摘要： 以牡丹品种赵粉为试材,采用RT-PCR和RACE方法从雄蕊中获得了一个牡丹苹果酸脱氢酶基因cDNA全长,命名为PsMDH,GenBank登录号为HQ449567。其cDNA全长1 283 bp,包含80 bp的5’非编码区、203 bp的3’非编码区和一个长度为999 bp编码332个氨基酸的开放阅读框。序列比对和系统进化分析表明,PsMDH与毛果杨的亲缘关系最近,相似性达93.37%以上。相对荧光定量PCR分析表明,PsMDH在心皮中的表达量最高,在雄蕊中表达量最低。成功构建植物表达载体,为牡丹品种改良提供基础。

关键词: 牡丹, 苹果酸脱氢酶基因克隆, 表达, 载体构建

Abstract: In this work,a full-length cDNA sequence of MDH gene was obtained from petals of tree peony(Paeonia suffruticosa L.cv.Zhao Fen)using RT-PCR and RACE,named PsMDH(GenBank accession No.HQ449567).The full length of PsMDH cDNA is 1 283 bp,containing a 5′-untranslated region(5′-UTR) of 80 bp,a 3′-UTR of 203 bp,and an opening reading frame(ORF) of 999 bp encoding a 332 predicted amino acids.Subsequently,sequence comparison and phylogenetic analysis revealed that PsMDH shared more than 93.37% homology with Populus trichocarpa.Relative Real-time PCR analysis indicated that PsMDH showed the highest transcript abundance in carpels and the lowest levels in stamens.Construction of plant expression vector lay a foundation for variety improvement in tree peony.

Key words: Tree peony, Cloning of malate dehydrogenases, Bioinformatics analysis , Expression , Construction of vector

中图分类号: 

• Q785