作者：Favero P, Mourao FDA, Stipp LCL, Mendes BMJ.
作者单位：Univ Sao Paulo
摘要：The difficulty in adult tissue genetic transformation in woody species is still an obstacle to be overcome, including in most sweet orange cultivars of the Brazilian citrus industry. This work reports that, after in vitro culture adjustments, transgenic adventitious buds of 'Hamlin', 'Pra', and 'Valencia' sweet oranges (Citrus sinensis L. Osbeck) were recovered using adult material as explant source, in genetic transformation experiments via Agrobacterium tumefaciens. The transgenic buds were identified by the GUS histochemical analysis and confirmed by PCR analysis, which indicated the presence of an amplified fragment of 817 bp corresponding to the uidA gene sequence. The efficiencies of genetic transformation for 'Hamlin', 'Pra', and 'Valencia' sweet orange cultivars were 2.5, 1.4, and 3.7%, respectively. Media supplemented with auxins and cytokinins during co-culture, and media with high concentrations of cytokinins (3 mg L-1) during transgenic selection led to the transformation and, consequently, the regeneration of adequate number of adventitious buds for the three cultivars. The use of sonication during the explant disinfection was not effective to reduce endophytic contamination and reduced transformation efficiency.
关键词：Citrus sinensis; Tissue culture; Biotechnology; Genetic improvement; Agrobacteri
Authors: Favero P, Mourao FDA, Stipp LCL, Mendes BMJ
Abstract: The difficulty in adult tissue genetic transformation in woody species is still an obstacle to be overcome, including in most sweet orange cultivars of the Brazilian citrus industry. This work reports that, after in vitro culture adjustments, transgenic adventitious buds of \'Hamlin\', \'Pra\', and \'Valencia\' sweet oranges (Citrus sinensis L. Osbeck) were recovered using adult material as explant source, in genetic transformation experiments via Agrobacterium tumefaciens. The transgenic buds were identified by the GUS histochemical analysis and confirmed by PCR analysis, which indicated the presence of an amplified fragment of 817 bp corresponding to the uidA gene sequence. The efficiencies of genetic transformation for \'Hamlin\', \'Pra\', and \'Valencia\' sweet orange cultivars were 2.5, 1.4, and 3.7%, respectively. Media supplemented with auxins and cytokinins during co-culture, and media with high concentrations of cytokinins (3 mg L-1) during transgenic selection led to the transformation and, consequently, the regeneration of adequate number of adventitious buds for the three cultivars. The use of sonication during the explant disinfection was not effective to reduce endophytic contamination and reduced transformation efficiency.
Key words: Citrus sinensis; Tissue culture; Biotechnology; Genetic improvement; Agrobacterium tumefaciens
文献注录：Favero P, Mourao FDA, Stipp LCL, Mendes BMJ.. Genetic transformation of three sweet orange cultivars from explants of adult plants [J]. Acta Physiologiae Plantarum. 2012, 34(2): 471-477.
报/刊名：《Acta Physiologiae Plantarum》，发表于2012 / 第 34(2) 期。
文献类型： [J]
页码：第 471-477 页 / 共7 页