| Oleic acid, because of its special biological characteristicsand great economic value, has become the hot spot of rape breeding. Currently, the low erucic and glucosinlate oil seed rape has been widely planted. The elongation route of oleic acid for double-zero rapeseed has been cut off gradualy during the process of conventional breeding, the linoleic acid and linolenic acid, which are the products of the elongation of oliec acid, however, still have reletively high proportion of the fatty acid content in rapeseed. So these non-oleic fatty acids should be reduced for enriching the content of oleic acid in the new high-oleic germplasm of rapeseed.Three conservative sequences.one was347bp from Fad2gene, one394bp from Fad3gene and the other359bp from Fatb gene, were cloned. Recombinant PCR method was used to vitro fusion the conserved sequence of the three genes to of an1100bp interference DNA fragment. The fragment was with pros and cons inserted to the interference carrier of PFGC5941, which was guided by seed-specific RNA Napin, then transformed to Brassica Rape9. The three genes were expressed by silencing suppressor to cut off its biosynthetic pathway. A total of757resistant seedlings were selected by16mg/L PPT selection pressure, all of them were measured by PCR analysis, and64of them were transgenic plants. The transgenic oilseed rape has poor growth power, and most of them died with the growth period, and accompanied by the flowerless and sterile phenomenon, only five transgenic TO rape seed were obtained. The Fad2. Fad3. Fatb gene expression analysis and fatty acid composition of the seed were determined by the method of quantitative PCR and meteorological chromatography. The results showed that the gene expression inhibition rate of the Fad2, Fad3, Fatb were99%or more, and the oleic acid content increased21.77%compared with the control. This study is interesting and will be helpful to the study of fatty acid synthesis and metabolism mechanism. |
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