黄秋葵八氢番茄红素脱氢酶基因的克隆与分析
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引用本文:李永平,王 彬,陈敏氡,刘建汀,朱海生,温庆放.黄秋葵八氢番茄红素脱氢酶基因的克隆与分析[J].西北植物学报,2019,39(7):1195~1202
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李永平,王 彬,陈敏氡,刘建汀,朱海生,温庆放* (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,福州 350013) 
基金项目:福建省属公益类科研院所基本科研专项(2019R1026-9);
中文摘要:该研究根据黄秋葵(Hibiscus esculentus L.)转录组测序获得的八氢番茄红素脱氢酶基因(HePDS)序列(GenBank登录号为MG372370)设计引物,克隆验证得到1条HePDS基因全长为2 020 bp cDNA,开放阅读框(ORF)包含1 686个碱基;预测其编码561个氨基酸,理论分子量为62.62 kD,等电点为8.155;编码的蛋白与海岛棉(Gossypium barbadense)、雷蒙德氏棉(Gossypium raimondii)、陆地棉(Gossypium hirsutum)同源蛋白的相似性均在93%以上,均含有1个保守的二核苷酸结合域和1个类胡萝卜素结合域,显示其高度的保守性。荧光定量PCR 分析表明,HePDS基因在黄秋葵根、茎、叶、花和果荚中均有表达;叶发育过程以嫩叶中表达最高,果实发育中以花后2 d表达量最高。类胡萝卜素含量随着叶、果实发育逐渐升高,成熟叶的含量最高,果实以花后4 d含量最高,且HePDS基因的表达与类胡萝卜素含量存在密切的相关性。该研究结果为进一步探讨HePDS基因的功能和调控机制,以及采用VIGS和CRISPR/Cas9技术开展黄秋葵基因功能的反向遗传学研究奠定了基础。
中文关键词:黄秋葵  八氢番茄红素脱氢酶  类胡萝卜素
 
Cloning and Expression Analysis of a Phytoene Desaturase Gene in Hibiscus esculentus L.
Abstract:In this study, a PDS gene was cloned from Hibiscus esculentus L. by transcriptome sequencing. The full length cDNA sequence of PDS was 2 020 bp, which contained a 1 686 bp open reading frame (ORF) that encoded 561 amino acids, with a predicted molecular weight (mol.wt) of 62.62 kD and a hypothetical isoelectric point (pI) of 8.155. It shared over 93% identity with the homologous proteins from Gossypium barbadense, Gossypium raimondii and Gossypium hirsutum All have a NAD(P) binding rossmann like domain and a carotenoid binding domain, showing that it was highly conservative. This gene was named HePDS and the GeneBank accession number was MG372370. Real time PCR analysis revealed that HePDS could be expressed in different tissues of Hibiscus esculentus L., including roots, stems, leaves, flowers and fruits, and the expression level was the highest in tender leaves and 2 days after flowering during the process of leaf and fruit development, respectively. The content of carotenoids increases gradually with the development of leaves and fruits, while the content in mature leaves was the highest and that of fruit was the highest after flowering 4 d. These results revealed the expression of HePDS genes closely related to carotenoid content, and provided a base for the study of carotenoid molecular regulation mechanism and reverse genetics in Hibiscus esculentus L..
keywords:Hibiscus esculentus L.  phytoene desaturase  carotenoid
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