Department of Horticultural Biotechnology, Kyung Hee University, Yongin-si 1732, Korea.
Int J Mol Sci. 2022 May 3;23(9):5062. doi: 10.3390/ijms23095062.
Chinese cabbage, a major crop in Korea, shows self-incompatibility (SI). SI is controlled by the type 2A serine/threonine protein phosphatases (PP2As). The gene is controlled by regulatory subunits that comprise a 36 kDa catalyst C subunit, a 65 kDa regulatory A subunit, and a variety of regulatory B subunits (50-70 kDa). Among them, the PP2A 55 kDa B regulatory subunit () gene located in the A05 chromosome has 13 exons spanning 2.9 kb, and two homologous genes, and , were found to be present on the A06 and A08 chromosome, respectively. In this study, we performed a functional analysis of the / gene using clustered regularly interspaced short palindromic repeats/CRISPR-associated system 9 (CRISPR/Cas9)-mediated gene mutagenesis. CRISPR/Cas9 technology can be used to easily introduce mutations in the target gene. Tentative gene-edited lines were generated by the -mediated transfer and were selected by PCR and Southern hybridization analysis. Furthermore, pods were confirmed to be formed in flower pollination (FP) as well as bud pollination (BP) in some gene-edited lines. Seed fertility of gene-edited lines indicated that the gene plays a key role in SI. Finally, self-compatible T-DNA-free T gene-edited plants and edited sequences of target genes were secured. The self-compatible Chinese cabbage developed in this study is expected to contribute to Chinese cabbage breeding.
白菜是韩国的主要作物，表现出自交不亲和性（SI）。SI 由 2A 型丝氨酸/苏氨酸蛋白磷酸酶（PP2As）控制。 基因受调节亚基控制，包括 36 kDa 的催化 C 亚基、65 kDa 的调节 A 亚基和多种调节 B 亚基（50-70 kDa）。其中，位于 A05 染色体上的 PP2A 55 kDa B 调节亚基（）基因包含 13 个外显子，跨度为 2.9 kb，发现两个同源基因 和 分别存在于 A06 和 A08 染色体上。在这项研究中，我们使用簇状规则间隔短回文重复序列/CRISPR 相关系统 9（CRISPR/Cas9）介导的基因诱变对 / 基因进行了功能分析。CRISPR/Cas9 技术可用于轻松在靶基因中引入突变。通过 -介导的转移产生了暂定的基因编辑系，并通过 PCR 和 Southern 杂交分析进行了选择。此外，在一些基因编辑系中，在花授粉（FP）和芽授粉（BP）中证实形成了豆荚。基因编辑系的种子育性表明 基因在 SI 中起关键作用。最后，获得了无 T-DNA 的自交亲和 T 基因编辑植物和目标基因的编辑序列。本研究开发的自交亲和白菜有望为白菜育种做出贡献。
