A transient expression system in soybean mesophyll protoplasts reveals the formation of cytoplasmic GmCRY1 photobody-like structures
- 121 Downloads
Soybean (Glycine max (L.) Merr.), grown for its plant oils and proteins, is one of the most important crops throughout the world. Generating stable and heritable transgenic soybeans is relatively inefficient; therefore, there is an urgent need for a simple and high-efficient transient transformation method by which to enable the investigation of gene functions in soybeans, which will facilitate the elucidation and improvement of the molecular mechanisms regulating the associated agronomic traits. We established a system of transient expression in soybean mesophyll protoplasts and obtained a high level of protoplast transfection efficiency (up to 83.5%). The subcellular activity of the protoplasts was well preserved, as demonstrated by the dynamic formation of GmCRY nucleus photobodies (NPs) and/or cytoplasmic photobody-like structures (CPs) in response to blue light. In addition, we showed that GmCRY1b CPs colocalized with GmCOP1b, a co-ortholog of Arabidopsis thaliana CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1), which provided new insight into the potential roles of GmCRY1s in the cytoplasm.
Keywordssoybean protoplast transfection subcellular localization cryptochrome photobodies
Unable to display preview. Download preview PDF.
This work was supported by the National Key Research and Development Plan (2016YFD0101005), the National Natural Science Foundation of China (31871705 and 31422041), and the Central Public-Interest Scientific Institution Basal Research Fund (Y2016JC13).
- Li, S., Cong, Y., Liu, Y., Wang, T., Shuai, Q., Chen, N., Gai, J., and Li, Y. (2017). Optimization of Agrobacterium-mediated transformation in soybean. Front Plant Sci 8, 246.Google Scholar
- Shen, J., Fu, J., Ma, J., Wang, X., Gao, C., Zhuang, C., Wan, J., and Jiang, L. (2014). Isolation, culture, and transient transformation of plant protoplasts. Curr Protoc Cell Biol 63, 2. 8. 1–17.Google Scholar
- Wu, F., and Hanzawa, Y. (2018). A simple method for isolation of soybean protoplasts and application to transient gene expression analyses. J Vis Exp 131.Google Scholar