CAJ | 학술논문

马铃薯Y病毒是近年来危害烟草生产的重要病毒之一,严重影响烟草的产量与品质.本研究利用DNAMAN软件对GenBank数据库中已登录的马铃薯Y病毒 (Potato virus Y, PVY) 全基因组序列进行序列比对,设计引物,以烟草肌动蛋白基因为内参,建立了烟草PVY的实时定量检测体系.获得的real-time PCR扩增基线平整,指数扩增明显,斜率大;稳定性和重现性好,变异系数小;循环阈值与PCR起始模板量对数之间存在良好的线性关系.与DAS-ELISA相比,该方法具有高效、灵敏、特异性强等优点,为从分子生物学水平上检测烟草中PVY提供了新的技术手段.
마령서Y병독시근년래위해연초생산적중요병독지일,엄중영향연초적산량여품질.본연구이용DNAMAN연건대GenBank수거고중이등록적마령서Y병독 (Potato virus Y, PVY) 전기인조서렬진행서렬비대,설계인물,이연초기동단백기인위내삼,건립료연초PVY적실시정량검측체계.획득적real-time PCR확증기선평정,지수확증명현,사솔대;은정성화중현성호,변이계수소;순배역치여PCR기시모판량대수지간존재량호적선성관계.여DAS-ELISA상비,해방법구유고효、령민、특이성강등우점,위종분자생물학수평상검측연초중PVY제공료신적기술수단.
Potato virus Y (PVY) is one of the most important viruses with huge damage to tobacco yields and quality in recent years.A quantitative method using real-time PCR technique basing on fluorescence dye SYBR Green was employed to detect PVY in tobacco in this study.PVY genome sequence alignments logged in Genebank were analyzed using DNAMAN software.Primer 5.0 software was used to design specific primers of PVY gene and actin gene.Results showed that the amplification curve had flat baseline, distinct exponential area, large and stable slope and the coefficient of variation was very small.There was a linear relationship between threshold cycle values where samples crossed threshold and the logarithmic values of template concentration.Compared with DAS-ELISA, real-time PCR can be used as a new method to detect PVY components in tobacco quantificationally, which is faster, more sensitive and specific.