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利用18S rDNA和ITS分子指标,结合序列比对和系统发育树分析,对两株微藻进行分子鉴定、总脂提取以及脂肪酸各组分测定,这两株微藻分离自距我国山东省东营广饶县小码头44.08 km的渤海沿岸地下海水中。结果表明,D12 Strain为盐生微拟球藻(Nannochloropsis salina)(命名:Nannochloropsis salina12),而另一株D2 Strain为寻常小球藻(Chlorella vulgar-is)(命名:Chlorella vulgaris D2)。获得培养至平台期的N.salina D12和C.vulgaris D2油脂含量分别为44%和35%。气质联用色谱分析结果显示,N.salina D12中,脂肪酸成分主要为C16∶0,C16∶1和C18∶1,占总脂肪酸含量约80%,C20以上的长链多不饱和脂肪酸约15%,其中EPA(C20∶5)约10%。C.vulgaris D2中,脂肪酸以C16∶0,C18∶1和C18∶2为主,占总脂肪酸含量90%,不含C20以上的长链多不饱和脂肪酸。以上结果显示,本文利用分子生物学方法及脂肪酸检测所鉴定出的一株N.salina D12和一株C.vulgaris D2脂肪酸类型主要为C16和C18,符合生物柴油分子特点,暗示了这两株微藻具有作为优质产能微藻原料的巨大潜力。
Using 18S rDNA and ITS molecular markers, combined with sequence alignment and phylogenetic tree analysis, molecular identification, total lipid extraction and fatty acid determination of two strains of microalgae were carried out. The two microalgae were isolated from Guangrao, Dongying, Shandong Province, China County small dock 44.08 km in the Bohai Sea coast of the sea. The results showed that D12 Strain is Nannochloropsis salina (named: Nannochloropsis salina12) and the other D2 Strain is Chlorella vulgaris (named: Chlorella vulgaris D2). The oil content of N.salina D12 and C. vulgaris D2 which reached the plateau reached 44% and 35%, respectively. GC-MS analysis showed that the main fatty acid components of N.salina D12 were C16: 0, C16: 1 and C18: 1, accounting for about 80% of the total fatty acids, about 15% of the long chain polyunsaturated fatty acids above C20, , Of which EPA (C20: 5) is about 10%. Among C.vulgaris D2, the fatty acids are mainly C16: 0, C18: 1 and C18: 2, accounting for 90% of the total fatty acids, without long-chain polyunsaturated fatty acids above C20. The above results show that in this paper, molecular biology methods and fatty acid detection identified a N.salina D12 and a C.vulgaris D2 fatty acids mainly C16 and C18, in line with the characteristics of biodiesel molecules, suggesting that the two strains of micro Algae have great potential as raw materials for producing high-quality microalgae.