Jung Yu-Jin, Go Jiyun, Kim Jin-Young, Lee Hyo-Ju, Kim Jong-Hee, Lee Hye-Mi, Cho Yong-Gu, Kang Kwon-Kyoo
Division of Horticultural Biotechnology, Hankyong National University, Anseong 17579, Republic of Korea.
Institute of Genetic Engineering, Hankyong National University, Anseong 17579, Republic of Korea.
Plants (Basel). 2025 Jan 23;14(3):345. doi: 10.3390/plants14030345.
We generated and characterized knockout mutant lines of the gene using the CRISPR/Cas9 system. The knockout lines of the gene showed that 1 bp and 7 bp deletion, early termination codons were used for protein production. Agronomic characteristics of knock-out lines were reduced in plant height, culm diameter, panicle length, seed size and weight, except for the number of tillers. In addition, we analyzed the expression levels of carotenoid biosynthesis genes by qRT-PCR. Among the genes encoding carotenoid metabolic pathway enzymes, the level of transcripts of , , , and were higher in the KO lines than in the WT line. In contrast, transcription of the ε, - and genes were downregulated in the KO lines compared to the WT line. Also, the KO lines decreased carotenoid content and ABA amount compared to WT, while preharvest sprouts increased. These results suggested that they would certainly help explain the molecular mechanisms of PHS in other crops, such as wheat and barley, which are susceptible to PHS.
我们使用CRISPR/Cas9系统构建并鉴定了该基因的敲除突变体系。该基因的敲除系显示有1个碱基对和7个碱基对的缺失，蛋白质合成使用了提前终止密码子。除了分蘖数外，敲除系的农艺性状在株高、茎粗、穗长、种子大小和重量方面均有所降低。此外，我们通过qRT-PCR分析了类胡萝卜素生物合成基因的表达水平。在编码类胡萝卜素代谢途径酶的基因中，敲除系中 、 、 、 和 的转录本水平高于野生型系。相反，与野生型系相比，敲除系中ε、 - 和 基因的转录下调。此外，与野生型相比，敲除系的类胡萝卜素含量和脱落酸含量降低，而收获前发芽增加。这些结果表明，它们肯定有助于解释小麦和大麦等其他易发生收获前发芽的作物中收获前发芽的分子机制。
