Method for obtaining cole with high oleic acid and low erucic acid by dual-approach gene expression synchronous suppression

Abstract

The invention belongs to the field of biotechnology and discloses a method which utilizes the improved RNA interference technology and the genetic engineering measure to carry out interference silence simultaneously to the gene expression of two metabolic pathways (1) and (2) in which the oleic acid is converted into other fatty acid, prevents the oleic acid from being converted into the fatty acids of other types to the most extent and achieves the object of greatly improving the content of the oleic acid and greatly reducing the content of the erucic acid. The embodiment connects the interference fragment of the FAD2 gene on the desaturation pathway (1) and the interference fragment of the FAE1 gene on the chain extension pathway (2) into a syncretized interference fragment and establishes the corresponding interference carrier according to the syncretized interference fragment. By the mediation of agrobacterium LAB4404, the brassica napus is converted, thus obtaining the brassica napus strains which can inherit stably, wherein the oleic acid content of the brassica napus strain is more than 80% and the erucic acid content thereof can not be detected.

Description

technical field [0001] The invention relates to using RNA interference technology to simultaneously inhibit the gene expression of two pathways of oleic acid chain elongation and desaturation, prevent oleic acid from being further metabolized into other types of fatty acids through these two pathways, and further make the fatty acid components in rapeseed The method for greatly increasing the content of oleic acid and decreasing the content of erucic acid belongs to the field of biotechnology. Background technique [0002] Oleic acid is a monounsaturated fatty acid (18:1 Δ9 ), high stability, such as not easy to smoke at high temperature; can reduce total cholesterol and blood lipids in the human body, reduce harmful low-density lipoprotein cholesterol (LDC), and maintain beneficial high-density lipoprotein cholesterol (HDL) levels at the same time. While linolenic acid reduces harmful low-density lipoprotein cholesterol, it also reduces the level of beneficial high-density...

AI Technical Summary

Problems solved by technology

The fundamental reason for this is that there are two metabolic pathways for the conversion of oleic acid into other fatty acids: the desaturation pathway ① and the chain elongation pathway ② ( figure 1 ), the existing methods only inhibited the pathway ①, but did not block the pathway ② at the same time. As a result, it was difficult to obtain stable genetically high oleic acid materials

In addition, we reported the construction of FAD2 gene and FAE1 gene RNA interference vectors in 2007 (Peng Qi, Liu Chunlin, etc., Biotechnology Bulletin, 2007, 5: 263-266), but we did not report how to use RNA interference through genetic engineering. The technology inhibits the gene expression of these two pathways at the same time, and finally obtains high oleic acid and low erucic acid transgenic rapeseed materials

The use of genetic engineering to reduce erucic acid content has only been reported to inhibit the construction of FAE1 gene expression interference vectors (Tian Baoming et al., Journal of Oil Crops, 2007, 2: 27-31), and there is no further research on transgenic plants. to report

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