新型耐热绿藻(Scenedesmus sp.)稳定遗传转化条件的建立

IF 1.5 4区 农林科学 Q3 FISHERIES Turkish Journal of Fisheries and Aquatic Sciences Pub Date : 2023-07-15 DOI:10.4194/trjfas23044
Dilan Akin, Cagla Sonmez, Meral Yucel, H. Oktem
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引用次数: 0

摘要

近年来,微藻成为脂肪酸、类胡萝卜素和其他营养素等高价值化合物的一个有吸引力的来源。在之前的一项研究中,对一种新的耐热淡水微藻Scenedesmus sp.ME02进行了表征,该微藻具有高酚含量,特别是富含黄酮类化合物。本文报道了两种不同的方法来提高该菌株生长参数的效率。首先,研究了缺磷对Scenedesmus sp.ME02生物量和脂质特性的影响。尽管如此,磷的消耗导致微藻的生物量生产力略有下降,对脂质含量和生产力的影响最小;脂肪酸甲酯的分布发生了显著变化。特别是,当细胞从P-补充(对照)条件切换到P-耗尽条件时,油酸百分比(18:1)增加了两倍以上,而亚麻酸百分比(C18:2n6t)和亚麻酸百分比)(C18:3n6)显著降低。总体而言,多不饱和脂肪酸含量显著降低。其次,利用两种不同的转化载体,优化了Scenedesmus sp.ME02的电穿孔遗传转化条件。通过RT-PCR在用两种载体转化的菌落中检测GFP基因表达。通过共聚焦显微镜在初始转化一年后的转基因系中检测到GFP蛋白的表达。
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Establishment of Stable Genetic Transformation Conditions for Novel Thermotolerant Scenedesmus sp. (Chlorophyta)
Microalgae became an attractive source of high value compounds such as fatty acids, carotenoids and other nutrients in recent years. In a previous study, the characterization of a novel thermo-tolerant freshwater microalga, Scenedesmus sp. ME02 with high phenolic content, particularly rich in flavonoids was done. Here, two different approaches are reported to increase the efficiency of the growth parameters of this strain. Firstly, the effects of phosphorus (P) depletion on biomass and lipid characteristics of Scenedesmus sp. ME02 were investigated. Although, P depletion resulted in a slight reduction in biomass productivity of microalgae and had a minimal effect on the lipid content and productivity; fatty acid methyl ester profile changed substantially. Particularly, percent oleic acid (18:1) increased by more than two-fold, whereas percent linolelaidic acid (C18:2n6t) and linolenic acid (C18:3n6) decreased significantly when cells were switched from P-replete (control) to P-deplete conditions. Overall, the polyunsaturated fatty acid content was significantly reduced. Secondly, stable genetic transformation conditions for Scenedesmus sp. ME02 via electroporation were optimized using two different transformation vectors. GFP gene expression was detected in colonies transformed with both vectors by RT-PCR. GFP protein expression was detected in the transgenic lines one year after initial transformation by confocal microscopy.
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来源期刊
Turkish Journal of Fisheries and Aquatic Sciences
Turkish Journal of Fisheries and Aquatic Sciences FISHERIES-MARINE & FRESHWATER BIOLOGY
CiteScore
3.10
自引率
0.00%
发文量
43
审稿时长
3 months
期刊介绍: Turkish Journal of Fisheries and Aquatic Sciences" (TrJFAS) is a refereed academic journal has been published by Central Fisheries Research Institute of Turkey and Japan International Cooperation Agency (JICA), and published in English. It aims to address research and needs of all working and studying within the many varied areas of fisheries and aquatic sciences. The Journal publishes English language original research papers, critical review articles, short communications and technical notes on applied or scientific research relevant to freshwater, brackish and marine environments. TrJFAS was published biannually (April & November) between 2001 and 2009. A great number of manuscripts have been submitted to the journal for review from acceptance of the SCI index. Thereby, the journal has been published quarterly (March, June, September and December) from 2010 to 2017. The journal will be published monthly in 2018.
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