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| Detection and Viral Sequence Analysis of Four Lily-infecting Viruses in Beijing |
| LU Jing-Tong, KONG Xiang-Feng, LI Jie-Wen, JIA Gui-Xia* |
| School of Landscape Architecture/Urban and Rural Ecological Environment Beijing Laboratory/Ministry of Education Forest and Flower Breeding Laboratory/Beijing Key Laboratory of Flower Germplasm Innovation and Molecular Breeding/National Flower Engineering Technology Research Center, Beijing Forestry University, Beijing 100083, China |
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Abstract Lilies (Lilium spp.) are susceptible to complex viral infections. Virus detection and sequence analysis are helpful in identifying the types of infecting viruses and revealing viral variations. In this study, based on the conservative gene sequences of Lily symptomless virus (LSV), Lily mottle virus (LMoV), Cucumber mosaic virus (CMV), and Plantago asiatica mosaic virus (PLAMV), 4 pairs of primers were used to establish a multiplex RT-PCR detection method for these 4 viruses by optimizing the PCR extension time. This method was used to sample virus infections in lilies grown in the fields in Beijing. Furthermore, the 4 viral products were isolated for sequencing and alignment to construct a systematic evolutionary tree. The results showed that in the quintuplex RT-PCR reaction, extending the extension reaction time to 90 s resulted in the amplification of all 4 viruses and the 18S rRNA (reference gene). Sampling results indicated that lilies grown in the fields were often co-infected with 2 or more viruses, with detection rates in the order of LSV (100%)>CMV (92.31%)>LMoV (30.77%)>PLAMV (15.38%). Sequence analysis revealed that the average similarity of the isolated viral products to their respective viral sequences was above 84.4%, confirming the identification of the 4 target viruses. Variability analysis of the 4 viruses showed that when CMV was hosted by lilies, it formed a distinct branch, indicating host specificity. LMoV could be divided into 2 branches in the evolutionary relationship, with the experimental isolates belonging to branch Ⅱ. No clear branching patterns were observed for LSV and PLAMV. This study provides technical support for the routine molecular diagnosis and research of lily viral diseases, offering a foundational reference for disease prevention and control.
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Received: 11 October 2023
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Corresponding Authors:
* gxjia@bjfu.edu.cn
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