Highly Efficient Biolistic Transformation of Embryogenic Cell Suspensions of Banana Via a Liquid Medium Selection System

文献类型： 外文期刊

第一作者： Dong, Tao

作者： Dong, Tao;Bi, Fang-cheng;He, Wei-di;Deng, Gui-ming;Gao, Hui-jun;Sheng, Ou;Li, Chun-yu;Yang, Qiao-song;Yi, Gan-jun;Hu, Chun-hua;Huang, Yong-hong

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关键词： ECS; embryogenic cells; liquid screening system; Musa; transgenesis

期刊名称：HORTSCIENCE （ 影响因子：1.455； 五年影响因子：1.617 ）

ISSN： 0018-5345

年卷期： 2020 年 55 卷 5 期

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收录情况： SCI

摘要： An efficient biolistic transformation system of banana combined with a liquid medium selection system was developed during this study. An embryogenic cell suspension (ECS) of Musa acuminata cv. Baxi (AAA) was bombarded with a particle delivery system. After 7 days of restoring culture in liquid M2 medium, embryogenic cells were transferred to a liquid selection M2 medium supplemented with 10 mu g/mL hygromycin for resistance screening. The untransformed cell clusters were inhibited or killed, and a small number of transformants proliferated in the liquid selection medium. After the 0th, first, second, and third generation of antibiotic screening, there were 0, 65, 212, and 320, respectively, vitality-resistant buds obtained from a 0.5-mL packed cell volume (PCV) of embryogenic cell suspension. The beta-glucuronidase (GUS) staining, polymerase chain reaction (PCR) analysis, and Southern blot hybridization results all demonstrated a 100% positive rate of regenerated resistant seedlings. Interestingly, the number of buds obtained through third-generation screening was almost equal to that obtained from the original ECS in M2 medium without antibiotics. These results suggested that the liquid medium selection system facilitated the proliferation of a positive transgenic ECS, which significantly improved the regeneration rate of transformants. This protocol is suitable for the genetic transformation of all banana genotypes and is highly advantageous to varieties with low callusing potential.

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