| 摘要: |
| 海南龙血树是国产血竭的主要基源植物,其血竭主要化学成分为类黄酮化合物。为进一步了解DcWD40-1在类黄酮生物合成中的潜在功能和作用机制,该研究根据海南龙血树转录组数据,利用RT-PCR技术在海南龙血树中克隆了一个WD40基因DcWD40-1,该基因全长1 550 bp, 包含一个1 353 bp的开放阅读框,编码450个氨基酸,蛋白质分子量50.77 kD,理论等电点5.71。生物信息学分析显示,DcWD40-1属于WD40蛋白家族成员,具有5个保守的WD40结构域,和其他植物WD40蛋白同源性高,保守性强。利用Genome Walking方法分离了1 503 bp的DcWD40-1启动子序列,该区域具有典型真核生物启动子结构特征,并含有多个应答激素和胁迫的响应元件。表达分析显示,血竭诱导剂能够诱导DcWD40-1的表达,DcWD40-1的变化与血竭形成及类黄酮积累正相关。此外,DcWD40-1也能对茉莉酸甲酯、细胞分裂素、油菜素内酯和UV-B处理做出积极响应。 |
| 关键词: 海南龙血树, 血竭, 类黄酮, WD40转录因子, 基因表达 |
| DOI：10.11931/guihaia.gxzw201808015 |
| 分类号:Q949.45 |
| 文章编号:1000-3142(2020)01-0136-07 |
| Fund project:海南省自然科学基金(318MS093); 国家自然科学基金(81773845)[Supported by Natural Science Foundation of Hainan Province(318MS093); the National Natural Science Foundation of China(81773845)]。 |
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| Cloning and expression analysis of WD40 transcription factor gene DcWD40-1 from Dracaena cambodiana |
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ZHU Jiahong1, XIA Dongnan2, WANG Ying1, GUO Dong1,
LI Huiliang1, MEI Wenli1, PENG Shiqing1*
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1.1. Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou
571101, China;2.2. Institute of Tropical Agriculture and Forestry, Hainan University, Haikou 570228, China
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| Abstract: |
| Hainan Dragon trees(Dracaena cambodiana)are the main plant resources of dragon's blood in China, and the main chemical constituents of the dragon's blood are flavonoids. The recent studies about dragon's blood mainly focus on the chemical constituents and pharmacological activities, while the molecular mechanisms of dragon's blood formation remain unknown. WD40 transcription factor plays an important role in flavonoid accumulation. In this study, a WD40 gene named DcWD40-1 was cloned in Dracaena cambodiana based on transcriptome data and RT-PCR techno-logy. The full-length of cDNA of DcWD40-1 was 1 550 bp, containing 1 353 bp opening reading frame(ORF), and encoding 450 amino acids with the calculated molecular weight of 50.77 kD and calculated pI 5.71. Bioinformatics analysis showed that DcWD40-1 belonged to a member of WD40 superfamily, had five conserved WD40 domains, and shared high identities to WD40 proteins with other plants. A 1 503 bp-length promoter region of DcWD40-1 was isolated by Genome Walking method, which had structural characteristics of typical eukaryotic promoters. The promoter region of DcWD40-1 contained lots of hormone responsible elements, such as abscisic acid-responsive element, auxin-responsive element, salicylic acid-responsive element and jasmonic acid-responsive element; also had many cis acting elements related stress such as light, cold, hot and anaerobic inducer. Expression analysis showed that DcWD40-1 was induced by dragon's blood inducers, positively related to flavonoids accumulation and formation of dragon's blood. In addition, DcWD40-1 can also respond positively to jasmonic acid, cytokinin, brassinosteroid and UV-B treatment. These results will lay the foundation for further study of the potential functions and mechanisms of DcWD40-1 in flavonoid biosynthesis in Dracaena cambodiana. |
| Key words: Dracaena cambodiana, dragon's blood, flavonoids, WD40 transcription factor, gene expression |
