Zang Yiming, Gong Qiang, Xu Yanhao, Liu Huiyun, Bai Hao, Li Na, Du Lipu, Ye Xingguo, Lan Caixia, Wang Ke
College of Plant Science & Technology, Huazhong Agricultural University, Wuhan, China.
Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, China.
Front Genet. 2022 May 5;13:873850. doi: 10.3389/fgene.2022.873850. eCollection 2022.
The () gene controls the caryopsis type of cereal crops by regulating lipid biosynthetic pathways. Based on the sequence and its homologous gene sequences in wheat, a conserved sgRNA was designed to obtain the mutants from the barley variety "Vlamingh" and the wheat variety "Fielder" -mediated transformation. A total of 19 and 118 transgenic plants were obtained, and 11 and 61 mutant plants were identified in T transgenic plants in barley and wheat after PCR-RE detection, and the editing efficiencies of the targeted gene were 57.9 and 51.7% in barley and wheat, respectively. The grain shape of the barley mutants was naked. Five different combinations of mutations for wheat genes were identified in the T generation, and their homozygous-edited plants were obtained in the T generation. Interestingly, the conjoined plants in which one plant has different genotypes were first identified. The different tillers in an individual T plant showed independent transgenic or mutant events in both barley and wheat, and the different genotypes can stably inherit into T generation, indicating that the T transgenic plants were the conjoined type. In addition, we did not find any off-target mutations in both barley and wheat. A candidate method for detecting putative-edited wheat plants was suggested to avoid losing mutations in this investigation. This study provides not only materials for studying the function of the gene in barley and wheat but also a system for detecting the mutants in wheat.
（）基因通过调控脂质生物合成途径来控制谷类作物的颖果类型。基于该基因序列及其在小麦中的同源基因序列，设计了一个保守的sgRNA，通过大麦品种“Vlamingh”和小麦品种“Fielder”介导的转化来获得突变体。共获得19株和118株转基因植株，经PCR-RE检测后，在大麦和小麦的T代转基因植株中分别鉴定出11株和61株突变体植株，目标基因在大麦和小麦中的编辑效率分别为57.9%和51.7%。大麦突变体的籽粒形状为裸粒。在T代中鉴定出小麦基因的五种不同突变组合，并在T代中获得了它们的纯合编辑植株。有趣的是，首次鉴定出一株植物具有不同基因型的连体植株。在大麦和小麦中，单个T植株的不同分蘖均表现出独立的转基因或突变事件，且不同基因型可稳定遗传至T代，表明T代转基因植株为连体类型。此外，在大麦和小麦中均未发现任何脱靶突变。本研究提出了一种检测推定编辑小麦植株的候选方法，以避免在本研究中丢失突变。该研究不仅为研究该基因在大麦和小麦中的功能提供了材料，也为检测小麦中的突变体提供了一个系统。
