黄秋葵LCYE基因的克隆与表达分析

发布时间：2018-03-20 15:02

  本文选题：黄秋葵　切入点：叶黄素　出处：《农业生物技术学报》2017年10期 　论文类型：期刊论文

【摘要】：ε-番茄红素环化酶(lycopene epsilon-cyclase,LCYE)是参与类胡萝卜素合成的关键酶,在类胡萝卜素合成中发挥着重要作用。为了探究黄秋葵(Hibiscus esculentus)中LCYE基因的功能,本研究成功克隆了黄秋葵类胡萝卜素合成途径中下游LCYE,其cDNA全长1 845 bp,具有1 605个碱基的完整开放阅读框(ORF),编码534个氨基酸,理论分子量(Mw)为59.73 kD,等电点(pI)为6.22。黄秋葵LCYE与亚洲棉(Gossypium arboretum)、可可(Theobroma cacao)及麻风树(Jatropha curcas)同源蛋白的相似性均在80%以上,具高度的保守性,将基因命名为HyLCYE,GenBank登录号为:KX257999。荧光定量分析表明,LCYE在黄秋葵成熟叶中表达最高,果实发育中以花后3 d高表达。应用超高效液相色谱法(ultra performance liquid chromatography,UPLC)对叶黄素含量进行分析,在不同发育时期的叶中,成熟叶中叶黄素含量最高;在果实中,以花后3 d的果实含量最高。结果表明,HyLCYE基因表达与叶黄素含量成正相关。不同基因型秋葵品种同一时期的类胡萝卜素以绿色品种最高,红色秋葵品种次之,白色秋葵品种总含量最低。该实验结果为进一步开发利用黄秋葵提供了科学依据。
[Abstract]:蔚 -lycopene epsilon-cyclase (LCYE) is a key enzyme involved in carotenoid synthesis and plays an important role in carotenoid synthesis. In order to explore the function of LCYE gene in Hibiscus esculentus, In this study, we cloned the middle and lower reaches of the anemone carotenoid synthesis pathway LCYE, its cDNA length was 1 845 BP, with 1 605 bases complete open reading frame, encoding 534 amino acids. The theoretical molecular weight was 59.73 KD and the isoelectric point (Pi) was 6.22. The homologous proteins of LCYE, Gossypium arboretum, Theobroma cacaoand Jatropha curcasa were all more than 80%, which were highly conserved. The gene was named HyLCYEN GenBank accession number: KX257999.The fluorescence quantitative analysis showed that the highest expression of LCYE was found in the mature leaves of Okra, and the high expression was observed 3 days after anthesis in fruit development. The content of lutein was analyzed by ultra performance liquid chromatographyof UPLC. The content of lutein in mature leaves was the highest in the leaves of different developmental stages, and in the fruit, the content of lutein was the highest in mature leaves. The results showed that the expression of HyLCYE gene was positively correlated with lutein content. The carotenoids of different genotypes of okra were the highest in the same period, followed by red anemones. The results provided scientific basis for further development and utilization of Okra.
【作者单位】： 福建省农业科学院作物研究所/蔬菜研究中心/福建省蔬菜工程技术研究中心;
【基金】：福建省属公益类科研院所基本科研专项(No.2015R1026-9) 福建省自然科学基金(No.2015J01118) 福建省农业科学院创新团队PI项目(No.2016PI-40)
【分类号】：Q943.2;S649
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