CAJ | 학술논문

马铃薯X病毒（PVX）、马铃薯Y病毒（PVY）、马铃薯S病毒（PVS）、马铃薯卷叶病毒（PLRV）、马铃薯M病毒（PVM）和马铃薯A病毒（PVA）是马铃薯生产田中发生率较高、危害较严重的病毒，因此，建立特异性强、灵敏度高、检测速度快的RT-PCR分子检测体系对保障马铃薯种薯质量具有非常重要的意义。试验筛选了6种病毒的特异引物、优化了PCR部分的试剂以及确定了退火温度。结果表明，当Mg2+浓度为2.6 mmol/L、dNTPs浓度为0.1 mmol/L、Taq DNA聚合酶浓度为0.03 U/μL，以及退火温度为55.5℃时反应体系最稳定。最终，建立了一套通用RT-PCR检测体系，只需要变换每种病毒的引物，就可以实现对PVX、PVY、PVS、PLRV、PVM和PVA病毒的RT-PCR检测。每种病毒检测灵敏度均能达到pg以上。
마령서X병독（PVX）、마령서Y병독（PVY）、마령서S병독（PVS）、마령서권협병독（PLRV）、마령서M병독（PVM）화마령서A병독（PVA）시마령서생산전중발생솔교고、위해교엄중적병독，인차，건립특이성강、령민도고、검측속도쾌적RT-PCR분자검측체계대보장마령서충서질량구유비상중요적의의。시험사선료6충병독적특이인물、우화료PCR부분적시제이급학정료퇴화온도。결과표명，당Mg2+농도위2.6 mmol/L、dNTPs농도위0.1 mmol/L、Taq DNA취합매농도위0.03 U/μL，이급퇴화온도위55.5℃시반응체계최은정。최종，건립료일투통용RT-PCR검측체계，지수요변환매충병독적인물，취가이실현대PVX、PVY、PVS、PLRV、PVM화PVA병독적RT-PCR검측。매충병독검측령민도균능체도pg이상。
The higher incidence and more serious viruses in potato fields are potato virus X (PVX), potato virus Y (PVY), potato virus S (PVS), potato leafroll virus (PLRV), potato virus M (PVM), and potato virus A (PVA), so it is very important to guarantee the quality of seed potato by establishment of RT-PCR molecular detection system with specificity, high sensitivity and fast detection speed for these viruses. In this research, the primer for each virus was screened, PCR reagents were optimized, and annealing temperature was determined. The results indicated that when the concentration of Mg2 + was 2.6 mmol/L, dNTPs was 0.1 mmol/L, Taq DNA polymerase was 0.03 U/μL, and annealing temperature was 55.5℃, the reaction system was the most stable. Finally, a universal RT-PCR detection system was established, which only need to change primer for each virus when the RT-PCR detection for PVX, PVY, PVS, PLRV, PVM and PVA was needed. The detection sensitivity of each virus could reach the level of pg.