GRF-GIF 嵌合蛋白提高了转基因植物的再生效率。
A GRF-GIF chimeric protein improves the regeneration efficiency of transgenic plants.
机构信息
Department of Plant Sciences, University of California, Davis, CA, USA.
Howard Hughes Medical Institute, Chevy Chase, MD, USA.
出版信息
Nat Biotechnol. 2020 Nov;38(11):1274-1279. doi: 10.1038/s41587-020-0703-0. Epub 2020 Oct 12.
The potential of genome editing to improve the agronomic performance of crops is often limited by low plant regeneration efficiencies and few transformable genotypes. Here, we show that expression of a fusion protein combining wheat GROWTH-REGULATING FACTOR 4 (GRF4) and its cofactor GRF-INTERACTING FACTOR 1 (GIF1) substantially increases the efficiency and speed of regeneration in wheat, triticale and rice and increases the number of transformable wheat genotypes. GRF4-GIF1 transgenic plants were fertile and without obvious developmental defects. Moreover, GRF4-GIF1 induced efficient wheat regeneration in the absence of exogenous cytokinins, which facilitates selection of transgenic plants without selectable markers. We also combined GRF4-GIF1 with CRISPR-Cas9 genome editing and generated 30 edited wheat plants with disruptions in the gene Q (AP2L-A5). Finally, we show that a dicot GRF-GIF chimera improves regeneration efficiency in citrus, suggesting that this strategy can be applied to dicot crops.
基因组编辑提高农作物农艺性能的潜力通常受到植物再生效率低和可转化基因型少的限制。在这里,我们表明,表达一种融合蛋白,该融合蛋白将小麦生长调节因子 4(GRF4)与其共因子 GRF 相互作用因子 1(GIF1)结合在一起,可显著提高小麦、小黑麦和水稻的再生效率和速度,并增加可转化的小麦基因型数量。GRF4-GIF1 转基因植物可育且无明显发育缺陷。此外,GRF4-GIF1 在没有外源细胞分裂素的情况下诱导有效的小麦再生,这有利于在没有选择标记的情况下选择转基因植物。我们还将 GRF4-GIF1 与 CRISPR-Cas9 基因组编辑相结合,生成了 30 株在 Q(AP2L-A5)基因中发生突变的编辑小麦植株。最后,我们表明,双子叶植物的 GRF-GIF 嵌合体提高了柑橘的再生效率,这表明该策略可应用于双子叶作物。