Song Jiejie, Luo Hongmei, Xu Zhichao, Zhang Yuxi, Xin Hua, Zhu Dan, Zhu Xunzhi, Liu Mengmeng, Wang Weiqing, Ren Hongwei, Chen Hongyu, Gao Ting
Key Laboratory of Plant Biotechnology in Universities of Shandong Province, College of Life Sciences, Qingdao Agricultural University, Qingdao 266109, People's Republic of China.
J Genet. 2020;99.
The endangered medicinal plant is one of the important natural source of furanocoumarin, which has been used as mucolytic, antitussive, antitumour and antibacterial. However, the genetic information of furanocoumarin biosynthesis in is scarce at present. The objective of this study was to mine the putative candidate genes involved in the biosynthesis pathwayof furanocoumarin and provide references for gene identification, and functional genomics of . We carried out the transcriptome analysis of leaves and roots in , which provided a dataset for gene mining. Psoralen, imperatorin and isoimperatorin were detected in by high performance liquid chromatography analysis. Candidate key genes were mined based on the annotations and local BLAST with homologous sequences using BioEdit software. The relative expression of genes was analysed using quantitative real-time polymerase chain reaction. Further, the CYP450 genes were mined using phylogenetic analyses using MEGA 6.0 software. Atotal of 156,949 unigenes were generated, of which 9021 were differentially-expressed between leaves and roots. A total of 82 unigenes encoding eight enzymes in furanocoumarin biosynthetic pathway were first obtained. Seven genes that encoded key enzymes in the downstream furanocoumarin biosynthetic pathway and expressed more in roots than leaves were screened. Twenty-six candidate CYP450 unigenes expressed abundantly in roots and were chiefly concentrated in CYP71, CYP85 and CYP72 clans. Finally, we filtered 102 differentially expressed transcription factors (TFs) unigenes. The transcriptome of was characterized which would help to elucidate the furanocoumarin biosynthetic pathway in and provide an invaluable resource for further study of furanocoumarin.
这种濒危药用植物是呋喃香豆素的重要天然来源之一,呋喃香豆素已被用作黏液溶解剂、镇咳药、抗肿瘤药和抗菌药。然而,目前该植物中呋喃香豆素生物合成的遗传信息稀缺。本研究的目的是挖掘参与呋喃香豆素生物合成途径的假定候选基因,并为该植物的基因鉴定和功能基因组学提供参考。我们对该植物的叶和根进行了转录组分析,这为基因挖掘提供了一个数据集。通过高效液相色谱分析在该植物中检测到了补骨脂素、欧前胡素和异欧前胡素。使用BioEdit软件基于注释和与同源序列的本地BLAST挖掘候选关键基因。使用定量实时聚合酶链反应分析基因的相对表达。此外,使用MEGA 6.0软件通过系统发育分析挖掘CYP450基因。共产生了156,949个单基因,其中9021个在叶和根之间差异表达。首次获得了总共82个编码呋喃香豆素生物合成途径中八种酶的单基因。筛选出七个在呋喃香豆素生物合成途径下游编码关键酶且在根中表达高于叶的基因。26个候选CYP450单基因在根中大量表达,主要集中在CYP71、CYP85和CYP72家族。最后,我们筛选出102个差异表达的转录因子(TFs)单基因。对该植物的转录组进行了表征,这将有助于阐明该植物中呋喃香豆素的生物合成途径,并为呋喃香豆素的进一步研究提供宝贵资源。