CAJ | 학술논문

根据 GenBank 中已发表的建兰花叶病毒（Cymbidium mosaic virus ， CyMV ）、齿兰环斑病毒（Odontoglossum ringspot virus ， ORSV ）和菜豆黄花叶病毒（Bean yellow mosaic virus ， BYMV ）外壳蛋白（CP）基因序列保守区域分别设计特异性引物，通过优化多重PCR反应条件，建立能同时检测文心兰3种病毒的多重PCR检测体系。该体系能够一次扩增出CyM V、ORSV和BYM V的特异片段，其大小分别是551、325和212 bp。测序结果表明，3种病毒序列与相应的参考序列相似性均达98％以上。灵敏度测定结果表明，从相当于或大于10－2 m g的感病植物组织中能够检测到这3种病毒。
근거 GenBank 중이발표적건란화협병독（Cymbidium mosaic virus ， CyMV ）、치란배반병독（Odontoglossum ringspot virus ， ORSV ）화채두황화협병독（Bean yellow mosaic virus ， BYMV ）외각단백（CP）기인서렬보수구역분별설계특이성인물，통과우화다중PCR반응조건，건립능동시검측문심란3충병독적다중PCR검측체계。해체계능구일차확증출CyM V、ORSV화BYM V적특이편단，기대소분별시551、325화212 bp。측서결과표명，3충병독서렬여상응적삼고서렬상사성균체98％이상。령민도측정결과표명，종상당우혹대우10－2 m g적감병식물조직중능구검측도저3충병독。
By using specific primers designed on the basis of conserved sequences of coat protein gene ,a multiplex PCR protocol for the detection of 3 commonly found disease‐causing viruses ,i .e .,Cymbidium mosaic (CyMV) , Odontoglossum ringspot (ORSV) and bean yellow mosaic (BYMV) ,in Oncidium was developed .Three specific fragments were simultaneously amplified in a single PCR reaction system .Their molecular weights were determined to be 571 bp for CyM V ,325 bp for ORSV ,and 212 bp for BYM V .Sequence analysis indicated that the 3 viruses were at least 98% homogenous with their respective reference viruses .The sensitivity test on this newly developed methodology showed that it was capable of accurately detecting the 3 viruses simultaneously in a bulbous iris tissue at the level of 10-2 mg .