舟形藻生长、品质特性及转化筛选标记研究

Analysis of growth and quality of Navicula teneraand determination of its selectable makers in genetic engineering

研究了舟形藻(Navicula tenera)细胞表型和积累模式,利用索式抽提法和气质联用(GC-MS)方法对其进行总油脂含量和脂肪酸组分的分析,并研究了舟形藻对氨苄青霉素(Amp)、卡那霉素(Kan)、草胺膦(PPT)、庆大霉素(Gen)、链霉素(Str)、遗传霉素(G418)、潮霉素(Hyg)以及氯霉素(Cm)这8种常用抗生素的敏感性。结果显示,舟形藻细胞以沉积底部的生长模式积累;其总油脂含量占其干物质重量的8.31%,脂肪酸种类主要有C16∶0,C16∶3(n-4)和C20∶5(n-3)(EPA),分别占总脂肪酸的64.13%、15.87%和19.62%;舟形藻对Cm和G418非常敏感,对Hyg较敏感,而对不同浓度的Amp、Kan、Gen、Str以及PPT极不敏感。为舟形藻基因工程筛选标记的确立,进而建立其遗传转化系统奠定了基础。

The cell phenotype and accumulation pattern of Navicula tenera, a kind of marine unicellular eukaryotic microalgae, was investigated. The total lipid and fatty acids composition was analyzed by Soxhlet extraction and GC-MS. The sensitivities of Navicula tenera on eight kinds of antibiotics including Ampicilin （Amp）, Kanamycin （Kan）, Phosphinothricin （PPT）, Gentamycin （Gen）, Strep- tomycin （Str）, ttygromycin （Hyg） , Geneticin （G418） and Chloramphenicol （Cm） were measured. The results showed that the cell accumulation pattern of Navicula tenera was settling to the bottom of the medium. The dry mass of Navicula tenera contained 8.31% of total lipid that was mainly constituted by 64. 13% of C16：0, 15.87% of C16：3 （n-4） and 19.62% of C20：5 （ n-3 ） （EPA）. Navicula tenera was extremely sensitive to Cm and G418 and was relatively sensitive to Hyg. However, it was not sensitive to Amp, Kan, PPT, Gen, and Str whether their test concentrations high or not. The results provided important data in developing selectable makers in genetic engineering of Navicula tenera.