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首页> 外文期刊>Revista Brasileira de Fruticultura >Eficiência de transforma??o genética de citrange 'carrizo' com duas constru??es gênicas
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Eficiência de transforma??o genética de citrange 'carrizo' com duas constru??es gênicas

机译：两个基因构建体对'carrizo'柑桔的遗传转化效率

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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 work aimed to evaluate the transformation efficiency 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 antibiotic kanamycin, 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
机译：遗传转化被认为是用于柑橘遗传改良的重要工具。但是，转化效率可能会根据多种因素而变化，包括所利用的基因构建。这项工作旨在评估'Carrizo'柑桔[Poncirus trifoliata（L.）Raf。的转化效率。 x Citrus sinensis（L.）Osbeck]，包含两个不同的基因构建，包括由启动子拟南芥韧皮部蛋白质2（AtPhP2）和拟南芥蔗糖转运蛋白2（AtSuT2）控制的uidA（GUS）基因。来自体外发芽幼苗的Epicotil片段用作外植体。赋予对卡那霉素抗生素抗性的nptII基因在结构中用作再生转基因植物的选择剂。为了验证uidA基因的表达，对所有再生的芽进行了X-GLUC组织化学分析。从利用的4790个上胚轴片段中，有366个芽在组织化学分析中具有阳性反应，并进一步嫁接到体外生长的砧木上。从每种基因的构建中，从这些芽中选择5个进行PCR分析，并使用特定引物进行uidA基因序列扩增。通过PCR评估的所有选择的植物证实了基因整合。通过组织化学分析评估的转化效率和未转化芽的数量在所利用的基因构建中有所不同

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