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黄花蒿(Artemisia annua L.)是菊科(Asteraceae)蒿属(Artemisia)一年生草本植物。WD40转录因子基因与植物叶片表面腺毛发育、次生代谢相关。本研究利用RACE技术从黄花蒿中克隆出Aa WD40基因(GenBank登录号:KY411922),并对其进行了生物信息学分析。AaW D40基因的cDNA全长1 498 bp,其中5’末端非翻译区为209 bp,3’末端非翻译区为183 bp。开放阅读框(ORF)的长度为1 106 bp,编码368个氨基酸残基,DNAMAN分析预测其等电点为4.64,分子量为41.29 kD。对AaW D40氨基酸序列进行同源性分析,发现其与多种植物具有较高的同源性。生物信息学分析发现AaW D40氨基酸序列含有4个典型的WD40重复区域,符合WD40基因的特征,所以命名为AaW D40。利用Swiss-Model对AaW D40蛋白进行三级结构分析,AaW D40蛋白空间结构错综复杂,与多种植物WD40空间结构域非常相似。系统发育树分析AaW D40与其他WD40均源于同一祖先,并且AaW D40与DpWDR、GhTTG4的亲缘关系最为接近。荧光实时定量PCR分析显示,AaW D40在根、茎、叶中都可以检测到表达,其中根的表达量最高,其次为叶,再次为茎。本研究为进一步研究AaW D40与黄花蒿次生代谢的关系提供了理论基础。
Artemisia annua L. is an Artemisia annual herb of the Asteraceae family. The gene of WD40 is related to the development of glandular trichomes and the secondary metabolism of plant leaves. In this study, Aa WD40 gene (GenBank accession number: KY411922) was cloned from Artemisia annua using RACE technology and bioinformatics analysis was performed. The full-length cDNA of AaW D40 gene was 1 498 bp in length, 209 bp in the 5 ’untranslated region and 183 bp in the 3’ untranslated region. The length of open reading frame (ORF) was 1 106 bp, encoding 368 amino acid residues. The DNAMAN analysis predicted the isoelectric point was 4.64 and the molecular weight was 41.29 kD. AaW D40 amino acid sequence homology analysis found that with a variety of plants have high homology. Bioinformatics analysis revealed that the AaW D40 amino acid sequence contains four typical WD40 repeat regions, which are consistent with the characteristics of the WD40 gene, so named AaW D40. Using the Swiss-Model to analyze the tertiary structure of the AaW D40 protein, the spatial structure of the AaW D40 protein is intricate and very similar to the WD40 spatial domain of many plants. Phylogenetic tree analysis AaW D40 and other WD40 are derived from the same ancestor, and AaW D40 and DpWDR, GhTTG4 the closest genetic relationship. Real-time quantitative PCR analysis showed that the expression of AaW D40 was detectable in roots, stems and leaves, among which the root expression was the highest, followed by the leaves and again the stems. This study provides a theoretical basis for further study on the relationship between AaW D40 and secondary metabolism of Artemisia annua.