title: [Suppression of apple polyphenol oxidase by double-stranded RNA (RNAi)].
abstract: Antisense and sense gene fragments (710 base pairs) of apple polyphenol oxidase (APPO) gene were obtained by RT-PCR amplification, using the total RNAs isolated from ripen apple fruit as the template. These two fragments were ligated with a 1000 bp spacer, YYT (crtW+crtY fusion) gene, which is relative to carotenoid synthesization in subcocci. The full-length 2446 bp-target gene was then inserted into plant binary vector pYPX145 to generate the recombinant plasmid pYF7704, which carried the expression unit, of APPO dsRNA. pYF7704 was transformed to apple (Malus x domestica) var. Red Fuji via agrobacterium tumefaciens mediated leaf disc transformation. With the selection of Karamycin and GUS detection assays, transgenic shoots of A PPO dsRNA were obtained. The results of FQ-RT-PCR indicated that APPO mRNA level was suppressed to 91.69% in transgenic shoots compared to wide shoots. The data suggested that dsRNAi technology on apple polyphenol oxidase is feasible to be utilized in transgenic shoots.
authors.affiliation: Nanjing Agriculture University, Nanjing 210095, China.
authors.forename: Yan Hong
authors.initials: YH
authors.lastname: Cao
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zhang
authors.forename: Quan Hong
authors.initials: QH
authors.lastname: Yao
authors.forename: Ri He
authors.initials: RH
authors.lastname: Peng
authors.forename: Ai Sheng
authors.initials: AS
authors.lastname: Xiong
authors.forename: Xian
authors.initials: X
authors.lastname: Li
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D004156:Catechol Oxidase / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics; D027845:Malus / Q000201:enzymology* / Q000235:genetics*; D008957:Models, Genetic; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics; D034622:RNA Interference / Q000502:physiology; D012330:RNA, Double-Stranded / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014170:Transformation, Genetic / Q000235:genetics
chemical_list: D018744:DNA, Plant; D012330:RNA, Double-Stranded; D004156:Catechol Oxidase