广藿香PatTIFY6b基因克隆与功能分析  

Cloning and Functional Analysis of PatTIFY6b Gene in Pogostemon cablin

作　　者：黄焕超 王瑜桑 夏快飞[3,4] 李俭 张明永[3,4] 何梦玲 Huang Huanchao;Wang Yusang;Xia Kuaifei;Li Jian;Zhang Mingyong;He Mengling(School of Chinese Materia Medica,Guangdong Pharmaceutical University,Guangzhou,510006;Key Laboratory of Lingnan Medicinal Materials Production and Development,National Administration of Traditional Chinese Medicine,Guangzhou,510006;Key Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement,South China Botanical Garden,Chinese Academy of Sciences,Guangzhou,510650;Guangdong Provincial Key Laboratory of Applied Botany,South China Botanical Garden,Chinese Academy of Sciences,Guangzhou,510650)

机构地区：[1]广东药科大学中药学院,广州510006 [2]国家中医药管理局岭南药材生产与开发重点研究室,广州510006 [3]中国科学院华南植物园,华南农业植物分子分析与遗传改良重点实验室,广州510650 [4]中国科学院华南植物园,广东省应用植物学重点实验室,广州510650

出　　处：《分子植物育种》2023年第8期2554-2561,共8页Molecular Plant Breeding

基　　金：广东省科技厅项目(2017A030303082)资助。

摘　　要：茉莉酸甲酯能诱导广藿香叶片中百秋李醇含量升高,但是其中的分子机制不清楚。本研究从茉莉酸甲酯处理的广藿香叶片转录组数据中筛选到一个表达量显著增加的基因,经过序列比对发现该基因与唇形科丹参转录因子Sm TIFY6b具有高度同源性,因此将其命名为PatTIFY6b。通过生物信息学对PatTIFY6b编码蛋白的结构和功能进行分析和预测,并以广藿香cDNA为模板,克隆得到PatTIFY6b。PatTIFY6b定位于细胞核,在广藿香不同组织中均有表达,叶片中表达量最高。茉莉酸甲酯处理下,PatTIFY6b和百秋李醇合成途径基因具有相似的表达模式,说明茉莉酸甲酯可能通过PatTIFY6b调控百秋李醇的合成。利用原核表达系统,成功表达了PatTIFY6b蛋白。将PatTIFY6b过表达在双子叶模式植物拟南芥中,筛选到纯合株系PatTIFY6b-ox-17。本研究首次克隆广藿香PatTIFY6b基因并对其功能进行初步探究,为后期研究广藿香百秋李醇生物合成的分子调控机制提供理论基础。MI ethyl jasmonate can promote the accumulation of patchouli alcohol,but molecular mechanism is unclear.In this study,we screened an up-regulated gene from transcriptome of Pogostemon cablin treated with methyl jasmonate,named PatTIFY6b due to its sequence similarity to SmTIFY6b of Salvia miltiorrhiza.The protein encoded by PatTIFY6b were analyzed and predicted by bioinformatics,and full length of PatTIFY6b was cloned.PatTIFY6b localized in the nucleus and expressed in different tissues of Pogostemon cablin.In leaves,Pat-TIFY6b has the highest level of expression compared with that in other tissues.Upon methyl jasmonate treatment,PatTIFY6b and the genes related with patchouli alcohol synthesis have similar patterns of expression,suggesting that methyl jasmonate could regulate the synthesis of patchouli alcohol through PatTIFY6b.PatTIFY6b encodes 18.6 kD protein and can be successfully induced in prokaryotic system.Overexpressed PatTIFY6b in Arabidopsis thaliana,and screened the homozygous line PatTIFY6b-ox-17.In this study,we cloned the PatTIFY6b and studied its function firstly,which provided a theoretical basis for the molecular mechanism of patchouli alcohol biosynthesis.

关 键 词：广藿香 PatTIFY6b 茉莉酸甲酯 表达模式 百秋李醇 

分 类 号：S567.239[农业科学—中草药栽培]