马尾松PmCYP720B1基因的克隆及表达分析  

Cloning and Expression Analysis of Pm CYP720B1 Gene from Pinus massoniana

作　　者：刘佳禾 镐青青 姚圣 陈佩珍[1] 季孔庶[1] Liu Jiahe;Hao Qingqing;Yao Sheng;Chen Peizhen;Ji Kongshu(Key Laboratory of Forest Genetics&Biotechnology of the Ministry of Education,Co-Innovation Center for Sustainable Forestry in Southern China,Nanjing Forestry University,Nanjing,210037)

机构地区：[1]南京林业大学,林木遗传与生物技术教育部重点实验室,南方现代林业协同创新中心,南京210037

出　　处：《分子植物育种》2025年第1期80-91,共12页Molecular Plant Breeding

基　　金：国家重点研发计划项目(2017YFD0600304);江苏高校优势学科建设工程资助项目(PAPD)共同资助。

摘　　要：CYP720Bs在针叶树的松柏油树脂防御系统中起到重要的作用。本研究从马尾松(Pinus massoniana)中克隆Pm CYP720B1基因,并进行生物信息学分析;分析其密码子偏好性;利用烟草叶片瞬时转化法对Pm CYP-720B1进行亚细胞定位;用RT-qPCR分析Pm CYP720B1在马尾松不同组织中的相对表达量。结果表明:PmCYP720B1的ORF区域全长1446 bp,编码481个氨基酸;密码子偏好性分析发现该基因适合使用烟草、拟南芥进行异源表达。亚细胞定位发现Pm CYP720B1表达蛋白定位于内质网中。RT-qPCR分析结果表明,Pm CYP720B1在植物体内的根、茎、叶等组织中均有表达,表达量在新叶中最高,在茎中最低,且在幼嫩的新叶中明显高于老叶。本研究认为Pm CYP720B1基因具有典型的P450结构域,并推测其与松脂合成、植物光和代谢有关。CYP720Bs play an important role in the oleoresin defense system of conifers.Pm CYP720B1 gene was cloned from Pinus massoniana,its bioinformation and codon preference were analyzed in this study.The subcellular localization of Pm CYP720B1 was conducted using tobacco leaf transient transformation method,and the relative expression level of Pm CYP720B1 in different stages and tissues was analyzed by RT-qPCR.The results demonstrated that the ORF region of Pm CYP720B1 is 1446 bp in length,encoding 481 amino acids,and codon preference analysis revealed that the gene is more suitable for heterologous expression using the Nicotiana tabacum or Arabidopsis thaliana.After performing subcellular localization,the Pm CYP720B1 expression protein is found to be distributed in endoplasmic reticulum.RT-qPCR analysis indicated that Pm CYP720B1 is expressed in root,stem and leaf tissues in vivo,with the highest expression in tender leaves and the lowest in stems,and significantly higher in tender leaves than in mature leaves.The conclusion is,the Pm CYP720B1 gene has a typical P450 domain,which may be related to rosin synthesis and plant photosynthesis.

关 键 词：马尾松 Pm CYP720B1 密码子偏好性 亚细胞定位 实时荧光定量PCR 

分 类 号：S791.248[农业科学—林木遗传育种]