自然生物技術｜GRF–GIF嵌合蛋白可提高轉基因植物再生效率

Debernardi, J.M., Tricoli, D.M., Ercoli, M.F. et al. A GRF–GIF chimeric protein improves the regeneration efficiency of transgenic plants. Nat Biotechnol (2020).

https://doi.org/10.1038/s41587-020-0703-0

基因組編輯改善作物農藝性能的潛力通常受到植物再生效率低和可轉化基因型很少的限制。

在這裡，我們表明結合小麥生長調節因子4（GRF4）及其輔因子GRF相互作用因子1（GIF1）的融合蛋白的表達大大提高了小麥，黑小麥和水稻的再生效率和再生速度，並增加了可轉化的小麥基因型。

GRF4-GIF1轉基因植物可育且沒有明顯的發育缺陷。

此外，在沒有外源細胞分裂素的情況下，GRF4-GIF1誘導了小麥的高效再生，這有助於選擇無選擇標記的轉基因植物。我們還將GRF4–GIF1與CRISPR–Cas9基因組編輯結合在一起，並生成了30個經編輯的Q基因（AP2L-A5）被破壞的小麥植株。

最後，我們顯示了雙子葉植物GRF-GIF嵌合體可提高柑橘的再生效率，表明該策略可應用於雙子葉植物。

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.

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