Li Xueyuan, Mei Desheng, Liu Qing, Fan Jing, Singh Surinder, Green Allan, Zhou Xue-Rong, Zhu Li-Hua
Department of Plant Breeding, Swedish University of Agricultural Sciences, Alnarp, Sweden.
CSIRO Food, Nutrition & Bioproducts Flagship, Canberra, ACT, Australia.
Plant Biotechnol J. 2016 Jan;14(1):323-31. doi: 10.1111/pbi.12386. Epub 2015 May 21.
High oleic oil is an important industrial feedstock that has been one of the main targets for oil improvement in a number of oil crops. Crambe (Crambe abyssinica) is a dedicated oilseed crop, suitable for industrial oil production. In this study, we down-regulated the crambe fatty acid desaturase (FAD) and fatty acid elongase (FAE) genes for creating high oleic seed oil. We first cloned the crambe CaFAD2, CaFAD3 and CaFAE1 genes. Multiple copies of each of these genes were isolated, and the highly homologous sequences were used to make RNAi constructs. These constructs were first tested in Arabidopsis, which led to the elevated oleic or linoleic levels depending on the genes targeted, indicating that the RNAi constructs were effective in regulating the expression of the target genes in nonidentical but closely related species. Furthermore, down-regulation of CaFAD2 and CaFAE1 in crambe with the FAD2-FAE1 RNAi vector resulted in even more significant increase in oleic acid level in the seed oil with up to 80% compared to 13% for wild type. The high oleic trait has been stable in subsequent five generations and the GM line grew normally in greenhouse. This work has demonstrated the great potential of producing high oleic oil in crambe, thus contributing to its development into an oil crop platform for industrial oil production.
高油酸油是一种重要的工业原料,一直是许多油料作物油脂改良的主要目标之一。海甘蓝(Crambe abyssinica)是一种专用油料作物,适合用于工业油脂生产。在本研究中,我们下调了海甘蓝脂肪酸去饱和酶(FAD)和脂肪酸延长酶(FAE)基因,以生产高油酸种子油。我们首先克隆了海甘蓝CaFAD2、CaFAD3和CaFAE1基因。分离出这些基因的多个拷贝,并使用高度同源序列构建RNA干扰(RNAi)载体。这些载体首先在拟南芥中进行测试,根据靶向基因的不同,导致油酸或亚油酸水平升高,这表明RNAi载体在调控非同一但密切相关物种中靶基因的表达方面是有效的。此外,使用FAD2-FAE1 RNAi载体下调海甘蓝中的CaFAD2和CaFAE1,导致种子油中油酸水平显著提高,与野生型的13%相比,最高可达80%。高油酸性状在随后的五代中保持稳定,转基因株系在温室中正常生长。这项工作证明了海甘蓝生产高油酸油的巨大潜力,从而有助于将其发展成为一个用于工业油脂生产的油料作物平台。