偃松PpDIV1基因克隆及生物信息学分析  

Full-Length cDNA Cloning and Bioformatic Analysis of PpDIV1 genes from Pinus pumila

作　　者：王思瑶 于宏影 刘亭亭[3] 刘洋 崔嵘 杨兰 WANG Si-yao;YU Hong-ying;LIU Ting-Ting;LIU Yang;CUI Rong;YANG Lan(Harbin Research Institute of Forestry Machinery,National Forestry and Grassland Administration,Harbin 150086;Research Center of Cold Temperate Forestry,CAF,Harbin 150086;Heilongjiang Provincial Center For Disease Control And Prevention,Harbin,150030)

机构地区：[1]国家林业和草原局哈尔滨林业机械研究所,哈尔滨150086 [2]中国林业科学研究院寒温带林业研究中心,哈尔滨150086 [3]黑龙江省疾病预防控制中心,哈尔滨150030

出　　处：《温带林业研究》2021年第3期38-43,共6页Journal of Temperate Forestry Research

基　　金：科技基础资源调查专项(2019FY100802-06);林木遗传育种国家重点实验室开放基金(K2019105)。

摘　　要：【目的】挖掘偃松种子萌发过程关键基因,为揭示偃松种子难于萌发的生理学和遗传学机制提供理论依据,并为进一步进行偃松资源开发利用提供技术保障。【方法】利用偃松种子萌发RNA-Seq数据及加权基因共表达网络分析结果筛选偃松DIV基因并克隆,通过生物信息网站及软件对其进行生物信息学分析。【结果】从偃松种子中克隆1个响应种子萌发的DIVARICATA(DIV)基因,该基因属于MYB家族,此基因命名为PpDIV1,并对其进行生物信息学及表达特性分析。结果表明,该基因具有完整的开放读码框(ORF),ORF长885 bp,编码294个氨基酸。该基因编码的氨基酸序列与月季DIV蛋白(GenBankID:XP_024190861)相似度最高,可达66.13%。PpDIV1基因存在两个SANT超家族位点,分别为SWI3、ADA2、N-CoR和TFIIIB结合位点及MYB结合位点。根据实验室前期对种子不同萌发时期转录组的测序结果显示,PpDIV1基因在此过程中不断上调表达。【结论】PpDIV1为偃松MYB家族中的新基因,该基因的获得为进一步挖掘其在偃松种子萌发过程中的调控机制研究奠定了基础。【Objective】In order to provide a theoretical basis for revealing the physiological and genetic mechanism of the difficult germination of the seeds of Pinus pumila,and provide technical support for development and utilization of the resources of Pinus pumila,we detected the key genes of seed germination of Pinus pumila.【Method】Screening and clone DIV gene of Pinus pumila base on the RNA-Seq data of seed germination of Pinus pumila and results of WGCNA analysis,then analyzed the gene through biological information website and software.【Result】A DIVARICATA(DIV)gene from the seed of Pinus pumila in response to seed germination was cloned,named PpDIV1,and the gene belongs to MYB family.In the same time,bioinformatics and expression characteristics of the gene was analyzed in this study.The results showed that the gene contain complete ORF(opening reading frame),ORF was 885 bp,encoding 294 amino acids,the amino acid sequence has the highest similarity with the DIV protein of Rosa chinensis(GenBankID:XP_024190861),up to 66.13%.There are two SANT superfamily locis in PpDIV1 gene,they are SWI3,ADA2,N-CoR and TFIIIB binding sites and MYB binding sites,respectively.According to the sequencing results of transcripts of different germination stages of seeds in the early stage of the laboratory,it showed that the expression of PpDIV1 gene was upregulated in this process.【Conclusion】Bioinformatics analysis showed that PpDIV1 gene was new the MYB family member from Pinus pumila.The gene laid the foundation of the regulatory mechanism of seed germination process for further study.

关 键 词：偃松 DIV MYB 克隆 

分 类 号：S791.24[农业科学—林木遗传育种]