Establishing a highly efficient Agrobacterium-mediated transformation system in sweet buckwheat

Buckwheat (Fagopyrum esculentum Moench) is a healthy cereal with a short lifecycle, and an efficient transgenic method for gene function analysis of sweet buckwheat is lacking. In this study, we developed a simple Agrobacterium-mediated transformation protocol using a floral-dip. For this transformation experiment, the Agrobacterium strain GV3101 harboring binary vector pHZM69 with enhanced green fluorescent protein (eGFP) as well as a resistance gene to the herbicide Basta was used. The effect of different cell densities (OD600 = 0.4, 0.6, 0.8 and 1.0) applied for infection was measured, and maximum transformation efficiency (12.42%) was obtained with cell density of OD600 = 0.6, followed by OD600 = 0.4 (8.48%), OD600 = 0.8 (4.39%) and OD600 = 1.0 (3.62%). Here, sweet buckwheat was successfully transformed by Agrobacterium-mediated floral dip method, which could be a useful tool for the production of transgenic sweet buckwheat and promote genomic function analysis in sweet buckwheat.

Automated summary

In this study, a simple Agrobacterium-mediated transformation protocol using a floral-dip was developed for gene function analysis of sweet buckwheat.