Open AccessSelective gene dosage by CRISPR ‐Cas9 genome editing in hexaploid Camelina sativa
Author(s) -
Morineau Céline,
Bellec Yannick,
Tellier Frédérique,
Gissot Lionel,
Kelemen Zsolt,
Nogué Fabien,
Faure JeanDenis
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.12671
Subject(s) - biology , camelina , camelina sativa , polyploid , gene , genetics , genome editing , crispr , mutagenesis , genome , oleic acid , allele , mutant , cas9 , microbiology and biotechnology , botany , crop , ecology
In many plant species, gene dosage is an important cause of phenotype variation. Engineering gene dosage, particularly in polyploid genomes, would provide an efficient tool for plant breeding. The hexaploid oilseed crop Camelina sativa, which has three closely related expressed subgenomes, is an ideal species for investigation of the possibility of creating a large collection of combinatorial mutants. Selective, targeted mutagenesis of the three delta‐12‐desaturase ( FAD 2 ) genes was achieved by CRISPR ‐Cas9 gene editing, leading to reduced levels of polyunsaturated fatty acids and increased accumulation of oleic acid in the oil. Analysis of mutations over four generations demonstrated the presence of a large variety of heritable mutations in the three isologous Cs FAD 2 genes. The different combinations of single, double and triple mutants in the T3 generation were isolated, and the complete loss‐of‐function mutants revealed the importance of delta‐12‐desaturation for Camelina development. Combinatorial association of different alleles for the three FAD 2 loci provided a large diversity of Camelina lines with various lipid profiles, ranging from 10% to 62% oleic acid accumulation in the oil. The different allelic combinations allowed an unbiased analysis of gene dosage and function in this hexaploid species, but also provided a unique source of genetic variability for plant breeding.