紫山药黄酮醇合成酶DaFLS1基因的克隆和表达分析  

Cloning and expression analysis of Da FLS1 gene in purple yam(Dioscorea alata L.)

作　　者：闫瑞霞[1,2] 龚义勤[1] 殷剑美[2] 韩晓勇[2] 张培通[2] 柳李旺[1] 

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室/农业部华东地区园艺作物生物学与种质创制重点实验室/园艺学院,江苏南京210095 [2]江苏省农业科学院经济作物研究所,江苏南京210014

出　　处：《江苏农业学报》2014年第6期1448-1455,共8页Jiangsu Journal of Agricultural Sciences

基　　金：江苏省农业科技自主创新基金项目[CX(13)5008]

摘　　要：为了解黄酮醇在紫山药中的合成特点,利用3'-和5'-RACE技术从紫山药中分离到黄酮醇合成酶Da FLS1基因编码序列(Gen Bank登录号:KJ022640)。Da FLS1基因编码序列全长为1 113 bp,其开放阅读框长度为1 005 bp,编码334个氨基酸。Da FLS1蛋白质属于水溶性胞质不稳定蛋白质,无跨膜区和信号肽结构。保守区域分析结果显示,Da FLS1蛋白质属于α-酮戊二酸和二价铁离子依赖型的加氧酶家族[2OG-Fe(II)Oxygenase superfamily];Da FLS1氨基酸序列与水仙(AFS63900)同源性最近(75%),其次是洋葱(AAT68476,74%),与金鱼草(ABB53382)的同源性仅为66%。系统进化分析结果显示,紫山药与其他作物亲缘关系较远。Da FLS1在紫山药幼嫩叶片中表达水平最高,并且在紫山药幼叶中的表达有2个高峰期,而在其他组织中的表达量极低或不表达,即Da FLS1在紫山药中的表达情况符合黄酮醇合成酶基因(FLS)的特征。上述结果说明,得到的基因为紫山药黄酮醇合成酶基因Da FLS1,具有FLS基因的生物信息学特征和表达特征。The full-length coding sequence of flavonol synthase gene（Da FLS1） in purple yam was obtained by using3＇-and 5＇-RACE methods.Da FLS1（Gen Bank acc.no： KJ022640）,is 1 113 bp in length,containing an open reading frame of 1 005 bp encoding 334 amino acids.The Da FLS1 protein is water soluble without signal peptide and transmembrane domain,localized in cytoplasm.Conserved domain search revealed Da FLS1 protein belonged to the 2OG-Fe（II） oxygenase superfamily,whose activity depended on the co-existence ofα-ketoglutaric acid and ferrousion.Drthology of the deduced amino acid sequence showed Da FLS1 was highly homologous with Narcissus tazetta（AFS63900,75% similarity）,and Allium cepa（AAT68476,74% identity）.The phylogenetic tree constructed based on the deduced amino acid sequence indicated yam had far relationships with other plants.Da FLS1 exhibited the highest expression level in young leaf with two peaks over the whole growth period.There was little or no expression in other tissues.The expression of Da FLS1 gene in purple yam shared the same features of FLSs expression in other plants.

关 键 词：紫山药 黄酮醇合成酶 黄酮醇 DA FLS1基因 

分 类 号：S632.1[农业科学—蔬菜学]