Genome editing during apomictic reproduction in self-incompatible Hieracium produces high knockout frequencies in T1 progeny

Hieracium subgenus Pilosella species undergo facultative apomixis, which is asexual seed formation. We assessed the efficacy of CRISPR/Cas9 editing in this apomict by targeting the endogenous phytoene desaturase (PDS) gene, which induces a visible albino phenotype when mutated. High-throughput screening of Cas9-mediated editing events was carried out using amplicon deep sequencing of two PDS gene fragments from T0 and T1 independent transformants. Results from T0 and T1 lines are shown here.

山柳菊属（Hieracium）毛莲亚属（Pilosella）物种具有兼性无融合生殖特性，即通过无性途径形成种子。本研究以该无融合生殖物种为材料，靶向其内源八氢番茄红素脱氢酶（phytoene desaturase, PDS）基因，以评估CRISPR/Cas9系统的编辑效率——该基因发生突变后可产生肉眼可见的白化表型。我们通过对T0代与T1代独立转化体的两段PDS基因片段进行扩增子深度测序，完成了Cas9介导编辑事件的高通量筛选。本文展示了T0代及T1代株系的编辑结果。