苹果茎沟病毒部分分离物的生物学特性与分子鉴定

The biological characteristics and molecular identification of some Apple stem grooving virus isolates

通过昆诺藜接种鉴定和 ELISA 检测,从梨和苹果上分离获得苹果茎沟病毒(Apple stem groo-ving virus,ASGV)23个分离物。采用 TC-RT-PCR 对这些分离物进行扩增,均获得特异的扩增片段,PCR 产物经5%PAGE 电泳,出现大小约500、530和600bp 的3种迁移率不同的泳动带型。根据PCR 产物电泳迁移率的差异,选取3个来源于梨的分离物 P-L4、P-6-1-17和 P-3-2-67的 PCR 产物进行克隆与序列测定。经 BLAST 搜索,3个分离物的扩增片段与苹果分离物 P-209的 CP 基因3′端核苷酸序列同源性分别为92.2%、90.4%和88.4%。3个分离物间的核苷酸序列也有较大差异,P-L4/P-6-1-17为95.5%、P-L4/P-3-2-67为90.4%、P-6-1-17/P-3-2-67为88.6%。

Twenty-three isolates of Apple stem grooving virus were obtained from pear and apple by mechanically inoculating on Chenopodium quinoa and ELISA confirmation. Specific fragments were amplified from all these samples by tube capture reverse transcription-polymerase chain reaction （TC-RT-PCR）. These PCR products showed three type bands with different migration rates （500bp, 530bp and 600bp） on 5% PAGE. The PCR products of three isolates P-L4, P-6-1-17 and P-3-2-67 from pear with different migration rates were cloned and sequenced. A BLAST search result revealed that these three isolates had 92.2%, 90.4% and 88.4% nucleotide sequence similarity with the 3＇ends of P-209, an isolates from apple. And the nucleotide sequences among these three isolates also showed some different. The similarities of P-L4 and P-3-2-67, P-L4 and P-6-1-17, P-6-1-17 and P-3-2-67 were 90.4%, 95.5% and 88.6% , respectively.