RESULTS: CrPINMa possessed a long central hydrophilic loop and characteristic motifs within it, which indicated that it belonged to the canonical rather than the non-canonical PINs. CrPINMa was positioned in the lineage leading to Arabidopsis PIN6 but not that to its PIN1, and it had undergone numerous gene duplications. CRISPR/Cas9 genome editing had been performed in ferns for the first time, producing diverse mutations including local frameshifts for CrPINMa. Plants possessing disrupted CrPINMa exhibited retarded leaf emergence and reduced leaf size though they could survive and reproduce at the same time. CrPINMa transcripts were distributed in the shoot apical meristem, leaf primordia and their vasculature. Finally, CrPINMa proteins were localized to the plasma membrane rather than other cell parts.
CONCLUSIONS: CRISPR/Cas9 genome editing is feasible in ferns, and that PINs can play a role in fern leaf development.
结果:CrPINMa具有长的中央亲水环和其中的特征图案,这表明它属于规范的而不是非规范的PIN。CrPINMa位于导致拟南芥PIN6的谱系中,而不是PIN1,并且经历了许多基因重复。CRISPR/Cas9基因组编辑首次在蕨类植物中进行,产生不同的突变,包括CrPINMa的局部移码。具有破碎的CrPINMa的植物尽管可以同时存活和繁殖,但却表现出叶片出苗迟缓和叶片大小减小。CrPINMa转录本分布在茎尖分生组织中,叶原基和它们的脉管系统。最后,CrPINMa蛋白定位于质膜而不是其他细胞部分。
结论:CRISPR/Cas9基因组编辑在蕨类植物中是可行的,并且PIN可以在蕨类植物叶片的发育中发挥作用。