建立糖皮质激素诱导系统,调节 Liriodendron 杂交种的体细胞胚胎发生。

Forestry research Pub Date : 2024-02-29 eCollection Date: 2024-01-01 DOI:10.48130/forres-0024-0003
Xinying Chen, Ye Liu, Lu Lu, Siqin Liu, Yuhao Weng, Jisen Shi, Zhaodong Hao, Jinhui Chen
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摘要

转录因子(TF)的精确表达对植物的生长和发育至关重要,尤其是在体细胞胚胎发生过程中。然而,功能遗传学常用的传统过表达方法可能会导致有害影响。因此,在提高体细胞胚胎发生效率时,必须确保 TFs 以可控和及时的方式表达。本研究采用地塞米松/糖皮质激素受体(DEX/GR)诱导表达系统来调节Liriodendron杂交种体细胞胚胎发生过程中细胞核内目标TFs蛋白的表达水平。我们选择了WUSCHEL(WUS)基因作为评估该系统有效性的模型,WUSCHEL是一种功能性TF,在体细胞胚胎发生过程中发挥着重要作用。通过DEX处理,我们诱导LhWUS-GR/LhWUS-GFP-GR融合蛋白从细胞质转位到细胞核,从而导致WUS驱动的体细胞胚胎发生。随着 DEX 浓度的增加,LhWUS-GFP-GR 融合蛋白向细胞核的迁移也相应增加。此外,我们还观察到,当暴露于不同浓度的 DEX 时,表达 LhWUS-GR 的胼胝体的增殖率更高。值得注意的是,在最佳 DEX 浓度下,体细胞胚胎发生的效率有显著提高。总之,我们的研究成功地在Liriodendron杂交种中利用了DEX/GR诱导系统,为在所需水平上精确控制和利用TFs提供了有价值的工具。这种创新方法有望促进我们对 TF 功能的了解,并通过对 TF 表达的调控促进植物发育。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Establishment of a glucocorticoid inducible system for regulating somatic embryogenesis in Liriodendron hybrids.

The precise expression of transcription factors (TFs) is crucial for plant growth and development, especially during somatic embryogenesis. However, conventional overexpression approaches, commonly used for functional genetics, can lead to deleterious effects. Therefore, it is imperative to ensure that TFs are expressed in a controlled and timely manner when aiming to enhance the efficiency of somatic embryogenesis. In this study, a dexamethasone/glucocorticoid receptor (DEX/GR) inducible expression system was employed to modulate the protein expression levels of target TFs within the nucleus during somatic embryogenesis in Liriodendron hybrids. We selected the WUSCHEL (WUS) gene, a well-established functional TF known for its vital role in somatic embryogenesis, as a model to assess the effectiveness of this system. Through DEX treatment, we induced the translocation of LhWUS-GR/LhWUS-GFP-GR fusion proteins from the cytoplasm to the nucleus, consequently leading to WUS-driven somatic embryogenesis. As the DEX concentration increased, there was a corresponding increase in the migration of the LhWUS-GFP-GR fusion protein into the nucleus. Additionally, we observed a higher proliferation ratio of callus expressing LhWUS-GR when exposed to varying DEX concentrations. Notably, the efficiency of somatic embryogenesis exhibited significant improvement under optimal DEX concentration. In conclusion, our study successfully utilizes the DEX/GR inducible system in Liriodendron hybrids, offering a valuable tool for the precise control and utilization of TFs at the desired levels. This innovative approach holds promise for advancing our understanding of TF function and enhancing plant development through the regulated manipulation of TF expression.

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