Development of Agrobacterium-mediated transient transformation in persimmon (Diospyros kaki Thunb.)

Abstract

The Agrobacterium-mediated transient gene expression is a powerful tool for the characterization of gene function in plants. Here, we have established in vivo transient transformation system with GFP as a reporter gene by Agrobacterium-mediated injection infiltration in persimmon (Diospyros kaki Thunb.) leaves. Transformation conditions (concentration of acetosyringone, bacterial density and days after agroinfiltration) were optimized to achieve higher transformation efficiency. This system is suitable for different types of persimmon cultivars such as ‘Mopanshi’ (PCA), ‘Eshi 1’ and ‘Baogai-tianshi’ (C-PCNA). Furthermore, the developed transient transformation system was successfully utilized for the overexpression of DkLAC1 gene. The results exhibited that the expression level of DkLAC1 increased obviously, which lead to remarkable accumulation of proanthocyanidin (PAs) in persimmon (‘Mopanshi’ and ‘Eshi 1’) leaves transformed with DkLAC1. In conclusion, based on the Agrobacterium-mediated injection infiltration, we have developed a simple and highly efficient transient transformation system that can quickly analyze the gene function in persimmon.