Sanchez Esther Rosales, Price R Jordan, Marangelli Federico, McLeary Kirsty, Harrison Richard J, Kundu Anindya
Crop Science Centre, University of Cambridge, Cambridge, CB3 0LE, UK.
NIAB, Cambridge, CB3 0LE, UK.
Plant Methods. 2024 Oct 18;20(1):160. doi: 10.1186/s13007-024-01270-8.
Plant breeding played a very important role in transforming strawberries from being a niche crop with a small geographical footprint into an economically important crop grown across the planet. But even modern marker assisted breeding takes a considerable amount of time, over multiple plant generations, to produce a plant with desirable traits. As a quicker alternative, plants with desirable traits can be raised through tissue culture by doing precise genetic manipulations. Overexpression of morphogenic regulators previously known for meristem development, the transcription factors Growth-Regulating Factors (GRFs) and the GRF-Interacting Factors (GIFs), provided an efficient strategy for easier regeneration and transformation in multiple crops.
We present here a comprehensive protocol for the diploid strawberry Fragaria vesca Hawaii 4 (strawberry) regeneration and transformation under control condition as compared to ectopic expression of different GRF4-GIF1 chimeras from different plant species. We report that ectopic expression of Vitis vinifera VvGRF4-GIF1 provides significantly higher regeneration efficiency during re-transformation over wild-type plants. On the other hand, deregulated expression of miRNA resistant version of VvGRF4-GIF1 or Triticum aestivum (wheat) TaGRF4-GIF1 resulted in abnormalities. Transcriptomic analysis between the different chimeric GRF4-GIF1 lines indicate that differential expression of FvExpansin might be responsible for the observed pleiotropic effects. Similarly, cytokinin dehydrogenase/oxygenase and cytokinin responsive response regulators also showed differential expression indicating GRF4-GIF1 pathway playing important role in controlling cytokinin homeostasis.
Our data indicate that ectopic expression of Vitis vinifera VvGRF4-GIF1 chimera can provide significant advantage over wild-type plants during strawberry regeneration without producing any pleiotropic effects seen for the miRNA resistant VvGRF4-GIF1 or TaGRF4-GIF1.
植物育种在将草莓从一种种植地域有限的小众作物转变为全球种植的经济重要作物的过程中发挥了非常重要的作用。但即使是现代的分子标记辅助育种，也需要经过多代植株，花费相当长的时间才能培育出具有理想性状的植株。作为一种更快的替代方法，可以通过组织培养进行精确的基因操作来培育具有理想性状的植株。之前已知在分生组织发育中起作用的形态发生调节因子、转录因子生长调节因子（GRFs）和GRF相互作用因子（GIFs）的过表达，为多种作物更轻松的再生和转化提供了一种有效策略。
与来自不同植物物种的不同GRF4 - GIF1嵌合体的异位表达相比，我们在此展示了在可控条件下二倍体草莓凤梨草莓夏威夷4号（草莓）再生和转化的综合方案。我们报告说，葡萄VvGRF4 - GIF1的异位表达在再次转化过程中提供了比野生型植株显著更高的再生效率。另一方面，VvGRF4 - GIF1或普通小麦（小麦）TaGRF4 - GIF1的miRNA抗性版本的失调表达导致了异常。不同嵌合GRF4 - GIF1株系之间的转录组分析表明，FvExpansin的差异表达可能是观察到的多效性效应的原因。同样，细胞分裂素脱氢酶/加氧酶和细胞分裂素响应调节因子也显示出差异表达，表明GRF4 - GIF1途径在控制细胞分裂素稳态中起重要作用。
我们的数据表明，葡萄VvGRF4 - GIF1嵌合体的异位表达在草莓再生过程中比野生型植株具有显著优势，且不会产生miRNA抗性VvGRF4 - GIF1或TaGRF4 - GIF1所见的任何多效性效应。
