Plant Cell Reports ( IF 5.3 ) Pub Date : 2023-01-25 , DOI: 10.1007/s00299-023-02980-4 Tawni Bull 1, 2, 3 , Juan Debernardi 4 , Megan Reeves 2 , Theresa Hill 5 , Lien Bertier 2, 6 , Allen Van Deynze 5, 7 , Richard Michelmore 2, 7
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Key message
GRF–GIF chimeric proteins from multiple source species enhance in vitro regeneration in both wild and cultivated lettuce. In addition, they enhance regeneration in multiple types of lettuce including butterheads, romaines, and crispheads.
Abstract
The ability of plants to regenerate in vitro has been exploited for use in tissue culture systems for plant propagation, plant transformation, and genome editing. The success of in vitro regeneration is often genotype dependent and continues to be a bottleneck for Agrobacterium-mediated transformation and its deployment for improvement of some crop species. Manipulation of transcription factors that play key roles in plant development such as BABY BOOM, WUSCHEL, and GROWTH-REGULATING FACTORs (GRFs) has improved regeneration and transformation efficiencies in several plant species. Here, we compare the efficacy of GRF–GIF gene fusions from multiple species to boost regeneration efficiency and shooting frequency in four genotypes of wild and cultivated lettuce (Lactuca spp. L.). In addition, we show that GRF–GIFs with mutated miRNA 396 binding sites increase regeneration efficiency and shooting frequency when compared to controls. We also present a co-transformation strategy for increased transformation efficiency and recovery of transgenic plants harboring a gene of interest. This strategy will enhance the recovery of transgenic plants of other lettuce genotypes and likely other crops in the Compositae family.
中文翻译：
GRF-GIF 嵌合蛋白增强生菜（Lactuca spp.）的体外再生和农杆菌介导的转化效率
关键信息
来自多个来源物种的 GRF-GIF 嵌合蛋白增强了野生和栽培生菜的体外再生。此外，它们还能增强多种生菜的再生能力，包括黄油生菜、生菜和脆头生菜。
抽象的
植物体外再生的能力已被用于植物繁殖、植物转化和基因组编辑的组织培养系统。体外再生的成功通常取决于基因型，并且仍然是农杆菌介导的转化及其用于改良某些作物物种的部署的瓶颈。对在植物发育中起关键作用的转录因子（如 BABY BOOM、WUSCHEL 和生长调节因子 (GRF)）的操纵，提高了多种植物物种的再生和转化效率。在这里，我们比较了来自多个物种的GRF-GIF基因融合在提高四种基因型野生和栽培生菜（ Lactuca spp. L.）的再生效率和出芽频率方面的功效。此外，我们还发现，与对照相比，具有突变 miRNA 396 结合位点的 GRF-GIF 提高了再生效率和射击频率。我们还提出了一种共转化策略，以提高转化效率和恢复含有目的基因的转基因植物。该策略将增强其他生菜基因型以及可能的其他菊科作物的转基因植物的恢复。
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