积雪草皂苷生物合成基因筛选与表达量分析  

Screening and Expression Analysis of Saponins Biosynthesis Genes from Centella asiatica

作　　者：李红金 黄鼎 姚绍嫦 李良波 黄荣韶 谭勇 明如宏 LI Hong-jin;HUANG Ding;YAO Shao-chang;LI-Liang-bo;HUANG Rong-shao;TAN Yong;MING Ru-hong(College of Pharmacy,Guangxi University of Chinese Medicine,Nanning 530200,China;Guangxi University of Chinese Medicine/Guangxi Key Laboratory of Zhuang and Yao Ethnic Medicine,Nanning 530200,China)

机构地区：[1]广西中医药大学药学院,广西南宁530200 [2]广西中医药大学/广西壮瑶药重点实验室,广西南宁530200

出　　处：《中药材》2023年第6期1376-1380,共5页Journal of Chinese Medicinal Materials

基　　金：广西科技基地和人才专项项目(桂科AD21220083);广西中医药大学博士启动基金项目(2021BS010);广西中医药大学桂派中医药传承创新团队项目(2022B005);民族药资源再生与生态保护适宜技术及产业化模式研究(2019YFC1712305)。

摘　　要：目的:筛选积雪草皂苷生物合成相关的关键酶基因,并利用荧光定量PCR(qRT-PCR)技术进行验证。方法:采用HPLC对积雪草不同部位进行皂苷含量测定;利用转录组学、KEGG富集分析等手段挖掘积雪草皂苷生物合成关键酶基因;采用qRT-PCR技术验证挖掘出的关键酶基因在不同组织中的表达模式,并使用皮尔逊(Pearson)法分析关键酶与皂苷类成分的相关性。结果:在根、茎、叶、叶柄、果实这5个不同组织中,叶的积雪草皂苷含量最高;挖掘遴选出积雪草皂苷生物合成相关的关键酶基因4个,分别为氧化鲨烯环化酶(CaOSC1、CaOSC2)、鲨烯环化酶(CaSE)、鲨烯合酶(CaSS);这4个关键酶基因均在叶中表达量最高。结论:积雪草不同组织的皂苷含量与4个关键酶基因表达量关系紧密,该结果为今后探究积雪草皂苷生物合成与积累的调控机制积累了必要的研究基础。Objective:To screen the key enzyme genes related to the biosynthesis of Centella asiatica saponins and verify them by quantitative fluorescence PCR(qRT-PCR).Methods:The saponins content of Centella asiatica in different parts was determined by HPLC.The key enzyme genes related to Centella asiatica saponin biosynthesis were excavated by transcriptomics and KEGG enrichment analysis.qRT-PCR was used to verify the expression patterns of the extracted key enzyme genes in different tissues.Pearson method was used to analyze the correlation between key enzyme and saponins.Results:The saponins content in leaves was the highest among the five different tissues(root,stem,leaf,petiole and fruit).Four key enzyme genes related to the biosynthesis of Centella asiatica saponins were extracted and selected,which were oxidized squalene cyclase(CaOSC1,CaOSC2),squalene cyclase(CaSE)and squalene synthase(CaSS).The expression levels of these four key enzyme genes were the highest in leaves.Conclusion:The saponins content in different tissues of Centella asiatica is closely related to the expression levels of four key enzyme genes,which accumulate the necessary research basis for the future exploration of the regulation mechanism of the biosynthesis and accumulation of saponins in Centella asiatica.

关 键 词：积雪草 皂苷 转录组 关键酶基因 表达模式 

分 类 号：R282.71[医药卫生—中药学]