木薯品种(系)类胡萝卜素代谢相关基因和蛋白表达水平分析
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国家自然科学基金项目(面上项目,重点项目,重大项目)


Analysis of genes and enzymes associated with carotenoid metabolism in cassava
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    摘要:

    摘 要:以白心木薯华南6068 (SC6068)、蛋黄木薯华南9号 (SC9)、紫叶黄心木薯(BGM019)和粉红木薯(Mirasol)为材料,探究木薯块根膨大期和成熟期与类胡萝卜素代谢通路相关的14个基因和4种蛋白质表达水平变化。用HPLC检测块根?-胡萝卜素含量的变化,分别用qRT-PCR和Western blot方法对类胡萝卜素代谢通路相关基因和蛋白酶的表达水平进行分析。以SC6068为对照,研究结果表明:SC9和BGM019块根膨大期表达水平显著高于成熟期的基因为DXR、DXS、PSY1、PDS、ZEP、CCD1和NCED3 (p<0.05),而表达量显著低于成熟期的基因包括PSY2、ZDS、LCYB、HYD和NCED2 (p<0.05)。Mirasol块根表达水平在膨大期显著高于成熟期的基因包括DXR、DXS、PSY1、PSY2、PDS、ZDS、LCYE和CCD1 (p<0.05),而显著低于成熟期的基因为HYD、ZEP、NCED2、NCED3和OR (p<0.05)。其中,ZDS、NCED2 和PSY2基因分别在SC9成熟期、Mirasol成熟期和BGM101成熟期表达水平最高,这些高表达基因有可能成为指示黄心和粉红木薯的标记基因,通过分析不同木薯品种(系)在膨大期和成熟期块根类胡萝卜素代谢途径相关基因的表达水平,有助于解析?-胡萝卜素积累的分子机理。此外,Western blot 结果显示抗坏血酸过氧化物酶、谷胱甘肽还原酶、超氧化歧化酶和HSP70虽然和块根类胡萝卜素代谢途径没有直接关联,但它们在木薯膨大期和成熟期块根表达水平有显著差异(p<0.05)。

    Abstract:

    Abstract The expressions of genes and enzymes related with carotenoid metabolic pathway in the storage roots of cassava swelling and mature periods was validated using white cassava SC6068, yellow cassava SC9, purple-leaf and yellow cassava BGM019 and pink cassava Mirasol as research materials. HPLC method was used to detect the changes of β-carotene contents; qRT-PCR and Western blot methods were applied to analyze the expressed changes of the genes and proteins regarding carotenoid pathway, respectively. In the present study, SC6068 was used as a control, the results showed that the expressed levels of DXR、DXS、PSY1、PDS、ZEP、CCD1 and NCED3 genes in SC9 and BGM019 swelling period were significantly higher than those in the mature period (p<0.05), while the expressions of genes including PSY2、ZDS、LCYB、HYD and NCED2 in swelling period were significantly lower than those in the mature period (p<0.05). The expressions of DXR、DXS、PSY1、PSY2、PDS、ZDS、LCYE and CCD1 genes in Mirasol swelling period were significantly higher than those in the mature period (p<0.05), and the expressions of HYD、ZEP、NCED2、NCED3 and OR genes in swelling period were significantly lower than those in the mature period (p<0.05). Of those, the expressions of ZDS、NCED2 and PSY2 genes were the highest in the mature periods of SC9, Mirasol and BGM101, respectively. These genes may use as the marker genes to determine yellow cassava and pink cassava varieties. These data, regarding the analysis of the gene expressions related with carotenoid metabolic pathway of storage roots in swelling and mature periods, will be helpful to understand the mechanism of ?-carotene accumulation. Furthermore, the Western blot results showed that there was no correlation relationship between ?-carotene accumulation and ascorbate peroxidase, glutathione reductase, FeSOD and HSP70; However, their expression is differed significantly during swelling and mature periods (p<0.05).

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邓昌哲,姚慧,安飞飞,等.木薯品种(系)类胡萝卜素代谢相关基因和蛋白表达水平分析[J].植物遗传资源学报,2017,18(3):555-563.

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  • 收稿日期:2016-07-07
  • 最后修改日期:2016-11-23
  • 录用日期:2016-11-23
  • 在线发布日期: 2017-05-19
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