利用CRISPR/Cas9基因编辑技术构建番茄SlGATA1突变体

GATA 作为一类广泛存在于真核生物中的转录因子，在植物各生长发育阶段和逆境胁迫响应过程中发挥着至关重要的作用。为探究转录因子 SlGATA1 在番茄植株生长发育和逆境胁迫响应中的具体生物学功能，文章通过实时荧光定量聚合酶链式反应(polymerase chain reaction, PCR)对 SlGATA1 基因表达模式进行分析，并利用 CRISPR/Cas9 基因编辑技术对 SlGATA1 基因进行编辑。结果表明：SlGATA1 基因在各组织中均有表达，在种子中的表达量最高；SlGATA1 基因的表达受高盐、干旱、高温及病原菌丁香假单胞菌(Pseudomonas syringae pv. tomato DC3000, Pst DC3000)的影响；通过根癌农杆菌介导的遗传转化体系和CRISPR/Cas9 基因编辑技术获得 4 株稳定遗传的 SlGATA1 基因敲除突变体，为番茄品种改良提供新的遗传材料。该研究为探究 SlGATA1 基因的功能奠定了基础。

GATA transcription factors are widely present in eukaryotes that play a crucial role in various biological processes, including growth, development, and environmental stress response. To investigate the specific biological functions of the SlGATA1 in tomatoes, quantitative real-time polymerase chain reaction (qRT-PCR) and CRISPR/Cas9 editing technology were applied to analyzing the expression pattern of the SlGATA1 and constructing the SlGATA1 knocked-out transgenic lines. The results revealed that the SlGATA1 was expressed in all tissues, with the highest expression level observed in seeds. Moreover, the expression of the gene was affected by salt stress, drought stress, heat stress, and the pathogen Pseudomonas syringae pv. tomato DC3000(Pst DC3000) infection. Two dual-target CRISPR/Cas9 knockdown vectors were successfully constructed, and four stable genetic knockout plants of the SlGATA1 were obtained by using Agrobacterium-mediated transformation system. The findings of this study provide valuable insights into the function of the SlGATA1 and offer new genetic materials for the improvement of tomato varieties.