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

JOURNAL OF CONSUMER PROTECTION AND FOOD SAFETY(2023)

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摘要
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 (OD 600 = 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 OD 600 = 0.6, followed by OD 600 = 0.4 (8.48%), OD 600 = 0.8 (4.39%) and OD 600 = 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.
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关键词
Transgenic,Buckwheat,Agrobacterium-mediated transformation,Gene function analysis,Floral dip
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