苦荞COP1基因的生物信息学分析及参与花色苷合成研究  

Bioinformatics analysis of COP1 gene in tartary buckwheat and its involvement in anthocyanin synthesis

作　　者：王丽娟[1] 赵佳利 杨姣 黄娟[1] 陈庆富[1] 邓娇[1] WANG Li-juan;ZHAO Jia-li;YANG Jiao;HUANG Juan;CHEN Qing-fu;DENG Jiao(Research Center of Buckwheat Industry Technology,Guizhou Normal University,Guiyang 550001,China)

机构地区：[1]贵州师范大学荞麦产业技术研究中心,贵阳550001

出　　处：《西南农业学报》2022年第9期1977-1985,共9页Southwest China Journal of Agricultural Sciences

基　　金：国家自然科学基金委员会-贵州省人民政府喀斯特中心项目(U1812401);贵州省科技计划项目[黔科合基础(2020)1Y095];国家现代农业产业技术体系荞麦育种岗位科学家专项资金(CARS-07-A5);贵州师范大学学术新苗基金项目(A类)[黔师新苗(2021)A16号]。

摘　　要：【目的】研究苦荞(Fagopyrum tartaricum Gaertn)COP1(Constitutive Photomorphogenic1)基因(FtCOP1)的生物信息学特征及其在花色苷生物合成中的表达模式,为进一步研究COP1基因参与调控苦荞花色苷生物合成的分子机制奠定基础,并为利用FtCOP1改良苦荞品质提供参考。【方法】利用PCR技术克隆苦荞COP1基因,利用生物信息学工具对其基因结构和编码蛋白的理化性质、二级和三级结构、亚细胞定位、保守结构域以及其它物种的COP1蛋白进化关系进行分析。利用实时荧光定量PCR(RT-qPCR)分析该基因在正常光照和黑暗处理苦荞芽菜中与花色苷生物合成结构基因的表达水平及花色苷含量的相关性。【结果】FtCOP1基因完整的CDS全长2082 bp,该基因结构由12个内含子与13个外显子构成,共编码693个氨基酸,其理论pI值为6.23,蛋白大小为75.876 kD,推测其具有亲水性,定位在细胞核中,含有1个跨膜螺旋结构域,无信号肽;二、三级结构主要元件为无规则卷曲和α螺旋,苦荞的COP1蛋白与藜麦COP1蛋白的亲缘关系最近,与其他物种COP1蛋白具有相似的motif和保守结构域,基因结构也相似。FtCOP1基因在黑暗培养的苦荞芽菜中的表达水平显著高于正常光照培养的苦荞芽菜,与花色苷生物合成结构基因的表达量及花色苷的积累量呈负相关。【结论】苦荞与其它物种COP1蛋白在植物进化中的功能比较保守,推测FtCOP1可能负调控光依赖型花色苷的生物合成。【Objective】The bioinformatics characteristics of FtCOP1 gene(Fagopyrum tartaricum Gaertn)fromCOP1(Constitutive Photomorphogenic 1)of Fagopyrum tartaricum Gaertn and the expression pattern in anthocyanin biosynthesis of tartary buckwheat are studied,which provides a basis for further studying the molecular mechanism of the involvement of COP1 gene in anthocyanins biosynthesis,and provides reference for improving the quality of tartary buckwheat by using FtCOP1.【Method】The COP1 gene of tartary buckwheat was cloned by PCR and analyzed by bioinformatics.The physicochemical properties,secondary and tertiary structure,prediction of subcellular localization,conserved domain and evolutionary relationship with COP1 protein of other species were analyzed.RT-qPCR was used to analyze the relationship between the gene and the expression level of anthocyanin biosynthesis structure genes and the anthocyanins content in tartary buckwheat under normal light and dark conditions.【Result】The complete length of CDS of FtCOP1 gene was 2082 bp,which consisted of 13 exons and 12 introns,encodings 693 amino acids.The theoretical pI value was 6.23 and the protein size was 75.876 kD.It was supposed to be hydrophilic,located in the nucleus and contained a transmembrane helical domain without signal peptide.The main components of the secondary and tertiary structures were random coil andαhelix.The COP1 protein of tartary buckwheat was most closely genetic related to the COP1 of quinoa.The COP1 protein has similar motif and conserved domains as that of other species,and the gene structure were also similar.The expression level of FtCOP1 in dark cultured sprouts was significantly higher than that in normal light cultured,and was negatively related to the expression of anthocyanin biosynthesis structural genes and anthocyanins accumulation.【Conclusion】COP1 proteins of tartary buckwheat and other species in plant evolution is conservative,and it is speculated that FtCOP1 may negatively regulate light-dependent anthocyanin biosyn

关 键 词：苦荞 FtCOP1基因 基因克隆 生物信息学分析 花色苷生物合成 

分 类 号：S517[农业科学—作物学]