Leaf-chewing insects are important pests that cause yield loss and reduce seed quality in soybeans (Glycine max). Breeding soybean varieties that are resistant to leaf-chewing insects can minimize the need for insecticide use and reduce yield loss. The marker gene for QTL-M, Glyma.07g110300 (LOC100775351) that encodes a UDP-glycosyltransferase (UGT) is the major determinant of resistance against leaf-chewing insects in soybean; it exhibits a loss of function in insect-resistant soybean germplasms. In this study, Agrobacterium-mediated transformation introduced the CRISPR/Cas9 expression vector into the soybean cultivar Tianlong No. 1 to generate Glyma.07g110300-gene mutants. We obtained two novel types of mutations, a 33-bp deletion and a single-bp insertion in the GmUGT coding region, which resulted in an enhanced resistance to Helicoverpa armigera and Spodoptera litura. Additionally, overexpressing GmUGT produced soybean varieties that were more sensitive to H. armigera and S. litura. Both mutant and overexpressing lines exhibited no obvious phenotypic changes. The difference in metabolites and gene expression suggested that GmUGT is involved in imparting resistance to leaf-chewing insects by altering the flavonoid content and expression patterns of genes related to flavonoid biosynthesis and defense. Furthermore, ectopic expression of the GmUGT gene in the ugt72b1 mutant of Arabidopsis substantially rescued the phenotype of H. armigera resistance in the atugt72b1 mutant. Our study presents a strategy for increasing resistance against leaf-chewing insects in soybean through CRISPR/Cas9-mediated targeted mutagenesis of the UGT genes.

咀嚼叶昆虫是导致大豆产量损失和种子质量下降的重要害虫（大豆）。培育抗咬叶昆虫的大豆品种可以最大限度地减少杀虫剂的使用并减少产量损失。QTL-M的标记基因，格莱玛.07g110300(LOC100775351) 编码 UDP-糖基转移酶 (UGT)，是大豆抗咬叶昆虫的主要决定因素；它表现出抗虫大豆种质的功能丧失。在这项研究中，农杆菌-介导转化将CRISPR/Cas9表达载体导入大豆品种天龙1号，生成Glyma.07g110300-基因突变体。我们获得了两种新型突变，即 33 bp 缺失和单 bp 插入。葡萄糖耐量试验编码区，从而增强了抗性棉铃虫和斜纹夜蛾。此外，过度表达葡萄糖耐量试验生产的大豆品种对这些因素更加敏感H。棉铃虫和斜纹夜蛾。突变株系和过表达株系均未表现出明显的表型变化。代谢物和基因表达的差异表明葡萄糖耐量试验通过改变类黄酮含量以及与类黄酮生物合成和防御相关的基因的表达模式，参与赋予咀嚼叶昆虫抗性。此外，异位表达葡萄糖耐量试验基因在ugt72b1的突变体拟南芥基本上挽救了表型棉铃虫电阻在阿图格特72b1突变体。我们的研究提出了一种通过 CRISPR/Cas9 介导的定向诱变来增强大豆对咀嚼叶昆虫的抗性的策略。UGT基因。