在Quercus ilex L.育种方面的进展:通过核糖蛋白蛋白的CRISPR/Cas9技术
Vera Pavese1, Andrea Moglia1, Anna Maria Milani1
1Dipartimento di Scienze Agrarie, Forestali e Alimentari-Department of Agricultural, Forest and Food Sciences (DISAFA), Università degli Studi di Torino, Torino, Italy.
Frontiers in plant science
|March 5, 2024
概括
CRISPR/Cas9核糖蛋白 (RNP) 技术首次应用于树 (Quercus ilex) 上. 这种方法在原生质中实现了基因编辑,使得对潜在的树衰退综合征耐药性的转基因无修改成为可能.
科学领域:
- 植物生物技术 植物生物技术
- 基因编辑 基因编辑
- 森林遗传学 森林遗传学
背景情况:
- 荷尔姆树 (Quercus ilex) 在地中海地区在生态和经济上至关重要,但受到树衰退综合征的威胁.
- 基于CRISPR/Cas9核糖蛋白 (RNP) 的技术提供了无转基因的植物修饰.
- 开发高效的遗传工具对于像树这样的反抗性物种至关重要.
研究的目的:
- 报告CRISPR/Cas9-RNP技术在Holm Oak的首次应用.
- 为了优化原生细胞的隔离和转移,霍姆树.
- 为了证明在Holm树原生体中成功编辑基因.
主要方法:
- 评估了从体外叶子和胚胎前质量中分离的原生质.
- 通过使用绿色光蛋白标记物,评估原生质转染效率.
- 交付的CRISPR/Cas9 RNPs针对植物脱酶 (pds) 基因进入树木原体.
- 培养了转染的原始质体,以发展胚胎性细胞.
主要成果:
- 亲胚性质量产生了高质原体数量 (11 x 10^6/ml),具有良好的生存能力.
- 在24小时内实现了62%的转化效率在树原塑中.
- 在PDS目标位点证明了5.6%±0.5%的基因编辑效率.
- 在45天后,成功地从编辑的原生体中再生了胚胎性calli.
结论:
- 克里斯普尔/Cas9-RNP技术是可行的基因编辑树原生体.
- 亲胚性质量是高产原生质隔离的合适来源.
- 这项研究为树的基因改进提供了基础,可能解决树衰退综合征.
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