关键词: 平邑甜茶, MhVPE, RNA干扰, ihpRNA, 拟南芥

Abstract: Based on the cDNA sequence of MhVPE gene（GenBank accession number：FJ891065）from Malus hupehensis（Pamp.）Rehd.，two pairs of specific primers containing different enzyme sites，named F1/F2 and R1/R2 respectively，were designed. With the template of pMD-MhVPE plasmid constructed previously，positive-sense strand pMD-F and antisense strand pMD-R were obtained，which were inserted into specific sites of the expression vector pART27，separately. The RNAi vector of pART-RNAi-MhVPE containing a hairpin structure was confirmed by the digestion of restriction enzymes. pART-RNAi-MhVPE vector was then transformed into Arabidopsis thaliana col-0 using the floral dip method by Agrobacterium-mediated transformation system to verification its efficacy. Seventeen transgenic plants were confirmed by kanamycin resistant selection and PCR testing. Semi RT-PCR results suggested that the expression level of AtVPE homologous gene in transgenic Arabidopsis thaliana was decreased，which proved that the expression of the targeted gene was inhibited efficiently and the ihpRNA vector of the MhVPE gene was constructed successfully，building a solid foundation for further study on the function of vacuolar processing enzyme（VPE）gene.

Key words: Malus hupehensis（Pamp.）Rehd., MhVPE, RNA interference, ihpRNA, Arabidopsis thaliana