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目的研究葡萄糖对三角褐指藻细胞生长、岩藻黄素含量和岩藻黄素生物合成相关基因表达的影响。方法本实验以分光光度法研究葡萄糖对三角褐指藻生长影响,HPLC研究藻细胞岩藻黄素含量,荧光定量聚合酶链式反应(PCR)研究岩藻黄素生物合成相关基因表达。结果在平台期末,葡萄糖处理组藻细胞密度均比对照组高,说明葡萄糖促进三角褐指藻细胞生长。HPLC结果表明,随着葡萄糖浓度的升高,三角褐指藻细胞中岩藻黄素含量呈逐渐下降的趋势。其中,葡萄糖为50mg·L-1时,岩藻黄素含量最低,为0.26 mg·g-1干重,比对照组降低了67.5%,说明葡萄糖抑制了三角褐指藻岩藻黄素的生物合成。荧光定量PCR结果表明,10~50 mg·L-1葡萄糖处理三角褐指藻时,岩藻黄素生物合成相关基因zep、pys、zds、lcyb、crtiso和pds的表达量均比对照组低。该结果与岩藻黄素含量变化一致。结论葡萄糖抑制三角褐指藻岩藻黄素的生物合成是通过下调岩藻黄素生物合成相关基因表达来实现的。
Objective To study the effects of glucose on the growth of Phaeodactylum tricornutum, the content of fucoxanthin and the gene expression of fucoxanthin biosynthesis. Methods The effect of glucose on the growth of Phaeodactylum tricornutum was studied spectrophotometrically. The content of fucoxanthin in algae cells was investigated by HPLC and the expression of genes related to the biosynthesis of fucoxanthin by fluorescence quantitative polymerase chain reaction (PCR). Results At the end of the platform, the density of algal cells in the glucose-treated group was higher than that in the control group, indicating that glucose promoted the growth of Phaeodactylum tricornutum. The results of HPLC showed that the content of fucoxanthin decreased gradually with the increase of glucose concentration. Among them, when the concentration of glucose was 50 mg · L-1, the content of fucoxanthin was the lowest, which was 0.26 mg · g-1 dry weight, which was 67.5% lower than that of the control group, indicating that glucose inhibited the growth of Phaeodactylum fucoxanthin synthesis. The result of real-time PCR showed that the expression of fucoxanthin biosynthesis genes zep, pys, zds, lcyb, crtiso and pds were lower than that of the control group when treated with 10-50 mg · L-1 glucose. This result is consistent with the change of fucoxanthin content. Conclusion Glucose inhibits the biosynthesis of fucoxanthin by down-regulating the gene expression of fucoxanthin biosynthesis.