【摘要】：為深入研究RNA干擾尿苷二磷酸葡萄糖焦磷酸化酶(UGPase)基因的表達對三角褐指藻(Phaeodactylum tricornutum Bohlin)碳流分配的影響,本研究以RNA干擾UGP表達的三角褐指藻為對象,測定了野生和轉基因藻株的葉綠素熒光參數和生物量,分析了它們的脂肪酸組成和金藻昆布多糖、總脂及蛋白質的含量。研究顯示,轉基因藻株的最大光能轉化效率(Fv/Fm)從指數生長后期開始顯著低于野生型,但野生和轉基因藻株的終生物量無明顯差別。轉基因藻株(AS、IR)的C16與C18之和分別占總脂肪酸含量的66.90%和68.89%,比野生藻株分別提高了19.40%和22.97%,說明抑制UGP表達改變了三角褐指藻的脂肪酸組成。轉基因藻株的金藻昆布多糖、總脂及蛋白質含量與野生型藻株差別明顯:藻株AS的金藻昆布多糖含量降低25.70%,蛋白質含量上升3.80%,總脂含量提高了13.2%;藻株IR的金藻昆布多糖含量降低40.70%,蛋白質含量上升8.30%,總脂含量增加了25.20%。研究結果表明,RNA干擾UGP的表達改變了三角褐指藻的脂肪酸組成,使得在最終生物量無明顯減少的情況下,光合作用固定的碳更多轉向油脂合成。本研究進一步明確了RNA干擾UGP的表達對三角褐指藻碳流分配的影響機制,為通過基因工程改造藻種提高微藻的生物柴油生產潛力提供了新思路。
[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.
【作者單位】： 海水養(yǎng)殖教育部重點實驗室(中國海洋大學);中國海洋大學海洋生命學院;青島海洋科學與技術國家實驗室海洋漁業(yè)科學與食物產出過程功能實驗室;
【基金】：國家高技術研究發(fā)展計劃項目(2013AA065801;2014AA022001)資助~~
【分類號】：Q949.2

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