2种化学型的黄花蒿萜类代谢物及其相关基因表达分析

Expression Analysis of Two Chemical-Types of Artemisia Annua L.Terpenoids and Their Related Genes

目的分析2种化学型黄花蒿萜类代谢物及其转录组数据信息,探讨2种化学型黄花蒿差异代谢物及其可能形成原因。方法采集江苏盱眙县、河南中牟县两地黄花蒿种子于同一条件下种植,通过顶空-气相色谱-三重四极杆串联质谱(HS-GC-QQQ-MS/MS)分析黄花蒿的挥发性成分,超快速高效液相色谱-三重四级杆飞行时间串联质谱(UFLC-Triple TOF-MS/MS)非靶向分析2种化学型黄花蒿的代谢物,并采用转录组测序分析挥发性成分相关萜类生物合成基因的表达。结果依据挥发性主成分类别分型,江苏盱眙县种源黄花蒿为蒿酮型,河南中牟县种源黄花蒿为樟脑型。UFLC-Triple TOF-MS/MS检测到的差异代谢物通过KEGG数据库分析可知蒿酮型和樟脑型黄花蒿分别在倍半萜、三萜生物合成和二萜生物合成路径显著上调。转录组数据中通向萜类骨架生物合成的甲基赤藓糖醇-4-磷酸(MEP)途径和甲羟戊酸(MVA)途径注释到11个关键酶的23个候选基因具有显著差异。单萜合成路径中,1,8-桉树脑合酶(TPS-Cin)共检测到10个候选基因。此外,还发现2个冰片脱氢酶候选基因在樟脑型黄花蒿中高表达,3个黄花蒿醇脱氢酶2(ADH2)候选基因在蒿酮型黄花蒿中高表达。结论该研究为揭示黄花蒿化学型形成分子机制提供科学数据。

OBJECTIVE To investigate the differential metabolites of two chemical-types of Artemisia annua and their possible causes by analyzing the data of terpenoid metabolites and their transcriptome of two chemical-types of Artemisia annua.METHODS The seeds of Artemisia annua were collected from Xuyi County,Jiangsu Province,and Zhongmou County,Henan Province,in China,and grown under the same conditions.The volatile components of Artemisia annua were analyzed by headspace-gas chromatography-triple quadrupole mass spectrometry(HS-GC-QQQ-MS/MS),non-targeted analysis of the differential metabolites of 2 chemical-types of Artemisia annua were performed by ultra-fast liquid chromatography-triple quadrupole-time of flight tandem mass spectrometry(UFLC-Triple TOF-MS/MS),and the expression of the biosynthetic genes of the volatile components was analyzed by transcriptome sequencing.RESULTS Based on the typing of volatile components,Artemisia annua from Xuyi County,Jiangsu Province was classified as artemisia ketone type,and Artemisia annua from Zhongmou County,Henan Province was classified as camphor type.The differential metabolites detected by UFLC-Triple TOF-MS/MS were significantly up-regulated in sesquiterpene,triterpene,and diterpene biosynthesis pathways by Kyoto encyclopedia of genes and genomes(KEGG)database analysis for artemisia ketone type and camphor types of Artemisia annua,respectively.The 2-methyl-D-erythritol-4-phosphate(MEP)pathway leading to terpene skeleton biosynthesis and the mevalonate(MVA)pathway annotated 23 candidate genes for 11 key enzymes that were significantly differential in the transcriptome data.In the monoterpene synthesis pathway,10 candidate genes were detected for 1,8-cineole synthase(TPS-Cin).In addition,two borneol dehydrogenase candidates were found to be highly expressed in camphor-type Artemisia annua,and three Artemisia annua alcohol dehydrogenase(ADH2)candidates were highly expressed in artemisia ketone-type Artemisia annua.CONCLUSION This study provides scientific data to reveal the molecular mechanism of Artemisia annua chemical-types formation.