大豆BBX32基因生物信息学分析及基因编辑靶点设计

Bioinformatics Analysis and Gene Editing Target Design of BBX32 Gene in Soybean

BBX32基因是一种含锌指结构的转录因子,广泛的参与植物生长和发育过程,但在大豆中,BBX32基因的具体生物学功能还尚未被解析。为研究大豆BBX32基因在大豆中的分子机理和生物学功能,本研究通过同源比对分析发现,大豆中含有两个与拟南芥BBX32同源的基因,分别命名为GmBBX32a和GmBBX32b。同时,通过检索RNA-seq数据库,对GmBBX32a和GmBBX32b在大豆中不同组织的表达部位进行分析,结果发现GmBBX32a在花、叶和豆荚中具有较高的表达量,而GmBBX32b在豆荚和花中具有较高的表达量,说明GmBBX32a和GmBBX32b可能参与大豆的开花和籽粒大小形成。此外,利用CRISPR/Cas9技术,构建GmBBX32a和GmBBX32b基因的敲除载体,并转化到大豆毛根中进行靶点检测,最终获得了有编辑效率的基因编辑载体,为接下来获得稳定转基因材料提供有效编辑靶点信息。

BBX32 genes belong zinc finger transcription factor, which are widely involved in plant growth and development.However, the biological function of BBX32 genes had not been resolved in soybean.To investigate the molecular mechanism of BBX32 genes in soybean, homology alignment analysis was performed.The result showed that the two homologous of Arabidopsis BBX32 were found in soybean, and named GmBBX32 a and GmBBX32 b.The expression levels GmBBX32 a and GmBBX32 b in different tissues of soybean were analyzed by using the RNA-seq database.The result showed that the expression level of GmBBX32 a was higher in flower, leaf and pod, while the expression level of GmBBX32 b was higher in pod and flower, indicating that GmBBX32 a and GmBBX32 b might be involved in flowering and seed size formation in soybean.In addition, the knockout vectors of GmBBX32 a and GmBBX32 b were constructed and transformed into soybean hair roots for target detection by using CRISPR/Cas9 technology, which should be provide efficient editing target information for the next generation of stable transgenic materials.Our work provides a theoretical basis for in-depth analysis of the complex molecular mechanisms in regulating soybean yield traits.