Improved soybean oil quality by targeted mutagenesis of the fatty acid desaturase 2 gene family.
Soybean oil is high in polyunsaturated fats and is often partially hydrogenated to increase its shelf life and improve oxidative stability. The trans-fatty acids produced through hydrogenation pose a health threat. Soybean lines that are low in polyunsaturated fats were generated by introducing mutations in two fatty acid desaturase 2 genes (FAD2-1A and FAD2-1B), which in the seed convert the monounsaturated fat, oleic acid, to the polyunsaturated fat, linoleic acid. Transcription activator-like effector nucleases (TALENs) were engineered to recognize and cleave conserved DNA sequences in both genes. In four of 19 transgenic soybean lines expressing the TALENs, mutations in FAD2-1A and FAD2-1B were observed in DNA extracted from leaf tissue; three of the four lines transmitted heritable FAD2-1 mutations to the next generation. The fatty acid profile of the seed was dramatically changed in plants homozygous for mutations in both FAD2-1A and FAD2-1B: oleic acid increased from 20% to 80% and linoleic acid decreased from 50% to under 4%. Further, mutant plants were identified that lacked the TALEN transgene and only carried the targeted mutations. The ability to create a valuable trait in a single generation through targeted modification of a gene family demonstrates the power of TALENs for genome engineering and crop improvement.
Cellectis plant sciences Inc., New Brighton, MN, USA.
William
W
Haun
Andrew
A
Coffman
Benjamin M
BM
Clasen
Zachary L
ZL
Demorest
Anita
A
Lowy
Erin
E
Ray
Adam
A
Retterath
Thomas
T
Stoddard
Alexandre
A
Juillerat
Frederic
F
Cedrone
Luc
L
Mathis
Daniel F
DF
Voytas
Feng
F
Zhang
D001483:Base Sequence; D044943:Fatty Acid Desaturases / Q000235:genetics*; D005227:Fatty Acids / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D009753:Nutritive Value / Q000235:genetics; D019301:Oleic Acid / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000737:chemistry / Q000378:metabolism; D012639:Seeds / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013024:Soybean Oil / Q000737:chemistry*; D013025:Glycine max / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism
D005227:Fatty Acids; D010940:Plant Proteins; D019301:Oleic Acid; D013024:Soybean Oil; D044943:Fatty Acid Desaturases
fatty acid desaturase; linoleic acid; metabolic engineering; oleic acid; targeted mutagenesis