白花泡桐纤维合成酶PfCesA4基因克隆及初步表达分析  

Cloning and Preliminary Expression Analysis of PfCesA4 from Paulownia fortunei

作　　者：苏江[1] 何金祥[1] 付传明[1] 冼康华[1] 黄宁珍[1] Su Jiang;He Jinxiang;Fu Chuanming;Xian Kanghua;Huang Ningzhen(Guangxi Key Laboratory of Plant Conservation and Restoration Ecology in Karst Terrain,Guangxi Institute of Botany,The Chinese Academy of Sciences,Guilin,541006)

机构地区：[1]广西壮族自治区中国科学院广西植物研究所,广西喀斯特植物保育与恢复生态学重点实验室,桂林541006

出　　处：《分子植物育种》2020年第11期3530-3536,共7页Molecular Plant Breeding

基　　金：广西科技创新能力与条件建设计划项目(桂科能1598025-36);桂林市重点研发计划项目(20170108-2,20170108-5);桂林市科学研究与技术开发计划(20180107-6);广西植物研究所基本业务费(桂植业17011);广西喀斯特植物保育与恢复生态学重点实验室自主课题项目(17-259-23)共同资助

摘　　要：本研究的目的是克隆白花泡桐纤维素生物合成途径关键酶基因--纤维素合成酶家族基因。采用PCR和RACE技术,利用Oligo、Primer Premier 5等软件进行引物设计,从白花泡桐中克隆出纤维素合成酶基因一条,NCBI分析后,命名为PfCesA4(NCBI登录号:MK340936)。PfCesA4基因的cDNA全长为3735 bp,其开放阅读框(ORF)长度为3138 bp,能够编码含有1045个氨基酸的蛋白质,此蛋白质的相对分子质量为119081.91 u,等电点为7.24。二级结构分析表明其以α-螺旋和无规则卷曲为主。结构域和跨膜区分析表明其在N端含有锌指结构域及2个跨膜区,在C端含有6个跨膜区,这些结构域能够表现纤维素合成酶的特征。系统进化树分析表明在所选取的物种当中,白花泡桐与芝麻的亲缘关系较近。组织特异性分析结果表明,其在茎中的表达量最高,根部次之,叶部最少;且在茎中,木质部的表达量高于韧皮部,说明其可能主要参与次生壁的建成。通过对PfCesA4基因的克隆及分析,能够为泡桐纤维素生物合成途径及后期利用分子手段对泡桐木材进行遗传改良提供基因资源。The purpose of this study was to clone cellulose synthase family genes which are the key enzyme genes of cellulose biosynthesis pathway from Paulownia fortunei.Using PCR and RACE techniques and primers designed with softwares like Oligo and Primer Premier 5,one cellulose synthase gene was cloned from Paulownia fortunei.After NCBI analysis,it was named PfCesA4(NCBI accession number:MK340936).The cDNA of PfCesA4 gene is3735 bp in length and its open reading frame(ORF)length is 3138 bp,capable of encoding a protein containing1045 amino acids.The relative molecular mass of this protein is 119081.91 u and the isoelectric point is 7.24.Secondary structure analysis showed that it was dominated byα-helix and random coil.Meanwhile domain and transmembrane region analysis indicated that it contain a zinc finger domain at the N-terminus and six transmembrane regions at the C-terminus,which characterize cellulose synthase.Phylogenetic tree analysis showed that among the selected species,the relationship between Paulownia fortunei and Sesamum indicum was closer.The results of tissue specific analysis showed that the expression of PfCesA4 was the highest in the stem,the root was the second,and the leaf was the least.The expression in the xylem and stem were higher than that of the phloem indicating that PfCesA4 may mainly participate in the construction of the secondary wall.Through the cloning and analysis of PfCesA4,while enriching the research in this field,it can provide important genetic resource for genetic improvement of paulownia wood by genetic engineering.

关 键 词：白花泡桐(Paulownia fortunei) 纤维素合成酶 基因克隆 

分 类 号：S792.43[农业科学—林木遗传育种]