利用CRISPR/Cas9技术对蒙古冰草落粒相关基因Sh1的编辑

Editing of Fallen Grain Related Genes Sh1 of Agropyron mongolicum Keng Using CRISPR/Cas9 Technology

落粒性是禾本科植物在进化过程中为了抵御恶劣环境条件、繁衍后代,通过长期的生存竞争,逐渐形成的一种适应机制,是禾本科植物系统演化过程中的里程碑事件。本研究通过CRISPR/Cas9技术对蒙古冰草中的Sh1基因进行编辑,从而终止其对蒙古冰草的落粒性的调控。所构建的载体在原生质体水平靶向敲除蒙古冰草落粒相关基因Sh1,通过载体构建、验证、原生质体转化等方法,结果表明,3个靶位点的突变类型主要为点突变和小片段缺失(1 bp或5 bp缺失),说明CRISPR/Cas9系统在蒙古冰草原生质体水平发挥了作用,为后续进行蒙古冰草基因编缉提供实验依据。

Plant shattering is an adaptive mechanism gradually evolved from long-term survival competition in order to resist harsh environmental conditions and breed offspring during the evolution process of Gramineae. In this study, the Sh1 gene in Agropyron mongolicum Keng was edited by CRISPR/Cas9 technology to terminate its regulation of the shattering of Agropyron mongolicum Keng. The constructed vector was targeted at the level of protoplasts to knock out the RNA-related gene Sh1, vector construction, validation, and protoplast transformation. The results showed that there were mutations in all three targeted sites include mainly to point mutation (C-T replacement) and small deletion (1 or 5 bp deletion), the CRISPR/Cas9 system can be genome editing in Agropyron mongolicum Keng, can provide theoretical research for the subsequent genetic compilation of Agropyron mongolicum Keng.