RNA干扰UGP基因表达对三角褐指藻碳流分配的影响

发布时间：2019-06-06 13:05

【摘要】：为深入研究RNA干扰尿苷二磷酸葡萄糖焦磷酸化酶(UGPase)基因的表达对三角褐指藻(Phaeodactylum tricornutum Bohlin)碳流分配的影响,本研究以RNA干扰UGP表达的三角褐指藻为对象,测定了野生和转基因藻株的叶绿素荧光参数和生物量,分析了它们的脂肪酸组成和金藻昆布多糖、总脂及蛋白质的含量。研究显示,转基因藻株的最大光能转化效率(Fv/Fm)从指数生长后期开始显著低于野生型,但野生和转基因藻株的终生物量无明显差别。转基因藻株(AS、IR)的C16与C18之和分别占总脂肪酸含量的66.90%和68.89%,比野生藻株分别提高了19.40%和22.97%,说明抑制UGP表达改变了三角褐指藻的脂肪酸组成。转基因藻株的金藻昆布多糖、总脂及蛋白质含量与野生型藻株差别明显:藻株AS的金藻昆布多糖含量降低25.70%,蛋白质含量上升3.80%,总脂含量提高了13.2%;藻株IR的金藻昆布多糖含量降低40.70%,蛋白质含量上升8.30%,总脂含量增加了25.20%。研究结果表明,RNA干扰UGP的表达改变了三角褐指藻的脂肪酸组成,使得在最终生物量无明显减少的情况下,光合作用固定的碳更多转向油脂合成。本研究进一步明确了RNA干扰UGP的表达对三角褐指藻碳流分配的影响机制,为通过基因工程改造藻种提高微藻的生物柴油生产潜力提供了新思路。
[Abstract]:In order to study the effect of RNA interfering with the expression of uridine diphosphate glucose pyrophosphate (UGPase) gene on the carbon flow distribution of (Phaeodactylum tricornutum Bohlin) in Phaeodactylus triangularis, the aim of this study was to investigate the effect of RNA interference on the expression of UGP in Phaeodactylus triangularis. The chlorophyll fluorescence parameters and biomass of wild and transgenic algae strains were determined, and their fatty acid composition and the contents of polysaccharides, total lipids and proteins were analyzed. The results showed that the maximum light energy conversion efficiency (Fv/Fm) of transgenic algae was significantly lower than that of wild type from the late stage of exponential growth, but there was no significant difference in the final biomass between wild and transgenic algae. The sum of C16 and C18 of transgenic algae strain (AS,IR) accounted for 66.90% and 68.89% of the total fatty acid content, respectively, which was 19.40% and 22.97% higher than that of wild algae strain, respectively. The results showed that inhibition of UGP expression changed the fatty acid composition of Phaeodactylus triangularis. The content of total fat and protein in transgenic algae strain AS decreased by 25.70%, protein content increased by 3.80%, and total fat content increased by 13.2%. The content of polysaccharide, protein and total fat increased by 40.70%, 8.30% and 25.20%, respectively. The results showed that the expression of UGP interfered with RNA changed the fatty acid composition of Phaeodactylus triangularis, and the carbon fixed by photosynthesis shifted to oil synthesis without significant decrease in final biomass. In this study, the mechanism of RNA interfering UGP expression on carbon flow distribution of Phaeodactylus triangularis was further clarified, which provided a new idea for improving the biodiesel production potential of microalgae by genetic engineering.
【作者单位】：海水养殖教育部重点实验室(中国海洋大学);中国海洋大学海洋生命学院;青岛海洋科学与技术国家实验室海洋渔业科学与食物产出过程功能实验室;
【基金】：国家高技术研究发展计划项目(2013AA065801;2014AA022001)资助~~
【分类号】：Q949.2

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