国产龙血竭基原植物肉桂酸-4-羟基化酶基因(C4H)分离比较  

Isolation and Comparison of C4H Gene from Original Plants of Chinese Dragon's Blood Li Shuang Yang Chunyong Li Ge Wang Yanfang Zhao Hongyou Peng Jianming Yan Zhen Li Er

作　　者：李爽 杨春勇[1] 李戈[1] 王艳芳[1] 赵宏友 彭建明[1] 严珍 里二[1] 张丽霞[1] 王延谦 Li Shuang;Yang Chunyong;Li Ge;Wang Yanfang;Zhao Hongyou;Peng Jianming;Yan Zhen;Li Er;Zhang Lixia;Wang Yanqian(Yunnan Key Laboratory of Southern Medicine Utilization,Yunnan Branch of Institute of Medicinal Plant Development,Chinese Academy of Medical Sciences&Peking Union Medical College,Jinghong,666100)

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所云南分所,云南省南药可持续利用研究重点实验室,景洪666100

出　　处：《分子植物育种》2025年第3期912-918,共7页Molecular Plant Breeding

基　　金：国家自然科学基金项目(81703638);云南省科技人才和平台计划项目(202105AD160054,202105AG070011);云南省基础研究计划面上项目(202101AT070015);云南省重大科技专项计划项目(202102AA100020)共同资助。

摘　　要：肉桂酸-4-羟基化酶(cinnamate 4-hydroxylase,C4H)是苯丙烷代谢途径的第2个酶和第1个细胞色素P450单加氧酶。为了揭示国产龙血竭基原植物中C4H基因序列及表达模式等信息,初步分析C4H与龙血竭生物合成的关系,本研究以剑叶龙血树、海南龙血树和岩棕为试验对象,采用同源克隆法克隆C4H基因序列并进行生物信息学分析,利用实时荧光定量PCR(RT-qPCR)分析其在不同组织的表达量。结果表明,3种龙血树C4H基因的编码区长度为1518 bp,且高度保守,编码505个氨基酸残基,形成1个包含血红素环活性中心的球蛋白。聚类分析显示龙血树属植物C4H基因聚为一支,且与百合目的百合科和石蒜科植物C4H基因亲缘关系较近。RT-qPCR分析发现3种龙血树中PAL和C4H以协同方式表达,剑叶龙血树中根和茎的表达量显著高于叶,海南龙血树各组织表达量差异不大,岩棕中根的表达量远高于茎和叶。分析表明C4H是龙血竭生物合成的潜在关键酶之一,本研究对龙血树C4H基因的分离比较将有利于龙血竭生物合成的分子机制。Cinnamate 4-hydroxylase(C4H)is the second enzyme and the first cytochrome P450 monooxygenase in phenylpropane metabolism.In order to reveal the information of C4H gene sequence and expression pattern in origi-nal plants of Chinese dragon's blood,and preliminarily analyze the relationship between C4H and dragon's blood bio-synthesis,this study took Dracaena cochinchinensis,Dracaena cambodiana and Aizong(Dracaena sp.)as experi-mental objects,cloned C4H gene sequence by homologous cloning method and conducted bioinformatics analysis.RT-qPCR was used to analyze the expression levels of the C4H gene in different tissues.A 1518 bp sequence of C4H gene encoding 505 amino acid residues was obtained from all three Dracaena species.The C4H gene of these three Dracaena trees is highly conserved,and the amino acid sequence forms a globulin containing heme ring.Phylogenetic analysis showed that the C4H genes of Dracaena were clustered into one group,which closely related to the C4H genes of Liliaceae and Lycoraceae in evolution.RT-qPCR analysis showed that PA L and C4H were expressed in a synergistic manner in Dracaena trees.The PA L and C4H expression levels of roots and stems in Dracaena cochinchinensis were significantly higher than those in leaves,and there was no significant difference in various tissues of Dracaena cambo-diana,while the expression levels of roots in Aizong(Dracaena sp.)were much higher than those in stems and leaves.The results showed that C4H was one of the potential key enzymes in dragon's blood biosynthesis.In this study,the iso-lation and comparison of C4H gene in Dracaena will be beneficial to the molecular mechanism analysis of dragon's blood biosynthesis.

关 键 词：龙血树 龙血竭 肉桂酸-4-羟基化酶 

分 类 号：Q943.2[生物学—植物学]