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Plant Biotechnology Reports

, Volume 13, Issue 5, pp 521–531 | Cite as

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

  • Yu Jin Jung
  • Hyo Ju Lee
  • Jong Hee Kim
  • Dong Hyen Kim
  • Hee Kyoung Kim
  • Yong-Gu Cho
  • Sangsu BaeEmail author
  • Kwon Kyoo KangEmail author
Original Article

Abstract

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 T2 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.

Keywords

Black rice Anthocyanin OsLDOX OsF3′H OsDFR CRISPR/Cas9 Gene editing 

Notes

Acknowledgements

This work was carried out with the support of “Cooperative Research Program for Agriculture Science & Technology Development (Project No. PJ01319302)” Rural Development Administration, Republic of Korea.

Supplementary material

11816_2019_579_MOESM1_ESM.docx (24 kb)
Supplementary file1 (DOCX 24 kb)
11816_2019_579_MOESM2_ESM.docx (206 kb)
Supplementary file2 (DOCX 205 kb)

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Copyright information

© Korean Society for Plant Biotechnology 2019

Authors and Affiliations

  • Yu Jin Jung
    • 1
    • 2
  • Hyo Ju Lee
    • 1
  • Jong Hee Kim
    • 1
  • Dong Hyen Kim
    • 1
  • Hee Kyoung Kim
    • 1
  • Yong-Gu Cho
    • 4
  • Sangsu Bae
    • 3
    Email author
  • Kwon Kyoo Kang
    • 1
    • 2
    Email author
  1. 1.Department of Horticultural Life ScienceHankyong National UniversityAnseongSouth Korea
  2. 2.Institute of Genetic EngineeringHankyong National UniversityAnseongSouth Korea
  3. 3.Department of ChemistryHanyang UniversitySeoulSouth Korea
  4. 4.Department of Crop ScienceChungbuk National UniversityCheongjuSouth Korea

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