Zhang Yongxing, Guo Wei, Chen Limiao, Shen Xinjie, Yang Hongli, Fang Yisheng, Ouyang Wenqi, Mai Sihua, Chen Haifeng, Chen Shuilian, Hao Qingnan, Yuan Songli, Zhang Chanjuan, Huang Yi, Shan Zhihui, Yang Zhonglu, Qiu Dezhen, Zhou Xinan, Cao Dong, Li Xia, Jiao Yongqing
Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture and Rural Affairs, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan, China.
National Key Laboratory of Crop Genetic Improvement, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, China.
Front Plant Sci. 2022 Feb 22;13:802716. doi: 10.3389/fpls.2022.802716. eCollection 2022.
Leaf-chewing insects are important pests that cause yield loss and reduce seed quality in soybeans (). 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, (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, -mediated transformation introduced the CRISPR/Cas9 expression vector into the soybean cultivar Tianlong No. 1 to generate gene mutants. We obtained two novel types of mutations, a 33-bp deletion and a single-bp insertion in the coding region, which resulted in an enhanced resistance to and . Additionally, overexpressing produced soybean varieties that were more sensitive to . and . Both mutant and overexpressing lines exhibited no obvious phenotypic changes. The difference in metabolites and gene expression suggested that 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 gene in the mutant of substantially rescued the phenotype of resistance in the mutant. Our study presents a strategy for increasing resistance against leaf-chewing insects in soybean through CRISPR/Cas9-mediated targeted mutagenesis of the genes.
咀嚼式口器昆虫是导致大豆产量损失和种子质量下降的重要害虫。培育抗咀嚼式口器昆虫的大豆品种可以减少杀虫剂的使用需求并降低产量损失。QTL-M的标记基因（LOC100775351）编码一种UDP-糖基转移酶（UGT），是大豆对咀嚼式口器昆虫抗性的主要决定因素；它在抗虫大豆种质中表现出功能丧失。在本研究中，通过农杆菌介导的转化将CRISPR/Cas9表达载体导入大豆品种天龙一号以产生基因突变体。我们在编码区获得了两种新型突变，一个33bp的缺失和一个单碱基插入，这导致对[具体昆虫1]和[具体昆虫2]的抗性增强。此外，过表达[基因名称]产生了对[具体昆虫1]和[具体昆虫2]更敏感的大豆品种。突变体和过表达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，[基因名称]通过改变类黄酮含量以及与类黄酮生物合成和防御相关基因的表达模式参与赋予对咀嚼式口器昆虫的抗性。此外，[基因名称]基因在[相关植物名称]的[突变体名称]中的异位表达显著挽救了[突变体名称]中对[具体昆虫1]的抗性表型。我们的研究提出了一种通过CRISPR/Cas9介导的对[基因名称]基因进行靶向诱变来提高大豆对咀嚼式口器昆虫抗性的策略。
