Open AccessSignificant enhancement of fatty acid composition in seeds of the allohexaploid, Camelina sativa , using CRISPR /Cas9 gene editing
Author(s) -
Jiang Wen Zhi,
Henry Isabelle M.,
Lynagh Peter G.,
Comai Luca,
Cahoon Edgar B.,
Weeks Donald P.
Publication year - 2017
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.12663
Subject(s) - camelina , camelina sativa , biology , linoleic acid , food science , genome editing , crispr , cas9 , gene , polyunsaturated fatty acid , fatty acid , linolenic acid , erucic acid , biochemistry , botany , genetics , agronomy , crop
Summary 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 FAD 2 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 ~16% to <4%) and linolenic acid (a decrease from ~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 FAD 2 alleles. In the allohexaploid, Camelina, guide RNA s were designed that simultaneously targeted all three homoeologous FAD 2 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 FAD 2 genes in each of the three Camelina subgenomes.