青海省部分地区马铃薯病毒种类与Y病毒株系类型研究

Potato virus species and Y virus strain types in part areas of Qinghai province

【目的】了解马铃薯主要病毒对青海省部分地区马铃薯生产的危害,明确青海省部分马铃薯产区主要病毒类型及PVY株系类别,对马铃薯病毒病的防治和品种选育工作提供科学依据。【方法】利用DAS-ELISA和RT-PCR结合生物信息学分析方法对采自青海省大通县和互助县的181份疑似马铃薯病毒病样品进行病毒种类检测和Y病毒株系类型分析。【结果】181份样品中,PVY、PVS、PVM和PLRV病毒的检出率分别为17.68%、19.34%、4.42%和4.42%,其中,PVY单独侵染率最高,达14.92%,其次PVS为12.71%,样品中未检出PVX、PVA和AMV病毒。复合侵染中以2种病毒复合侵染为主,其中PVS+PVM、PVY+PVS和PVS+PLRV发生情况较为常见。DAS-ELISA检测PVY单独侵染结合RT-PCR混检获得20份PVY阳性样品,对其进行马铃薯Y病毒株系血清学检测分析发现,青海省部分地区马铃薯Y病毒株系以PVY^(O+C)血清型为主,其中PVY^(O+C)血清型阳性株系19份,占比为95%,PVY^(N)与PVY^(C)血清型阳性株系各1份,均占比为5%。为进一步明确PVY亚株系,根据血清学检测结果,对获得的19个PVY分离物的基因序列进行分析,发现PVY病毒核苷酸一致率为97.0%~100%,氨基酸一致率为95.8%~100%,且未发现重组位点,种群基因较稳定,PVY分离物系统发育分析发现,16个PVY分离物与PVY^(N:O)株系亲缘关系较近,3个PVY分离物PVY^(NTN-NW)株系亲缘关系较近,青海省部分地区马铃薯Y病毒株系类型以PVY^(N:O)、PVY^(NTN-NW)株系为主,与血清学检测结果一致。【结论】青海省部分地区主要马铃薯病毒种类有PVY、PVS、PVM和PLRV,PVY和PVS是青海地区马铃薯病毒的主要流行种类,其中Y病毒株系类型以PVY^(N:O)和PVY^(NTN-NW)株系为主,且同源性很高,PVY的种群基因较稳定。

【Objective】The study aimed to understand the harm of main potato viruses to potato production in some areas of Qinghai province,to clarify the main virus types and PVY strain types in some potato production areas of Qinghai province,and to provide a scientific basis for the prevention and control of potato virus diseases and variety selection.【Method】181 samples of suspected potato virus disease collected from Datong county and Huzhu county of Qinghai province were used for virus species detection and Y virus strain type analysis using DAS-ELISA and RT-PCR combined with bioinformatics analysis methods.【Result】Among the 181 samples,the detection rates of PVY,PVS,PVM and PLRV viruses were 17.68%,19.34%,4.42% and 4.42%,respectively.Among them,PVY alone had the highest infection rate of 14.92%,followed by PVS at 12.71%.PVX,PVA and AMV viruses were not detected in the samples.In the compound infection,two kinds of viruses are the main ones,among which PVS+PVM,PVY+PVS and PVS+PLRV were more common.DAS-ELISA detection of PVY alone infection combined with RT-PCR mixed detection to obtain 20 PVY positive samples,the serological analysis of potato Y virus strains on them showed that the potato Y virus strains in some areas of Qinghai province were PVY^(O+C) serotypes.Among them,19 strains were positive for PVY^(O+C) serotypes,accounting for 95%,and 1 strain was positive for PVY^(N) and PVY^(C) serotypes,accounting for 5%.In order to further clarify the PVY sub-strains,the gene sequences of the 19 PVY isolates obtained were analyzed according to the serological test results,and it was found that the nucleotide identity rate of PVY virus was 97.0%-100%,and the amino acid identity rate was 95.8%-100%,and no recombination sites were found,and the population genes were relatively stable.The phylogenetic analysis of PVY isolates showed that 16 PVY isolates were closely related to PVY^(N:O) lines,and 3 PVY isolates were related to PVY^(NTN-NW) lines.The relationship was relatively close,and the strains of potato Y virus in some areas of Qinghai province were mainly PVY^(N:O) and PVY^(NTN-NW) strains,which were consistent with the serological test results.【Conclusion】The main potato virus species in some areas of Qinghai province were PVY,PVS,PVM and PLRV,PVY and PVS were the main epidemic species of potato virus in Qinghai area,among which the Y virus strains were mainly PVY^(N:O) and PVY^(NTN-NW) strains,with high homology,and the PVY population genes were relatively stable.