期刊
PLANT BIOTECHNOLOGY JOURNAL
卷 15, 期 5, 页码 648-657出版社
WILEY
DOI: 10.1111/pbi.12663
关键词
Camelina sativa; gene editing; CRISPR/Cas9; allohexaploid; oleic acid; fatty acid composition
资金
- U.S. National Science Foundation [EPSCoR-1004094]
- U.S. Department of Energy, Office of Science, OBER [DOEBER SC0012459]
- NSF Plant Genome Program [1444612, 13-39385]
- University of Nebraska Foundation [01133480]
- Direct For Biological Sciences [1444612] Funding Source: National Science Foundation
- Direct For Biological Sciences
- Division Of Integrative Organismal Systems [1339385] Funding Source: National Science Foundation
- Division Of Integrative Organismal Systems [1444612] Funding Source: National Science Foundation
The CRISPR/Cas9 nuclease system is a powerful and flexible tool for genome editing, and novel applications of this system are being developed rapidly. Here, we used CRISPR/Cas9 to target the FAD2 gene in Arabidopsis thaliana and in the closely related emerging oil seed plant, Camelina sativa, with the goal of improving seed oil composition. We successfully obtained Camelina seeds in which oleic acid content was increased from 16% to over 50% of the fatty acid composition. These increases were associated with significant decreases in the less desirable polyunsaturated fatty acids, linoleic acid (i.e. a decrease from similar to 16% to <4%) and linolenic acid (a decrease from similar to 35% to <10%). These changes result in oils that are superior on multiple levels: they are healthier, more oxidatively stable and better suited for production of certain commercial chemicals, including biofuels. As expected, A. thaliana T-2 and T-3 generation seeds exhibiting these types of altered fatty acid profiles were homozygous for disrupted FAD2 alleles. In the allohexaploid, Camelina, guide RNAs were designed that simultaneously targeted all three homoeologous FAD2 genes. This strategy that significantly enhanced oil composition in T-3 and T-4 generation Camelina seeds was associated with a combination of germ-line mutations and somatic cell mutations in FAD2 genes in each of the three Camelina subgenomes.
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