Du Hewei, Huang Min, Hu Jieyun, Li Jiansheng
College of Life Science, Yangtze University, Jingzhou, Hubei, 434025, Peoples's Republic of China.
Hubei Collaborative Innovation Center for Grain Industry, Yangtze University, Jingzhou, 434025, Peoples's Republic of China.
BMC Plant Biol. 2016 Jun 14;16(1):137. doi: 10.1186/s12870-016-0827-z.
Stearoyl-acyl carrier protein desaturase (SAD) is a key enzyme that catalyses the conversion of stearoyl-acyl carrier protein (ACP) to oleoyl-ACP, a precursor for the biosynthesis of polyunsaturated fatty acids. ZmSAD1 (GenBank: KU949326) is a major QTL for stearic acid content in maize seeds. To investigate the biological function and the application potential of maize ZmSAD1 in oil biosynthesis, we isolated the full-length ZmSAD1 cDNA from maize B73 and overexpressed it in Arabidopsis and maize.
Under seed-specific overexpression of ZmSAD1 in Arabidopsis, the stearic acid content and the ratio of saturated to unsaturated fatty acids in the seeds were significantly decreased relative to those in the control. Conversely, in transgenic ZmSAD1 RNAi Arabidopsis seeds, the contents of stearic acid and long-chain saturated acids and the ratio of saturated to unsaturated fatty acids were significantly increased; in addition, the oleic acid content was significantly decreased. More importantly, transgenic ZmSAD1 maize that expressed high levels of ZmSAD1 in its mature seeds showed reduced stearic acid content (1.57 %) and a lower saturated to unsaturated fatty acid ratio (20.40 %) relative to those (1.64 % and 20.61 %, respectively) of the control. Conversely, down-regulation of ZmSAD1 in maize resulted in increased levels of stearic acid (1.78 %), long-chain saturated acids (0.85 %) and the ratio of saturated to unsaturated fatty acids (21.54 %) relative to those (1.64 %, 0.74 %, and 20.61 %, respectively) of the control, whereas the oleic acid (32.01 %) level was significantly decreased relative to that (32.68 %) of the control.
Our work demonstrates that the contents of stearic acid, oleic acid, and long-chain saturated acids, and the ratio of saturated to unsaturated fatty acids, are modified in maize seeds by seed-specific overexpression or down-regulation of ZmSAD1. Therefore, the ZmSAD1 gene is a useful tool for engineering the seed oil composition in maize and other crops.
硬脂酰 - 酰基载体蛋白去饱和酶(SAD)是一种关键酶,催化硬脂酰 - 酰基载体蛋白(ACP)转化为油酰 - ACP,而油酰 - ACP是多不饱和脂肪酸生物合成的前体。ZmSAD1(GenBank:KU949326)是玉米种子中硬脂酸含量的一个主要数量性状位点(QTL)。为了研究玉米ZmSAD1在油脂生物合成中的生物学功能及应用潜力,我们从玉米B73中分离出ZmSAD1 cDNA全长,并在拟南芥和玉米中进行过表达。
在拟南芥中种子特异性过表达ZmSAD1时,种子中的硬脂酸含量以及饱和脂肪酸与不饱和脂肪酸的比例相对于对照均显著降低。相反,在转基因ZmSAD1 RNA干扰拟南芥种子中,硬脂酸和长链饱和酸的含量以及饱和脂肪酸与不饱和脂肪酸的比例显著增加;此外,油酸含量显著降低。更重要的是,在成熟种子中高水平表达ZmSAD1的转基因ZmSAD1玉米,其硬脂酸含量(1.57%)和饱和脂肪酸与不饱和脂肪酸的比例(20.40%)相对于对照(分别为1.64%和20.61%)有所降低。相反,玉米中ZmSAD1的下调导致硬脂酸(1.78%)、长链饱和酸(0.85%)水平以及饱和脂肪酸与不饱和脂肪酸的比例(21.54%)相对于对照(分别为1.64%、0.74%和20.61%)增加,而油酸水平(32.01%)相对于对照(32.68%)显著降低。
我们的研究表明,通过种子特异性过表达或下调ZmSAD1,玉米种子中硬脂酸、油酸和长链饱和酸的含量以及饱和脂肪酸与不饱和脂肪酸的比例会发生改变。因此,ZmSAD1基因是改良玉米和其他作物种子油成分的有用工具。