稀莶上双生病毒的分子鉴定技术研究

利用双生病毒的通用引物PA/PB对采于福建省漳州地区表现黄脉症状的稀莶（Siegesbeckia orientalis）样品的总DNA进行扩增,得到病毒分离物FJ12。对FJ12DNA-A（FJ12A）全长2.7kb,具有典型的双生病毒科病毒特征,即编码6个ORFs,其中病毒链编码AVl（CP）和AV2共2个ORFs,互补链编码AC1-AC4共4个ORFs。基因组序列分析表明,FJ12A与中国广东报道的稀莶黄脉病毒（Siegesbeckia yellow vein virus,SbYVV）同源性达95.7%,因此,FJ12应为SbYVV的一个新分离物。利用DNAβ的特异引物beta01/02从样品中扩增得到卫星DNAβ分子（FJ12β）,FJ12β全长l331nts,编码一个功能ORF（C1）。FJ12β与SbYVV DNAβ的同源性最高,为70.2%,因此,FJ12DNAβ是双生病毒卫星分子的一个新种,建议命名为福建稀莶黄脉病毒伴随卫星。FJ12样品中存在DNA-A和DNAβ的复合侵染。

Molecular Identification Technology of Geminiviruses on Siegesbeckia Orientalis

The virus isolate FJ12 has been amplified by means of PA /PB from the total DNA of Siegesbeckia orientalis plants showing yellow vein symptom in Zhangzhou,Fujian Province.FJ12 DNA-A(FJ12A) was 2770 nts,with the typical genomic organization of begomoviral DNA-A,two ORFs(AV1 and AV2) in the virion-sense DNA and four ORFs(AC1 to AC4) in the complementary-sense DNA.Pairwise percentage nucleotide identity showed that the complete nucleotide of FJ12A had the highest sequence identity(95.7%) with SbYVV-(CN:Gd13:04)(AM183224),an isolate of Siegesbeckia yellow vein virus(SbYVV) from Guangdong province.The DNAβ molecule associated with the FJ12A was found with primers β01 /02(FJ12β).FJ12β was 1331nts and had eapacity to code possible ORFs C1 as proposed to be functiona1.Pairwise percentage nucleotide identity showed that FJ12β shared the highest sequence identity(70.2%) with SibYVB-(CN:Gd24:04)(AM230644),which indicated that it is a distinct betasatellite and was suggested to be Siegesbeckia yellow vein Fujian virus.PCR results showed that these infected plants contained DNA-A and DNA-β.