 para explantes folha por durante 30 dias. Tratamentos mais prolongados (45 e 60 dias) afetaram negativamente a calogênese e foram inversamente proporcionais a concentração de nitrogênio na solução nutritiva. Palavras-chave: nutrição mineral, cultura de tecido NITROGEN TREATMENT OF CARNATION (Dianthus caryophyllus L., CARYOPHYLLACEAE) AND in vitro CALOGENESISABSTRACT: Efficient calogenesis induction and rapid cell multiplication are fundamental requirements in plant biotechnology. The success of calogenesis is dependent on the growth medium components and the quality of explants. This work is referred to the influence of Dianthus caryophyllus L. nitrogen treatment on calogenesis induction in vitro. Carnation cuts rooted in sand pots were treated with nutrient solutions containing 5 nitrogen levels. Leaves, internodes and node explants were collected and inoculated on callus induction culture media containing Murashige & Skoog (1962) salts and vitamins, supplemented with 1 g L -1 hidrolysed casein, 2 µmol L -1 kinetin and 3 µmol L -1 2,4-D. No plant deficiency and toxicity symptoms were apparent on the treated plants during the 60 day treatment. The nitrogen treatment affected calogenesis in relation to calli fresh and dry weights. Callus fresh weigth yield was proportional to nitrogen concentration up to 267 mg L -1 for leaf explant during 30 days. Longer treatments (45 and 60 days) affected calogenesis negatively which were inversely proportional to the nitrogen concentration of the nutrient solution.
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