Agrobacterium tumefaciens-mediated transformation is widely used for the genetic transformation of Lotus corniculatus. In this study, we optimized the genetic transformation system of L. corniculatus to improve the transformation efficiency. Firstly, besides the commonly used cotyledon, we successfully transformed and regenerated the hypocotyl as an additional explant. Secondly, we used DsRed2 instead of kanamycin as a screening marker to screen transgenic callus and shoots, and conducted PCR analysis to verify the selected plants based on the presence of red fluorescence conferred by DsRed2. The results confirmed the reliability and convenience of DsRed2 as a suitable reporter for the transgenic selection of L. corniculatus. Furthermore, we addressed the limitations of commonly used agar-based rooting media by introducing vermiculite as an alternative to agar. The results revealed that the vermiculite-based rooting medium effectively induced root initiation, and facilitated the growth of both roots and shoots in transformed seedlings. Thus, our vermiculite-based rooting medium could serve as an ideal substitute for traditional agar-based rooting medium. To sum up, we have developed a rapid, convenient, and efficient transformation system of L. corniculatus using both cotyledons and hypocotyls as explants, employing DsRed2 as a screening marker, and implementing a vermiculite-based MS medium as rooting medium.

根癌农杆菌介导的转化广泛用于百脉根的遗传转化。本研究对百脉根遗传转化体系进行了优化，以提高转化效率。首先，除了常用的子叶外，我们成功地转化并再生了下胚轴作为额外的外植体。其次，我们使用DsRed2代替卡那霉素作为筛选标记来筛选转基因愈伤组织和芽，并根据DsRed2赋予的红色荧光的存在进行PCR分析以验证所选植物。结果证实了DsRed2作为百脉根转基因选择合适报告基因的可靠性和便利性。此外，我们通过引入蛭石作为琼脂的替代品来解决常用的基于琼脂的生根介质的局限性。结果表明，蛭石生根培养基能有效诱导根的萌发，并促进转化幼苗根和芽的生长。因此，我们的蛭石生根培养基可以作为传统琼脂生根培养基的理想替代品。综上所述，我们以子叶和下胚轴为外植体，以DsRed2为筛选标记，以蛭石为基础的MS培养基作为生根培养基，开发了一种快速、方便、高效的百脉根转化体系。