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三角褐指藻岩藻黄素合成途径及其关键基因对高光照的响应
张南南，罗 玲，陈 卓，杨之帆，黄凤洪，万 霞，龚阳敏
中国油料作物学报 ›› 2017, Vol. 39 ›› Issue (1) : 128.
PDF(1881 KB)
PDF(1881 KB)
三角褐指藻岩藻黄素合成途径及其关键基因对高光照的响应
Biosynthesis pathway of fucoxanthin and change of expression levels of key genes for fucoxanthin synthesis in response to high irradiance in Phaeodactylum tricornutum
本研究以三角褐指藻为材料，以三角褐指藻岩藻黄素的合成途径中主要的六个酶：八氢番茄红素合成
酶、八氢番茄红素脱氢酶、ζ－胡萝卜素脱氢酶、胡萝卜素异构酶、番茄红素β－环化酶和玉米黄素环氧酶为研究对
象，研究高光照胁迫对三角褐指藻岩藻黄素合成代谢途径中关键基因表达量以及岩藻黄素含量的变化规律。以光
照强度为５００μｍｏｌｐｈｏｔｏｎｓ／（ｍ２·ｓ），分别照射处理三角褐指藻３ｈ，６ｈ，１２ｈ，采用ＨＰＬＣ对岩藻黄素含量进行定
量分析。随着光照时间的延长，岩藻黄素含量呈现先上升后下降的趋势；高光照６ｈ样品中岩藻黄素含量为１．８５ｍｇ
·ｇ－１ＤＣＷ （ＤｒｙＣｅｌｌＷｅｉｇｈｔ），比高光照０ｈ对照组中岩藻黄素含量增加了２．１６倍。ｑＲＴ－ＰＣＲ分析结果表明玉米
黄素环氧酶基因ｚｅｐ１和八氢番茄红素合成酶基因ｐｙｓ转录本在高光照胁迫６ｈ丰度最高，与对照组相比提高了近
两倍，基因表达量的变化表现出与岩藻黄素含量变化相一致的趋势。高光照处理３ｈ后，胡萝卜素异构酶基因ｃｒｔｉ
ｓｏ３，八氢番茄红素脱氢酶基因ｐｄｓ，ζ－胡萝卜素脱氢酶基因ｚｄｓ的表达水平相比于对照组明显降低，表现出应对高
光照胁迫的快速响应；番茄红素β－环化酶基因ｌｃｙｂ，胡萝卜素异构酶基因ｃｒｔｉｓｏ１的表达水平则在高光照处理１２ｈ
后明显提高；与ｚｅｐ１和ｐｙｓ相比，ｌｃｙｂ和ｃｒｔｉｓｏ１基因的表达对高光强胁迫处理的响应时间要滞后至少６ｈ。本研究揭
示了高光照胁迫处理下三角褐指藻中岩藻黄素含量及其合成途径中关键基因的表达模式，表明ｚｅｐ１和ｐｙｓ基因表
达水平与岩藻黄素合成量之间存在明显的线性关系。
In this study, we examined the changes of fucoxanthin content and transcript quantity of key genes encoding biosynthesis pathway of fucoxanthin containing six key enzymes，phytoene synthase, phytoene desaturase, ζ-carotene desaturase, carotenoid isomerase, lycopene β-cyclase, and zeaxanthin epoxidase (excluding the unidentified enzymes in diadinoxanthin cycle and violaxanthin cycle) in the diatom P. tricornutum. P. tricornutum cells were exposed to high light intensity of 500μE/(m2•s) for 3h, 6h and 12h; and fucoxanthin content was quantified by HPLC. Fucoxanthin content increased and then declined during 12 h of high irradiance, and peaked at 6h (1.85 mg·g-1DCW), which was 2.16-fold higher than that of control sample (0h for high irradiance). Quantitive RT-PCR showed that the transcript abundances of genes encoding zeaxanthin epoxidase (zep1) and phytoene synthase (pys) peaked at 6h of high irradiance, which was two-fold higher than that of control (0h for high irradiance) and consistent with the trend of change of fucoxanthin content. The transcript quantity of the genes encoding ζ-carotene desaturase3, (crtiso3), phytoene desaturase (pds) and ζ-carotene desaturase (zds) decreased greatly upon high irradiance for 3h compared to the control, and showed a rapid response to high irradiance stress. The expression levels of genes encoding lycopene β-cyclase (lcyb) and carotenoid isomerase (crtiso1) was only enhanced after high irradiance for 12 h. The response of the expression of lcyb and crtiso1 to high irradiance stress was at least 6 h later than that of zep1 and pys. This study revealed the change of fucoxanthin content and expression pattern of genes encoding fucoxanthin biosynthesis pathway under the condition of high irradiance stress in the diatom P. tricornutum, showing an evident linear relationship between fucoxanthin content and the expression levels of zep1 and pys.
高光照 / 三角褐指藻 / 岩藻黄素 / 转录本丰度 {{custom_keyword}} /
high irradiance / Phaeodactylum tricornutum / fucoxanthin / transcript abundance {{custom_keyword}} /
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中国农业科学院科技创新工程项目（ＣＡＡＳ－ＡＳＴＩＰ－２０１３－ＯＣＲＩ）
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