利用小RNA测序技术检测贵州西番莲病毒

Identification of Viruses Infecting Passiflora edulis in Guizhou Province by Small RNA Sequencing

病毒病害是西番莲重要病害之一,明确病毒种类可为病害防治提供理论依据。本研究应用小RNA测序技术对6份疑似病毒侵染的西番莲叶片样本进行病毒种类鉴定,发现混合样本中含有黄瓜花叶病毒(Cucumbermosaicvirus,CMV)和夜来香花叶病毒(Telosma mosaic virus, TeMV)。应用RT-PCR分别扩增6份样本中2种病毒的外壳蛋白(coat protein, CP)基因序列,进一步证实6份样本中均含有CMV和TeMV。CMV的CP基因开放阅读框大小为657 bp,与已报道的其他分离物的一致性为82.3%~95.9%;TeMV的CP基因开放阅读框为816 bp,与其他分离物的一致性分别为87.8%~99.3%和87.8%~98.3%。上述结果表明,贵州西番莲中的病毒是CMV和TeMV的分离物,分别命名为CMV-GZ(CP基因登录号为MH623077)、TeMV-GZ1(MH623078)和TeMV-GZ2(MH623079)。

Virus disease is one of the main diseases of Passiflora edulis, and the identification of virus species can provide a theoretical basis for the disease prevention. In this study, six suspected virus-infected leaf samples were mixed and analyzed by small RNA deep sequencing. Cucumber mosaic virus (CMV) and Telosma mosaic virus (TeMV) were detected from the mixed sample. Viruses identified by small RNA sequencing were validated by virus coat protein gene (CP) specific RT-PCR from the above six samples. CMV and TeMV was detected from all six samples, respectively. The open reading frame of CP gene of CMV and TeMV obtained by RT-PCR was 657 bp and 816 bp, correspondingly. The nucleotide sequence identity of CMV-GZ (accession number: MH623077), TeMV-GZ1 (accession number: MH623078) and TeMV-GZ2 (accession number: MH623079) was 82.3%-95.9%, 87.8%-99.3% and 87.8%-98.3%, respectively, with the reported CMV and TeMV isolates. CMV and TeMV in P. edulis was first reported Guizhou. The combined infection of the two viruses may be a potential threat to P. edulis.