广西局地西番莲病毒病的病原鉴定及优势病毒分析

 在广西采集表现斑驳、花叶症状疑似病毒病的西番莲叶片样品,利用小RNA测序技术,结合生物信息学分析,鉴定侵染西番莲的病毒种类。参考测序结果采用DAS-ELISA和RT-PCR方法对2015~2018年间采集的385份疑似病毒病样品进行检测,分析引发广西西番莲病毒病的优势病毒种类。结果显示:小RNA共获得20 921 061 clean reads,拼接获得560个contigs,其中99个contigs被注释为夜来香花叶病毒(telosma mosaic virus,TeMV),97个contigs被注释为黄瓜花叶病毒(cucumber mosaic virus,CMV),69个contigs被注释为东亚西番莲病毒(East Asian passiflora virus,EAPV),12个contigs被注释为大豆花叶病毒(soybean mosaic virus,SMV)。提取送测序的同批样品叶片的总RNA进行RT-PCR验证,可检测出TeMV、EAPV和CMV 3种病毒,未检出SMV。采用DAS-ELISA和RT-PCR对采集的样品进行检测,结果发现284份样品为阳性样品,检出率为73.76%,其中TeMV的检出率最高为64.16%,其次为EAPV和CMV,检出率分别为41.30%和11.43%;3种病毒存在复合侵染现象,其中TeMV+EAPV的检出率最高为24.94%,TeMV+CMV的检出率为4.16%,EAPV+CMV的检出率为0.26%,3种病毒复合侵染的检出率为4.94%。

Identification of pathogens and analysis of the dominant viruses of passion fruit virus disease in some areas of Guangxi

Leaves of passion fruit with suspected virus symptoms were collected in Guangxi for RNA extraction, the small RNA library was constructed and sequenced by high-throughput sequencing. Bio-informatics was performed to identify the viruses. Primers were designed according to the sequencing results for identification the viruses in passion fruit.The result showed that 560 contigs were produced from the 20 921 061 clean reads by library sequencing and assembly. Among them, 99, 97, 69 and 12 contigs were annotated as telosma mosaic virus(TeMV), cucumber mosaic virus(CMV), East Asian passiflora virus(EAPV) and soybean mosaic virus(SMV), respectively. All the samples were confirmed positive by RT-PCR for TeMV, EAPV and CMV, but negative for SMV. During 2015 to 2018, a total of 385 passion fruit samples with virus-like symptoms were collected from the main producing areas in Guangxi. 284 out of the 385 samples (73.76%) were positively for TeMV, EAPV and CMV infection by DAS-ELISA and RT-PCR. Among these three viruses, TeMV show the highest detection rate of 64.16%, and then EAPV with 41.30%, CMV with 11.43%. Mixed infections rate of TeMV + EAPV was 24.94%, TeMV+CMV was 4.16%, and EAPV+CMV was 0.26%. Mixed infection rate of all the above three viruses was 4.94%.