A simple and efficient in planta transformation method for pommelo (Citrus maxima) using Agrobacterium tumefaciens

A simple and efficient transformation method is important for plant genetic breeding and gene function analysis. In this study, we developed a in planta transformation approach for pommelo (Citrus maxima), which includes seed germination, seedling decapitation, Parafilm funnel-type-winding, Agrobacterium infection, co-culture, selective-culture, dark treatment for buds induction, transgene integration confirmation and transgenic plant propagation. The highest transformation efficiency (20.41%) was obtained when using hygromycin as selection marker, followed by Basta (1937%) and Kanamycin (3.21%). Trans genic plants obtained using this method could be graft-propagated in 3 months post transformation, which is shorter than the tissue-culture-dependent transformation. The transformation protocol presented here has great potential to be used for citrus genetic breeding and improvement. (C) 2016 Elsevier B.V. All rights reserved.