马铃薯Y病毒云南昭通烟草分离物的检测及年度间差异比较

Detection and variation analysis of Potato virus Y tobacco isolates in Zhaotong,Yunnan Province from 2009 to 2011

利用DAS-ELISA检测试剂盒检测昭通烟草脉斑病样品,结果表明存在马铃薯Y病毒O株系和马铃薯Y病毒N株系两个不同株系病毒。利用Sprimer和M4引物扩增、克隆、测序,得到长度为1 771bp目的片段,该片段包含病毒的外壳蛋白基因序列、3′UTR序列以及部分Nib基因序列;序列分析表明与湖南HN-2分离物(GenBankNo.GQ200836)和美国NE-11分离物(GenBank No.DQ157180)核苷酸序列相似性均为98%,系统进化分析表明其具有较近的亲缘关系。

The samples with tobacco vein necrosis symptoms were isolated from Zhaotong, Yunnan Province from 2009 to 2011, and detected by DAS-ELISA. The results showed that the samples contained two strains of PVY, namely PVY° and PVYN. The expected PCR fragments were obtained by PCR with Sprimer and M4 primers. The fragments were cloned and sequenced. The sequence analysis showed that the length of the nucleotide sequence of different isolates were 1 771 bp, which contained PVY cp gene sequence, 3rUTR sequence and part of Nib gene sequence. Phylogenetic analysis showed that PVY tobacco isolates from Zhaotong had close relationships with Hu- nan HN-2 isolates （GenBank No. GQ200836） and the United States NE-11 isolate （GenBank No. DQ157180）. The homology of nucleotide sequences between them was above 98%.