An efficient Agrobacterium-mediated soybean transformation method using green fluorescent protein as a selectable marker
2019
Yang, Songnan | Hu, Yanfeng | Cheng, Zongming | Rice, John Hollis | Miao, Long | Ma, Jing | Hewezi, Tarek | Li, Yan | Gai, Junyi
Genetic transformation plays a vital role in gene functional study and molecular breeding of soybean. Conventional soybean transformation methods using chemical selectable markers, such as antibiotic or herbicide resistance genes, rely on the identification of positive transgenic lines at advanced developmental stages, making selection procedure labor intensive and time consuming. Utilization of a visual maker to track the transgene would avoid the uncertainty and blindness in the transformation process. In this research, we used green fluorescent protein (GFP) as the selectable marker to detect transgenics at early stages of soybean development. Positive transformants were detected recurrently during each stage of the process based on visualization of the green fluorescence signal, which help us to discard the non-transgenic ones in each stage to reduce the unnecessary experimental cost and lab space. In addition, the positive transgenic seeds can be identified before planting for early detection of transgene and obtain homozygous lines in advance. The method established in this study is also a useful reference for other plant species.
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