Efficient breeding of high oleic rice cultivar by editing OsFAD2-1 via CRISPR/Cas9

Yingying Wu, Zhihui Chen, Chan Wang, Yang Xu, Xia Li, Jianping Zhu, Xiaoli Tan, Jie Yang

Journal of Integrative Agriculture; 25 April 2025;

Highlight:

• - CRISPR-editing OsFAD2-1 boosts oleic acid and stabilizes rice bran oil.
• - Edited lines maintain key agronomic traits of breeding-ready germplasm.
• - Gene editing accelerates high-oleic breeding in Jiangsu japonica rice

In summary, this study demonstrates the enormous potential of gene editing for crop qualityimprovement and validates the crucial function of the OsFAD2 gene in controlling fatty acid metabolisminrice. Future studies could examine how OsFAD2 interacts with other genes linked to fatty acid metabolismand how gene editing techniques could be used to further raise oleic acid levels. Combining gene editingtechnology with contemporary molecular breeding techniques can be successfully used to create premiumricevarieties with exceptional nutritional value and wide adaptability, which will improve the nutrition valueofrice grains and contribute to global food security. The OsFAD2-1 gene was edited using CRISPR/Cas9 in this study, significantly increasing the amount ofoleic acid and decreasing linoleic acid in rice grains. The potential of gene editing to improve rice qualitywas demonstrated by the edited lines’ increased nutritional value and stable agronomic traits. Future studiesshould attempt to combine this technique with advanced breeding practices aiming for sustainable agriculture.

See https://www.sciencedirect.com/science/article/pii/S2095311925001224

Figure 1

Knockout of OsFAD2 by CRISPR/Cas9 method produces rice grains with high oleic content. A, OSFAD2-1 converts oleic acid (18:1) into linoleic acid (18:2). B, gene structure of OsFAD2. The blackbox represents the coding regions, and the red box indicates the mutation site. C, DNA sequences flankingthe target site of sgRNA in Suken118 (SK118) and its two mutants (SK118-1 and SK118-2). Insertedsequences are shown in red letter(s), and blue nucleotides indicate PAM (protospacer adjacent motif). Thenumber of inserted/substituted (‘+’) and deleted (‘-’) nucleotides are shown on the right of the sequence. Onthe far right is a peak plot of DNA sequencing flanking the target site. D, fatty acid compositionintheSK118 and mutant lines. Data are presented as mean±SD (n=3). ns, not significant; ** indicates significant difference at P≤0.01 (Student’s t-test). E, morphology of SK118, SK118-1, and SK118-2 plants at themature stage. Scale bar=5 cm. F, grain appearance of wild-type and mutant lines, showing both dehulledand brown grains. Scale bar=1 cm. G, comparison of rapid viscosity analyzer spectra of rice floursbetween SK118 and mutant lines. H–N, agronomic traits of wild-type and mutant plants, includingpaniclenumber per plant (H), grain number per panicle (I), seed-setting rate (J), plant height (K), 1,000-grainweight (M), and panicle length (N). Data are presented as mean±SD (n=5). ns, not significant; ** indicatessignificant difference at P≤0.01 (Student’s t-test).