作者：Miyata LY, Mourão Filho FAA, Scarpare Filho JA, Zambon F, Bassan MM, Mendes BMJ, Harakava R.
摘要：Genetic transformation is considered an important tool to be applied in citrus genetic improvement. However, transformation efficiency may vary according to several factors, including the genetic construction utilized. This study was carried out to evaluate the efficiency of genetic transformation of ‘Carrizo’ citrange [Poncirus trifoliata (L.) Raf. x Citrus sinensis (L.) Osbeck], containing two different gene constructions including the uidA (GUS) gene controlled by the promoters Arabidopsis thaliana phloem protein 2 (AtPhP2) and Arabidopsis thaliana sucrose transporter 2 (AtSuT2). Epicotil segments from in vitro germinated seedlings were used as explants. The nptII gene, which confers resistance to the kanamycin antibiotic, was used in the constructions as a selection agent to regenerate transgenic plants. X-GLUC histochemical analysis was performed in all regenerated shoots in order to verify the expression of the uidA gene. From the 4790 epicotyl segments utilized, 366 shoots had a positive reaction in the histochemical analysis, and were further grafted on in vitro grown rootstocks. Five out of these shoots, from each gene construction, were selected to perform PCR analysis, with specific primers for uidA gene sequence amplification. All selected plants evaluated by PCR confirmed gene integration. Transformation efficiency and number of non-transformed shoots, evaluated by histochemical analysis, varied among gene constructions utilized.
关键词：biotechnology; Citrus spp.; uidA gene (GUS); genetic improvement; tissue culture
文献注录：Miyata LY, Mourão Filho FAA, Scarpare Filho JA, Zambon F, Bassan MM, Mendes BMJ, Harakava R.. Genetic transformation efficiency of \'carrizo\' citrange with two gene constructions [J]. Revista Brasileira de Fruticultura. 2011, 01: 311-315.
报/刊名：《Revista Brasileira de Fruticultura》，发表于2011 / 第 01 期。
文献类型： [J]
页码：第 311-315 页 / 共5 页