天香百合、药百合黄酮醇合成酶FLS基因克隆和表达分析  

Cloning and expression analysis of FLS gene of flavonol synthetase in Lilium auratum and L.speciosum var. gloriosoides

作　　者：刘筱琳 孙婷婷 杨捷 何恒斌 LIU Xiaolin;SUN Tingting;YANG Jie;HE Hengbin(Beijing Key Laboratory of Ornamental Plants Germplasm Innovation and Molecular Breeding,National Engineering Research Center for Floriculture,Beijing Laboratory of Urban and Rural Ecological Environment,College of Landscape Architecture,Beijing Forestry University,Beijing 100083,China)

机构地区：[1]北京林业大学园林学院,花卉种质创新与分子育种北京市重点实验室,国家花卉工程技术研究中心,城乡生态环境北京实验室,北京100083

出　　处：《浙江农业学报》2024年第2期344-357,共14页Acta Agriculturae Zhejiangensis

基　　金：国家自然科学基金(31672191);“十三五”国家重点研发计划(2019YFD1001002)。

摘　　要：以天香百合(Lilium auratum)和药百合(L.speciosum var.gloriosoides)为研究材料,分别克隆获得黄酮醇合成酶(flavonol synthase, FLS)基因,命名为LaFLS和LsFLS。实验结果表明,LaFLS和LsFLS基因均含完整的开放阅读框1 035 bp,均编码344个氨基酸,氨基酸序列高度保守,均具有DIOX-N结构域和2-酮戊二酸和铁(Ⅱ)依赖性双加氧酶结构域,属于2-酮戊二酸和铁(Ⅱ)依赖性双加氧酶超家族;系统进化分析表明,LaFLS和LsFLS除与东方系百合西伯利亚和索邦的FLS亲缘关系最近外,与百合科郁金香(Tulipa fosteriana)等亲缘关系较近;生物信息学分析显示,LaFLS和LsFLS蛋白无信号肽序列和跨膜结构域,均为亲水性蛋白,亚细胞定位结果显示二者主要定位在细胞质中。基因表达分析结果表明,在花蕾发育过程中,LaFLS和LsFLS随花蕾发育出现先上升后下降再上升的趋势,而且在花被片无色区的表达量显著高于有色区域。The flavonol synthetase gene FLS were cloned from Lilium auratum and L.speciosum var.gloriosoides,respectively,and named as LaFLS and LsFLS.The results showed that the LaFLS and LsFLS genes contained 1035 bp complete open reading frame and encoded 344 amino acids.The amino acid sequences were highly conserved,and both genes had the DIOX-N domain and the 2-ketoglutaric acid and iron(Ⅱ)dependent dioxygenase domain,belonging to the 2-ketoglutaric acid and iron(Ⅱ)dependent dioxygenase superfamily.Phylogenetic analysis showed that LaFLS and LsFLS were closely related to Tulipa fosteriana,except for the FLS of oriental hybrids Siberia and Sorbonne.Bioinformatics analysis showed that LaFLS and LsFLS were hydrophilic proteins without signal peptide sequences and transmembrane domains.Subcellular localization results showed that they were mainly located in the cytoplasm.The results of gene expression analysis showed that LaFLS and LsFLS increased first,then decreased and then increased with the development of flower bud,and the expressions of LaFLS and LsFLS were significantly higher in the colorless region than in the colored region.

关 键 词：东方百合 黄酮醇合成酶基因 黄酮醇 克隆 基因表达 

分 类 号：S682.265[农业科学—观赏园艺]