The effect of carbon source on in vitro organogenesis of chrysanthemum thin cell layers

The effect of carbon source on in vitro organogenesis of chrysanthemum thin cell layers

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The effect of carbon on in vitro organogenesis of chrysanthemum 165 Bragantia, Campinas, v.63, n.2, p.165-177, 2004 ( 1 ) Received in March 26 th and approved in December 10 th , 2003. ( 2 ) Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa, 761-0795, Japan. E-mail: jaimetex@lycos.com ÁREAS BÁSICAS THE EFFECT OF CARBON SOURCE ON IN VITRO ORGANOGENESIS OF CHRYSANTHEMUM THIN CELL LAYERS (1) JAIME A. TEIXEIRA DA SILVA (2) ABSTRACT Carbon source is an indispensable factor for the development of an in vitro morphogenic program of chrysanthemum micropropagation. The choice of carbon source affects the qualitative and quantitative outcome, and also the response of thin cell layers when these are placed onto morphogenic (callus, root, shoot, somatic embryo) media. Threshold survival levels (TSLs) could not be obtained for sucrose, fructose or glucose. TSLs together with organ differentiation were, however, obtained for mannose (60-80 g.L -1 ), xylose and lactose (40 g.L -1 ) and cellulose (60-80 g.L -1 ), making these carbon sources suitable as potential carbon sources for positive selection systems for chrysanthemum genetic transformation. Key words: Dendranthema X grandiflora , phytotoxicity, regeneration capacity, threshold survival level. RESUMO EFEITO DE CARBONO NA ORGANOGÊNESE DE CAMADAS CELULARES FINAS DE CRISÂNTEMO A fonte de carbono é factor indispensável no sucesso de um programa de desenvolvimento morfogênico de crisântemo. A escolha da fonte de carbono afeta a resposta qualitativa e quanti- tativa de camadas celulares finas, quando são colocadas em meios de desenvolvimento morfogênico (calo, raiz, eixo caulinar, embrião somático). Não se estabeleceram níveis de sobrevivência quando sucrose, frutose ou galactose foram usados, mas sim para manose (60-80 g.L -1 ), xilose e lactose (40 g.L -1 ), e celu- lose (60-80 g.L -1 ), sugerindo o potencial dessas fontes de carbono em sistemas de seleção positivas em programas de transformação genética de crisântemo. Palavras-chave: Dendranthema X grandiflora , fitotoxicidade, capacidade regenerativa, nível de sobrevivência. Abbreviations: 2,4-D, 2,4-dichlorophenoxyacetic acid; BA, 6-benzyladenine; ES, explant survival; IAA, indole-3-acetic acid; NAA, α -naphthalene acetic acid; PGR, plant growth regulator; SE, somatic embryo; tTCL, transverse thin cell layer; TDZ, thidiazuron; TSL, threshold survival level.
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J.A.T. Silva Bragantia, Campinas, v.63, n.2, p.165-177, 2004 166 1. INTRODUCTION Sugars control the expression of many plant genes and their connection to metabolic and developmental processes is unequivocal (K OCH , 1996). Plant genetic transformation systems still require the use of antibiotics for the selection of cells or tissues that contain an antibiotic-degrading gene, such as nptII (kanamycin and other aminoglycoside antibiotics) and hptII (hygromycin), inter alia . This selection system is gradually being replaced by a positive (R EED et al., 2001) selection system, since in the former there is the risk of resistance build-up to a selective substance by microbes, and a concern for potential negative environmental impacts (USFDA, 2002). Xylose, like mannose (a glucose epimer at the
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The effect of carbon source on in vitro organogenesis of chrysanthemum thin cell layers

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