菘蓝IiCYP79F1基因的克隆与表达分析
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陕西省社会发展科技攻关(2016SF390);


Cloning and Expression Analysis of IiCYP79F1 Gene from Isatis indigotica Fort.
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    摘要:

    该研究以菘蓝叶片为材料,采用RTPCR方法克隆菘蓝IiCYP79F1基因,并对其进行生物信息学与表达模式分析。结果表明:(1)成功获得IiCYP79F1基因的gDNA全长(2 109 bp),包含3个外显子及2个内含子,ORF全长为1 626 bp,编码541个氨基酸(GenBank登录号为KY774689.1);生物信息学分析显示,IiCYP79F1蛋白的二级结构主要由α螺旋(43.81%)、无规则卷曲(35.49%)、延伸链(13.68%)和β转角(7.02%)组成;其氨基酸序列与西兰花、欧洲油菜、芜菁和芝麻菜的相似性较高,其蛋白与芝麻菜的亲缘关系最近。(2)qRTPCR分析显示,IiCYP79F1基因呈时空特异性表达,且在茎中与幼苗期高表达;MeJA、Ag+、葡萄糖和机械损伤处理均能有效促进IiCYP79F1基因的表达,而SA和低温处理则具有明显的抑制作用。该研究结果为进一步探讨IiCYP79F1在菘蓝芥子油苷生物合成中的作用奠定了基础,也为培育高芥子油苷含量的菘蓝新种质提供了新思路。

    Abstract:

    IiCYP79F1 gene was cloned from Isatis indigotica Fort. by RTPCR method, and its bioinformatics and expression analysis were performed. The results showed as follows: (1) the full length of gDNA for IiCYP79F1 was 2 109 bp, including two introns and three extrons; and the full length of ORF was 1 626 bp, encoding 541 amino acids (GenBank accession Number: KY774689.1). Bioinformatics analysis indicated that the secondary structure of IiCYP79F1 protein was mainly composed of αhelix (43.81%), random coil (35.49%), extended strand (13.68%) and βturn (7.02%); and its amino acid sequences had a high degree of identity with that of CYP79F1 in Brassica oleacea, Brassica napus, Brassica rapa and Eruca sativa. The phylogenetic analysis exhibited that IiCYP79F1 protein of I. indigotica Fort. had the closest relationship with Eruca sativa. (2) qRTPCR analysis demonstrated that IiCYP79F1 gene was expressed in spatiotemporal specificity and was highly expressed in stem and the seedling stage. MeJA, Ag+, glucose and mechanical injury promoted the expression of IiCYP79F1 gene, while SA and low temperature treatments had inhibiting actions. The results could lay the foundation for the further exploration of the function of IiCYP79F1 on the synthesis of glucosinolates, and provide a new idea for the new germplasm resources development.

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屈新运,秦苗苗,赵桂红,等.菘蓝IiCYP79F1基因的克隆与表达分析[J].西北植物学报,2018,38(10):1809-1816

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  • 在线发布日期: 2018-11-23
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