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依据NCBI基因库中公布的建兰花叶病毒(cymbidium mosaic virus,Cy MV)序列,利用Oligo7引物设计软件设计病毒特异性引物,以健康和感病的指示植物苋色藜及染病蝴蝶兰总RNA为模板,采用RT-PCR方法进行扩增,从感病的指示植物及染病蝴蝶兰中均扩增出与预期的431 bp大小完全一致的扩增产物,而健康的指示植物无此扩增产物。此扩增片段的测序进一步佐证了RT-PCR检测试验。利用BLAST及DNAMAN6.0.3.99软件与来自海南、南京、台湾以及韩国的病毒分离物进行同源性分析,其同源性均达92%以上。
According to Cymbidium mosaic virus (Cy MV) sequences published in NCBI gene bank, virus-specific primers were designed by using Oligo7 primer design software. The total RNAs of healthy and susceptible plants, The amplified products were amplified by RT-PCR. Amplification products corresponding to the expected size of 431 bp were amplified from susceptible plants and infected Phalaenopsis. However, healthy plants did not amplify the amplified products. The sequencing of this amplified fragment further confirmed the RT-PCR test. Homology analysis of the virus isolates from Hainan, Nanjing, Taiwan and South Korea using BLAST and DNAMAN6.0.3.99 software yielded over 92% homology.