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首页> 外文期刊>Journal of Rubber Research >Somatic Embryo, an Alternative Target Tissue for Agrobacterium-Mediated Transformation in Hevea brasiliensis
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Somatic Embryo, an Alternative Target Tissue for Agrobacterium-Mediated Transformation in Hevea brasiliensis

机译：体细胞胚，一种农杆菌介导的巴西橡胶树转化的替代目标组织

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Traditionally, embryogenic callus has been used as the only receptor source of genetic transformation in H. brasiliensis. In this study, we showed that somatic embryos were a viable alternative to embryogenic callus. The appropriate concentration of both decontamination and selection antibiotics was firstly screened. Visual and cytological observations found both epidermal and subepidermal cotyledon cells could be infected, indicating that somatic embryos were susceptible to antibiotics and Agrobacterium. Subsequently, five isolated consecutive transformation experiments were carried out to produce the transgenic plants using a total of 1158 somatic embryos, of which 47 (4.06 %) GUS-positive embryos were produced after cell differentiation and embryogenesis. Six were successfully multiplied and produced 33 plants. PCR and Southern blot analysis confirmed that the T-DNA was integrated into the recovered plants. Finally, three additional cycles of secondary embryogenesis were carried out for transgenic embryo proliferation efficiency assessment. The rate of proliferation tended to slightly increase with each proliferation generation, and one initial transgenic embryo-derived GUS-negative embryo were observed in the fifth multiplication cycle, indicating that transgenic plants could be propagated through secondary embryogenesis and some initial transgenic embryos might be chimeric. This is the first attempt on producing transgenic plants using somatic embryos as the target tissue in H. brasiliensis. It is demonstrated that somatic embryos can be used as an alternative target tissue for Agrobacterium-mediated transformation in Hevea brasiliensis.
机译：传统上，胚性愈伤组织已被用作巴西杆菌中遗传转化的唯一受体来源。在这项研究中，我们表明体细胞胚是胚性愈伤组织的可行替代品。首先筛选去污和选择抗生素的适当浓度。视觉和细胞学观察发现表皮和表皮下的子叶细胞均可被感染，这表明体细胞胚胎对抗生素和农杆菌敏感。随后，进行了五个分离的连续转化实验，以使用总共1158个体细胞胚产生转基因植物，其中47个（4.06％）GUS阳性胚在细胞分化和胚胎发生后产生。成功繁殖了六只，生产了33株植物。 PCR和Southern印迹分析证实T-DNA已整合到回收的植物中。最后，进行了三个额外的二次胚胎发生周期，以评估转基因胚胎的增殖效率。随着每一代的繁殖，增殖的速率趋于轻微增加，并且在第五个繁殖周期中观察到了一个初始的转基因胚衍生的GUS阴性胚，这表明转基因植物可以通过次级胚发生繁殖，并且一些初始的转基因胚可能是嵌合的。 。这是使用体细胞胚作为巴西嗜血杆菌的靶组织生产转基因植物的首次尝试。结果表明，体细胞胚可用作农杆菌介导的巴西橡胶树转化的替代靶组织。

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