Direct stacking of sequence-specific nuclease-induced mutations to produce high oleic and low linolenic soybean oil

BMC Plant Biology

Research Article

Benjamin M. Clasen¹  Maria Elena Gamo¹  Thomas J. Stoddard¹  Feng Zhang¹  Ann Yabandith¹  Andrew Coffman¹  Zachary L. Demorest¹  Jeff Bissen¹  Luc Mathis¹  Adam Retterath¹  Daniel F. Voytas¹  Nicholas J. Baltes¹  Song Luo¹ 

[1] Calyxt, Inc., 600 County Road D West Suite 8, 55112, New Brighton, MN, USA;

关键词: TALEN;    Genome editing;    Soybean;    High oleic acid;    Low linolenic acid;    Soybean oil;    Targeted mutagenesis;   

DOI  :  10.1186/s12870-016-0906-1

 received in 2016-04-29, accepted in 2016-09-26,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundThe ability to modulate levels of individual fatty acids within soybean oil has potential to increase shelf-life and frying stability and to improve nutritional characteristics. Commodity soybean oil contains high levels of polyunsaturated linoleic and linolenic acid, which contribute to oxidative instability – a problem that has been addressed through partial hydrogenation. However, partial hydrogenation increases levels of trans-fatty acids, which have been associated with cardiovascular disease. Previously, we generated soybean lines with knockout mutations within fatty acid desaturase 2-1A (FAD2-1A) and FAD2-1B genes, resulting in oil with increased levels of monounsaturated oleic acid (18:1) and decreased levels of linoleic (18:2) and linolenic acid (18:3). Here, we stack mutations within FAD2-1A and FAD2-1B with mutations in fatty acid desaturase 3A (FAD3A) to further decrease levels of linolenic acid. Mutations were introduced into FAD3A by directly delivering TALENs into fad2-1a fad2-1b soybean plants.ResultsOil from fad2-1a fad2-1b fad3a plants had significantly lower levels of linolenic acid (2.5 %), as compared to fad2-1a fad2-1b plants (4.7 %). Furthermore, oil had significantly lower levels of linoleic acid (2.7 % compared to 5.1 %) and significantly higher levels of oleic acid (82.2 % compared to 77.5 %). Transgene-free fad2-1a fad2-1b fad3a soybean lines were identified.ConclusionsThe methods presented here provide an efficient means for using sequence-specific nucleases to stack quality traits in soybean. The resulting product comprised oleic acid levels above 80 % and linoleic and linolenic acid levels below 3 %.

【 授权许可】

CC BY   
© The Author(s). 2016