Zhao Xu, Liu Zhiyuan, She Hongbing, Xu Zhaosheng, Zhang Helong, Gao Wujun, Qian Wei
State Key Laboratory of Vegetable Biobreeding, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
College of Life Sciences, Henan Normal University, Xinxiang 453007, China.
Int J Mol Sci. 2025 Jun 20;26(13):5946. doi: 10.3390/ijms26135946.
Spinach ( L.) is an important leafy vegetable but is vulnerable to viral infections that significantly affect its quality and yield. In this study, we identified virus-infected spinach exhibiting typical symptoms with yellowing, wrinkling, and mottling in Beijing. But conventional RT-PCR screening for twelve common plant viruses yielded negative results. Then, using transcriptome sequencing along with a de novo assembly approach, we obtained the complete viral genome, which consists of RNA1 (5916 nucleotides) and RNA2 (3576 nucleotides). BLASTN analysis against the NCBI viral genome database revealed high homology with broad bean wilt virus 2 (BBWV2), leading us to designate this isolate as BBWV2-SP (GenBank accession numbers PV102464 and PV102465). Phylogenetic analysis indicated that BBWV2-SP shares 96.69% nucleotide sequence identity with a Liaoning isolate from MN786955 and clusters within the Chinese evolutionary lineage. We developed primers targeting the conserved region of the RNA2 coat protein, amplifying a 478-base-pair product. All symptomatic spinach samples tested positive, while asymptomatic controls remained negative, confirming the causal relationship between BBWV2-SP and the observed disease symptoms. This study provides the complete genome assembly of the spinach isolate BBWV2-SP and establishes a molecular detection protocol for BBWV2 in spinach. These findings offer essential technical support for field monitoring, epidemiological surveillance, and disease control strategies, while also enhancing our understanding of BBWV2's genetic diversity and mechanisms of pathogenicity.
菠菜（L.）是一种重要的叶菜类蔬菜，但易受病毒感染，这会显著影响其品质和产量。在本研究中，我们在北京鉴定出表现出典型症状（发黄、起皱和斑驳）的病毒感染菠菜。但对十二种常见植物病毒进行的常规逆转录聚合酶链反应（RT-PCR）筛选结果为阴性。然后，我们采用转录组测序及从头组装方法，获得了完整的病毒基因组，其由RNA1（5916个核苷酸）和RNA2（3576个核苷酸）组成。针对美国国立生物技术信息中心（NCBI）病毒基因组数据库的BLASTN分析显示，该病毒与蚕豆萎蔫病毒2（BBWV2）具有高度同源性，因此我们将该分离株命名为BBWV2-SP（GenBank登录号PV102464和PV102465）。系统发育分析表明，BBWV2-SP与来自MN786955的辽宁分离株核苷酸序列同一性为96.69%，并聚类于中国进化谱系内。我们设计了针对RNA2外壳蛋白保守区域的引物，扩增出一个478碱基对的产物。所有有症状的菠菜样本检测均呈阳性，而无症状对照仍为阴性，证实了BBWV2-SP与所观察到的病害症状之间的因果关系。本研究提供了菠菜分离株BBWV2-SP的完整基因组组装，并建立了菠菜中BBWV2的分子检测方案。这些发现为田间监测、流行病学监测和病害控制策略提供了重要的技术支持，同时也增进了我们对BBWV2遗传多样性和致病机制的理解。
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