CRISPR/Cas9-targeted mutagenesis of F3′H, DFR and LDOX, genes related to anthocyanin biosynthesis in black rice (Oryza sativa L.)

• Published in: Plant biotechnology reports Vol. 13; no. 5; pp. 521 - 531
• Main Authors: Jung, Yu Jin, Lee, Hyo Ju, Kim, Jong Hee, Kim, Dong Hyen, Kim, Hee Kyoung, Cho, Yong-Gu, Bae, Sangsu, Kang, Kwon Kyoo
• Format: Journal Article
• Language: English
• Published: Singapore Springer Singapore 01.10.2019; Springer Nature B.V; 한국식물생명공학회
• Subjects: Agriculture; Biomedical and Life Sciences; Biosynthesis; Biotechnology; Cell Biology; CRISPR; Crop improvement; Deoxyribonucleic acid; DNA; Genes; Genetically modified organisms; Genomes; Life Sciences; Mutagenesis; Mutation; Offspring; Original Article; Oryza sativa; Plant Biochemistry; Plant Sciences; Progeny; Rice; Site-directed mutagenesis; T-DNA; 농학; Anthocyanin; CRISPR/Cas9; Black rice; Gene editing
• ISSN: 1863-5466; 1863-5474
• DOI: 10.1007/s11816-019-00579-4

Altering a trait by CRISPR-Cas9-targeted mutagenesis offers great advantages in identifying gene function and crop improvement. In the present study, three genes ( OsF3′H, OsDFR and OsLDOX ) in the anthocyanin biosynthesis pathway were successfully edited on the Heugseonchal or Sinmyungheugchal variety using the CRISPR/Cas9 system. As a result, the ratio of the edited plants in the transformed early generation was 56.7%. These edited mutant lines were observed with the changes of seed color and anthocyanin content. All mutations were stably inherited to the T 2 progeny. In addition, we could select edited homozygous mutant lines lacking the T-DNA already in the first offspring generation. Also the insertion of vector backbone sequences in f3′h-9 , dfr-4 and ldox-16 lines was not detected in the whole genome resequencing. These results demonstrated that the CRISPR/Cas9 system can induce clearly gene-specific mutations with a high efficiency in rice and null plants selected from these mutants cannot be distinguished from non-GMO plants even under strict GMO regulation.