Department of Plant Breeding, Swedish University of Agricultural Sciences, P.O. Box 190, SE-234 22 Lomma, Sweden.
Institute of Biotechnology, Addis Ababa University, Addis Ababa P.O. Box. 1176, Ethiopia.
Int J Mol Sci. 2024 Jun 7;25(12):6322. doi: 10.3390/ijms25126322.
Metabolic engineering enables oilseed crops to be more competitive by having more attractive properties for oleochemical industrial applications. The aim of this study was to increase the erucic acid level and to produce wax ester (WE) in seed oil by genetic transformation to enhance the industrial applications of . Six transgenic lines for high erucic acid and fifteen transgenic lines for wax esters were obtained. The integration of the target genes for high erucic acid ( and ) and for WEs ( and ) in the genome of cv. 'Derash' was confirmed by PCR analysis. The qRT-PCR results showed overexpression of and and downregulation of RNAi- in the seeds of the transgenic lines. The fatty acid profile and WE content and profile in the seed oil of the transgenic lines and wild type grown in biotron were analyzed using gas chromatography and nanoelectrospray coupled with tandem mass spectrometry. A significant increase in erucic acid was observed in some transgenic lines ranging from 19% to 29% in relation to the wild type, with a level of erucic acid reaching up to 52.7%. Likewise, the transgenic lines harboring and genes produced up to 25% WE content, and the most abundant WE species were 22:1/20:1 and 22:1/22:1. This study demonstrated that metabolic engineering is an effective biotechnological approach for developing into an industrial crop.
代谢工程使油籽作物更具竞争力，因为它们具有更吸引人的特性，适用于油脂化工工业应用。本研究的目的是通过遗传转化提高芥酸水平并在种子油中生产蜡酯（WE），以增强 的工业应用。获得了 6 条高芥酸和 15 条生产 WE 的转基因系。通过 PCR 分析证实了高芥酸（ 和 ）和 WE（ 和 ）靶基因在 cv. 'Derash' 基因组中的整合。qRT-PCR 结果表明，转基因系种子中 和 的表达上调，RNAi- 的表达下调。使用气相色谱和纳喷雾串联质谱法分析了在生物培养箱中生长的转基因系和野生型种子的脂肪酸谱和 WE 含量及谱。一些转基因系的芥酸含量显著增加，与野生型相比增加了 19%至 29%，芥酸水平高达 52.7%。同样，携带 和 基因的转基因系产生高达 25%的 WE 含量，最丰富的 WE 种类为 22:1/20:1 和 22:1/22:1。本研究表明，代谢工程是开发 成为工业作物的有效生物技术方法。
