作者：Gao, Erlin, Zhao, Yunju, Guo, Wenwu and Wang, Pengwei
摘要： For perennial woody plants, it is important to establish an efficient transformation method to facilitate gene functional studies and protein subcellular localization assays. Therefore, calluses are ideal materials for these purposes. Here, we optimized a series of over-expression vectors with different fluorescent protein tags and antibiotic selections, providing the possibility to generate transgenic callus expressing two different fusion proteins at the same time. On the other hand, a series of RNAi (RNA interference) vectors were also modified by introducing free eGFP or mCherry as reporters to facilitate visual screening. With the help of these improvements, stable transgenic lines expressing an autophagosome marker, GFP-FhATG8a (Autophagy 8a) were generated, and further studies showed that autophagosomes were more abundant during NaCl treatment or phosphate deficiency. By knocking down the expression of a key autophagy gene, FhATG5, the resulting callus was more susceptible to salt, indicating that autophagy is essential for stress response in citrus. In summary, our study provided an effective system for callus-base transformation to study gene function, and this new approach was successfully used to study the activity of autophagy in citrus as an exexample
文献注录：Gao, Erlin, Zhao, Yunju, Guo, Wenwu and Wang, Pengwei. Optimized plasmids for callus transformation and functional studies of autophagy in citrus [J]. . 2023, 3.
报/刊名：《》，发表于2023 / 第 3 期。
文献类型： [J]
页码：第 页 / 共 页