差速离心结合脉冲电泳分离枣疯病植原体DNA

Isolation of Phytoplasma Genomic DNA from Jujube Witches Broom by Differential Centrifugation Combined with PFGE

为了解枣疯病植原体的分子特征，明确植原体的致病机理及其与寄主的互作关系，以枣疯病组培苗为材料，利用差速离心结合脉冲电泳分离枣疯病植原体DNA。结果表明：感病组培苗在1600～2200kb之间出现一条明亮的条带，而健康组培苗没有，回收后经PCR检测，证明该目的条带是植原体DNA，实时定量PCR的分离结果表明：该实验方法能有效地将植原体基因组和植物基因组分离。

In order to know the molecular characteristics and pathogenic mechanism of jujube witches broom （JWB） phyto- plasma, as well as its interaction with the host, the author isolated the phytoplasma genomie DNA from JWB in tissue-culture ju- jube seedlings by using differential centrifugation combined with pulsed field gel electrophoresis （PFGE）. The resuhs indicated that the infected tissue-cuhure seedlings had a bright stripe in 1600~ 2200 kb, but the healthy tissue-culture seedlings had not. After recycling, PCR detection proved that this target stripe was the phytoplasma genomic DNA of JWB. The separation results of real-time quantitative PCR showed that this experimental method could effectively separate the phytoplasma genome and plant ge- nome.