Simultaneous silencing of FAD2 and FAE1 genes affects both oleic acid and erucic acid contents in Brassica napus seeds.
The fatty acid composition in the seed oil was significantly modified following the introduction of transgenes. To further enhance the desirable characteristics of rapeseed oil, it would be beneficial to develop a new approach for the simultaneous silencing of two or more target genes. Our goals in the current study were to (1) increase oleic acid to more than 75%, (2) reduce polyunsaturated fatty acids (PUFA) to about 10% and erucic acid to zero, and (3) accomplish these changes in a single-transformation event. In a single transformation, two fragments amplified from the fatty acid (Delta12)-desaturase 2 (BnaFAD2) and fatty acid elongase 1 (BnaFAE1) genes of Brassica napus were linked together to form a fusion fragment. The fusion fragment was then used to assemble unique intron-spliced hairpin interfering constructs. In the transgenic plant FFRP4-4, the expression of BnaFAD2 and BnaFAE1 genes was completely inhibited. The composition of oleic acid in FFRP4-4 rose to 85%, PUFA dropped to 10% and erucic acid was undetectable. All hybrid F(1) seeds obtained from the reciprocal crossing of FFRP4-4 and GX-parents (with different genetic backgrounds) contained more than 80% oleic acid, about 10% PUFA and very low, or undetectable, erucic acid. The results confirmed that the fusion fragment silencing construct can simultaneously and effectively silence the target genes on a consistent basis. The strategy provides a useful tool for detecting gene function and advancing genetic engineering techniques for the improvement of agricultural crops.
Peng
Qi
Q
Hunan Provincial Key Laboratory of Crop Germplasm Innovation and Utilization, Hunan Provincial Institute of Oil Crops, Hunan Agricultural University, Changsha 410128, China.
Hu
Yan
Y
Wei
Ran
R
Zhang
Yuan
Y
Guan
Chunyun
C
Ruan
Ying
Y
Liu
Chunlin
C
D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism*; D029688:Brassica napus / Q000201:enzymology / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D004883:Erucic Acids / Q000378:metabolism*; D044943:Fatty Acid Desaturases / Q000235:genetics / Q000378:metabolism*; D000081031:Fatty Acid Elongases; D005231:Fatty Acids, Unsaturated / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D016254:Mutagenesis, Insertional; D019301:Oleic Acid / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics; D012639:Seeds / Q000201:enzymology / Q000235:genetics; D014170:Transformation, Genetic
D004269:DNA, Bacterial; D004883:Erucic Acids; D005231:Fatty Acids, Unsaturated; D010940:Plant Proteins; D018749:RNA, Plant; C036483:T-DNA; C049811:erucic acid; D019301:Oleic Acid; D044943:Fatty Acid Desaturases; D000123:Acetyltransferases; D000081031:Fatty Acid Elongases