Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan.
Sci Rep. 2013;3:1532. doi: 10.1038/srep01532.
The liverwort Marchantia polymorpha is an emerging model organism on account of its ideal characteristics for molecular genetics in addition to occupying a crucial position in the evolution of land plants. Here we describe a method for gene targeting by applying a positive/negative selection system for reduction of non-homologous random integration to an efficient Agrobacterium-mediated transformation system using M. polymorpha sporelings. The targeting efficiency was evaluated by knocking out the NOP1 gene, which impaired air-chamber formation. Homologous recombination was observed in about 2% of the thalli that passed the positive/negative selection. With the advantage of utilizing the haploid gametophytic generation, this strategy should facilitate further molecular genetic analysis of M. polymorpha, in which many of the mechanisms found in land plants are conserved, yet in a less complex form.
地钱是一种新兴的模式生物，因为除了在陆生植物的进化中占据关键地位外，它还具有适合分子遗传学的理想特征。在这里，我们描述了一种通过应用正/负选择系统来减少非同源随机整合的方法，将其应用于利用地钱孢子体进行高效农杆菌介导的转化系统，从而实现基因靶向。通过敲除 NOP1 基因（该基因影响气室的形成）来评估靶向效率。在通过正/负选择的组织中，观察到同源重组约占 2%。利用单倍体配子体世代的优势，该策略应该有助于进一步对地钱进行分子遗传学分析，因为在陆生植物中发现的许多机制在这种植物中都得到了保守，但形式更为简单。
