番茄褐色皱纹果病毒与凤果花叶病毒复合侵染的检测及病毒全基因组序列分析

Co-infection of tomato brown rugose fruit virus and pepino mosaic virus in tomato and analysis of their full genome sequences

中文关键词:番茄褐色皱纹果病毒  凤果花叶病毒  复合侵染  全基因组序列  系统发育分析

keywords:tomato brown rugose fruit virus  pepino mosaic virus  co-infection  full genome sequence  phylogenetic analysis

中文摘要:为明确田间番茄Solanum lycopersicum植株出现花叶、泡状突起和褐色坏死等症状是否由番茄褐色皱纹果病毒（tomato brown rugose fruit virus，ToBRFV）和凤果花叶病毒（pepino mosaic virus，PepMV）复合侵染所致，在我国北方3个农业园区采集疑似被ToBRFV和PepMV侵染的26份番茄样品，采用反转录PCR技术对其进行检测，选择代表性样品的PCR产物进行拼接后获得ToBRFV和PepMV全基因组序列，对获得的全基因组序列进行相似性、变异性和系统发育分析。结果显示：在26份样品中，有20份样品仅被ToBRFV单独侵染，侵染率为76.92%，未见PepMV单独侵染，有3份样品被ToBRFV和PepMV复合侵染，侵染率为11.54%。ToBRFV-240417与ToBRFV荷兰分离物ToBRFV-6166394_2（GenBank登录号为OM515237.1）的核苷酸序列相似性最高，达99.70%；ToBRFV-231025、ToBRFV-240304与ToBRFV中国分离物ToBRFV-XG（GenBank登录号为OR451555.1）的核苷酸序列相似性最高，分别为99.88%和99.90%；PepMV-231025与PepMV美国分离物Ch2（GenBank登录号为DQ000985.1）的核苷酸序列相似性最高，达97.43%；PepMV-240304与PepMV德国分离物DSMZ PV-0716（GenBank登录号为OR233202.1）的核苷酸序列相似性最高，达97.58%。该结果丰富了ToBRFV、PepMV全基因组序列信息，有助于进一步了解它们之间的遗传进化关系。

Abstract:In order to clarify whether the symptoms of tomato (Solanum lycopersicum) in the fields, such as mosaic, bubble-like protrusions and brown necrosis, are caused by co-infection of tomato brown rugose fruit virus (ToBRFV) and pepino mosaic virus (PepMV), 26 tomato samples suspected to be infected by ToBRFV and/or PepMV were collected from three agricultural parks in northern China in this study. Reverse transcription (RT)-PCR was used to detect the pathogenic virus. The full genomic sequences of ToBRFV and PepMV were obtained by assembly of sequences of the PCR products of representative samples, and then the analysis of similarity, variability and phylogenetic tree were performed. The results showed that among the 26 samples, 20 samples were infected by ToBRFV only, with an infection rate of 76.92%, while no single infection of PepMV, three samples were co-infected by ToBRFV and PepMV, with a co-infection rate of 11.54%. The genomic sequence of ToBRFV-240417 had the highest similarity with the Dutch isolate ToBRFV-6166394_2 (GenBank accession number: OM515237.1), reaching 99.70%. The genomic sequences of ToBRFV-231025 and ToBRFV-240304 had the highest similarity of 99.88% and 99.90%, respectively, with the China isolate ToBRFV-XG (GenBank accession number: OR451555.1). The genomic sequence of PepMV-231025 had the highest similarity with that of American isolate Ch2 (PepMV-Ch2) (GenBank accession number: DQ000985.1), which was 97.43%. The genomic sequence of PepMV-240304 had the highest similarity with PepMV German isolate DSMZ PV-0716 (GenBank accession number: OR233202.1), reaching 97.58%. These findings expand the full genomic sequence data of ToBRFV and PepMV, facilitating a deeper understanding of their genetic and evolutionary relationships.