野蔷薇(Rosa multiflora)抗白粉病基因RmMlo的克隆与表达分析

Cloning and Expression Analysis of Resistance Powdery Mildew Gene RmMlo in Rosa multiflora

以野蔷薇(Rosa multiflora)为试材,采用RT-PCR和RACE方法从叶片中获得了一个Mlo同源基因的cDNA全长,命名为RmMlo,GenBank登录号为JF310747。序列分析结果表明,RmMlo基因cDNA全长为1993bp,包含49bp的5′非编码区、243bp的3′非编码区和一个长度为1700bp编码567个氨基酸的开放阅读框。氨基酸序列分析显示其蛋白质的C末端具有两个保守的MLO模体。序列比对和系统进化分析表明,RmMlo与拟南芥等双子叶植物的Mlo亲缘关系较近,相似性达90%以上。半定量RT-PCR分析表明,RmMlo在感病叶片中的表达量最高,在根中不表达,其表达模式与拟南芥的AtMlo有一定差别。

In this research,a full-length cDNA sequence of Mlo gene,named RmMlo（GenBank accession No.JF310747）was obtained from leaves of Rosa multiflora by RT-PCR and RACE.Sequence analysis indicated that RmMlo was 1 993 bp in full length containing a 5＇-untranslated region（5＇-UTR）of 49 bp,a 3＇-UTR of 243 bp,and an opening reading frame（ORF）of 1 700 bp encoding 567 amino acids.Subsequently,amino acid sequence analysis revealed that RmMlo had two conserved MLO motifs in the protein C-terminal.Sequence alignment and phylogenetic analysis showed that RmMlo shared more than90%homology with Mlo from Arabidopsis thaliana and other dicotyledon.Finally,semiquantitative RT-PCR analysis indicated that RmMlo showed the highest expression abundance in infected leaves and no expression in root,which was different from the expression pattern of AtMlo.