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Somatic Embryo, an Alternative Target Tissue for Agrobacterium-Mediated Transformation in Hevea brasiliensis

文献类型： 外文期刊

作者： Huang, T. D. 1 ; Li, J. 1 ; Li, Y. T. 1 ; Huang, H. S. 1 ; Hua, Y. W. 1 ;

作者机构： 1.Chinese Acad Trop Agr Sci, Key Lab Biol & Genet Resources Rubber Tree, Rubber Res Inst, Minist Agr, Danzhou 571737, Hainan, Peoples R China

关键词： Hevea brasiliensis;somatic embryo;genetic transformation;Agrobacterium tumefaciens;secondary embryogenesis

期刊名称：JOURNAL OF RUBBER RESEARCH （ 影响因子：0.667； 五年影响因子：0.618 ）

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收录情况： SCI

摘要： 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.

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