Function Analysis of the PR55 / B Gene Related to Self-Incompatibility in Chinese Cabbage Using CRISPR/Cas9

• Published in: International journal of molecular sciences Vol. 23; no. 9; p. 5062
• Main Authors: Shin, Na-Ri, Shin, Yun-Hee, Kim, Han-Seul, Park, Young-Doo
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 03-05-2022; MDPI
• Subjects: Brassica - genetics; Brassica oleracea; Brassica rapa; Catalysts; China; Chinese cabbage; Chromosomes; CRISPR; CRISPR-Cas Systems; CRISPR/Cas9; Exons; Fertility; Function analysis; Functional analysis; Gene Editing; Gene expression; Genes; Genetic modification; Genomes; Genomics; Genotype & phenotype; Homology; Hybridization analysis; Incompatibility; Kinases; Mutagenesis; Mutation; Nucleotide sequence; Phosphorylation; Plant Breeding; Plant reproduction; Pollination; PR55/B gene; Proteins; Regulatory subunits; Seeds; Self-incompatibility; Signal transduction; T-DNA; China; self-incompatibility; Brassica rapa; CRISPR/Cas9; PR55/B gene
• ISSN: 1422-0067; 1661-6596; 1422-0067
• 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.