玉米COBRA基因家族分析及重要成员ZmCOBL03基因编辑

Analysis of COBRA Gene Family and Gene Editing of ZmCOBL03 in Maize

利用生物信息学的方法分析玉米COBRA家族10个成员的序列特征、与其他物种的进化关系和组织特异性表达模式。结果表明,10个COBRA家族基因都含有CCVS保守结构域,并且均定位于细胞膜上。系统进化分析结果显示,该家族可以分为2个亚族,每个亚族内的基因具有相似的基因结构和理化性质。以ZmCOBL03为候选基因开展基因编辑,采用双靶标位点的设计思路,利用农杆菌介导的玉米遗传转化技术和CRISPR/Cas9基因编辑技术,成功获得ZmCOBL03基因靶向突变的玉米株系。表型分析证实,ZmCOBL03基因突变后严重影响了玉米的正常发育,表现为植株极端矮化且生殖生长受到抑制,证实ZmCOBL03基因在植物细胞壁纤维素合成中发挥重要作用。

The sequence characteristics,evolutionary relationships with other species and tissue-specific expression patterns of 10 COBRA family members were analyzed by bioinformatics method.The results showed that all the 10 COBRA family genes contained CCVS conserved domains and were localized in the cell membrane.The results of phylogenetic analysis showed that the family could be divided into two subfamilies,and the genes in each subfamily had similar gene structure and physical and chemical properties.Further research focused on the significant member ZmCOBL03 as a candidate gene for gene editing.In this study,using a dual target site design strategy,maize lines with targeted mutations of the ZmCOBL03 gene were successfully obtained through maize genetic transformation mediated by Agrobacterium tumefaciens and the CRISPR/Cas9 gene editing technology.Phenotypic analysis confirmed that the mutation of ZmCOBL03 gene seriously affected the normal development of maize,which showed dwarf plants without female and male panicle,and confirmed that ZmCOBL03 gene played an important role in plant cell wall cellulose synthesis.