Agrobacterium-mediated genetic transformation of 'Hamlin' sweet orange

Pesquisa Agropecuária Brasileira

Beatriz Madalena Januzzi Mendes²  Raquel Luciana Boscariol²  Francisco De Assis Alves Mourão Filho¹  Weliton Antonio Bastos De Almeida¹ 

[1],Universidade de São Paulo Centro de Energia Nuclear na Agricultura Piracicaba SP

关键词: Citrus sinensis;    epicotyls;    seedlings;    transgenics;    breeding methods;    Citrus sinensis;    epicótilo;    plântulas;    transgênicos;    métodos de melhoramento;   

DOI  :  10.1590/S0100-204X2002000700009

来源: SciELO

【 摘 要 】

The development and optimization of efficient transformation protocols is essential in new citrus breeding programs, not only for rootstock, but also for scion improvement. Transgenic 'Hamlin' sweet orange (Citrus sinensis (L.) Osbeck) plants were obtained by Agrobacterium tumefaciens-mediated transformation of epicotyl segments collected from seedlings germinated in vitro. Factors influencing genetic transformation efficiency were evaluated including seedling incubation conditions, time of inoculation with Agrobacterium and co-culture conditions. Epicotyl segments were adequate explants for transformation, regenerating plants by direct organogenesis. Higher percentage of transformation was obtained with explants collected from seedlings germinated in darkness, transferred to 16 hours photoperiod for 2-3 weeks, and inoculated with Agrobacterium for 15-45 min. The best co-culture condition was the incubation of the explants in darkness, for three days in culture medium supplemented with 100 muM of acetosyringone. Genetic transformation was confirmed by performing beta-glucoronidase (GUS) assays and, subsequently, by PCR amplification for the nptII and GUS genes.

【 授权许可】

CC BY   
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