一个凤丹PoFAD2基因家族新成员PoFAD2-2的克隆及表达模式分析  

Cloning and bioinformatics analysis of a new PoFAD2 family member PoFAD2-2 from Paeonia ostii T. Hong & J. X. Zhang

作　　者：孙静[1] 陈明 孟家松[1] 张克亮 陶俊[1] SUN Jing;CHEN Ming;MENG Jia-song;ZHANG Ke-liang;TAO Jun(College of Horticulture and Plant Protection,Yangzhou University/Jiangsu Key Laboratory of Crop Genetics and Physiology,Yangzhou 225009,China)

机构地区：[1]扬州大学园艺与植物保护学院/江苏省作物遗传生理重点实验室,江苏扬州225009

出　　处：《江苏农业学报》2018年第6期1339-1346,共8页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(31600564);江苏省自然科学基金项目(BK20160460);江苏省林业三新工程项目[LYSX(2016)45]

摘　　要：脂肪酸脱氢酶2(FAD2)能将油酸催化成亚油酸,是单不饱和脂肪酸转化为多不饱和脂肪酸的关键酶。本研究根据筛选到的PoFAD2-2基因序列,克隆到凤丹PoFAD2-2基因全长的c DNA,命名为PoFAD2-2。其开放阅读框为1 155 bp,编码384个氨基酸,蛋白质分子量为44 000,等电点为8. 51。生物信息学分析结果显示,该基因编码的蛋白质偏亲水性,包含5个跨膜结构域,属于膜结合型的FAD超家族。多序列比对与系统进化树分析结果发现,PoFAD2-2基因与PoFAD2-1基因亲缘关系较远,对于PoFAD2-2基因,凤丹在进化上与油橄榄[Olea europaea(common olive)(KY652929.1)]、靛木[Wrightia tinctoria (GU190864.1)]的相似性较高。荧光定量分析结果表明,PoFAD2-2在根和子房中没有表达,在花中的相对表达量最高,随着种子逐渐成熟,PoFAD2-2在种子中的表达量先逐渐降低再升高然后再降低,表明此基因表达具有组织特异性。PoFAD2-2基因在种子发育进程中的表达趋势与PoFAD2-1相同。Fatty acid desaturase 2(FAD2)catalyzes the conversion of oleic acid to linoleic acid,and it is a key enzyme which converts monounsaturated fatty acids to polyunsaturated fatty acids.In this study,the full-length cDNA of PoFAD2-2 gene was cloned from PoFAD2-2 gene and named PoFAD2-2.Its open reading frame was 1 155 bp,384 amino acids were encoded,the molecular weight of protein was 44 000,the isoelectric point was 8.51.Bioinformatics analysis results showed that the protein encoded by the gene was hydrophilic,contained five transmembrane domains,and belonged to the membrane-bound FAD superfamily.Multiple sequence alignment and phylogenetic analysis found that,PoFAD2-2 gene was more distantly related to PoFAD2-1 gene.PoFAD2-2 was evolutionarily similar to Olea europaea and Wrightia tinctoria.Quantitative real-time PCR results showed that PoFAD2-2 was not expressed in roots and ovaries,and the expression level of PoFAD2-2 was highest in petals.With the gradual maturation of seeds,the expression of PoFAD2-2 in the seeds was gradually decreased,then increased and then decreased.This gene expression was tissue-specific.The gene expression trend of PoFAD2-2 in seed development was the same as that of PoFAD2-1.

关 键 词：凤丹 脂肪酸脱氢酶2 生物信息学 荧光定量 

分 类 号：S565.9[农业科学—作物学] S685.11