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采用已报道杆状DNA病毒属(Badnavirus)的通用检测引物BadnaFP/BadnaRP,从7份芋样品中扩增到Badnavirus病毒的RT/RNase H基因保守区域,获得12个克隆序列,长度为576 bp。序列分析结果显示,来自不同样品的克隆间核苷酸和氨基酸序列相似性分别为78.8%~99.5%和81.3%~99.5%;来自同一样品扩增产物的克隆间在序列上也存在较大差异,核苷酸和氨基酸序列相似性分别为89.6%~100%和92.7%~100%。在系统进化树中,本研究所获序列与Badnavirus病毒的亲缘关系相对较近,聚在同一簇,表明此病毒为Badnavirus属病毒,但与芋杆状病毒(Taro bacilliform virus,TaBV)序列相似性较低,核苷酸和氨基酸相似性分别为58.0%~62.2%和58.5%~64.1%,推测为杆状DNA病毒属的一个新种。根据所测定序列设计了用于该病毒检测的引物P197/P433,对51份芋样品检测结果表明,该引物可有效检测来源于我国芋的Badnavirus病毒。
The Badnavirus common detection primer BadnaFP / BadnaRP was used to amplify the RT / RNase H gene conserved region of Badnavirus from 7 samples. Twelve cloned sequences with a length of 576 bp were obtained. Sequence analysis showed that the nucleotide and amino acid sequence identities of the clones from different samples were 78.8% -99.5% and 81.3% -99.5%, respectively. There was also a significant difference in the sequences between the clones derived from the same sample The nucleotide and amino acid sequence identities were 89.6% -100% and 92.7% -100%, respectively. In the phylogenetic tree, the sequences obtained in this study are relatively close to the Badnavirus and clustered in the same cluster, indicating that the virus is a Badnavirus, but similar in sequence to the Taro bacilliform virus (TaBV) The similarity of nucleotides and amino acids were 58.0% -62.2% and 58.5% -64.1%, respectively, suggesting a new species of baculovirus. P197 / P433 was designed according to the determined sequence. The results of 51 samples of taro showed that this primer could detect Badnavirus effectively.