国产龙血竭基原植物类黄酮3'-羟化酶基因(F3'H)克隆及表达分析  

F3'H Gene Cloning and Expression Analysis of Original Plants of Dragon's Blood

作　　者：李爽 赵宏友 杨春勇[1] 王艳芳[1] 李戈[1] 彭建明[1] 严珍 王延谦 Li Shuang;Zhao Hongyou;Yang Chunyong;Wang Yanfang;Li Ge;Peng Jianming;Yan Zhen;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》2024年第19期6310-6317,共8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：类黄酮3'-羟化酶(flavonoid 3'-hydroxylase,F3'H)作为植物类黄酮代谢的关键酶之一,参与龙血树中龙血竭树脂的生物合成。为了揭示国产龙血竭基原龙血树中F3'H基因序列和表达模式等信息,本研究以剑叶龙血树、海南龙血树和岩棕为研究对象,采用同源克隆法克隆3种龙血竭基原植物的F3'H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)法分析其在不同组织的表达量。结果表明,在3种龙血树中均获得一条编码区长1533 bp的F3'H基因序列,编码510个氨基酸残基。生物信息学分析表明,它们的序列同源性达到99.57%,氨基酸序列的理化性质、二级结构和三级结构几乎完全一致。系统进化分析表明龙血树属植物F3'H基因聚为一支且进化上非常接近,在F3'H进化树上龙血树属并未与同科植物聚为一类,单子叶植物和双子叶植物亦并未分别聚为一类。RT-qPCR分析发现,剑叶龙血树中F3'H在茎中的表达量远高于根和叶,海南龙血树和岩棕中F3'H在茎中的表达量低于根和叶。本研究有利于揭示F3'H在龙血竭生物合成中的潜在作用,为龙血竭人工结脂技术开发提供一定分子基础。Flavonoid 3'-hydroxylase(F3'H)is one of the key enzymes in plant flavonoid metabolism,and it is involved in the biosynthesis of dragon's blood in Dracaena trees.In order to reveal the F3'H gene sequence and expression pattern of Dracaena trees,the F3'H gene sequences of three Dracaena(Dracaena cochinchinensis,Dracaena cambodiana,and Aizong)were cloned and bioinformatics analysis was conducted in this study.RT-qPCR was used to analyze the expression levels of the F3'H gene in different tissues.The result showed that a 1533 bp sequence of F3'H gene encoding 510 amino acid residues was obtained from all three Dracaena species.Bioinformatics analysis showed that their sequence homology reached 99.57%,and the physicochemical properties,secondary structure and tertiary structure of amino acid sequences were almost identical.Phylogenetic analysis showed that the F3'H genes of Dracaena trees were clustered into one branch.Interestingly,the F3'H genes of genus Dracaena does not cluster with the Liliaceae.In addition,the F3'H genes of monocotyledons and dicotyledons were not clustered separately in phylogenetic tree.RT-qPCR analysis showed that the expression level of F3'H in stem of Dra-caena cochinchinensis was much higher than that in root and leaf.However,the expression level of F3'H in root and leaf was higher than that in stem for Dracaena cambodiana and Aizong(Dracaena sp.).This study would be helpful to reveal the potential role of F3'H in dragon's blood biosynthesis of Dracaena and provide a molecular basis for the development of artificial lipid formation technology of dragon's blood.

关 键 词：龙血树 龙血竭 类黄酮3'-羟化酶 

分 类 号：S567.19[农业科学—中草药栽培]