Cell and Developmental Biology, John Innes Centre, Colney Lane, Norwich, NR4 7UH, UK.
University of Cambridge, Sainsbury Laboratory, Bateman Street, Cambridge, CB2 1LRUK.
J Exp Bot. 2024 Sep 27;75(18):5585-5591. doi: 10.1093/jxb/erae256.
Plant macroevolutionary studies leverage the phylogenetic position of non-flowering model systems like the liverwort Marchantia polymorpha to investigate the origin and evolution of key plant processes. To date, most molecular genetic studies in Marchantia rely on hygromycin and/or chlorsulfuron herbicide resistance markers for the selection of stable transformants. Here, we used a sulfonamide-resistant dihydropteroate synthase (DHPS) gene to enable sulfadiazine-based transformation selection in M. polymorpha. We demonstrate the reliability of sulfadiazine selection on its own and in combination with existing hygromycin and chlorsulfuron selection schemes through transgene stacking experiments. The utility of this system is further demonstrated through confocal microscopy of a triple transgenic line carrying fluorescent proteins labelling the plasma membrane, cortical microtubules, and the nucleus. Collectively, our findings and resources broaden the capacity to genetically manipulate the increasingly popular model liverwort M. polymorpha.
植物宏观进化研究利用非开花模式系统（如地钱属植物）的系统发育位置来研究关键植物过程的起源和进化。迄今为止，地钱属的大多数分子遗传学研究依赖于潮霉素和/或氯磺隆除草剂抗性标记物来选择稳定的转化体。在这里，我们使用磺胺抗性二氢喋呤合成酶（DHPS）基因来实现地钱属的磺胺嘧啶基转化选择。我们通过转基因堆叠实验证明了磺胺嘧啶单独选择以及与现有的潮霉素和氯磺隆选择方案相结合的可靠性。通过携带标记质膜、皮质微管和细胞核的荧光蛋白的三转基因系的共聚焦显微镜观察进一步证明了该系统的实用性。总的来说，我们的发现和资源拓宽了遗传操作日益流行的模式地钱属的能力。
